Day :
- Track 9: Clinical Medicine
Track 10: Regenerative Medicine
Track 11: Applications of Tissue Engineering & Regenerative Medicine
Location: Rome, Italy
Chair
Lewis K. Clarke
Bay Area Rehabilitation Medicine Associates
USA
Co-Chair
Ion N. Mihailescu
National Institute for Lasers
Romania
Session Introduction
Lewis K. Clarke
Bay Area Rehabilitation Medicine Associates
USA
Title: Optimizing intrinsic mechanisms of neuroregeneration in the CNS: Utilizing mitochondrial and neurosteroid chemistry
Time : 09:00-09:20
Biography:
Lewis Clarke obtained his Master of Science from University of Texas at Dallas in 1977 in Human Development with Biostatistics. In 1986, he completed his Doctor of Medicine degree from Texas Tech School of Medicine. In 1987, he received his PhD from the Department of Cell Biology and Neurobiology at University of Texas Health Sciences Center at Dallas. He completed his medical internship at Emory University and Baylor College of Medicine in 1986 and finished his residency training at Baylor College of Medicine in Physical Medicine and Rehabilitation. He has a clinical and research practice in the Houston Texas area and has started two rehabilitation hospitals.
Abstract:
Neuronal repair in the central nervous system through neuroregenerative processes has always been considered to be beyond the recuperative capability of the human body. It is now understood that neurogenesis does indeed occur in the CNS and can result in functional as well as structural restoration. However, the heterogeneity of ischemic, traumatic, and degenerative brain injury obviates the implementation of any single intervention or therapy. A parsimonious beginning would be the augmentation of mitochondrial number and energy production as this would address all cellular function including stem cell production and migration. While much is known about the contribution of CoQ10, carnitine, and pyrroloquinoline quinone, an integrative approach should include bioactive lipids in the mitochondrial membrane, sigma 1 receptors, and neurosteroids such as DHEA, pregnenolone, and progesterone produced de novo in the glia. These neurosteroids can be considered catalysts and promote neurogenesis and neurite outgrowth through their activation of sigma 1 receptors probably in the mitochondrial membrane lipid rafts of the endoplasmic reticulum. Activated sigma 1 receptors increase calcium in the mitochondria resulting in activation of the TCA cycle, increasing mitochondrial hypermetabolism ultimately resulting in neurite outgrowth.
Increasing neurosteroids through exogenous administration, membrane maintenance with cholesterol and phospholipids, and augmentation of mitochondrial function would optimize intrinsic mechanisms of neuroregeneration.
Fumio Arai
Kyushu University
Japan
Title: Pot1a regulates self-renewal activity of hematopoietic stem cells
Time : 09:20-09:40
Biography:
Fumio Arai has completed his Ph.D at the age of 28 years from Meikai University and postdoctoral studies from Keio University School of Medicine. He is a Professor of Department of stem cell biology and Medicine, Graduate School of Medical Sciences, Kyushu University. His research interest is in studying the mechanisms of the cell fate regulation of HSCs at the single cell level for the establishment of the system that is able to expand HSCs.
Abstract:
Aging or repeated cell division induces the accumulation of DNA damage, which impairs hematopoietic stem cell (HSC) function. Protection of telomeres 1 (Pot1), a component of shelterin, contributes to the suppression of unnecessary DNA damage response (DDR) at telomeres. We identified that high levels of Pot1a was expressed in HSCs, and that this expression decreased shapely with age. Knockdown of Pot1a increased telomeric DDR and the frequency of symmetric differentiation divisions in cultures, and significantly reduced long-term reconstitution (LTR) activity. In contrast, overexpression of Pot1a or treatment with exogenous Pot1a protein Pot1a protein prevented telomeric DDR and maintained symmetric self-renewing divisions and LTR activity in HSCs, indicating that Pot1a rejuvenated stem cell activity of HSCs. Human POT1 protein also increased the number of cord blood HSCs. These data suggest that the protection of telomeric DNA from DDR signaling is critical for sustained self-renewal of HSCs and that Pot1a is a novel target for ex vivo expansion of HSCs.
Ion N. Mihailescu
National Institute for Lasers
Romania
Title: Soft transfer pulsed laser technologies for aplications in regenerative medicine
Biography:
Ion N. Mihailescu is Senior Scientific Researcher 1st degree in LSPI – INFLPR and Professor and PhD supervisor at Physics Department, University of Bucharest. Current fields are biophysics and biomedicine; plasma and laser physics and theory and current research interests go to surface studies with lasers; surface processing; deposition and modification of thin solid structures by high intensity laser irradiation; and biomaterials. He received the 1994 Galileo Galilei Award of the International Commission for Optics (ICO) “for outstanding contributions to the field of optics". He authored more than 400 publications, Hirsch index 27, total citations over 3000. http://www.researcherid.com/rid/A-5403-2011.
Abstract:
Recent results in organic/inorganic composite nanostructured layers synthesized by MAPLE and LDW are reviewed. The optimum deposition regime was reached based upon the results of investigations by SEM, TEM, SAED, XTEM, AFM, XRD, XPS or FTIR methods. Biocompatibility, bioactivity and biodegradation were assessed by dedicated in-vitro tests. Urease immobilized by MAPLE in form of thin films was shown to preserve its activity in breaking down and diagnose of urea content in blood. The application of MAPLE was extended to the transfer and immobilization of IgG molecules. We studied the effect of the lipid addition in the initial solution upon the protein thin films adhesion to substrate. We showed that the composite PMMA-bioglass films efficiently protects metal implants against the action of human fluids. The MAPLE obtained nanocomposites Ag:HA-organosolv lignin proved noncytotoxic, supporting the normal development and promoting the proliferation of the adhered human mesenchymal cells. The lignin addition potentiated the anti-microbial activity of HA doped with silver ions against either bacterial or fungal biofilms. Mesotetraphenylporphyrin clean and liquid-free micropatterns on Si substrates were fabricated by LDW. The propulsor metal film thickness was found to be a key parameter, which determines the laser fluence range allowing the clean transfer, predominant mechanism of the blister formation and laser-induced heating of the transferred material. We conclude that the thin films prepared by MAPLE and LDW techniques were identical in chemical composition, structure, morphology, and most likely functionality resembling the base material, as proved by physical-chemical characterization and in-vitro assays.
Roberto Ebensperger
Pontificia Universidad Católica de Chile
Chile
Title: Nanoencapsulation of stem cells and regenerative medicine
Time : 10:35-10:55
Biography:
Roberto Ebensperger is Associate Professor of Clinical Pharmacology and Director of the Laboratory of Cellular Therapy and Regenerative Medicine in the Pontificia Universidad Catolica de Chile. He is Pharmacist and PhD in Biochemistry from the Universidad de Chile. He has had several postdoctoral training in molecular cardiology at Medizinische Hochschule Hannover, hair follicle biology and pathology at Centre for Skin Sciences, University of Bradford, hematology and aging biology at Université Pierre et Marie Curie Paris VI. He was invited professor at Equipe Biologie Cellulaire du Vieillissement, Université Pierre et Marie Curie. Currently, he is interested in mesenchymal stem cell applications in wound healing. In 2013 he founded Plasticel, a spin-off company that initiated R&D activities in applications for cellular therapy for plastic surgery and baldness.
Abstract:
Cell nanoencapsulation is a novel delivery system based on a self-assembly technique mediated by electrostatic interactions called Layer-by-Layer (LbL) deposition, without an increase in volume implant because of the nanometric thickness of its layers. LbL coats the entire surface of individual cells, providing mechanical resistance to cells against manipulation and storage conditions prior to implantation in the patient. LbL is formed by polymeric layers or film depositions that are mediated by alternating opposite electrostatic interactions on a charged template. Using this technology, single-cell nanocapsule formation using human adipose-derived mesenchymal stem cells (ADSC) was assessed to determine the experimental factors to successfully preserve viability and functionality of cells in order to be used in regenerative medicine applications. For example, nanoencapsulation of mesenchymal stem cells (MSC) could be useful for cell targeting by conjugating a specific antibody, tissue engineering by incorporating a matrix protein that could be used as a scaffold and immuneisolation when using multilayer nanoencapsulation. Thus MSC may serve as a promising platform for cell-based encapsulation in regenerative medicine, both for cellular therapy as well as for tissue engineering.
Akiko Mammoto
Harvard Medical School
USA
Title: The role of LRP5 in lung angiogenesis and regeneration
Time : 10:55-11:15
Biography:
Akiko Mammoto received her Ph.D from Osaka University in Japan and completed her postdoctoral studies at Boston Children’s Hospital/Harvard Medical School. She is currently an Instructor in the Vascular Biology Program at Boston Children’s Hospital, has published more than 65 papers in high impact journals and serves as an editorial board member of Scientific Reports.
Abstract:
Chronic lung diseases, including chronic obstructive pulmonary disease, pulmonary fibrosis and asthma, are the third leading cause of death in the United States. To date, lung transplantation is the only way to save patients with end-stage chronic lung diseases. However, because of the shortage of transplant donors, high cost, and serious complications, lung transplantation is not an optimal approach. It has been recognized that adult human lungs may have the potential to regenerate after pneumonectomy (PNX). Thus, stimulating regeneration of adult lungs could be a good therapeutic strategy for chronic lung diseases. Angiogenesis, the growth of new blood vessels, plays a key role in organ development, homeostasis, and regeneration. We have reported that the Wnt co-receptor, low-density lipoprotein receptor-related protein 5 (LRP5), controls neonatal lung vascular and alveolar development. Here we demonstrate that LRP5 also controls adult lung regeneration through angiopoietin-Tie2 signaling. Compensatory lung growth after PNX is inhibited in Lrp5 knockout mice. We have also developed a unique method to implant hydrogel on the mouse lung, which enables us to clearly visualize lung-specific angiogenesis in mice. Using this method, we found that host lung-derived angiogenesis and alveolar cell recruitment are inhibited in hydrogels implanted on the lungs of Lrp5 knockout mice or mice treated with Tie2 inhibitor. Modulation of LRP5 signaling may therefore lead to the development of novel therapeutic interventions for various lung diseases and the improvement of lung organ engineering.
Pedro G. Morouco
Polytechnic Institute of Leiria
Portugal
Title: Direct digital manufacturing trends for regenerative medicine
Time : 11:15-11:35
Biography:
Pedro G. Morouco is PhD Research Fellow and Head of the CDRsp BioFabrication Research Group. Co-edited 2 books, authored and co-authored more than 70 papers published in books (n=12), international journals (n=31) and international conferences (n=36). Member of the Editorial Board in several international peer-review journals and was distinguished with the New Investigator Award 2014 from ISBS. Chairman of the CDRsp Advanced Courses on Regenerative Medicine and Workshops on DDM, Pedro is engaged in several national and international projects focusing on biomechanics and/or DDM.
Abstract:
Direct digital manufacturing (DDM) is not a futuristic expertise. Actually, its use has been employed throughout a wide spectrum of areas/industries. One of the areas where DDM is having a major impact is on the development of implants for tissue science and regenerative medicine. To do so, both anatomical as functional characteristics must be addressed. In fact, a combination of scaffolds, cells and/or growth factors should aim the production of implants (temporary or permanent), so that damaged or degenerated tissues can be fully restored. The DDM ability to join different composites layer-by-layer, enriches hybrid 3D structures that aim to mimic the native tissue and adds a new degree in freedom of design. However, the available methodologies lack some levl of expertise regarding biocompatibility, bioactivity and/or biodegradation (e.g. the lack of porosity and architecture control on 3D scaffolds). In this presentation we will show new technologies that we have been developing for 3D printing (both hard and soft scaffolds), as well as present major achievements that we have accomplished regarding bone and cartilage regeneration. For instance, the printing of multiple (up to 3) hydrogels with, or without loaded cells, that intend to generate layered mechanically stable implants through the double-printing of hydrogels with thermoplastics. This technology has shown promising results to overcome identified gaps in cartilage regeneration.
Wanda Lattanzi
Università Cattolica del Sacro Cuore
Italy
Title: Biobanking of subcutaneous adipose tissue: effect of site, cryopreservation, age and BMI
Time : 11:35-11:55
Biography:
Wanda Lattanzi, is an MD PhD, specialized in Medical Genetics, and an Assistant Professor of Anatomy at the Università Cattolica S. Cuore, Rome. Her research activity focuses mostly on molecular and cellular biology of bone and connective tissues, including the study of congenital malformations and the development of regenerative strategies based on mesenchymal stem cells. She coordinates national and international research projects funded by the NIH, the EU commission and the Italian Ministry of Health. She is author of 35 peer-reviewed manuscripts, member of the Editorial board of diverse international journals and reviewers in national grant programs.
Abstract:
The preferred setting for regenerative medicine applications is the availability of clinically acceptable off-the-shelf tissue and cell-based products. Cryopreserved adipose tissue (AT) represents a valid approach to this aim. Our previous study demonstrated that superficial subcutaneous AT is endowed with increased yield and stem properties Here we compare adipose-derived stem cells (ASCs) isolated from cryopreserved AT harvested from superifical and deep sites. Cellular yield, viability, immunophenotype, proliferative and multilineage potential were analyzed. The influence of BMI and donor age was also taken into account. To this aim, lipoaspirates were collected through manual liposuction from female patients undergoing elective lipofilling. The tissue was cryopreserved at -80°C for a month and after that in liquid nitrogen cylinder at −196 C for long-term storage. After fast thawing, cell morphology was analyzed histologically, hence adipose-derived stem cells (ASCs) were isolated by plastic-adherence and immunophenotyped. The expression of genes involved in stemness, trophic features and commitment, was analyzed. Cryopreservation reduced significantly the cellular yield, as expected, especially in deep harvests. It had no effect on cell viability, proliferation and multilineage potential of cells. Donor age and BMI significantly affected cell viability and gene expression, more markedly in deep than in superficial subcutaneous fat harvests. Our data suggest that: 1. superficial subcutaneous AT may be cryopreserved and potentially banked without affecting cellular viability and biological properties; 2. the stem properties of superficial AT lipoaspirates are less affected by age and BMI, compared to deep harvests.
Samin Eftekhari
Ryerson University
Canada
Title: Artificial bone substitutes for bone repair purposes
Time : 11:55-12:15
Biography:
Samin Eftekhari is a PhD candidate in the Chemical engineering Department of Ryerson University, who will defend her PhD dissertation on August 2015. She is collaborating with Mechanical Engineering Department and St. Michael's hospital as well. Her PhD research focuses on design and development of innovative biodegradable nanocomposites projected for bone repair applications. She received her M.Sc. in Biomedical Engineering from Amirkabir Polytechnic University (Tehran Polytechnic), and her B.Sc. in Material Science & Engineering from Sahand Polytechnic University. She has over 10 years of experience in biomaterials research and commercialization, which resulted several scientific papers, international presentations, and a patent.
