4^th International Conference on Tissue Science and Regenerative Medicine July 27-29, 2015 Rome, Italy

Zogopoulos Panagiotis is a resident of Neurosurgery at the General Hospital of Nikaia-Piraeus “Agios Panteleimon”, Athens, Greece. He has received a 6-month advanced clinical training (clinical fellow) at the Neurosurgery Department of Osaka University Hospital in Japan, where an ongoing clinical trial about spinal cord transplantation with olfactory mucosa autograft is being conducted. Several of his papers have been published in reputed peer-review journals and he has presented various researches in international conferences.

Transplantation of embryonic neural tissues was introduced as a technique of damaged neuronal circuitries repair since the late 1970s, however due to ethic issues, research has recently been focused on stem cells transplantation, such as olfactory mucosa neurons. Patients with complete paraplegia (ASIA grade A or B) due to spinal cord injury more than 6 months previously and with less than 3cm vertical extent of cord injury on MRI, have been included in a relevant clinical trial at the Neurosurgery Department of Osaka University Hospital. The patient\'s olfactory mucosa graft is taken endoscopically and then processed into small pieces. Afterwards, spinal cord\'s posterior medial sulcus is opened, the scar is removed and the autograft is transplanted into the cavity. Patients have demonstrated electromyographic signals in response to voluntary effort as early as 3 months after the transplantation, while emergence of motor evoked potential has also been observed, indicating the recovery of electrophysiological conductivity of the corticospinal tract. Moreover, some patients have shown improvement in motor function below the level of injury. The neural condition of the severed caudal spinal cord seems to influence the potential for motor function recovery, while the time interval between injury and transplantation does not seem to be an important factor since improvement from ASIA grade A to grade D has been achieved in one patient even when the transplantation was performed ten years after the injury. Involuntary muscle spasm before transplantantion seems to be a predictor of success of regenerative treatment in chronic spinal cord injury patients.

Mahboubeh Nabavinia is a Ph.D. student from Iran. She is working on her Ph.D. thesis in Tissue Engineering and Stem Cell Research Lab. The objective of her Ph.D. dissertation is study on generation of modular bone tissue in fluidized bed bioreactor using core-shell microcapsule technique. She used electrostatic technique for entrapping MG63 cells or ADMSCs in core- shell microcapsule. She published some papers in these fields such as modeling bioreactor for tissue engineering application in 9th Royan international congress on stem cell biology and technology.

Three dimensional matrices that encapsulate stem cells are promising for cell growth and tissue formation in tissue engineering and regenerative medicine. The aim of this study was utilizing a simple one-stage technique which offers numerous attractive features for manufacturing multi applicable cell-laden hydrogel microcapsule for cell therapy and tissue engineering. Co-axial nozzle was applied to produce uniform core- shell microcapsule by electro spraying technique. Alginate (0.75 – 1.5 %w/v) was chosen as shell and various galantine-alginate blends (2.5, 5, 10, 20, 40 w/v gelatin to alginate) were selected as core materials. Some of important operation factors such as diameter of nozzles, voltage, flow rate of extrusion, and distance of gelling bath and nozzle were considered. The co-axial nozzle had a core tube with an inter diameter of 0.45 mm and a shell tube with an inter diameter of 0.9 mm. The results showed that increasing the flow rate of core to shell decreased shell thickness while uniformity of microcapsules was improved. In addition, the size of microcapsules decreased by rising voltage and reducing flow rate of core and shell extrusion. The results showed that the flow ratio of 4, applied voltage of 5.5 KV and distance of 2.5 cm could produce the uniform microcapsules with average diameters of 800 and 700 for shell and core, respectively. Cell growth analysis using osteoblast cells cultured by initial cell density 1.5 * 106 cell/ml in the cores depicted a 12.9 fold increase after 14 days, indicating that the core- shell microcapsules improved microenvironment for cell expansion. The present study demonstrated that core- shell microcapsules have high potential for using in tissue engineering bottom- up approach as building block for manufacturing organs and complicated tissue.

