Day 3 :
- 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.