Abstract:
The second most common transplantable tissue after blood is bone. Bone fractures are one of the most common forms of injury. In order to encourage the bone healing process, bone grafts are often needed. The patient’s own bone is the gold standard for bone grafting, but the lack of the bone supply raised the need for artificial bone substitutes. The novel biomaterials with tailorable biodegradation rate and mechanical strength have been developed. Using this we are in the process of developing orthopedic implants with customizable shapes based the patient’s age and the location of the damaged bone in the body. This novel material can also be used in a 3-D printer in order to obtain custom-fit implants. Constituents of these implants closely mimic the properties of real bone such as chemical and thermal properties. These implants will revolutionize current ways of bone repair treatments, since they provide three advantages over other metallic and non-metallic implants: (1) elimination of the second surgery to extract the implant (2) prevention of complications such as immunogenic responses caused by using other individual’s or animal’s bone powder, (3) reduction of long term adverse effect such as pain from thermal contraction of metallic impacts in cold weather, and (4) tailorable biodegradation rate, mechanical strength, and shape customizable for each patient, Such a product will give patients in need of bone grafts a speedy and efficient recovery.
- Track 6: Scaffolds
Track 7: Whole Organ Engineering & Approaches
Location: Rome, Italy
Chair
Joseph Purita
Institute of Regenerative and Molecular Orthopedics USA
Co-Chair
Andrea Barbetta
Sapienza University of Rome
Italy
Session Introduction
Joseph Purita
Institute of Regenerative and Molecular Orthopedics
USA
Title: Cutting edge concepts in the use of stem cell and PRP injections in an office setting
Time : 10:40-11:00
Biography:
Purita is Director of Institute of Regenerative and Molecular Orthopedics (www.stemcellorthopedic.com) in Boca Raton, Florida. The Institute specializes in the use of Stem Cells and Platelet Rich Plasma injections. Dr. Purita is a pioneer in the use of Stem Cells and Platelet Rich Plasma. The Institute has treated some of the most prominent professional athletes from all major sports in both the U.S.A. and abroad. He received a B.S. and MD degree from Georgetown Univ. Dr. Purita is board certified in Orthopedics by ABOS. He is a Fellow American College of Surgeons, Fellow American Academy Orthopedic Surgeons, and a Fellow American Academy of Pain Management. He is also certified in Age Management Medicine. He has lectured and taught extensively throughout the world on the use of Stem Cells and Platelet Rich Plasma. He has been instrumental in helping other countries in the world establish guidelines for the use of Stem Cells in their countries. He has been invited to lecture on these techniques throughout the world as a visiting professor.
Abstract:
The presentation concerns PRP and Stem Cell (both bone marrow and adipose) injections for musculoskeletal conditions in an office setting. Indications are given as to which type of cell and technique to use to accomplish repair. Stem cells, both bone marrow derived (BMAC) and adipose, are used for the more difficult problems. PRP injections are utilized for the less severe problems. Indications are given when to use Stem Cells verses PRP and when to use both. The newest concepts in stem cell science are presented. These concepts include the clinical use of MUSE cells, exosomes, and Blastomere like stem cells. Basic science of both PRP and stem cells are discussed. This presentation defines what constitutes an effective PRP preparation. Myths concerning stem cells are dispelled. One myth is that mesenchymal stem cells are the most important stem cell. This was the initial interpretation of Dr. Arnold Caplan the father of mesenchymal stem cell science. Dr. Caplan now feels that MSCs have an immunomodulation capacity which may have a more profound and immediate effect on joint chemistry and biology. We now learn in the talk that the hematopoietic stem cells are the drivers of tissue regeneration. Also discussed are adjuncts used which enhance the results. These therapies include supplements, LED therapy, lasers, electrical stimulation, and cytokine therapy. The scientific rationale is presented for each of these entities as to how they have a direct on stem cells.
Andrea Barbetta
Sapienza University of Rome
Italy
Title: Bioprinted 3D vascularized network tissue constructs using cell-laden bioink
Time : 11:00-11:20
Biography:
Andrea Barbetta has completed his Ph.D. in 2001 from University of Durham (UK) and postdoctoral studies from University of Kyoto (Japan) and Sapienza University of Rome. At present, he is lecturer at the Department of Chemistry of Sapienza University of Rome. He has published 40 papers in reputed journals and 1 patent. His research interests are focused mainly in the development of methods for the fabrication of scaffolds for tissue engineering, in particular by means of microfluidics and rapid prototyping.
Abstract:
Microfabrication technologies have been proposed as methods to create vascularized tissues. However, despite significant advances, insufficient aligned cellular organization and limited hierarchical architecture has impeded progress toward mimicking the highly vascularized tissue in 3D. To address these challenges, we introduce a new paradigm of vascularization that uses bioprinting as a robust method for fabricating 3D tissues constructs. This approach is based on a cell-laden fiber deposition technique that uses low-viscous solutions of biocompatible materials and cells and can form 3D, interconnected hydrogel fiber grids with high fidelity and reproducibility. The described method uses calcium-alginate as sacrificial templating polymer during the 3D printing process, and produces methacrylated gelatin cell-laden constructs with features in the order of 100 micrometer.We used this technology to produce 3D pre-vascular networks to be used as scaffold for a second, post-seeded cellular type. Endothelial cells (HUVECs) have been 3D printed in interconnected fiber meshes and spread and matured in tubular structures. Cardiomyocytes have been seeded on top of the endothelial network, giving rise to a pre-vascularized, 3D cellular construct that showed strong spontaneous beating behavior. This methodology, that combines bioprinting and scaffold-based approaches, can represent a new paradigm for the in vitro vascularization of 3D tissues.
Paola Brun
University of Padova
Italy
Title: Increasing concentration of self-assembling peptides finely tunes properties of electrospun scaffolds and improves osteoblast cells adhesion
Time : 11:20-11:40
Biography:
Paola Brun is an Assistant Professor at the Department of Molecular Medicine, University of Padova. She obtained her Ph.D. in 2005. Her scientific research activity covers a range of multidisciplinary topics mainly focused on cell biology, bioengineering and molecular medicine. She actively collaborates with the Departments of Industrial Engineering and Physics to find out innovative materials and techniques able to promote proliferation and differentiation of different cell populations. She has published 55 papers in peer-reviewed journals and she is an editorial board member of seven scientific journals.
Abstract:
Hydrogels are water-swollen insoluble polymeric networks, structurally similar to the extracellular matrix of many tissues. We previously reported that hydrogels tailored using particular physiological fibrils of self-assembling peptides (SAPs, e.g. EAK, RGD-EAK, EAbuK) specifically support adhesion and proliferation of osteoblast cells (Acta Biomaterialia, 2011). However, since SAPs hydrogels are characterized by relatively modest moduli (up to 10 kPa), they are hardly applicable as scaffolds for the treatment of bony defects. In the effort to optimize the extracellular matrix mimetic biocompatibility and stiffness, in this study different SAPs at increasing concentrations were electrospun with poly-É›-caprolactone (PCL). By electron microscopy, FT-IR, XPS, and contact angle measurements we found that the percentages of peptides embedded in the scaffolds directly correlates with the diameters of the electrospun fibers and the hydrophilicity of the matrix surfaces. Moreover the enrichment of PCL with SAPs at different concentrations resulted in different trends in tensile modulus, stress and strain. Biological assays reported that matrices characterized by increased hydrophilicity and proper fiber diameter promoted human osteoblast cells adhesion, supported calcium deposition and increased osteoblast-related genes expression. Thus, 10% RGD-EAK motif induced expression of integrin-binding sialoprotein, osteonectin, and runt-related transcription factor 2 mRNA specific transcript levels as determined by quantitative RT-PCR. In conclusion, increasing concentrations of SAPs modulated the physical properties of PCL scaffolds and in particular rectified the softness of SAPs hydrogels.
Ignazio Castagliuolo
University of Padova
Italy
Title: Hydrogel scaffolds in tissue engineering of the enteric nervous system
Time : 11:40-12:00
Biography:
Ignazio Castagliuolo is an Associate Professor at the Department of Molecular Medicine, University of Padova. He graduated in 1987 in Medicine. He spent 7 years at Boston University and Harvard Medical Center in Boston performing scientific research in the field of Gastrointestinal inflammation. Then he moved back to Padua were he has performed research mainly focused on cell biology, bioengineering and molecular medicine. He actively collaborates with the Departments of Industrial Engineering of University of Padua to develop innovative scaffolds to support colonization and differentiation of different cell populations. He has published 150 papers in peer-reviewed journals and he is an active reviewer for several scientific journals.
Abstract:
The enteric nervous system (ENS) is the complex network of neurons and glial cells embedded within the gut wall that autonomously regulates most gastrointestinal functions. In the ENS, distinct neuronal subpopulations encode excitatory and inhibitory neurotransmitters. Indeed, interruptions in the neuronal network and derangement in the architecture of the myenteric ganglia have been linked to gastrointestinal dismotility and visceral sensory disorders. Since the extracellular matrix proteins have been long known to provide cues for migration and regeneration of neuronal cells, in this study we designed novel hydrogel scaffolds for intestinal tissue engineering. Peptides mimicking the extracellular matrix of the ENS were synthesized by Fmoc chemistry and differently conjugated. Enteric neurons were isolated from the longitudinal muscle myenteric plexus of young mice and seeded onto different decorated scaffolds. After 5 days on culture, cell adhesion, neuronal marker expression and neurochemical coding were evaluated by confocal microscopy and quantitative RT-PCR. Hydrogels containing laminin adhesive motif greatly promoted adhesion of neuronal cells relatively to other cells of the myenteric ganglia. Arg-Gly-Asp (RGD) motif supported the adhesion of glial cells and also induced expression of mRNA specific for neurotrophin-3, a soluble factor involved in neurogenesis. Ionic-complementary peptide EAbuK fostered axonal outgrowth. Finally, conjugation of RGD and laminin enhanced the expression of choline acetyltransferase, thus inducing cholinergic neurons. In conclusion, our study demonstrated that neuronal subpopulations are modulated by varying the composition of scaffolds. These observations will be helpful in the designed of engineered scaffolds personalized for the treatment of defects of the ENS.
Valeria Chiono
Politecnico di Torino
Italy
Title: Biomimetic polyurethane scaffolds for a stem cell based therapy in myocardial regeneration
Time : 12:00-12:20
Biography:
Valeria Chiono has a Master Degree in Chemical Engineering summa cum laude (2001) and a Ph.D in Chemical and Materials Engineering from the University of Pisa (2006). She is currently associate professor at the Department of Mechanical and Aerospace Engineering, Politecnico di Torino. She is coordinator of the national FIRB Project “Bioartificial materials and biomimetic scaffolds for a stem cells-based therapy for myocardial regeneration” and of the project “Smart Injectable Drug-Delivery systems for Parkinson’s and Alzheimer’s Disease Treatment”. She has collaborated in several national and international project. She is currently author of 40 manuscripts in international journals and 4 patents.
Abstract:
Scaffolds for myocardial Tissue Engineering (TE) should display biomimetic properties respect to cardiac extracellular matrix (ECM), including elastomeric properties [1]. Cardiac regeneration depends on cardiac progenitor cells (CPCs) as well as the milieu in contact with them. Laminin-1 (LN1), typical of developing heart and over-expressed in pathological heart, promotes CPC proliferation and viability [1]. In this work, a thermoplastic polyurethane (PU) was synthesized from poly(ε-caprolactone) diol (Mn = 2000 Da), 1,4-budandiisocyanate and L-lysine ethyl ester dihydrochloride [2]. Bi-layered scaffolds with 0°/90° lay-down pattern were prepared by additive-manufacturing technique [2]. Functionalisation with LN1 or gelatin (G) was performed in two steps: 1) acrylic acid grafting/polymerization by Argon plasma treatment; 2) carbodiimide-mediated coupling of proteins. Scaffolds with mean fibre diameter of 1525 ïm and mean spacing of 5055 ïm were prepared. FITR-ATR analysis of protein-coated scaffolds showed higher intensity of the absorption bands at 3370 cm-1 (-OH and –NH stretching) and 1650 cm-1 (amide I). Contact angle decreased from 90° for PU to 60-65° after G- or LN1-grafting. XPS analysis confirmed acrylic acid grafting/polymerization and protein conjugation. Scaffolds were degraded in vitro by lipase (0.3 mg/ml) in 3 weeks. CPC proliferation on PU-LN1 scaffolds was higher than on PU and PU-G scaffolds, increasing from 8,18% on day 7 to 11,8% on day 14. LN1-functionalization stimulated CPC differentiation into cardiomyocytes and endothelial cells.
Marzena Zychowicz
Polish Academy of Sciences
Poland
Title: Reciprocal interactions between iPS-derived neural cell-seeded modified collagen scaffolds and nerve tissue
Time : 12:20-12:40
Biography:
Marzena Zychowicz, in 2012 in the Mossakowski Medical Research Centre, Polish Academy of Sciences, has accomplished her PhD in the field of neural stem cells and bioengineering surfaces. In the recent project she was interested in the standardization of the biomimetic microenvironment for the regenerative medicine application or toxicity testing. She is also involved in the project concerning mesenchymal stem cells and their application in the neurodegenerative diseases. She is co-author of 13 publications.
Abstract:
Induced pluripotent stem (iPS) cells are promising candidates as patient specific cell sources for transplantation or for modeling of diseases. By combining the human iPS-derived neural cells seeded on 3-D collagen scaffolds with dissociated or organotypic neural tissue we are attempting to create a biomimetic microenvironment for studying mutual interactions. To create the microstructured 3D niche, we have used type-I collagen scaffolds. Since collagen scaffolds can exhibit poor mechanical properties and rapid degradation in vivo, depending of cross linking , we have focused on modification of collagen physicochemical properties with improved mechanical and thermal stability without loss of its bioactivity. The human iPS cells, cultured in feeder free conditions, express markers of pluripotency (Oct, Sox2, Nanog), while after neural commitment and differentiation the cells exhibit nestin, GFAP, PDGFRα, β-tubulin III, Map-2, Dcx, GalC. The differentiated population of cells was seeded onto collagen scaffolds and co-cultured with rat organotypic spinal cord (OSC) slices or dissociated dorsal root ganglion (DRG) neurons. The collagen scaffold alone promoted neurite outgrowth from dissociated DRG neurons and when seeded with iPS-derived cells supported migration of cells from the OSC, however no migration of IPS-derived cells from the scaffold to the OSC could be detected. Such a biomimetic environment could be applied in regenerative medicine for testing directed migration from the nerve tissue into the seeded collagen scaffold.
- Track 1: Regeneration & Therapeutics
Location: Rome, Italy
Chair
Todd K. Rosengart
Baylor College of Medicine
USA
Co-Chair
Kristin Comella
Bioheart Inc.
USA
Session Introduction
Todd K. Rosengart
Baylor College of Medicine
USA
Title: Myocardial regeneration via cellular reprogramming
Biography:
Todd K. Rosengart is Professor and the DeBakey-Bard Chair of the Michael E. DeBakey Department of Surgery at the Baylor Medical College in Houston, TX. He received his MD degree through the Northwestern University Honors Program, obtained his general surgical training at New York University and served his cardiothoracic surgery fellowship at The New York Hospital. He was a Medical Staff Fellow in the Surgery Branch of the National Heart, Lung and Blood Institute. He holds twelve U.S. patents and is the author of over 150 peer-reviewed articles, editorials, reviews and book chapters. As a physician-entrepreneur, Dr. Rosengart is co-founder of Vitals.com and XyloCor Therapeutics.