Jalil Nourisa graduated his BSc and MSc both from Amirkabir University of Technology (Tehran Polytechnic), and now is research assistant at Orthopaedic & Dental Biomechanics Lab (ODBL). He currently and actively focused his research on the biomechanical investigation of fixation devices for distal tibial fracture, mechano-regulation algorithms, and bone scaffolds

Bone secondary healing process consists two different ossification mechanisms, i.e. intramembranous ossification and endochondral ossification (EO). EO is distinguished by differentiation of mesenchymal stem cells (MCs) into chondrocytes and sequential tissue differentiation of: fibrous tissue (FT); cartilage (CT); immature bone (IB); and mature bone (MB). Several mechano-regulation (MR) algorithms have been proposed in the literature to date in order to predict the healing process exclusively followed EO. Some clear differences were observed among possible pathways of tissue development proposed in different algorithms. The aim of this study was to address the question of which pathways can predict EO the best by investigating the impact of different pathways on the EO. For This purpose, a 2D model of a broken tibia, with a 3mm fracture gap, was constructed, and differentiation of stem cells and development of tissues in the determined pathways was implemented as a biofeedback loop through using Python scripting in Abaqus software. Results of this study showed that the algorithm which restricts tissue differentiation from granulation tissue (GT) to CT, failed to predict bony bridge and some small islands of bone were left in some area of internal and peripheral callus. On the other hand, healing patterns predicted by algorithms which allow pathway of jumping from GT and FT to IB without passing through the CT, showed no differences in comparison with the original theory in which no constraints were applied in tissue differentiation processes. However, by considering the constraint that in order to develop BT, CT should already exists, it significantly improves our prediction, which is in agreement with experimental evidence showing that EO can take place though calcification of CT. Based on this investigation, two pathways should be considered crucial for correct prediction of EO: first, GT to CT, and second, GT and FT should not be allowed to directly differentiate into bone phase. Results of this work suggest new regulatory algorithm by defining development pathways according to biological events, and is able to correctly predict EO.rnKeywords: Endochondral ossification, Mechano-regulation algorithm, Fluid velocity, Octahedral shear strain, Finite element method, Callus formationrn

Hamid Kahkouei graduated his BSc from IAUCTB and MSc from SRBIAU, Iran. He focused his research on the biomechanical investigation of the tissue scaffold, and currently investigate the impact of internal architecture on the mechanical properties of the scaffolds.

Biodegradable scaffolds are widely used in the tissue engineering in order to provide a temporary structural template for cell seeding and extracellular matrix production. The porosity and pore size are two crucial parameters for scaffold success from both of mechanical and biological views. Although the influence of porosity on the mechanical behavior of scaffold was well-determined, studies about the impact of pore sizes are rare. This study was an attempt to determine the influence of variation in the pore size on the mechanical properties of a scaffold. For this purpose, finite element models with different pore sizes in the range of 40-350 μm were constructed in Abaqus software by use of Python scripting considering three different porosities of 55%, 70%, and 85%. Material properties of Poly (D, L-lactide) - PDLLA were assigned for entire models. Results show that by increase in the scaffold pore size, the effective Young’s modulus decreases with second-degree polynomial functions. On the contrary, the calculated maximum von Mises stress on the scaffold showed reduction by increase of pore size with the same trend. The increase of the poro size caused about 13% reduction in the effective Young’s modulus, and 12% to 22% increase in the stress. Results of this study can be used to better understanding of internal architecture impact on the mechanical behavior of scaffold. rnrnKeywords: Biodegradable, Scaffold, Porosity, Pore size, Finite element, PDLLA rn

Luciana Barros Sant’ Anna obtained her Master in Buco Dental Biology and Pathology (2001) and PhD in Buco Dental Biology- focus on Histology (2004) from Campinas University , SP, Brazil. In 2009 she completed her Postdoctoral studies in the field of human adult stem cells, with focus on cells and tissues derived from placenta at Centro di Ricerca E.Menni, Fondazione Poliambulanza, Istituto Ospedalieiro, Brescia, Italy. Currently, she is Researcher of Imunology Laboratory of the Institute of Research & Development, from University of Vale do Paraíba, São José dos Campos, SP, Brazil and Professor of Embriology, Histology and Anatomy of the same University. Since 2009 she is member of International Placenta Stem Cell Society (IPLASS).

