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Hypoxic Preconditioning Improved Neuroprotective Effect of Bone Marrow-Mesenchymal Stem Cells Transplantation in Acute Glaucoma Models

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Journal of Biomedical Science and Engineering 2016年第4期9卷 245-257页

作者： Titiek Ernawati Gatut Suhendro I Ketut Sudiana Suhartono Taat Putra Harjanto JM Sunarjo Agus Turchan fedik abdul rantam Department of Ophthalmology School of Medicine Widya Mandala University Surabaya Indonesia PHC Hospital Surabaya Indonesia Department of Ophthalmology School of Medicine Airlangga University Surabaya Indonesia Dr. Soetomo General Hospital Surabaya Indonesia Department of Pathology Anatomy School of Medicine Airlangga University Surabaya Indonesia Department of Physiology School of Medicine Airlangga University Surabaya Indonesia Department of Public Health School of Medicine Airlangga University Surabaya Indonesia Department of NeuroSurgery School of Medicine Airlangga University Surabaya Indonesia Laboratory of Stem Cell Institute of Tropical Disease Airlangga University Surabaya Indonesia Regenerative Medicine & Stem Cell Centre Airlangga University Surabaya Indonesia Laboratory of Virology and Immunology Department of Microbiology School of Veterinary Medicine Airlangga University Surabaya Indonesia

This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs) in Acute Glaucoma Models. The methods of this research were isolated mesenchymal stem cells from the bone marrow of adult wild-type Sprague-Dawley (SD) rats. BM-MSCs were cultured under normoxic or hypoxic (1% oxygen for 24 hours) conditions. Normoxic or hypoxic BM-MSCs were transplanted intravitreally 1 week after ocular hypertension induction by acutely increasing IOP to 100 - 120 mmHg for 60 minutes. Rats were killed 4 weeks after transplanted. Apoptosis was examined by tunnel assay and expression Brn3b (Brn3b = RGCs marker) by immunohistochemical analysis of the retina. Results showed that transplantation of hypoxic preconditioning BM-MSCs in acute glaucoma models resulted in a significant apoptosis decreasing (p < 0.05) and an significant increasing in RGCs (p < 0.05), as well as enhanced mor-phologic and functional benefits of stem cell therapy versus normoxic BM-MSCs transplantation. Conclusions: Hypoxic preconditioning enhances the capacity of BM-MSCs transplantation to improve neuroprotective effects of RGCs in Acute Glaucoma Models.

关键词： Hypoxic Preconditioning Transplantation Bone Marrow-Mesenchymal Stem Cells BM-MSCs Glaucoma Neuroprotective

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Study Biocompatibility and Osteogenic Differentiation Potential of Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) with Gelatin Solvent

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Journal of Biomedical Science and Engineering 2015年第7期8卷 420-428页

作者： Nike Hendrijantini Utari Kresnoadi Sherman Salim Bambang Agustono Endang Retnowati Iwan Syahrial Pungky Mulawardhana Manggala Pasca Wardhana Coen Pramono fedik abdul rantam Department of Prosthodontic Faculty of Dental Medicine Airlangga University Surabaya Indonesia Department of Clinical Pathology Airlangga University & Dr. Soetomo General Hospital Surabaya Indonesia Department of Pathology Faculty of Veterinary Medicine Airlangga University Surabaya Indonesia Department of Obstetric & Gynecology Airlangga University & Dr. Soetomo General Hospital Airlangga University Surabaya Indonesia Department of Oral and Maxillofacial Surgery/Dental Hospital Faculty of Dental Medicine Airlangga University & Dental Hospital Surabaya Indonesia Laboratory of Stem Cell Institute of Tropical Disease Airlangga University Surabaya Indonesia Regenerative Medicine & Stem Cell Centre Airlangga University & Dr. Soetomo General Hospital Surabaya Indonesia

The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) phenotypic characterization, 2) The in-vitro osteogenic differentiation potential of hUCMSCs, 3) The cytotoxicity of gelatin solvent to hUCMSCs using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. As a result, through characterization of hUCMSCs, the majority of target cells expressed specific MSCs markers, Cellular Differentiation (CD)73, smaller number of subpopulation expressed CD90 with only minimal subpopulation expressed CD105 and all negative MSCs markers. Osteoblastic differentiation was found in a significantly high number of cells when in vitro osteogenic differentiation of hUCMSCs with Alizarin Red staining was done. Biocompatibility analysis using the MTT assay showed that gelatin solvent and Alpha modification of minimum essential medium Eagle (α-MEM) was non-toxic for hUCMSCs in vitro. The study concluded that hUCMSCs isolated from human umbilical cord was capable of undergoing in vitro osteogenesis, indicating its potential as allogeneic stem cells for clinical application in bone tissue engineering.