Abstract:
Congestive heart failure (CHF), typically occurring as a result of myocardial infarction, is the leading cause of cardiac mortality in the West, afflicting over 5 million individuals and costing over $10 billion in medical care in the US alone. Cardiac stem cell therapy, which theoretically repopulates otherwise permanently scarred myocardium with contractile cells, has offered some promise in clinical trials. This strategy remains inherently constrained, however, by the hurdles of achieving adequate exogenous implant survival and integration into host myocardium. Potential risks of induced pluripotent stem (iPS) cell tumorigencity and immunogenicity may likewise limit these as stem cell implants. The recent discovery that “induced cardiomyocyte” (iCM) cells could be generated directly from somatic cells, for example using cocktails of transcription factors such as GATA4, MEF2c and TBX5, offers the remarkable new possibility of bypassing stem cell staging completely in the generation of autologous cardiomyocytes from cardiac fibroblasts. “Cellular reprogramming” likewise presents the intriguing possibility of converting scar fibroblasts in situ into functional iCMs, obviating entirely the challenges of exogenous cell implantation into a host myocardium. By converting scar directly back into functional myocardium, this transformational advance would eliminate the challenges of exogenous stem cell delivery that have slowed this field since its inception. While new evidence suggests that human cells may be more resistant to reprogramming than those transdifferentiated in small animal models, a number of methodologies are being developed to enable reprogramming even of human cells. Taken together, these new studies suggest an entirely new paradigm in CHF treatment that could potentially replace conventional therapies for end stage heart disease, include transplant and artificial heart implant strategies.
Kristin Comella
Bioheart Inc.
USA
Title: Adipose stem cells in vasculogenesis and angiogenesis
Time : 10:55-11:15
Biography:
Comella has over 15 years’ experience in corporate entities with expertise in regenerative medicine. She was recently named number 24 according to Terrapin’s list of the Top 50 Global Stem Cell Influencers. Ms. Comella has pioneered a variety of stem cell therapies including cord blood derived cells, bone marrow cells, muscle cells and adipose cells for use in many different applications. She has developed a wide range of regenerative products and techniques that have been successfully implemented into the clinic. She also led the team that gained the first ever FDA approval for clinical trials using a combined cell and gene therapy product. Ms. Comella has been a member of the Bioheart Inc. senior management team since 2004 and is currently serving as the Chief Scientific Officer and board member. Bioheart is a publically traded company focusing on the discovery, development and commercialization of autologous cell therapies for the treatment of degenerative diseases. Since joining Bioheart, she has played a major role in managing the product development, manufacturing and quality systems of cellular products. In addition, Ms. Comella is currently and actively serving on multiple boards in the stem cell arena. She was co-founder and Chief Executive Officer of Stemlogix, LLC for veterinary medicine. Ms. Comella has years of cell culturing experience including building and managing the stem cell laboratory at Tulane University's Center for Gene Therapy. Previously, she worked as a research engineer for Osiris Therapeutics developing stem cell therapies for osteoarthritis. Ms. Comella holds an M.S. in Chemical Engineering from The Ohio State University and a B.S. in Chemical Engineering from the University of South Florida.
Abstract:
The ready accessibility of adipose stem cells (ASC) make them a feasible and attractive form of autologous cell therapy requiring either no ex vivo expansion or relatively limited expansion. Our team has been working with ASCs for approximately 8 years. We have studied the effects of these cells in multiple indications in both animals and humans. ASCs are known to secrete angiogenic and anti-apoptotic factors which can increase tissue perfusion and limit ischemic tissue damage in several circumstances, including skeletal muscle, myocardial, and cerebral ischemia, as well as in cutaneous wound healing. ASCs may also be effective in orthopedic indications as they possess the potential to differentiate into bone and cartilage. This lecture will present the data that has been collected from thousands of patient treatments and discuss the potential of the cells in future applications.
Hyo Ihl Chang
Korea University
Republic of Korea
Title: Functions of natural pigments on gastric ulcer and cancer
Time : 11:15-11:35
Biography:
Hyo-Ihl Chang has completed his PhD in 1987 from North Carolina State University. He has been Professor of College of Life Sciences and Biotechnology, Korea University since 1988. He was a Dean of College of Life Sciences and Biotechnology, Korea University. Also he is a President of the Korean Society for Microbiology and Biotechnology in 2014. He has published more than 80 papers in reputed journals.
Abstract:
The Natural pigments have many applications in inflammatory, and oxidative related damage as well as in cancer chemotherapy. Recently, precise cellular roles of natural pigments, such as modulator of key cellular signaling pathway on variety diseases, are elucidated. On based on antioxidant, anthocyanins reduced naproxen-induced gastric ulcer. Anthocyanins reduced the level of lipid peroxidation and increased the level of the antioxidant enzymes. Anthocyanins increased the expression of Nuclear factor E2-related factor 2 (Nrf2) which is transactivator for cellular defense genes. Interestingly, anthocyanins induced gastrointestinal-glutathione peroxidase expression via Nrf2 that bind to regions of antioxidant response element (ARE) in GI-GPX promoter. Otherwise, Shikonin, and genipin stimulates production reactive oxygen species (ROS) in gastric cancer cells. They induced apoptotic cell death in gastric cancer cells in a caspase dependent manner. They also induced cell cycle arrest at G2/M phase via regulation of p21 by early growth response1 (Egr1). The p21 contains promoter region of Egr1 binding motif. Transient expression of Egr1 in AGS cells enhanced shikonin and genipin-induced p21 promoter activity, whereas suppression of Egr1 expression by small interfering RNA attenuated the ability of shikonin and genipin induced p21 promoter activity. Anthocyanins improve gastric ulceration through Nrf2 associated with antioxidant enzymes, such as GI-Gpx pathways. And, shikonin and genipin induced cell damage in AGS cells through the Egr1/p21 pathways.
Ana Maria Blanco Martinez
Federal University of Rio de Janeiro
Brasil
Title: Strategies to improve regeneration after peripheral nervous system trauma
Time : 11:35-11:55
Biography:
Ana Maria Blanco Martinez has completed her PhD at the age of 29 years from University of London, England, and postdoctoral studies from Rochester University, USA. She is an Associate Professor at the Federal University of Rio de Janeiro, Brasil, since 1985. She has published more than 100 papers in reputed journals and serving as an editorial board member of “Restorative Neurology and Neuroscience” and “International Journal of Neuropathology”. She has supervised 26 and 16 Master and PhD students, respectively, all in the field of Peripheral and Central Nervous System trauma.
Abstract:
Strategies aiming to enhance peripheral nervous tissue (PNS) regeneration after traumatic lesions are socially important because this condition is usually associated with permanent disabilities and low quality of life. Even in the PNS where axon regeneration is a well known process, only about 10% of adults who receive a surgical repair display full functional recovery. To evaluate strategies to improve regeneration, we have established models of trauma in the peripheral nervous system. In general, our projects are preclinical studies, in which we test a regenerative strategy. Our strategies include transection and tubulization with or without cell grafts and transection and tubulization with or without VEGF/GCSF gene therapy. Each one of these strategies is not capable of giving a significant result by itself, therefore there is a need for additional interventions that would have synergistic effects and maximize repair and functional outcomes. For that we use a combination of therapies, such as cell and gene therapy and rehabilitation procedures (treadmill training, for example). The results of our strategies are compared with autografts, the gold standard technique used by surgeons to repair a nerve lesion with tissue loss in humans. The tubulization technique mimics a lesion with tissue loss and is a manner to challenge the nerve to regenerate. The methods used to assess nerve regeneration and functional improvements include: Sciatic Functional Index, grasping test, eletroneuromiography, light and electron microscopy (transmission and scanning), immunohistochemistry, and enzyme activity of muscles. Our results clearly show that the strategies used are capable of enhancing peripheral nerve regeneration in terms of morphology and function.
Karin Schütze
CellTool GmbH
Germany
Title: Quality assurance for cell based therapeutics in 2D and 3D products
Time : 11:55-12:15
Biography:
Schuetze is a biologist and expert in non-contact cell handling and label-free cell analysis using innovative photonic technologies. She did her PhD work at the Institute for Applied Physical Chemistry in Heidelberg and her postdoc research at the University of California in Berkeley where she assembled her first Optical Trap. In 1993 she and her husband founded the PALM company, which was successfully sold to Zeiss. In 2008, they both founded their second company, the CellTool GmbH, where they develop Raman microscope systems for biomedical applications, providing physicians and biologists with a label-free and non-invasive tool for cell analysis.
Abstract:
In modern biomedicine there is an increasing demand for alternative, marker-free and non-destructive cell recognition and characterization methods. Raman Spectroscopy (RS) is a technology increasingly used for biomedical applications like identification of specific cell types. RS reveals detailed information on the metabolomic state of cells and tissues with the advantage that there is no need of labeling procedures. Here we demonstrate the feasibility of RS for quality control of 2D/3D cell based therapeutics exemplified for blood products and engineered human skin samples. In a first application RS was used to monitor erythrocytes and thrombocytes in donor blood over a time period of 30 and 8 days, respectively. Furthermore, Raman spectra were taken to identify and analyze fibroblasts, keratinocytes and melanocytes in a 3D human skin graft model. Raman Spectroscopy enabled quality control of blood products by allowing discrimination between young and old erythrocyte and thrombocyte samples. Thereby, differences found amongst the fresh and older samples were mainly associated with cells death. In 3D skin model it was possible to discriminate between different cell types and determine their allocation, also in a penetration depth up to 200µm, using RS. These two examples show that RS is a photonic marker for gentle yet highly specific detection and analysis of cells in biomedical products. It provides information about the entire metabolome of single cells even in a matrix setup with a depth of 200µm. Most importantly, RS can be used for quality assessment of cell cultures or engineered tissue without impairing cell viability.
Vicky Yamamoto
Keck School of Medicine of USC
USA
Title: Protein phosphatase/Smek complex negatively regulate Par3 and promote neuronal differentiation.
Time : 12:15-12:35
Biography:
Vicky Yamamoto is a cancer scientist at Keck School of Medicine of USC in the Department of Head and Neck Surgery with more than 10 years of research experience ranging from developmental neurobiology and stem cell to molecular targeted therapy. Prior to joining Keck School of Medicine of USC, Yamamoto worked at Mount St. Mary’s College, The Scripps Research Institute, the Cedars-Sinai Medical Center, and California Institute of Technology. Yamamoto received a PhD in biochemistry and molecular biology from Keck School of Medicine of USC. She has significant teaching experience and mentored numerous students.
Abstract:
Neural progenitor cells (NPCs) are self-renewing multipotent cells that are capable of differentiating into neurons and glial cells. Mechanisms that control the fate decisions of NPCs are not well understood. SMEK homolog 1, suppressor of mek1 (Smek-1) is a regulatory subunit of the serine/threonine protein phosphatase PP4. We found that Smek-1 is expressed in NPCs, promotes neuronal differentiation, and suppresses the proliferation of NPCs. Mass spectrometry analysis identified one of Smek-1’s binding partners, Par3. Par3, a cell polarity protein, is a negative regulator of neuronal differentiation. We demonstrate that Par3 is a substrate of Smek-1 and Smek-1 can negatively regulate its activity in neurogenesis. Interestingly, Smek-1 is expressed mainly in the nucleus but is exclusively localized in the cytoplasm during mitosis. We show that the cytoplasmic Smek-1 can interact with cytoplasmic Par3 and thereby mediate de-phosphorylation by the catalytic subunit PP4C. Collectively, our results show that the PP4/Smek-1 complex is likely a key regulator of neurogenesis.
Aysegul Batioglu-Karaaltin
Istanbul University Cerrahpasa School of Medicine
Turkey
Title: New approach for demaged fasial nerve regeneration based on stem cells
Time : 13:20-13:40
Biography:
Aysegul Batioglu-Karaaltin has completed her MD degree at the age of 24 years from Hacettepe University and her residency at the Department of ENT and Head and Neck at Ministry of Health Education and Research Hospital. She is working at Department of Otolaryngology Head and Neck Surgery, Istanbul University Cerrahpasa School of Medicine. She is carrying out more then 10 research about regenerative medicine and tissue engineering in the Head and Neck field. She has published more than 15 papers in reputed journals.
Abstract:
In facial nerve damage especially full-cut paralysis which can occur due to several conditions (congenital, tumor, iatrogenic-operation dependent), there is no spontaneous regeneration. Hence, several treatments are used to attempt regeneration of facial nerves such as end-to-end anastomosis, direct coaptation, artificial nerve guide conduit, autologous or decellularized grafts, anastomosis of distal facial nerve to the proximal cranial nerves. Since these techniques used have some disadvantages such as the limitation in source of Schwann cells, surgery intervention resulting in functional loss and difficulties in preparing nerve grafts, there is need for effective alternative techniques to be developed. Though the experiments associated with peripheral nerve injury in which adipose tissue, bone marrow, and hippocampal neural stem cell are used have obtained positive results, The usage of stem cells isolated from the olfactory region that is the only attainable neuronal stem cell in nerve regeneration if not the periferic nerve regeneration may provides an alternative treatment technique for facial nerve injury. Our group were used OSCs that isolated from olfactory mucosa tissue of human participants. 2mm excision was performed on right facial nerve for all rats. Reconstruction was performed with conduit in first group; conduit and phosphate-buffer saline in second group; conduit and labeled OSCs in third group. Rats were followed for whisker-movements and electroneuronography (ENoG) analyses. Recovery for third group were significantly different from first and second group. This study suggests that OSC may be used as a potent cellular therapy for accelerating regeneration of peripheral nerve injuries. A new study was designed for compared the effectiveness of adipose tissue, bone marrow and olfactory stem cells for facial nerve regeneration.
Ruiz-Navarro F
Austrian Society of Regenerative Medicine
Austria
Title: Neuron Point-Of-Care Stem cell therapy: Intrathecal transplant of autologous bone marrow-derived stem cells in patients with cerebral palsy. Preliminary Results
Time : 13:40-14:00
Biography:
Francisco Ruiz-Navarro (M.D) is Research Associate in the Austrian Society of Regenerative Medicine. Before, he was working as researcher at the Mexican Institute for Neurology and Neurosurgery in the Cerebrovascular Department focused in multi-centric population studies with Hispanic stroke patients. He was research assistant in the Center for Research and Advanced Studies of the National Polytechnic institute (CINVESTAV) in Mexico City at the Brain Bank and Physiology, Biology and Neuroscience Department. He was Research Assistant in La Raza Medical Center in Mexico City in the pediatric nephrology department. He obtained the Medical Degree in Anahuac University in Mexico City and became USMLE board certified in United States of America with outstanding grades. During his carrier he had been attending physician in different Mexican hospitals in Mexico City as part of general medicine, anesthesiology and neurosurgical teams. He performed clinical rotations at Jackson´s memorial Hospital
Abstract:
Background: Cerebral palsy (CP) encompasses the largest group of childhood movement disorders, the patterns and severity varies widely. Today, the management focuses only on a rehabilitation therapy that tries to secure the functions remained and prevents complications. However the treatments are not aimed to cure the disease. Stem cells (SCs) transplant via intrathecal is a new approach to the disease. Method: Our aim was to performed a pilot study under the condition of unproven treatment on clinical practice to assessed the safety and efficacy of Neuron Point-of-care Stem cell Therapy (N-POCST), an ambulatory procedure of autologous bone marrow derived SCs (BM-SCs) harvested from the posterior superior iliac crest undergo an on-site cell separation for intrathecal infusion via lumbar puncture. Results: 82 patients were treated in a period of 28 months, with a follow-up after 6 months. They had a mean age of 6,2 years old and male predominance (65,9%). Our preliminary results show that: A. No patient had any major side effects, B. Only 20% presented mild headache due to LP, C. 53% of the patients had an improvement in spasticity, D. 61% improved the coordination abilities, 23% improved the motor function, 15% improved the speech, 23% reduced the number of convulsive events with the same doses or less doses of anti-convulsive medication and 94% of the patients report a subjective general improvement. Conclusions: These results support previous worldwide publications that described the safety and effectiveness of autologous BM-SCs transplant for patients with CP.