Human placenta at term, which is usually discarded as medical waste, has attracted much attention as a source of tissues and stem / progenitor cells for research and potential clinical applications. The anti-inflammatory, antiscarring, and wound healing properties of amniotic membrane (AM) could make her useful for development of new therapeutic approaches in regenerative medicine and warrant studies of their effects in animal models of inflammatory and fibrotic pathologies. Biliary fibrosis and resultant cirrhosis are among the most common outcome of chronic liver disease. Currently, liver transplantation remains the only effective treatment. Here, we investigated if the intact hAM has the capacity to reduce a well-established liver fibrosis induced in rat by bile duct ligation (BDL) model. After 2 weeks from BDL, the liver was covered with a fragment of hAM, or left untreated. Six weeks later, the fibrosis was first assessed by semiquantitative scoring systems and by CellProfiler digital image analysis to quantify the area occupied by ductular reaction, activated myofibroblasts, collagen deposition, and by transforming growth factor (TGF)-1, the main profibrogenic factor in chronic liver disease. After 6 weeks of BDL, AM rats showed a significant reduction in the severity of fibrosis. The amount of collagen deposition, the ductular reaction and the myofibroblasts were all reduced to about 50% of levels observed in BDL rats. AM was able to markedly attenuated TGF-1immunoexpression in liver. These findings suggest that hAM patching, is useful for treating cholestatic fibrosis and the mechanism was partly by reduction in the TGF-1expression. \r\nAcknowledgments: This study was supported by grant # 2011/879-8, São Paulo Research Foundation (FAPESP).\r\n

Luciana Barros Sant’ Anna obtained her Master in Buco Dental Biology and Pathology (2001) and PhD in Buco Dental Biology- focus on Histology (2004) from Campinas University , SP, Brazil. In 2009 she completed her Postdoctoral studies in the field of human adult stem cells, with focus on cells and tissues derived from placenta at Centro di Ricerca E.Menni, Fondazione Poliambulanza, Istituto Ospedalieiro, Brescia, Italy. Currently, she is Researcher of Imunology Laboratory of the Institute of Research & Development, from University of Vale do Paraíba, São José dos Campos, SP, Brazil and Professor of Embriology, Histology and Anatomy of the same University. Since 2009 she is member of International Placenta Stem Cell Society (IPLASS).

\r\nHuman placenta at term, which is usually discarded as medical waste, has attracted much attention as a source of tissues and stem / progenitor cells for research and potential clinical applications. The anti-inflammatory, antiscarring, and wound healing properties of amniotic membrane (AM) could make her useful for development of new therapeutic approaches in regenerative medicine and warrant studies of their effects in animal models of inflammatory and fibrotic pathologies. Biliary fibrosis and resultant cirrhosis are among the most common outcome of chronic liver disease. Currently, liver transplantation remains the only effective treatment. Here, we investigated if the intact hAM has the capacity to reduce a well-established liver fibrosis induced in rat by bile duct ligation (BDL) model. After 2 weeks from BDL, the liver was covered with a fragment of hAM, or left untreated. Six weeks later, the fibrosis was first assessed by semiquantitative scoring systems and by CellProfiler digital image analysis to quantify the area occupied by ductular reaction, activated myofibroblasts, collagen deposition, and by transforming growth factor (TGF)-1, the main profibrogenic factor in chronic liver disease. After 6 weeks of BDL, AM rats showed a significant reduction in the severity of fibrosis. The amount of collagen deposition, the ductular reaction and the myofibroblasts were all reduced to about 50% of levels observed in BDL rats. AM was able to markedly attenuated TGF-1immunoexpression in liver. These findings suggest that hAM patching, is useful for treating cholestatic fibrosis and the mechanism was partly by reduction in the TGF-1expression. \r\nAcknowledgments: This study was supported by grant # 2011/879-8, São Paulo Research Foundation (FAPESP).\r\n

Marek Konop is a PhD student at Mossakowski Medical Research Centre PAS. My supervisor is prof. Lidia Rudnicka MD, PhD. His PhD thesis concerns of the influences of natural derived structural proteins in healing process (especially in diabetic wound model). During my PhD studies Marek Konop obtained KAP dressing (according method proposed by Lipkowski) and diabetic wound model.