关键词： Human Umbilical Cord Mesenchymal Stem Cells Mesenchymal Stem Cells Phenotypic Characterization Osteogenic Differentiation Bone Tissue Engineering

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The Osteogenic Capacity of Human Amniotic Membrane Mesenchymal Stem Cell (hAMSC) and Potential for Application in Maxillofacial Bone Reconstruction <i>in Vitro</i>Study

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Journal of Biomedical Science and Engineering 2014年第8期7卷 497-503页

作者： David B. Kamadjaja Purwati fedik A. rantam Ferdiansyah Coen Pramono Department of Oral & Maxillofacial Surgery Faculty of Dentistry Airlangga University Surabaya Indonesia Laboratory of Stem Cell Institute of Tropical Disease Airlangga University Surabaya Indonesia Regenerative Medicine & Stem Cell Centre Airlangga University & Dr. Soetomo General Hospital Surabaya Indonesia

Amniotic membrane of human placenta is a source of abundant mesenchymal stem cell (hAMSC) which makes it a potential source of allogeneic multipotent cell for bone healing. However, much has to be explored about its isolation procedure and the osteogenic differentiation potential. The aims of this study are to establish the procurement procedure of human amniotic membrane, the isolation and culture of hAMSC, the MSC phenotypic characterization, and the in vitro osteogenic differentiation of hAMSC. Results of the study are as follows. The quality of human amniotic membrane would be best if procured from Caesarean operation under highly aseptic condition to avoid fungal and bacterial contamination on the culture. Isolation procedure using modified Soncini protocol yielded large amount of MSC with high proliferative capacity in culture medium. Characterization of hAMSC showed that the majority of the target cells exhibited specific MSC markers (CD105 and CD90) with a small number of these cells expressing CD45, the marker of hematopoeitic cells. The in vitro osteogenic differentiation of hAMSC followed by Alizarin Red staining showed that osteoblastic differentiation was detected in a significantly high number of cells. This study concludes that hAMSCs isolated from human amniotic membrane have the capacity for in vitro osteogenesis which makes them be one of the potential allogeneic stem cells for application in maxillofacial bone reconstruction.

关键词： Human Amniotic Membrane Mesenchymal Stem Cells Modified Soncini Protocol MSC Phenotypic Characterization Osteogenic Differentiation Allogeneic Stem Cell Maxillofacial Bone Reconstruction

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Healing Mechanism and Osteogenic Capacity of Bovine Bone Mineral—Human Amniotic Mesenchymal Stem Celland Autogenous Bone Graft in Critical Size Mandibular Defect

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Journal of Biomedical Science and Engineering 2015年第10期8卷 733-746页

作者： David B. Kamadjaja Purwati fedik A. rantam Ferdiansyah D. Coen Pramono Department of Oral & Maxillofacial Surgery Faculty of Dentistry Airlangga University Surabaya Indonesia Laboratory of Stem Cell Institute of Tropical Disease Airlangga University Surabaya Indonesia Regenerative Medicine & Stem Cell Centre Airlangga University & Dr. Soetomo General Hospital Surabaya Indonesia Department of Internal Medicine Dr. Soetomo General Hospital Surabaya Indonesia Department of Orthopaedic & Traumatology Dr. Soetomo General Hospital Surabaya Indonesia