- Track 4: Tissue Regeneration
Track 5: Materials & Designs for Tissue Engineering
Chair
Shiva Akbarzadeh
Monash University
Australia
Co-Chair
Christopher L. Antos
Technische Universität Dresden
Germany
Session Introduction
Shiva Akbarzadeh
Monash University
Australia
Title: Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold
Time : 14:00-14:20
Biography:
Shiva Akbarzadeh obtained her PhD from the Ludwig Institute for Cancer Research, University of Melbourne in 2002. She has over 13 years of experience in cell biology and signalling in a variety of systems. In 2009 she established the Skin Culture Laboratory at the Victorian Adult Burns Service- the Alfred, focusing on translational research in skin regeneration. She leads the team responsible for manufacturing Cultured Epithelial Autograft for cellular based therapy of major burns and research in developing novel skin substitutes for clinical application. In addition, she provides lab based training on skin tissue engineering for scientists and surgeons from developing countries.
Abstract:
Tissue-engineered composite skin is a promising therapy for treatment of chronic and acute wounds, including burns. Providing the wound bed with a dermal scaffold populated by autologous dermal and epidermal cellular components can further entice host cell infiltration and vascularisation to achieve permanent wound closure in a single-stage. However, the high porosity and the lack of a supportive basement membrane in most commercially available dermal scaffolds hinders organized keratinocyte proliferation and stratification in vitro and may delay re-epithelization in vivo. The objective of this study was to develop a method to enable the in vitro production of a human skin equivalent (HSE) that included a porous scaffold and dermal and epidermal cells expanded ex vivo, with the potential to be used for definitive treatment of skin defects in a single procedure. A collagen- glycosaminoglycan dermal scaffold (Integra®) was populated with adult fibroblasts. A near normal skin architecture was achieved by addition of coagulated human plasma to the fibroblast-populated scaffold prior to seeding cultured keratinocytes. This resulted in reducing scaffold pore size and improving contact surfaces. Skin architecture and basement membrane formation was further improved by the addition of aprotinin (a serine protease inhibitor) to the culture media to inhibit premature clot digestion. Histological assessment of the novel HSE revealed expression of keratin 14 and keratin 10 similar to native skin, with a multi-layered neo-epidermis morphologically comparable to human skin. Furthermore, deposition of collagen IV and laminin-511 were detected by immunofluorescence, indicating the formation of a continuous basement membrane at the dermal-epidermal junction. The proposed method was efficient in producing an in vitro near native HSE using the chosen “off the shelf” porous scaffold (Integra®). The same principles and promising outcomes should be applicable to other biodegradable porous scaffolds, combined with autologous cells, for use in wound treatment.
Shuanhu Zhou
Brigham and Women Hospital Harvard Medical School
USA
Title: Adult stem cell aging and skeletal regenerative medicine
Time : 14:20-14:40
Biography:
Shuanhu Zhou, Ph.D. is an Associate Scientist at Brigham and Women's Hospital and Assistant Professor at Harvard Medical School. Dr. Zhou received his B.S. degree from Sichuan University, China and Ph.D. from Hebrew University of Jerusalem, Israel, and had his postdoctoral training in Harvard University, USA. Dr. Zhou has published more than 60 peer-reviewed papers and book chapters, and served as an evaluation expert for journals and organizations. He received several awards, including 2001 Rector Award of Hebrew University for excellent Ph.D. student and 2006 John Haddad Young Investigator Award from American Society for Bone and Mineral Research.
Abstract:
Regenerative potentials of many tissues are determined by the presence and functionality of adult stem and progenitor cells, which respond to exogenous cues to initiate tissue repair when needed. However, in many instances, resident adult stem/precursor cells suffer declining activity in response to aging, leading to reduced repair potential and chronic degenerative diseases. Human mesenchymal stem cells (MSCs) have been shown to be precursors of several different cellular lineages, including cartilage-forming chondrocytes and bone-forming osteoblasts. We reported that human marrow-derived MSCs show reproducible declines in proliferative and osteoblast potential with the age of the subject from whom the cells were obtained. Chemicals or drugs, which modulating endogenous MSCs, e.g. PTH and Vitamin D, have the potential to rejuvenate aged MSCs and to prevent or restore skeletal tissue degeneration and loss in the aging population. Tissue engineering approaches with adult stem cells have been applied clinically in cartilage and bone regeneration; however, more basic and translational researches as well as interactions between bench scientists and bedside clinicians are needed before tissue engineered biological organ replacement approaches with adult stem cells become standard clinical practice.
Lucie Bacakova
Institute of Physiology
Czech Republic
Title: Adhesion, growth and osteogenic differentiation of human adipose stem cells isolated by liposuction under low and high negative pressure
Time : 14:40-15:00
Biography:
Lucie Bacakova, MD, PhD, Assoc. Prof. has graduated from the Faculty of General Medicine, Charles University, Prague, Czechoslovakia in 1984. She has completed her Ph.D at the age of 32 years from the Czechoslovak Academy of Sciences, and became Associated Professor at the 2nd Medical Faculty, Charles University. She is the Head of the Department of Biomaterials and Tissue Engineering, Institute of Physiology, Academy of Sciences of the Czech Republic. She is a specialist for studies on cell-material interaction and vascular, bone and skin tissue engineering. She has published more than 150 papers in reputed journals (h-index 26).
Abstract:
Adipose-derived stem cells (ASCs) are promising for cell therapies and tissue engineering. Adipose tissue with these cells can be obtained by a relative non-invasive method, e.g. liposuction. The quality and quantity of ASCs can be influenced by the amount of negative pressure during liposuction. In this study, we focused on ASCs isolated from lipoaspirates taken from the same patient (a 43-year-old woman) under low negative pressure (-200 mmHg, LP) or high negative pressure (-700 mm Hg, HP). The ASCs were isolated by a method described by Estes et al. (Nat Protoc 5: 1294-311, 2010). Flow cytometry, performed in the 2nd passage, revealed that the cells contained markers typical for ASCs; only the population obtained under LP was more heterogeneous. The number of isolated ASCs and their subsequent proliferation activity in vitro was higher in cells obtained under HP. These differences persisted in passaged cells (tested up to 3 passages), after freezing and thawing of cells and also after seeding on Ti-Al-4V samples with various surface modifications (e.g., grinding, polishing, grit blasting, tarnishing), developed for construction of bone implants. However, when confluent ASCs were exposed to osteogenic medium (containing ascorbic acid, dihydroxyvitamin D3, dexamethasone, beta-glycerol phosphate and L-glutamine) for 5 days, the osteogenic cell differentiation, measured by intensity of fluorescence of collagen I, alkaline phosphatase and osteocalcin, was more pronounced in cells obtained under LP. Thus, ASCs obtained under both pressures have specific advantages, and their choice depends on their application, i.e. if their rapid growth or early differentiation is needed.
Christopher L. Antos
Technische Universität Dresden
Germany
Title: Calcineurin instructs when to stop regenerating once the correct size of zebrafish appendages is reached
Time : 15:00-15:20
Biography:
Christopher Antos, a Group Leader (Ph.D. Cell and Molecular Biology), who heads his laboratory at the DFG-Center for Regenerative Therapies Dresden, an Excellence Cluster at the Technische Universität Dresden. He got his Ph.D. at Southwestern Medical Center at Dallas, University of Texas (UTSW) and subsequently pursued a post-doctoral fellowship at the Max-Planck Institute for Developmental Biology in Tübingen, Germany, where he started researching regeneration biology using zebrafish as a model. Current research in Dr. Antos’s lab is focused on the molecular biology that regulates progenitor cell behavior during regeneration of zebrafish structures: principally, what is involved in inducing cells in the residual stump tissues to become progenitor cells and what is involved in instructing these progenitor cells to grow and pattern correctly. From this work, his lab uncovered that the previously uncharacterized molecule Smp is required for progenitor cell participation in regeneration and embryonic development is involved in the Wnt signal transduction by regulating β-catenin nuclear localization.
Abstract:
Animals that regenerate organs and appendages control the growth of stem and progenitor cells to reform lost structures to the same dimensions as the original structures. This proportional regeneration involves coordinating rapid allometric (disproportional) growth with the restoration of isometric (proportional) cell proliferation once the correct tissue dimensions are reached. It is unknown what executes this coordinated control. We show that the calcium-dependentphosphatase calcineurin regulates this control. Calcineurin inhibition results in continued allometric outgrowth of regenerating fins beyond their original dimensions. Congruent with these results, calcineurin activity is low when the rate of progenitor cell proliferation is highest, and its activity increases as the regeneration rate decreases. Furthermore, inhibition of calcineurin in uninjured adult fins switches isometric growth into allometric growth, demonstrating that calcineurin regulates appendage allometry. Previous results show that the rate of regenerative outgrowth is controlled by position along the proximodistal axis, but it is unknown what this positional control is. Our growth rate measurements and morphometric analysis of proximodistal asymmetry indicate that calcineurin inhibition shifts fin regeneration from a distal isometric growth program to an allometric proximal program. This shift is associated with the promotion of retinoic acid signaling, a signal transduction mechanism that affects positional information along the proximodistal axis. Furthermore, we provide evidence that calcineurin regulates potassium conductance via a potassium leak channel that has been shown to promote allometric growth. In summary, we identified a calcineurin-mediated mechanism that operates as a molecular switch between distal isometric growth and proximal allometric growth.
Urszula Stachewicz
AGH University of Science and Technology
Poland
Title: 3D imaging of cell interactions with nanofibers scaffolds
Time : 15:35-15:55
Biography:
Urszula Stachewicz is an assistant professor at AGH University of Science and Technology in Krakow, Poland, She is also working in the International Centre of Electron Microscopy for Materials Science. She received a doctorate from Delft University of Technology in the Netherlands and completed her postdoctoral study at the Queen Mary, University of London, UK. She worked in Philips Research Laboratories and the university spin-out company Nanoforce Technology Ltd. She is a peer reviewer for several funding agencies and journals and she is author of 20 peer reviewed publications.
Abstract:
The use of electrospun nanofibres for guided bone regeneration or bone scaffolds align with next-generation healthcare, especially as electrospun nanofibres are highlighted as being particularly effective in tissue engineering. However, optimization of the electrospinning process for cell growth and their interaction with nanofibre surfaces is yet to be determined. We develop a biodegradable electrospun nanofibre membrane for guided bone regeneration for bone scaffold applications. The interaction between osteoblasts and osteoblast-derived mineralized nodule formation on the nanofibre membrane is visualized using 3D imaging based on ‘slice-and-view’ combinations of ion beam and scanning electron microscopies (FIB-SEM). The presented 3D imaging technique therefore shows s a new approach in high resolution visualization the cell growth on electrospun nanofibers, and potentially other biomaterials, that will develop and design new biomaterials for a range of clinically important applications including orthopaedics of this work.
Paola Brun
University of Padova
Italy
Title: Hydrogel scaffolds in tissue engineering of the enteric nervous system.
Time : 15:55-16:15
Biography:
Paola Brun is an Assistant Professor at the Department of Molecular Medicine, University of Padova. She obtained her Ph.D. in 2005. Her scientific research activity covers a range of multidisciplinary topics mainly focused on cell biology, bioengineering and molecular medicine. She actively collaborates with the Departments of Industrial Engineering and Physics to find out innovative materials and techniques able to promote proliferation and differentiation of different cell populations. She has published 55 papers in peer-reviewed journals and she is an editorial board member of seven scientific journals.
Abstract:
The enteric nervous system (ENS) is the complex network of neurons and glial cells embedded within the gut wall that autonomously regulates most gastrointestinal functions. In the ENS, distinct neuronal subpopulations encode excitatory and inhibitory neurotransmitters. Indeed, interruptions in the neuronal network and derangement in the architecture of the myenteric ganglia have been linked to gastrointestinal dismotility and visceral sensory disorders. Since the extracellular matrix proteins have been long known to provide cues for migration and regeneration of neuronal cells, in this study we designed novel hydrogel scaffolds for intestinal tissue engineering. Peptides mimicking the extracellular matrix of the ENS were synthesized by Fmoc chemistry and differently conjugated. Enteric neurons were isolated from the longitudinal muscle myenteric plexus of young mice and seeded onto different decorated scaffolds. After 5 days on culture, cell adhesion, neuronal marker expression and neurochemical coding were evaluated by confocal microscopy and quantitative RT-PCR. Hydrogels containing laminin adhesive motif greatly promoted adhesion of neuronal cells relatively to other cells of the myenteric ganglia. Arg-Gly-Asp (RGD) motif supported the adhesion of glial cells and also induced expression of mRNA specific for neurotrophin-3, a soluble factor involved in neurogenesis. Ionic-complementary peptide EAbuK fostered axonal outgrowth. Finally, conjugation of RGD and laminin enhanced the expression of choline acetyltransferase, thus inducing cholinergic neurons. In conclusion, our study demonstrated that neuronal subpopulations are modulated by varying the composition of scaffolds. These observations will be helpful in the designed of engineered scaffolds personalized for the treatment of defects of the ENS.
Kara E. McCloskey
UC Merced
USA
Title: Vascularized cardiac tissue from induced pluripotent-derived cardiomyocytes and endothelial cells
Time : 16:15-16:35
Biography:
Kara E. McCloskey, PhD, is a Founding Associate Professor in the School of Engineering at the University of California, Merced. She received her BS and an MS in Chemical Engineering from The Ohio State University and her PhD through a joint program with Cleveland Clinic Foundation’s Biomedical Engineering Department and Ohio State University. She then completed her postdoctoral training in vascular stem cell and tissue engineering with Dr. Robert Nerem at the Georgia Institute of Technology. McCloskey is the founder and first chair of the Biological Engineering and Small-scale Technologies (BEST) graduate program at UC, Merced and serves as the university liaison for the UC Systemwide Bioengineering Multicampus Research Unit. Her research is in the field of cardiovascular tissue engineering with a specific focus on deriving functional cell products from stem cells. As a young investigator McCloskey earned a highly competitive $1.7 million New Faculty Award from the California Institute for Regenerative Medicine (CIRM) for studies towards developing cardiac tissue from stem cells, and another recent CIRM-funded Basic Biology award on directing specialized endothelial cell from stem cells. Dr. Kara McCloskey has over 14 years of experience in the area of endothelial cell (EC) fate from both human and mouse embryonic stem cells (ESC), and 9 years in cardiac fate, and now serves on the editorial board for the International Journal of Stem Cell Research & Therapy.
