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There is a summary for the study of MSCs in the area of osteonecrosis of the femoral head.It discusses the bionomics, isolation and culture, and osteogenesis of MSCs to expose its internal character, which has the potentiality to treat the osteonecrosis; Through the discussion of clinical application of MSCs in treating osteonecrosis of the femoral head, it provides the feasibility to treat the osteonecrosis with MSCs. At last, it analyzes and prospects the problems, which would be faced in the near future.
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[Objective]To discuss the tissue engineering method of treating bone defects through rabbit bone marrow mesenchymal stem cells(MSCs)'culture in vitro and make composite grafts with anti-extracted bovine cancellous bone(BCB).The composite grafts were implanted into segmental radial defects in rabbits was compared on the curative effect of repairing segmental bone defects in rabbits with the composite grafts.[Method]The purified,culture-expanded MSCs were combined with BCB in vitro according to condition of cell culture.The composite grafts were tested by scanning electron microscope.An bone defect(15mm in length) was created at two radii in each rabbit.The composite grafts were implanted into segmental radial defects in rabbits through open operation.The curative effect was evaluated by radiographic examination,histologic analysis,biomechanical test after surgery in experimental groups and control's and blank's.[Result]Roentgenographically,the bone defects that had been treated with grafts exhibited new bone formation increased with time.But the result of experimental groups were apparently superior to that of the control groups′s at 2、4、8、12、16 week after operation(P
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[Objective]To evaluate whether transplanted marrow mesenchymal stem cells interfered in TGF-?1 can differentiate to nucleus pulposus cells and increase the amount of proteoglycan and collagenase Ⅱ content in intervertebral discs.[Method]We used an in vivo model to investigate the feasibility of marrow mesenchymal stem cells that cultured in vitro and interfered in TGF-?1 delivery,retention,and survival in the degeneratived disc space.In 2,4,6,8 weeks we used immunohistochemical staining to determine the change of collagenase Ⅱ content;spectrophotometry to determine the change of amount of proteoglycan with Phlorglucinol;the experiment date were analyzed by SPSS 11.5 soft ware.[Result]We found MSCs could maintain viability and proliferate within the rabbit inter vertebral disc.The amount of proteoglycan and collagen Type Ⅱ content of the intervertebral in matrix synthesis in the experiment group was increased in 8 weeks.We found no changes in the modle group.[Conclusion]Our data suggest that transplanted marrow mesenchymal stem cells in vivo can survive and increase proteoglycan and collagen Type Ⅱ amount interfered in TGF-?1 in some periods,which support its potential use as a treatment of intervertebral disc degeneration.
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Objective To construct the adenoviral vector carrying Mus-?NGF gene and to study the expression of the gene in the marrow mesenchymal stem-cells(MSCs).Methods Mus-?NGF cDNA was cloned from mice glandulae maxillaries whole mRNA by reverse-transcription polymerase-chain-reactions.The Mus-?NGF cDNA was positively cloned into the adenoviral shuttle vector pAdtrack carrying cytomegalovirus promoter(CMV)and tag-protein(GFP),and then homologous recombined into competent BJ5 183 cells with the adenoviral backbone plasmid pAdeasy-1.The recombinant adenoviral plasmid was identified by restriction enzymes and transfected into HEK293 cells to package and amplify recombinant adenoviral particles which was capable of infecting and would express ?NGF and GFP proteins.After the adenovirus infected the MSCs,the expression was observed via fluorescent microscrope and detected using immunocytochemical method.The secretion of NGF was detected by WB.Results The recombinant adenoviral vector carrying Mus-?NGF gene was constructed and confirmed by restriction endonuclease enzyme analysis.The relucent green fluor-light would be observed in the MSCs.IHC showed that Mus-?NGF gene was exactly transcripted and expressed in the MSCs,and the secretion of NGF was confirmed.Conclusion The exogenous gene,such as ?NGF,an be imported into the MSCs efficiently by recombinant adenoviral vector,and can be expressed and secreted successfully in the cells.It improves foundation for the celluar transplant and gene intervention of MSCs expressing NGF in inner ear.
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@#ObjectiveTo observe the treatment possibility and effects on rats models with experimental femoral artery obliterans(EFAO) by transplantation of endothelial progenitor cells(EPC) from bone marrow. MethodsEFAO rat models were successfully made. Bone marrow mesenchymal stem cells(MSCs) were isolated from allogeneic male Wistar rat,cultured in DMEM in vitro. EGF,bFGF and IGF-1 were added into culture medium on the 10th day and the active EPC were labled with BrdU. 5×106 of the above cells were transplanted into the right hindlimbs i.m. as Group A in the 16 rats,and the same volume of normal saline into the opposite as Group B as control. Laser Doppler perfusion imaging was taken on the 0 day、the 14th day and the 28th day after transplantation. On the 14 day and the 28 day 8 rats were sacrificed respectively and muscles of all hindlimbs were extracted for immunohistochemical examinations (by FⅧ and BrdU).Results1. In group A the skin blood perfusion in hindlimb were significantly increased as compared with group B(P<0.05). 2. Some positive stained EPC by BrdU were found in Group A but not in Group B. 3. The numbers of blood vessels with positive staining of FⅧ in hindlimb on the 28 day in Group A were more obviously than that in Group B (P<0.01). ConclusionTransplantation of EPC from bone marrow can significantly increase the skin blood perfusion and capillary density in ischemic hindlimbs in EFAOrats.