Experiments on maxillofacial bone tissue engineering showed the promising result;however, its healing mechanisms and effectiveness had not been fully understood. The aim of this study is to compare the bone healing mechanism and osteogenic capacity between bovine bone mineral loaded with hAMSC and autogenous bone graft in the reconstruction of critical size mandibular bone defect. Critical size defects were made at the mandible of 45 New Zealand white rabbits reconstructed with BBM-hAMSC, BBM alone, and ABG, respectively. At the end of first, second, and twelfth weeks, five rabbits from each experimental week were sacrificed for histology and immunohistochemistry staining. Expressions of vascular endothelial growth factor (VEGF), bone mor-phogenic proteins-2 (BMP2), Runx2 and the amount of angiogenesis were analyzed in the first and second week groups, while expressions of Runx2, osteocalcin, collagen type-I fibres, trabecular area and bone incorporation were analyzed in the twelfth week groups. The result showed that expressions of VEGF, BMP2 and Runx2 as well as the amount of angiogenesis were higher in ABG compared with BBM-hAMSC group in the first and second weeks of healing. The result of twelfth week of healing showed that expressions of Runx2 and osteocalcin as well as the thickness of collagen type-I fibres were significantly higher in BBM-hAMSC compared to ABG group, while there was no statistically difference in trabecular area and bone incorporation between BBM-hAMSC and ABG group. This study concluded that early healing activities were higher in auto-genous bone graft than in BBM-hAMSC, while osteogenic activities in the late stage of healing were higher in BBM-hAMSC compared to autogenous bone graft. It was also concluded that the osteo-genic capacity of BBM-hAMSC was comparable to autogenous bone graft in the reconstruction of critical size defect in the mandible.

关键词： Bone Healing Mechanism Osteogenic Capacity Human Amniotic Mesenchymal Stem Cell Bovine Bone Mineral Autogenous Bone Graft Critical Size Mandibular Bone Defect

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Induced Monocytes-Derived HSCs (CD34+) with LPS Accelerated Homing Rat Bone Marrow-Mesenchymal Stem Cell (BM-MSCs, CD105) in Injured Pancreas

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Journal of Biomedical Science and Engineering 2015年第5期8卷 333-344页

作者： fedik A. rantam Purwati Budi Setiawan Sony Wibisono Ferdiansyah Joni Wahyuhadi Edward Mouli Dwikora N. Utomo Heri Suroto Candra Bumi Stem Cell Laboratory Institute of Tropical Disease Airlangga University Surabaya Indonesia Virology and Immunology Laboratory Department of Microbiology Faculty of Veterinary Medicine Airlangga University Surabaya Indonesia Tropic and Infection Division of Internal Medicine School of Medicine Airlangga University Surabaya Indonesia Animal Surgery Animal Hospital Faculty of Veterinary Medicine Airlangga University Surabaya Indonesia Regenerative Medicine Department of Orthopedic Dr. Soetomo Teaching Hospital/School of Medicine Airlangga University Surabaya Indonesia Department of Neurosurgery Dr. Soetomo Teaching Hospital/School of Medicine Airlangga University Surabaya Indonesia

Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration of homing process mechanism in injured pancreas. Mononucleated stem cells were isolated from aspirated whole rat BM using ficoll and cultured in α-MEM complete growth medium in 10 cm petridish. After two days, adherent cells after washing twice in petridish were added α-MEM growth medium and then mesenchymal cells were characterized using CD105 marker in third passage and labeled PKH26. Then haemopoietic stem cells (HSCs) were isolated with magnetic beads CD34+ and differentiated in vitro, and then induced monocytes with LPS. Animal experiment used 28 male Wistar rats, and divided them into 4 groups. After transplantation combined, both cells between monocyte derived HSc (mHSCs) and rat BM-MSC were analyzed expression of pair box gen 4 (Pax4), pancreatic and duodenal homeobox (Pdx1), C-peptide using immunohistochemistry, then secretion of insulin and C-peptide analyzed using indirect ELISA. Results showed that the expressions of Pax4, Pdx1, C-peptide found in the surface membrane cell of pancreatic cell, and secreted C-peptide and insulin were shown significant (P < 0.05) in transplanted group 2, 3 and 4, but in group 3 were transplanted with combined cells more dominant than non-combined cells. Conclusions suggested that combining of induced monocytes-derived HSCs and rat BM-MSCs has accelerated homing MSCs into injured pancreatic tissue.

关键词： Induced Monocyte Derived HSCs Rat BM-MSCs Homing Injured Pancreas

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