Abstract:
According to the American heart association, myocardial infarctions (MI) occur once every forty-four seconds in the United States. The chronic conditions that can develop from the damaged tissue requires the use of daily medications, pacemakers, and/or organ replacement in order to prevent congestive heart failure. Cardiovascular tissue engineering holds promising solutions to replace the need for whole heart transplants, and has made substantial progress towards repairing heart function after a MI. However, key challenges facing tissue engineering are 1) the source, 2) the packaging/delivery of cardiomyocytes (CM) in a manner that enables both cell survival and host integration, and vascularization of the new tissue. Our laboratory has developed a number of cell differentiation methods and engineering strategies addressing these key challenges in building functional heart tissue in vitro.
- Workshop on New frontiers in hard tissue regeneration: State of art
Location: Rome, Italy
Chair
Roberta Di Pietro
G. d’Annunzio University of Chieti-Pescara
Italy
Co-Chair
Ivo Kalajzic
University of Connecticut Health Center
USA
Session Introduction
Gianpaolo Papaccio
Seconda Università degli Studi Napoli
Italy
Title: Dental pulp stem cells: State of the art and suggestions for a true translation of research into therapy
Time : 13:30-13:50
Biography:
Gianpaolo Papaccio got the degree in Medicine cum Laude in 1979 and the Master Degree in Legal Medicine cum Laude in 1982, University of Naples Federico II, Italy. He worked as visiting scientist in USA- Washington University, Department of Biostructures, Seattle; in Germany, at the Department of Molecular Medicine, J. Liebig University of Giessen; in Switzerland at the Department of Oral Biology, University of Zurich. He currently works at the Second University of Naples as a Full Professor of Medical Histology and Embryology, School of Medicine. He joined the Editorial Board of Stem Cells, PLOs ONE, among others and he’s a Registered Referee for several Journals of Stem cells, Regenerative Medicine, Cell and cancer biology. He is author of more than 150 impacted scientific publications plus international e-book chapters and Italian textbooks.
Abstract:
Stem cells have the ability to rescue and/or repair injured tissue. In humans, it is possible to isolate different types of stem cells from the body. Among these, Dental Pulp Stem Cells (DPSCs) are relatively easily obtainable and exhibit high plasticity and multipotential capabilities. In particular, they represent a gold standard for neural-crest-derived bone reconstruction in humans and can be used for the repair of body defects in low-risk autologous therapeutic strategies. An electronic search was conducted on PubMed databases and supplemented with a manual study of relevant references. All research described in this review highlights that DPSCs are mesenchymal stem cells that could be used in clinical applications. Unfortunately, very few clinical trials have been reported. Major obstacles imposed on researchers are hindering the translation of potentially effective therapies to the clinic. Both researchers and regulatory institutions need to develop a new approach to this problem, drawing up a new policy for Good Manufacturing Practice (GMP) procedures. We strongly suggest that only general rules be standardized rather than everything. Importantly, this would not have an effect on the safety of patients, but may very well affect the results, which cannot be identical for all patients, due to physiological diversity in the biology of each patient. Alternatively, it would be important to study the role of specific molecules that recruit endogenous stem cells for tissue regeneration. In this way, the clinical use of stem cells could be successfully developed. DPSCs are mesenchymal stem cells that differentiate into different tissues, maintain their characteristics after cryopreservation, differentiate into bone-like tissues when loaded on scaffolds in animal models, and regenerate bone in human grafts. In summary, all data reported up to now should encourage the development of clinical procedures using DPSCs. Key words: adult stem cells; bone regeneration; human grafts, scaffold, tissue engineering, stem cell therapy
Ivo Kalajzic
University of Connecticut Health Center
USA
Title: αSMA expressing perivascular and satellite cells contribute to muscle heterotopic ossification
Time : 13:50-14:10
Biography:
Ivo Kalajzic has completed his Ph.D studies on Identification of stages of mesenchymal lineage differentiation. He is currently Associate professor at Dept. of Reconstructive Sciences at University of Connecticut Health Center. His laboratory focuses on studies on regulation of mesenchymal progenitor cell differentiation during bone fracture healing, and the role of notch signaling during repair. Other research projects within his group include the studies on progenitor cells within the muscle, and other tissues, including developing transplantation approaches to treat osteogenesis imperfecta. He has published more than 50 papers with focus on bone biology and orthopedics.
Abstract:
Heterotopic ossification is formation of bone in atypical locations including muscle. Heterotopic ossification generally occurs as a result of aberrant BMP signaling. However, further studies are required to identify BMP-responsive cells in the muscle and understand how they contribute to heterotopic ossification. Alpha smooth muscle actin (αSMA) is a marker of perivascular cells and mesenchymal progenitors that contribute to bone growth and fracture healing. We aimed to evaluate whether αSMA+ cells can contribute to bone formation during heterotopic ossification. To identify and trace cells we used αSMA promoter-driven inducible Cre (αSMACreERT2) combined with a Cre-activated tdTomato reporter Ai9 to generate SMA9 mice. Mature osteoblast/osteocytes were labeled with the Col2.3GFP reporter. To label muscle satellite cells we utilized Pax7CreERT2/Ai9 mice. Pax7Cre/Ai9 labels cells below the basal lamina consistent with a satellite cell phenotype, while SMA9 labels similarly localized cells, in addition to perivascular cells. Pax7Cre/Ai9+ satellite cells are CD45/CD31-, Sca1- but SM/C2.6+. The SMA9+ cells are CD45/CD31- and ~60% express satellite cell marker SM/C2.6. 5% express MSC markers Sca1 and PDGFRα, and a larger proportion express PDGFRβ. During BMP2-induced heterotopic ossification, SMA9+ cells comprised 28% chondrocytes and 44% of Col2.3GFP+ osteoblasts, while Pax7Cre/Ai9+ satellite cells labeled only muscle fibers. Culture of sorted cells indicated that SMA9+ and Pax7Cre/Ai9+ cells differentiated into myotubes while the negative cells do not. SMA9+ cultured cells showed the highest upregulation of osteogenic gene expression after culture in the presence of BMP2. To clarify which subset of SMA9 cells possess osteogenic potential, we subdivided SMA9+ cells based on expression of SM/C2.6 and evaluated their ability to differentiate into osteoblasts after transplantation into muscle with BMP2. We observed osteogenic differentiation of SMA9/SM/C2.6+ cells suggesting that once the satellite cell population is removed from its niche they can exhibit osteogenic potential. In conclusion, αSMACreERT2 labels a population of mesenchymal progenitor cells in the muscle that contribute to heterotopic ossification induced by BMP2. Our data indicate that the perivascular fraction of cells is responsible for heterotopic ossification, however, muscle satellite cells are capable of osteogenic differentiation after removal from their niche. Further study of these cells may indicate mechanisms by which pathogenic heterotopic ossification occurs.
Gunter Lepperdinger
University Salzburg
Austria
Title: Triggering osteocytogenesis in vitro and in vivo
Time : 14:10-14:30
Biography:
Gunter Lepperdinger holds a PhD in Biochemistry, is a developmental biologist by training and has worked in Biogerontology with specialization on Stem Cell Ageing since 2002 at ‘The Inst. for Biomedical Aging Research’ in Innsbruck, Austria. He recently moved to Salzburg University to chair the Stem Cell Biology and Healthy Longevity Research Unit. He is Editor to Karger’s ‘Gerontology’, furthermore serving as an Editorial Board Member for many international scientific journals and engaged in European research projects on Aeging (e.g. MOPACT) and Tissue Engineering (e.g. VASCUBone).
Abstract:
Mesenchymal stromal cells (MSC) can be reliably isolated from human connective tissues. Naïve MSC can be culture-expanded and induced to form osteoblasts in a standardized manner. Further differentiation to gain osteocytes is however hard to achieve; also knowledge concerning the molecular mechanisms which specifically trigger and convey osteocyte differentiation is scarce. Through a serendipitous observation while investigating the role of hyaluronan in MSC biology, we were able to unveil molecular mechanisms of osteocytogenesis. These involve specific stress response mechanisms regarding detoxification by means of glucuronidation, upregulation of O-glycosylation which grossly changes the cellular phospho-proteome, induction of stress granule formation and concomitant mRNA-binding thereby yielding differential regulation of translation of osteogenic transcription factors and the formation of poly-ADP–ribose (PAR) in the cytoplasm otherwise known to coordinate the nuclear DNA repair machinery. We could further show in 3D-mesenspheres that provoking this particular stress response pathway in the presence of osteogenic inducers promotes the calcification of MSC-derived microorganoids resulting in the formation of trabecular-like hard tissue structure bearing osteoblastic linings which engulf marrow-like stroma. These observations could be further corroborated in histological sections of human bone in particular showing the presence of stress granule and PAR deposition in osteocytes residing in compact bone. We could further show accelerated healing of a bone defect model in rat calvaria by provoking this stress response pathway.
Roberta Di Pietro
G. d’Annunzio University of Chieti-Pescara
Italy
Title: Potential of human amniotic fluid stem cells (AFSC) and natural scaffolds in bone regeneration and repair
Time : 14:30-14:50
Biography:
Roberta Di Pietro got the degree in Medicine cum Laude in 1985 and the PhD in Sports Medicine cum Laude in 1988, University of Chieti, Italy. She worked as a visiting scientist in UK at the Biochemistry Department, AFRC, Cambridge; in USA at the Pathology Department, USUHS, Bethesda, and at the Institute of Human Virology, University of Maryland, Baltimore. She currently works at the University of Chieti as a Full Professor of Histology. She joined the Editorial Board of Current Pharmaceutical Design as an Executive Guest Editor and she was recognized as a Registered Referee for Archives of Ophthalmological Reviews and Reproductive Biology and Endocrinology. She is now author of 171 scientific publications plus international e-book chapters and Italian textbooks.
Abstract:
Every year over 2 millions bone replacement procedures requiring the use of bone graft materials are performed worldwide. This makes bone second only to blood in the list of transplanted materials. Tissue engineering strategies are a very promising option for the healing of critical-size bone defects. To this aim, various classes of natural scaffolds (including collagen and hyaluronic acid-based hydrogel) have been used in the human clinical setting. However, cell-free scaffolds are inadequate to repair large bone defects. Recent studies have focused on the use of stem cells rather than mature differentiated cell types due to their expansion potential and ease of access. In particular, human AFSCs represent an attractive cell model for transplantation therapy due to the lack of significant immunogenicity, tumorigenicity and ethical issues. Although AFSCs have been investigated for bone repair, the cellular distribution and post-implantation viability remain key issues. Our research group explored whether AFSCs could improve bone healing in a rat model of large bone defect. For in vivo experiments male rats were injured at the femoral diaphysis in order to produce a 5-7 mm diameter full thickness bone gap. Human AFSCs were transplanted in vivo at the lesion sites after being loaded on a HA (natural nanocrystalline carbonated hydroxyapatite-Orthoss®) scaffold. Both gross anatomy and histological observations revealed greater bone tissue remodelling and regenerative response in rat specimens treated with HA scaffold supplemented with AFSCs. Our findings support previous studies showing AFSCs angiogenic effects and indicate the potential of these cells in bone reconstruction.
- Track 2: Stem Cells-Tools to Battle Cancer
Track 3: Bone and Cartilage Tissue Engineering
Location: Rome, Italy
Chair
Norman L. Lehman
The Ohio State University
USA
Co-Chair
Mei Wan
Johns Hopkins University School of Medicine
USA
Session Introduction
Ching-Chang Ko
University of North Carolina- Chapel Hill
USA
Title: Introducing a polydopamine-laced hydroxyapatite-gelatin nanocomposite for customized scaffolding in bone tissue engineering
Time : 16:35-16:55
Biography:
Ching-Chang Ko, is Professor of Orthodontics in the School of Dentistry at the University of North Carolina (UNC). He received his PhD in bioengineering and biomaterials from the University of Michigan. Since 2014, he has served as Program Director and Vice Chair of the Department of Orthodontics at UNC, and in 2014 was name Hale Distinguished Professor. He has contributed more than 100 publications to the scientific literature. His research is funded by NIH to develop new biomaterials for bone tissue engineering and to study orthodontic biomechanics. He is a member of AADR, AAO, ORS, ASME, ASCER, and Biomaterials.
Abstract:
Polydopamine has recently been reported to improve osteoblast adhesion and proliferation in vitro. We also found that preosteoblasts express functional dopamine receptors that could regulate cell proliferation and mineralization via the dopamine monomer released from the polydopamine-laced hydroxyapatite-gelatin nanocompsoite (PDHG). Thus, PDHG not only possesses enhanced physical properties of hydroxyapatite-gelatin bioceramics, but also can facilitate bone tissue engineering. The interconnection of polydopamine network within PDHG increases both compressive and tensile strength, approaching to that of natural cortical bone. The present study determined dose ranges for dopaminergic effect in osteogenesis, which will be used to optimize the composition of PDHG. In vitro tests of MC3T3-E1 culture used soluble dopamine drops and the real-time polymerase chain reaction (qPCR). At the end of culture, the expression of osteogenic genes was quantified with and without addition of dopamine antagonists. In a pilot in vivo test, a thermodifferential process in conjunction with the indirect scaffolding technique was applied to fabricate a customize PDHG prosthesis for jaw bone regeneration in rats. The 3mm resection of rat mandible was created at the premolar and molar region. The PDHG porous prosthesis was used to fix the osteomized mandible. The result showed that osteogenic gene expression was significantly greater in the dopamine-treated than control group (p<0.05), but inhibited by dopamine receptor antagonists. The rats underwent PDHG reconstruction behaver normal chewing and life activities; histological retrieval is currently underway.
Norman L. Lehman
The Ohio State University
USA
Title: Glioma tumor stem cell based models for evaluation of anti-glioma therapeutics.
Time : 16:55-17:15
Biography:
Lehman has completed his MD and PhD from the University of Southern California and postdoctoral studies from Stanford University School of Medicine. He is currently the Director of Neuropathology at the Ohio State University. He has published more than 50 per reviewed papers and has served as an editorial board member for Acta Neuropathologica and is an Associate Editor for the Journal of Neuropathology and Experimental Neurology.
Abstract:
Experimental models for anti-glioma therapies are limited by two important factors. Appropriate modeling of the blood-brain tumor barrier and modeling of the tumor stem cell component. The latter is important because glioblastoma tumor stem cells are more resistant to chemotherapy and radiation compared to the general population of glioma tumor cells. These problems have been approached using orthotopic xenograft models in rodents. Similar to fetal neural precursors, glioma stem cells can be cultured from tumor samples in the form of neurospheres. Human glioblastoma tumor stem cell neurospheres can be stereotactically injected into rodent brains, e. g., nude mice, which allows for tumor formation. The animals can then be treated with drugs or other anti-glioma modalities allowing close modeling of the blood-brain tumor barrier. Such models have successfully been used to test the efficacy of several experimental therapeutics, including Aurora-A kinase inhibitors in our laboratory. The benefits and limitations of using glioblastoma tumor stem cells compared to conventional tumor models will be discussed.