Introduction\r\nWound repair is a mechanism whereby the body tries to restored the integrity of the tissues. However in some diseases such as diabetes mellitus the healing process is impaired. The main goal of tissue engineering and regenerative medicine is to assemble functional scaffolds that restore, maintain, or improve damaged tissues or whole organs.\r\nTo enhance healing processes we developed keratin associated proteins (KAPs) powder prepared from mice fur, which added to damaged site, could be easily colonized by migrated epithelial cells. \r\nAim of the study: was to investigate the effect of KAP on wound healing rate using surgical wound model in diabetic mice. \r\nMaterials & methods: Chemical & biological studies\r\nProcedure of KAP preparation was carried out according method proposed by Lipkowski et al.1 Obtained m-KAP were impregnated by silver nanoparticles as established earlier by Sileikaite et al.2 Simultaneously to the KAPs preparation, pharmacologically induced diabetes was developed in mice as described by Wu et al.3 When diabetes was stable two wounds were made by skin incision and covered by KAPs. Wound tissue samples were collected on day 4, 7 and 14 and processed for histopathological studies. \r\nConclusions & Results \r\nDeveloped KAPs were characterized by very low immunogenicity. Keratin covering has been absorbed by regenerating skin during healing process. Additionally keratin powder well absorbed exoduses, while maintaining moist environment within wound area. Our preliminary results confirmed that keratin powder could be used as a wound dressing well adsorbed in regenerating skin during healing process.\r\n

Micheu Miruna Mihaela has an experience for over 14 years in the Clinical Emergency Hospital of Bucharest, where she currently is working as a cardiologist and scientific researcher in the Cardiology Department. She has completed her Ph.D. from the \"Carol Davila\" University of Medicine and Pharmacy Bucharest in 2012 with the thesis: “New Therapeutic Methods in Acute Myocardial Infarction: Autologous Stem Cell Transplant”. Her expertise includes stem cell therapy in cardiovascular diseases, cell culture and flow cytometry.She has participated with studies at over 40 scientific events from various countries and has published papers in different scientific journals.\r\n

Background. Reduced left ventricular ejection fraction (LVEF) after ST-segment elevation myocardial infarction (STEMI) has been recognised as a predictor of morbidity and mortality. The aim of this study was to investigate the effect of autologous bone marrow stem cell (ABMSC) therapy on major adverse cardiovascular events (MACE) in patients with STEMI and impaired LVEF, during a 6 months follow-up.\r\nMethods and materials. Eighteen patients hospitalized for a first STEMI treated by successful primary percutaneous coronary intervention (PCI) and LVEF < 40% were included. Clinical exams and complete echocardiograms were performed at 1, 3 and 6 months after STEMI, comprising routine angiographic follow-up at 6 months. In the ABMSC group, 50 ml of bone marrow were harvested 7 to 13 days after STEMI, the cell suspension being delivered via intracoronary route during the same day. \r\nResults. Patients with ABMSC therapy had decreased readmission for heart failure and decreased coronary revascularization compared to control patients (14.3% vs 36.4%, p > 0.05). There were no deaths during 6 months follow-up period in experimental group, nor in control group.\r\nConclusions. We observed a positive effect of ABMSC therapy in STEMI patients with severe systolic function impairment regarding the occurrence of MACE, even though there were no significant differences between groups - probably due to small number of patients included and short follow-up period. \r\nAcknowledgement. This paper is supported by the Sectoral Operational Programme Human Resources Development (SOP HRD), financed from the European Social Fund and by the Romanian Government under the contract number POSDRU/159/1.5/S/137390.\r\n