Brian Mehling
Blue Horizon International
USA
Title: Safety study of intravenously administered human cord blood stem cells in the treatment of symptoms related to chronic inflammation
Time : 17:35-17:55
Biography:
Brian Mehling is a practicing American orthopedic trauma surgeon, researcher, and philanthropist. He is spearheading groundbreaking research in stem cell therapy through his company, Blue Horizon International, a healthcare consulting organization focused on treatment and research using stem cells. He started his path in medicine through undergraduate study at Harvard University, obtaining his Bachelor of Arts and Master of Science degrees in Biochemistry from Ohio State University. Completing his degree of medicine at Wright State University School of Medicine, he received post graduate education through residencies and fellowships at St. Joseph’s Hospital in Paterson, NJ and the Graduate Hospital in Philadelphia, PA, while pursuing a Ph.D. in Chemistry.An orthopedic trauma surgeon, Mehling operates his own practice, Mehling Orthopedics, in both West Islip, NY and Hackensack, NJ. He is also on call at Good Samaritan Hospital, Hackensack University Medical Center, Hackensack University Medical Center at Pascack Valley, St. Joseph’s Medical Center, and St. Joseph’s Wayne Medical Center. Mehling has traveled extensively throughout Asia and the Middle East observing firsthand the differences in healthcare standards. Identifying the need for a universal high quality standard, he founded Blue Horizon International. He and his colleagues have successfully treated and monitored more than 600 patients using stem cell and regenerative therapies. Patients have been treated for a wide range of diseases and disorders including spinal cord injury and cerebral palsy. Mehling founded the Blue Horizon Charitable Foundation to enhance the quality of health to those in need. Efforts are focused on helping human service and health organizations, while also providing resources benefitting children orphaned from war and disease. Apart from his organizations, Dr. Mehling personally contributes his time and resources to charitable causes. He has sponsored patients’ trips to China for stem cell treatments, as well as hosting charitable events from his own homes in New York, the Hamptons, and France. Notable beneficiaries include The Sean Kimmerling Testicular Cancer Foundation and the Children of Chernobyl.
Abstract:
Stem cell research plays an important role in orthopedic regenerative medicine today. Arthritis literally means "inflammation of a joint." Mesenchymal Stem Cells (MSCs) represent a valuable tool for therapy of symptoms related to chronic inflammatory diseases. Blue Horizon Stem Cell Therapy Program is a leading provider of adult and children’s stem cell therapies. The purpose of this study is primarily to monitor the immune response in order to validate the safety of intravenous infusion of human umbilical cord blood derived MSCs (UC-MSCs), and secondly, to evaluate effects on biomarkers associated with chronic inflammation. Our study of blood test markers of 9 patients with chronic inflammation before and within three months after MSCs treatment demonstrates that there is no significant changes and MSCs treatment was safe for the patients. Analysis of different indicators of chronic inflammation included in initial, 24-hours, two weeks and three months protocols showed that stem cell treatment was safe for the patients. Further close monitoring and inclusion of more patients are necessary to fully characterize the advantages of UC-MSCs application in treatment of symptoms related to chronic inflammation.
Luguang Luo
Roger Williams Hospital, Charter Care-Prospect RI LLC
USA
Title: The advantage of bioengineering and stem cell approach for immunorejection free human islet
Biography:
Luguang Luo has been trained by medical physician and molecular biology background (Umass Medical Center and Brown Medical School USA) for more than 30 years. He initiated his career with senior outstanding endocrinologist Dr. Ivor Jackson at Brown Medical School. Dr. Luo’s significant contribution is to identify Thyrotropine Releasing Hormone (TRH) expression in pancreatic β-cell and may contribute β-cell regeneration. Since he became independent investigator, he collaborates with Dr. Quesenberry, experts in hematology, his efforts have been focusing bone marrow stem cells and diabetes and published more than 50 articles in peer reviewed Journals and about 100 abstracts presentations in American Diabetes Association Annual Meeting and Endocrine Association Annual meeting and others which lead him to receive numerous funds from JDRF, NIH and local hospitals. Recently, Dr. Luo focuses on to establish a microenvironment in retaining a balance of β-cell apoptosis and regeneration, resulting in the support of pancreatic β-cell survival and function while developing bioengineering approach to create immunorejection free human islet tissue. Dr. Luo is a Director of stem cell and diabetes Research Center in Roger William Hospital and professor affiliated with BU and Brown Medical schools. Dr. Luo is a reviewer in NIH study section and cNSF and has also been invited to review articles for numerous Journals such as Molecular Endocrinology, Journal of Neurology, Endocrinology, et al and Editor Board members.
Abstract:
Islets transplantation holds promise as a long term treatment to Type I diabetes. We have previously reported that bone marrow cells (BM) co-cultured with human islets generate a microenvironment suitable for repairing islets and promoting longevity. Our work strongly supports that BM and its subpopulation creates a microenvironment which sustains human islet beta cell function and survival in long-term. In additional, we found that the role from BM derived populations is of diversity. We hypothesize that mechanism of BM support human islet includes repair human islet injury, initiating human isle regeneration through vascularization and initiating beta cell transcription factor activations. Coculture human allogeneic BM and islet generates a reinstituted human islet tissue which suitable for transplantation in vivo for diabetes therapy. However, in vivo immunorejection issue still yet overcome with this reinstituted tissue. We hypothesize that co-encapsulated BM will generate microenvironment for human islet longevity while preventing immunorejection. This work provides exciting results for supporting the hypothesis. APA encapsulation was established by coating gel beads with additional layers of poly-L-ornithine and alginate to create a 4-layered immunoisolatory membrane. Optimal condition was created and tested. After 4 weeks of culture, encapsulated human islets with BM formed a 3D structure while groups without encapsulation formed a 2D structure. The advantage of this new approach also approved in vivo by transplantation encapsulated human islets in immunologically competent STZ-induced diabetic rats. Our results show that encapsulated human islets with BM creates a microenvironment benefitting human islet function/longevity while preventing immunorejection. We will summarize the advantage from our and others to propose potential clinical application of this novel discovery.
Mei Wan
Johns Hopkins University School of Medicine
USA
Title: Mobilization and differentiation of multipotent nestin+ stem cells during arterial remodeling
Biography:
Mei Wan is an Associate professor of the Center for Musculoskeletal Research, Department of Orthopaedic Surgery at Johns Hopkins University School of Medicine. She obtained her Ph.D. in Pathophysiology at Hebei Medical University in 1997. Her research for the past 17 years focuses on characterizing the mechanisms by which bone marrow mesenchymal stem cells (MSCs) are regulated in various physiological and pathological conditions such as bone remodeling, cancer development, vascular disorders, and tissue repair/remodeling. At earlier years of her career, she demonstrated that the role of proteosome degradation pathway in the regulation of TGFβ signaling. She also identified the central mechanism through which parathyroid hormone stimulates bone formation, which had been the major unresolved question in bone field. In recent years, she found that active TGFβ can be released from tissue in response to perturbations to the local environment such as bone remodeling (Nat. Med. 2009, Cell Stem Cell 2011), arterial injury (Stem Cells 2012, Stem Cell Dev. 2014), and lung injury (J. Immunol. 2014). The released active TGFβ stimulates the migration of MSCs to participate in tissue repair or remodeling. Currently, Dr. Mei Wan is an editorial board member for Journal of Bone and Mineral Research and Bone Research.
Abstract:
Multipotent mesenchymal stem cells (MSCs) can mobilize into the circulating blood under many circumstances, such as serious disease, injury, or stress. MSCs then migrate to the remodeling sites and differentiate toward distinct lineages of cells. However, the primary factors and signaling pathways that control MSCs recruitment to the injured sites and their commitment/differentiation into lineage-specific local cells are largely unknown. Here we show that active TGFβ controls the mobilization of MSCs to circulating blood in response to arterial injury and their recruitment to the target sites, where the cells give rise to either endothelial cells to repair the damaged endothelium or smooth muscle cells (SMCs)/myofibroblasts contributing to intimal hyperplasia. In our animal models of arterial injury, about 50% of the neointimal cells were derived from MSC lineages. Using an ex vivo cell migration assay established in our laboratory, we found that TGFβ activated from the injured vessels induces MSC migration, and this effect is mediated by Smad-MCP1 signaling cascade. Moreover, active TGFβ produced from the injured vessels also activated RhoA-ROCK signaling in MSCs and induced their differentiation to SMC/myofibroblastic neointimal cells. Inactivation of ROCK maintains the stemness of MSCs and their differentiation capacity to endothelial cells. Importantly, treatment of the arterial injured mice with ROCK inhibitor promoted re-endothelialization and inhibited neointima formation of the vessels. Thus, pharmacotherapies that inhibit RhoA-ROCK signaling offer a new therapeutic target for treating cardiovascular disease by promoting endothelium repair and inhibiting pathological intimal hyperplasia.
Somaieh Kazemnejad
Avicenna Research Institute
Iran
Title: Menstrual blood derived stem cells as a renewable source of adult stem cells: Features and advantages
Biography:
Somaieh Kazemnejad is associate professor and director of Tissue Engineering department in Avicenna Research Institute, where she leads the Regenerative Medicine Group, a multidisciplinary team of researchers including engineers, cell biologists, polymer chemists, clinicians, and veterinary surgeons. She was born in 1979 and completed her Ph.D at the age of 27 years from Tarbiat Modarres University of Iran. She is a Clinical Biochemist who is one of very few Iranian academics in the field of regenerative medicine and tissue engineering to have taken a research programme from fundamental research through to clinical application utilizing research in-vitro, in-vivo, preclinical large animal studies and clinical trials with focus on menstrual blood stem cells. She has published more than 28 papers in reputed journals in the field of tissue engineering and regenerative medicine.
Abstract:
Menstrual blood derived stem cells (MenSCs) can be easily obtained from women’s menstrual blood in a non-invasive technique without ethical issues of other stem cell types. These multipotent cells have the ability to differentiate into various functional cells, including osteocytes, adipocytes, cardiomyocytes, respiratory epithelial cells, neurocytes, myocytes, endothelial cells, pancreatic cells and hepatocytes. No evidence of tumor and ectopic formation, or any immune response has been demonstrated after being transplanted into animal models. To determine whether MenSCs outline common features with bone marrow-derived stem cells (BMSCs) or have source-specific peculiarities, we have done a head-to-head comparison of MenSCs with BMSCs in aspect of immunophenotyping, proliferation and differentiation characteristics. The evidence presented here narrates MenSCs are unique stem cell population with higher rate of proliferation and different trans-differentiation potential compared to BMSCs. Probably, it can be attributed to the particular immunophenotypic pattern and special signaling molecules involving in MenSCs development. In addition, we reviewed the possibilities of using MenSCs for diagnosis of diseases and as a novel alternative to current cell sources for cell-based therapies of neurological disorders, myocardial infarction, type I diabetes mellitus, liver cirrhosis, etc. Therefore, this study presents broad insights about possible advantages of MenSCs and their safety/efficacy profile for clinical applications. Key words: Menstrual blood-derived stem cells (MenSCs), Regenerative medicine, cell therapy
- Track 8: Novel Approaches in Guided Tissue Regeneration
Location: Rome, Italy
Chair
Aysegul Batioglu-Karaaltin
Istanbul University Cerrahpasa School of Medicine
Turkey
Co-Chair
Livia Visai
University of Pavia
Italy
Session Introduction
Massimiliano Colombo
University of Milan
Italy
Title: Treatment of AVN using the chamber induction technique and biotechnologies: Indications and clinical results
Time : 14:50-15:10
Biography:
Massimiliano Colombo has completed Medicine and Surgery at University of Milan and specialization in School in Orthopaedics and Traumatology at University of Milan. He is at present working as orthopaedic surgeon and researcher at unit of Orthopaedic Reparative Surgery and Risk Management, G. Pini Orthopaedic Institute, piazza Cardinal Ferrari 1, University of Milan. He has good knowledge of the problems of bone tissue as regards its regeneration and healing processes. He performed numerous national and international publications on the topic of bone necrosis, delayed consolidation of the fractures, non union, bone defects with particular interest in the field of biotechnology, such as growth factors, bone substitutes and mesenchymal stem cells. He produced: 3 medical scientific monographs; 43 orthopaedic scientific publications in Italian and foreign journals, including many with relevant impact-factor and multiple citations; 17 scientific posters presented at conferences. He performed an economy drug study on the use of growth factors in traumatology entitled "Comparative analysis of costs and cost/effectiveness ratio of the treatment of persistent post-traumatic non-unions of the tibia with autologous transplant of the Iliac Crest or Osigraft® (Eptotermin alfa, rhBMP-7,rhOP-1)”. He is performing a study about the effectiveness of a new system for the insertion of distal screws in the implant of long femoral nails. He recently completed a comparative study on the efficacy of growth factors (rh-BMP-7) with respect to autologous bone graft from the iliac crest in the treatment of 150 nonunions and loss of substance of the long bones.
Abstract:
Objective: to determinate the efficacy of core decompression technique with the use of recombinant morphogenetic proteins, autologous mesenchimal stem cells (MSCs) and xenograft bone substitute into the necrotic lesion of the femoral head on clinical symptoms and on the progression of osteonecrosis of the femoral head. Methods: we studied 38 patients and 40 hips with early stages of osteonecrosis of the femoral head. Results: Core decompression technique with the use of recombinant morphogenetic proteins, autologous MSCs and xenograft bone substitute afforded a significant reduction in pain and in joint symptoms and reduced the incidence of fractural stages. At 34 months, 33 patient reach the clinical and radiographic healing. Conclusion. This long term follow-up study confirmed that core decompression technique with the use of recombinant morphogenetic proteins, autologous MSCs and xenograft bone substitute might be an effective treatment for patients with early stages of osteonecrosis of the femoral head.
Livia Visai
University of Pavia
Italy
Title: Scaffolds, stem cells, biophysical stimulations and nanotechnology in tissue engineering and regenerative medicine
Time : 15:10-15:30
Biography:
Visai received the Ph.D title in Biochemistry in 1989 and She is presently an Assistant Professor in Biochemistry at the Medicine Faculty of Pavia University. Her scientific experience were performed abroad initially at the Connective Tissue Laboratory, Alabama University in Birmingham (USA) and then to the Center for Infectious and Inflammatory Diseases (IBT), Houston University in Texas (USA).She has been recently a Visiting Professor at the Department of Nanomedicine and Nanotechnology of the Methodist Hospital in Houston, Texas, (USA). She is the vice-director of the Interdepartmental Center for Tissue Engineering in Pavia University and the Head of the Nanotechnology Laboratory at Salvatore Maugeri Foundation in Pavia. She has published more than 129 papers in reputed journals and has been serving as an editorial board member of repute.
Abstract:
Recent advances in tissue engineering and regenerative medicine have shown that controlling cells micro-environment during growth is a key element to the development of successful therapeutic system. Various polymeric scaffolds have been used to support cellular growth and, to a certain extent, favor cell organization and tissue structure. A large pool of stem cell lines such as multipotent and pluripotent stem cells lines are available for these type of studies. Furthermore, the exposure of stem cells to biophysical stimuli has been reported to favor early and rapid activation of the tissue repair process. But all of these approaches has appeared to be rather limited since they do not offer the fine control of the cell micro-environment in space and time. We will be presenting the biological effects exerted by electrospun nanocomposite materials on tuning the cell fate of human multipotent and pluripotent stem cells. Then, we will continue by presenting the effects related to the exposure of human multipotent stem cells to Pulsed Electromagnetic Field (PEMF) or Low Level Laser Intensity (LLLI) on cell proliferation and differentiation towards osteoblasts. At the end, we will show the recent data obtained in the design of 3D scaffolds based on hydrogels as well as polymeric fibers to maintain the self- renewal of human pluripotent stem cells in feeder-free conditions.
Nelson R. Pinto
University of the Andes
Chile
Title: Leukocyte-Platelet Rich Fibrin Membrane (L-PRF) alone or associated with a biomimetic implant surface act as a biological scaffold for hard and soft tissue regeneration
Time : 15:30-15:50
Biography:
Nelson R. Pinto: University of the Andes, Faculty of Dentistry, Santiago, CHILE. Visiting Professor, Department of Oral Health Sciences / Periodontology , University Hospitals, Catholic University Leuven, BELGIUM. Founder and Chairman of the Research Center of Regenerative Medicine and Tissue Engineering , Concepción, CHILE. Clinical Editor POSEIDO JOURNAL. World Leader on Clinical Applications of L-PRF in Hard and Soft Tissue Regeneration and Wound Healing. Developer of the Technique for the Treatment of Chronic Wounds with L-PRF.(Best Oral Research Presentation : 4th Congress of the World Union of Wound Healing Societies, Yokohama, Japan, 2012) National and International Lecturer on Implant Dentistry and Regenerative Medicine. (+200).
Abstract:
Leukocyte- and Platelet-Rich Fibrin (L-PRF) is one of the four main families of platelet concentrates for surgical use. L-PRF is used to improve healing and promote tissue regeneration. Blood sample is taken without anticoagulant and immediately centrifuged to obtain a L-PRF clot that can be used directly or compressed into a membrane without damaging the cells and growth factors content. The L-PRF clot or membrane contains most of the platelets and leukocytes present in the initial blood harvest plus the platelet growth factors and stem cells that are also trapped within the fibrin network. With this architecture, L-PRF is the source of a strong and slow release of growth factors such as TGF-b1, VEGF, IGF-1, FGF, EGF, PDGF-AB, IL-1ß for more than three weeks in vitro. Through the release of these growth factors trapped within the fibrin gel or through the production of new molecules by the leukocytes, the L-PRF membranes have strong effects on the stimulation of the proliferation of most cell types (fibroblasts, keratinocytes, pre-adipocytes, osteoblasts, bone mesenchymal stem cells) and on the differentiation of the bone cells.The possibility to use L-PRF as a biological scaffold by itself or associate with a biomimetic implant surface as open the opportunity to regenerate soft and hard tissue in such a way that was not possible before. The clinical, immunohistochemestry and histological findings (SEM, Confocal Laser, and Optical Microscopy) of our animals and humans studies over the last 14 years confirm the potential of L-PRF as a biological scaffold for hard and soft tissue regeneration in acute or chronic wounds. What we thought impossible yesterday, could be routine tomorrow… “Natural Guided Regeneration with L-PRF”.
Aysegul Batioglu-Karaaltin
Istanbul University Cerrahpasa School of Medicine
Turkey
Title: Invivo tissue engineering trachea using adipose derived mesenchimal stem cell and Cadaveric tracheal scaffold
Time : 15:50-16:10
Biography:
Aysegul Batioglu-Karaaltin has completed her MD degree at the age of 24 years from Hacettepe University and her residency at the Department of ENT and Head and Neck at Ministry of Health Education and Research Hospital. She is working at Department of Otolaryngology Head and Neck Surgery, Istanbul University Cerrahpasa School of Medicine. She is carrying out more then 10 research about regenerative medicine and tissue engineering in the Head and Neck field. She has published more than 15 papers in reputed journals
Abstract:
Tracheal transplantation appears promising, especially for the patients who have been followed in intensive care units for a long time and who have received recurrent operations after traffic accidents or malignancies and, as a result, have to live with a tracheostomy for the remainder of their lives because of the diagnosis of tracheal stenosis. The treatment of tracheal defects longer than 6 cm (approximately half of the total length of trachea) is still controversial. Autograft materials, cadaveric tracheal allotransplantations, vascularized allotransplantation, cadaveric trachea decellularisation were tried. Cadaveric trachea decellularisation followed by seeding with autologous stem cells of the recipient is the best method with no need to use immunosuppressive medication, no requirement for the second major surgical procedure. Successful human transplantations have also been performed using this technique with bone marrow-derived MSCs. We published first successful in vivo tissue engineered trachea regeneration from a decellularized cadaveric trachea matrix with seeded adult adipose tissue-derived MSCs which was integrated into the recipient tracheal sides. It is required to protect the three dimensional structure of the trachea and its extracellular matrix during decellularisation process. The previous studies mostly employed detergent enzymatic method (DEM) and its modifications for tracheal decellularisation. Although ionic detergents used for trachea decellularisation are very effective in removing cellular remnants, they damage the natural structure of the tissue by affecting the integration of the extracellular matrix. We used combined decellularisation method called protected matrix decellularisation (PMD) which was developed by our group. And we control test of decellularisation both for rabbit and human cadaveric tracheas. As a result new protected matrix decellularisation followed by seeding with autologous adipose tissue-derived MSCs which have same origine with chondrocytes and chondrogenic differansiation is known as higher than other stem cells is promising for trakeal reconstruction.
Srecko Gajovic
University of Zagreb School of Medicine
Croatia
Title: IN VIVO imaging modalities to visualize mouse brain regenerative response after stroke and stem cell and biomaterial applications
Time : 16:10-16:30
Biography:
Srećko Gajović is Head of Histology and Embryology at University of Zagreb School of Medicine. He is Coordinator of FP7 REGPOT project GlowBrain dedicated to unlock the research potential of Croatian Institute for Brain Research through stem cell and biomaterial applications in the mouse stroke model. He is Editor-in-Chief of Croatian Medical Journal, best general medical scientific journal in this part of Europe, and ex-Chair of European COST Domain Committee for Biomedicine and Molecular Biosciences. He was born in Zagreb, Croatia, where he completed his studies of medicine and get a PhD at University of Zagreb. His postdoctoral positions were at Max Planck Institute for Biophysical Chemistry in Göttingen, Germany, and at Institute for Genetic Engineering and Biotechnology in Trieste, Italy. In his current research, he tries to understand the role of innate immunity in brain repair after stroke, and to develop tissue engineering strategies by combining stem cells and biomaterials for the brain repair.
Abstract:
To achieve the beneficial response and brain regeneration after stroke one of the possibilities is to combine stem cells and biomaterials and to monitor their effects in the living animals. This is the main aim of the FP7 REGPOT project GlowBrain, and for this purpose several in vivo small animal imaging modalities are used, which includes magnetic resonance imaging (MRI) and light based imaging modalities - bioluminescence imaging (BLI), fluorescence imaging (FLI) and Cherenkov luminescence imaging (CLI). To visualise the molecular events related to the activity of TLR2, GAP43, and CASP3 in the brain of the living mouse the maging was performed by IVIS Spectrum Pre-clinical In Vivo Imaging System (Perkin Elmer, US) in genetically modified animals carring luciferase reporter TLR2, GAP43, and CASP3 were upregulated after stroke. To analyze the apoptosis in subset of GAP43 cells the imaging with DEVD-aminoluciferin was performed. The findings suggested that CASP3 activity, not necessarily associated with neuronal apoptosis, increased, and CASP3 and GAP43 might be part of a common molecular pathway involved in early stress response after stroke. This added neuronal stress in addition to inflammation, repair, and apoptosis as important process to be assessed by the bioluminescent imaging as the brain response to stroke. The same strategy of in vivo insight in brain molecular events was applied as well to monitor the biocompatibility of brain applications of the stem cells and biomaterials. The combination of neural stem cells and the polysaccharide biomaterial alginate was designed. The in vivo imaging confirmed that the inflammation after brain transplantation was comparable to the control injection by PBS, proving the biocompatibility of the procedure. The described experimental approach opens the way to design and preclinical testing of innovative therapies for brain diseases.
- Special Session on Developments, innovations and breakthroughs in regenerative medicine
Location: Rome, Italy
Chair
Jonathan Schwartz
Marvel Medicals
USA
Session Introduction
Joseph Purita
Institute of Regenerative and Molecular Orthopedics
USA
Title: Lipogems a game changer in the field of re-generative stem cell therapy
Time : 16:45-17:05
Biography:
Purita is Director of Institute of Regenerative and Molecular Orthopedics (www.stemcellorthopedic.com) in Boca Raton, Florida. The Institute specializes in the use of Stem Cells and Platelet Rich Plasma injections. Dr. Purita is a pioneer in the use of Stem Cells and Platelet Rich Plasma. The Institute has treated some of the most prominent professional athletes from all major sports in both the U.S.A. and abroad. He received a B.S. and MD degree from Georgetown Univ. Dr. Purita is board certified in Orthopedics by ABOS. He is a Fellow American College of Surgeons, Fellow American Academy Orthopedic Surgeons, and a Fellow American Academy of Pain Management. He is also certified in Age Management Medicine. He has lectured and taught extensively throughout the world on the use of Stem Cells and Platelet Rich Plasma. He has been instrumental in helping other countries in the world establish guidelines for the use of Stem Cells in their countries. He has been invited to lecture on these techniques throughout the world as a visiting professor.
Abstract:
The use of adipose tissue has taken on increasing im-portance in the field of regenerative stem cell therapy. This therapy can be divided into two major pathways. One path-way involves plastic and reconstruction surgery. The other major pathway involves orthopedics and its related fields. Other uses are being found in gynecology, urology, and general surgery. Until recently the main uses of adipose tissue centered around the use of enzymatic produced SVF suspensions. Typically these suspensions were made with the use of the enzyme collagenase. Although collagenase has worked well in the past there is now a complaint method of producing a fast, enzyme free lipoaspirate that is turned into a therapeutic product. This new method comes from the Lipogems company. The Lipogems technique has been utilized in over 5,000 patients. It has recently obtained FDA clearance. Lipogems makes use of the high stem cell density of adipose tissue thought to be 1 out of every 100 cells. Lipogems utilizes mild mechanical forces in a closed system to produce micro fractured and purified adipose tissue graft which is non ex-panded and ready to use in a variety of regenerative appli-cations. More importantly, Lipogems adipose product has a very well preserved vascular stroma with slit-like capillaries wedged between adipocytes and and vascular channels. The stem cell niche is preserved enabling the pericytes to have a much higher survivability rate. The intact niche pro-vides a perfect environment for reparative response of acti-vated MSCs. As Dr. A. Caplan has pointed out, pericytes are precursor mesenchymal stem cells. Activated MSCs are crucial for the immune modulation and regeneration. The lipoaspirate of Lipogems shows a higher percentage of ma-ture pericytes and hMSCs than enzymatically digested lipoaspirates. We are now aware that there are novel growth factors secreted by Lipogems lipoaspirate which may explain the therapeutic efficacy of Lipogems. Also im-portant, is the fact that the Lipogems technique may pre-serve the important MUSE cell. Muse cells are unique in that they be pluripotent, have a high survivability rate, and are quite small by stem cell standards. They are considered to be an embryonic like stem cell without any of the risks of regular embryonic stem cells since they are the patient's own. They may represent a quantum leap in regenerative medicine. Since Lipogems preserves these cells it too is a quantum leap.
Michael Weber
Webermedical Laser treatment and research centers
Germany
Title: Basics and clinical applications of laserneedle acupuncture, intravenous, interstitial, intraarticular and photodynamic laser therapy
Time : 17:05-17:25
Biography:
Michael Weber has completed his studies in chemistry and biochemistry at University Marburg and at University Goettingen he completed his PhD during 1983. He was researcher at Max-Planck-Institute for Experimentel Medicine in Göttingen. He is the Leader of 3 medical laser centers in Germany and Bangkok in Thailand. He is the President of the International Society for Medical Laser Applications (ISLA) and also Board Member of the North American Association for Laser Therapy (NAALT). He is author of many international publications in the field of Medical Laser Therapy. He established his own company (weber medical GmbH) in 2003 supported by the German government and the EU.
Abstract:
In this lecture new methods of laser therapy will be presented covering nearly the whole range of important diseases in medicine. Laserneedle acupuncture with different wavelengths and penetration depths can be used as a highly effective and pain free method for all indications of acupuncture and pain management. Laserneedles can be applied on the body, on the skull or the ear as well. Infrared, red and yellow lasers can penetrate the skull bone and used today for benefit on brain diseases. Intravenous laser therapy is a systemic application of laser light with infrared, red, green, blue and yellow wavelengths. This method stimulates the immune system, leads to an improved microcirculation and oxygen supply and improves endurance in sports people. Furthermore all wavelenghts stimulate the different complexes of the respiratory chain in the mitochondria leading to an increase of ATP-production and thus are well suited for all fields of regenerative medicine in mitochondrial degeneration. Especially blue laser disscociates nitric oxide from haemoglobin and complex IV in the mitochondria and so leads to improved microcirculation, biogenesis of mitochondria and prevents cell senescense. Nitric oxide also activates telomerase and so protects against degradation of telomers with possibly extended life expectation. We also know today that intravenous laser therapy can activate endogenous stem cells with improvement of organ function.Main therapeutic fields are angiopathies, neuropathies, fibromyalgia and chronic fatigue syndrome, autoimmune and metabolic diseases, depression, all diseases on basis of mitochondrial degeneration and antiaging in general. Interstitial laser therapy is a new method using a fiberoptic catheters for application of different lasers in the depth of the tissue close to the spot of injury. It’s a new method for treatment and regeneration of herniated disks and other nerve injuries. For intraarticular laser therapy a fiberoptic laser catheter will be placed directly in a joint and is a highly effective treatment of advanced osteoarthritis of the knee, shoulder and other joints. All laser colors including non penetrating blue laser can be used successfully. This therapy can be combined with injection of PRP or mesenchymal stem cells with light activation for achieving regenerative effects on the cartilage. New therapeutic approaches for photodynamic cancer therapy are presented as well using the different laser colors externally and interstitially with new photosensitizers for all type of cancers.
Sciarretta Fabio Valerio
Clinica Nostra Signora della Mercede
Italy
Title: A novel chondral defect repair strategy: biologic resurfacing through innovative bone marrow concentration system
Time : 17:25-17:45
Biography:
Fabio Valerio Sciarretta attended the Liceo Classico " De Sanctis, " earning a Classic baccalaureate diploma in 1982. In the same year he joined the Faculty ' of Medicine and Surgery of the University of Rome "La Sapienza" . From January 1990 to April 1991, he fulfilled his military obligations in quality ' of Lieutenant physician at the Military Hospital of Cagliari with the post of " Assistant of the Orthopaedics Department " reporting the following final judgment : "Officer with outstanding moral and character accomplishment as well as ' comprehensive and updated professional preparation. He has carried out the task to him entrusted with zeal and high sense of duty and responsibility' providing a valuable contribution to the smooth running of the Department . " During this service , the 7/11/90 he has obtained a written note of satisfaction of the Director of Health ' Military Region of Sardinia for " the results obtained by acting with great professional expertise to a qualified orthopedic surgery ." He is affiliate Member of the American Academy of Orthopaedic Surgeons ( AAOS ), International Society of Arthroscopy , Knee Surgery & Orthopaedic Sports Medicine ( ISAKOS ), European Society of Sports Traumatology Knee Surgery and Arthroscopy ( ESSKA ), International cartilage Repair Society ( ICRS ) also member of the Italian Society of Orthopaedics and Traumatology ( SIOT ) , Society of Surgery of the knee, arthroscopy , sports medicine, cartilage , orthopedic technologies ( SIGASCOT ),member of the Italian Society of Arthroscopy ( SIA). He has been the editor of many Italian edition journals. From 1994 to 2000 he served as director of the first level of health care at the Hospital of St. John the Baptist in Rome of the Association of Italian Knights of the Sovereign Military Order of Malta. Since September 2000 , as a senior health care executive for a specified period from 2000 to 2007 and as a senior health care executive for an indefinite period , he served at the Department of Orthopaedics and Traumatology Hospital of Velletri (ASL RMH ) . Currently practices his profession at the among in the private clinics of Rome
Abstract:
While performing an arthroscopy of the knee, cartilage injuries, of any grade, have a whole reported frequency between 57.3% and 66% of cases. These numbers make it clear that, more and more, young adults under the age of 65 years, present with an advanced disease to joint cartilage. in recent years, have increasingly found their way the regenerative medicine therapies, which allow to correct alterations of the biological basis of the disease's process, rather than addressing only the symptoms. Orthobiologics are based on the use of mesenchymal stem cells that can mainly be obtained from bone marrow or adipose tissue. Since no surgical technique has still been proven to be the gold standard of chondral repair, in order to increase the results of the recently developed single stage chondral repair techniques delivering to defect site a larger number of multipotent cells to improve defect filling and repair, the augmented single stage procedures have been introduced. These techniques represent an evolution of the standard AMIC procedure, where a collagen membrane is used to repair and resurface chondral defects. In the augmented techniques the use of PRP or bone marrow concentrate is added to the collagen membrane. In this paper we present the initial experience with a new system for bone marrow aspiration and concentration, perfectly adapting to the timing of one step cartilage repair AMIC technique. The surgical technique steps are: chondral defect debridement, bone marrow iliac crest aspiration through power driven aspiration cannula, defect's sizing through aluminum template use, microfractures, collagen membrane sizing and soaking with bone marrow concentrate obtained through completely closed centrifugation and, finally, closure of the chondral defect with collagen membrane, bone marrow concentrate and fibrin glue. The surgical technique is safe, lasts 15-20 minutes, is completed all inside the OR and doesn't need cellular expansion. Randomized controlled clinical trials will confirm the results.
Al Sears
Palm Beach Institute for Anti-Aging Medicine
USA
Title: Cracking the telomere code: The first step to reverse aging
Time : 17:45-18:05
Biography:
Al Sears, M.D. is the founder of the Center for Health and Wellness, a successful integrative medicine and anti-aging facility in Royal Palm Beach, Florida, with over 25,000 patients. His cutting-edge therapies and reputation for solving some of the most difficult-to-diagnose cases attract patients from around the world. Sears was one of the first to be board-certified in anti-aging medicine. As a pioneer in this new field of medicine, he is an avid researcher, published author, and enthusiastic lecturer.Sears is board-certified as a clinical nutrition specialist and a member of the American College of Sports Medicine (ACSM), the American College for the Advancement in Medicine (ACAM), the American Medical Association (AMA), the Southern Medical Association (SMA), the American Academy of Anti-Aging Medicine (A4M), and the Herb Research Foundation, (HRF). Dr. Sears is also an ACE-certified fitness trainer.Sears currently writes and publishes the monthly e-Newsletter, Health Confidential, and daily email broadcast, Doctor’s House Call, and contributes to a host of other publications in the field. He has appeared on over 50 national radio programs, ABC News, CNN, and ESPN.Since 1999, Dr. Sears has published 15 books and reports on health and wellness with a readership of millions spread over 163 countries.
Abstract:
This lecture explores the discovery of the telomerase enzyme and its role as a crucial indicator of health and longevity. By tracing the evolution of telomere research, including the landmark Harvard study that fully restored youthful function in mice by activating the telomerase enzyme, Dr. Sears unveils the true age-reversing, and health-transforming potential of telomere therapy and the capacity to influence gene expression through novel interventions. As one of the only medical doctors to administer the world’s first telomere therapy, Dr. Sears relates its effects on his own patients, and how his own research uncovered new, more effective ways of supporting telomere length and altering telomere biology. Three learner objectives. By the end of the lecture, participants will understand: • The evidence that supports the telomere’s role as the true cause of aging. • What factors cause the telomere to shorten, and the most reliable interventions to support telomere length. • How the search for telomerase activators uncovered new, more powerful and more affordable means of affecting telomere regulation.
- Young Research Forum
Location: Rome, Italy
Chair
Karin Schütze
CellTool GmbH
Germany
Session Introduction
Nihal AlMuraikhi
Imperial College London
UK
Title: E-BABE- A direct differentiation of hipscs towards erythroid cells under serum-free and hypoxia condition
Time : 18:05-18:20
Biography:
Nihal AlMuraikhi is a Ph.D candidate expected to be awarded by the end of this year from Imperial College London. She is a lecturer at the Stem Cell Unit of the Medical School of King Saud University, Riyadh, Saudi Arabia.
Abstract:
Current protocols used in the differentiation of human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) toward erythropoiesis utilize two main approaches; Embryoid Body (EB) formation, which influences heterogeneity of the produced population, and/or co-culture with mouse stromal cells, which makes the xeno-free culture requirement difficult to achieve. In this study, we designed a serum-free experiment on the direct differentiation of hiPSCs toward erythroid cells under hypoxia condition bypassing the EB formation step and no co-culture system. Our protocol involves three phases: phase-I) Hematopoietic induction (7days), where cells were exposed to specific cytokines to trigger the mesodermal formation. Phase-II) Erythroid differentiation (7days) with reduced number of cytokines; EPO, SCF and IL-3; and finally, phase-III) Maturation of erythroid progenitors (14days) by exposing cells to EPO only. All culture phases were performed in hypoxia conditions. Phase-I showed an expression of an early hematopoietic marker, CD34 in parallel to a high expression of CD45, a pan leukocyte marker. Both markers peaked by phase-II then decreased gradually during maturation. Phase-I also showed a high expression of an early erythroid marker, CD71 and a late erythroid marker, CD235a over the culture period. Around 10% of the cells were enucleated erythrocytes by the end of phase-III, which was confirmed by Wright-Giemsa and reticulocytes staining. Functional analysis using CFU showed that the cell population were able to form hematopoietic colonies, specifically, erythroid progenitor. Further studies on maturation and enucleation are required in order to achieve fully mature and functional RBCs phenotype.
Celine Aubrun
Université d’Artois
France
Title: Bioceramic filler nature influence on acrylic cement properties after immersion in biological fluid.
Time : 18:20-18:35
Biography:
Physical and chemistry graduated teacher, Céline Aubrun is preparing a PhD in mechanical engineering at the Équipe Biomatériaux Artois of the Laboratoire génie civil et géoenvironnement (Université d’Artois). She specialized in biomaterials in the field of bone acrylic cements.
Abstract:
Cements are used in bone surgery to fill gaps issue from pathology or traumatology. We classically distinguish calcium phosphate cements with hydraulic setting and acrylic cements with polymeric setting, mainly formulated with polymethylmethacrylate (PMMA) and possibly include a biocompatible inorganic filler. We studied a cement based on 2-hydroxyethylmethacrylate (HEMA), natural polysaccharides and a bioceramic filler (tricalcium phosphate or hemihydrated calcium sulphate, TCP and plaster, respectively) developed for its advantages: hydrophilicity, biodegradability and mechanical characteristics. It aims filling bone defects with analgesic and support functions. The main interest substituting HEMA to PMMA is due to an in situ quick water-uptake when contact with biological fluids. Induced swelling ensures gap total filling by preventing formation of fibrous tissue. However a deterioration of mechanical properties occurs. Our study so aimed improving formulation of cement in order to limit mechanical properties loss while maintaining its water-uptake ability. Two bioceramic fillers were compared. Studied curing parameters were temperature increase and setting time. Mechanical properties andwater-uptake were measured initially and after 21 days in simulated body fluid (SBF). The highest temperature ranges from 50°C to 80°C and setting time from 1min30s to 4min30s. We found that plaster significantly improved mechanical properties. After 21 days in SBF cement Young modulus was 13.6MPa for plaster and 1.4MPa for TCP (initially 470MPa and 362MPa, respectively), while maintaining equivalent water-uptake (32%). These results open the way to a cement formulation allowing curing parameters and in situbehavior improvement, so that we can consider its use as bone substitute.
Aysu Arslan
Hacettepe University
Turkey
Title: Polybutyleneadipate-co-terephthalate (PBAT) scaffolds for tissue engineering applications
Time : 18:35-18:50
Biography:
Aysu Arslan has completed her B.Sc. degree from Hacettepe University, Department of Chemical Engineering. Currently she is M.Sc. student at the same department and she is a member of Hacettepe University Cell and Tissue Engineering Research Group, being coordinated by Prof. Dr. MenemÅŸe GümüÅŸderelioÄŸlu. She has published 1 paper in a related journal.
Abstract:
Polybutyleneadipate-co-terephthalate (PBAT) is an aliphatic-aromatic copolyester based on the monomers 1,4-butanediol, adipic acid and terephthalic acid. Recently, it has been considered as a promising biomaterial for tissue engineering applications due to its non-toxicity, desired physical properties and complete biodegradability under enzymatic conditions. However, existing studies are limited to cytotoxicity and basic cell proliferation assays with non-functional PBAT membranes which is not enough to understand PBAT’s behaviour as scaffolds for tissue regenaration. In this study, 2D and 3D porous PBAT scaffolds were fabricated via solvent casting, electrospinning, solvent casting-particulate leaching and melt molding-particulate leaching methods. Fabricated scaffolds were characterized due to their morphologies, mechanical properties, water absorption capacities, wettabilities and pre-osteoblastic cell promotion in order to estimate its potential for tissue regeneration. PBAT scaffolds were successfully fabricated in various structural forms with up to 90% porosities. Attachment and proliferation of MC3T3-E1 pre-osteoblasts on PBAT surfaces was observed with scanning electron microscope (SEM) on the 4th, 24th hours, 3rd and 11th day of cell culture. Results show that cells favoured the neat PBAT surfaces in all fabricated structural forms. At the 11th day of the cell culture, PBAT surfaces were covered by cells and extracellular matrix secreted by the cells. As a result, its easy processability, good mechanical properties, biocompatibility and well-known biodegradability makes it a powerful candidate for tissue engineering applications. In further studies, tissue engineering potentials of neat and modified 3D electrospun PBAT scaffolds will be investigated.
Alena.O. Stepanova
Meshalkin Novosibirsk State Research Institute of Circulation Pathology
Russia
Title: Influence of electron beam irradiation onto physical and chemical properties of electrospun produced 3D matrices
Time : 18:50-19:05
Biography:
Alena Stepanova has graduated from the Novosibirsk State University with master’s degree in molecular biology. She did the research for her master’s degree at the Molecular medicine Laboratory at the Institute of chemical biology and fundamental medicine Russian academy of Science in Novosibirsk. She is working at the same department the PhD Studies with the project “The study fundamentals of the vascular prosthesis fabricated by electrospinning in vitro and in vivo”.
Abstract:
Electron beam irradiation (EBI) is widely used in industrial production for sterilization of medical products. We have evaluated the influence of EBI onto mechanical strength, chemical properties and biocompatibility of electrospun produced 3D matrices (EPM). Sheets of EPM with oriented fibers (fiber diameter 1µm, scaffold thickness 100 µm) or tubes (inner diameter 1.7 mm, wall thickness 100 µm) were prepared from nylon 6, polylactic-co-glycolic acid, polycaprolactone (PCL) and their mixtures with gelatin. Electron accelerator ILU-6 (2.2 MeV, 400 mÐ, 10 Hz) was used for EBI of the materials in doses 25÷150 kGy with increment 25 kGy. Mechanical strength/structure were tested using Zwick/Roell Z100 testing machine/JSM-6460 LV scanning electron microscopy and differential scanning calorimetry. Human primary umbilical vein endothelial cells (HUVEC) and gingival fibroblasts (HGF) were used to evaluate adhesion and proliferation of cells at the surface of EPM. It was observed, EPM irradiation in a dose of 25 kGy does not affect the mechanic properties of all matrices studied. Irradiation of EPM with a dose higher than 50 kGy leads to embrittlement of all matrixes except from those produced from PCL. A dose of 100 kGy increases the proportional limit of the PCL scaffold, significantly increase adsorption of the protein on the surface of PCL fibers and allowed to introduce durable regions by irradiating EP tubes through the template with open areas. EBI of the matrices does not interfere with the capacity of EPM to support adhesion, viability and proliferation rate of HUVEC and HGF at the surfaces of EPM as it was shown by cells labeling with calcein/propidium iodine and incorporation of ethylenedioxy uridine. Thereby an electron beam irradiation of EPM was show to be a useful instrument to modify mechanic/chemical properties of EPM.
Shima Salmasi
University College London
UK
Title: Nanohydroxyapatite/polyhedral oligomeric silsesquioxane poly(carbonate-urea) urethane (nHA/POSS-PCU): applications in bone tissue engineering and regenerative medicine
Time : 19:05-19:20
Biography:
Shima Salmasi is a PhD student and research associate at the Centre of Nanotechnology and Regenerative Medicine, University College London. She completed her Master of Science in Nanotechnology & Regenerative Medicine and now, as part of her PhD, is researching on an innovative translational project with the aim of improving the current standards of spinal fusion surgery. This is a very interesting project with great potentials to provide a clinically effective solution to overcome the shortcomings of the currently available techniques of spinal fusion surgery using tissue engineering and nanotechnology.
Abstract:
Polyhedral oligomeric silsesquioxane poly(carbonate-urea) urethane (POSS-PCU), a new breed of novel nanocomposite material developed by researchers at the University College London has been extensively tested in terms of its physiochemical, in vitro, and in vivo properties, all of which have shown that its enhanced biocompatibility, superior mechanical engineering properties, and augmented degradative resistance renders POSS-PCU capable of functioning as a scaffold for bioartificial organs, nanoparticles for biomedical applications, and a coating for medical devices. The novelty of POSS-PCU further arises from the fact that it has been used in 3 first-in-human studies as a bypass graft, lacrimal duct, and the world's first synthetic trachea. Bone, as a living tissue, has the ability to constantly remodel, renew and regenerate to repair itself. However, large bone defects are considered as major problem for the clinician and society. Recently, we have investigated incorporation of different ratios of nanohydroxyapatite, the main inorganic component of the natural bone, into POSS-PCU nanocomposite for bone tissue engineering applications. This nanocomposite material could be used to better mimic the mineral component and the micro- & nano-structure of the natural bone and so far we have found promising results in terms of the scaffold supporting and enhancing material-cell interaction; i.e. attachment, proliferation and bone mineralisation of oestrogenic cell lines as well as osteoinductive properties for bone deposition and regeneration.