The Research Of BMSCs Transplantation Combined With TGF-β2/geneX Into Pulp Core Transplantation On Rabbit ONFH

[Objective] To investigate BMSCs composite transforming growth factor-beta2 and the geneX stents pulp core decompression transplantation for femoral head necrosis recovery promoting effect[Methods] BMSCs cell cultivation: take the experiment animal bone marrow for BMSCs primary culture and subculture.The healthy adult New Zealand white rabbit as experimental animals, hydrocortisone hormone to 40 mg/kg body weight, gluteal muscle injection for three days in a row,1 times a day.The models are successfully after 2 months, femoral head necrosis.Experimental animals were randomly divided into:normal group, control group, pure pulp core decompression group, pure geneX stent group, pulp core decompression and stent group, pulp bracket of core decompression+ BMSC+ TGF-beta2 group, each group of 5 animals. Lentivirus rLV-ZsGreen transfection cultivate good cells, pulp by core decompression after transplantation, GFP observation of BMSCs in animal models of distribution, survival situation.Give BMSCs after 2 months of treatment, groups of animals of the femoral head, immunohistochemical staining observation of the Cbfal, BMP2 in protein localization, Q - quantitative PCR detection Cbfal, BMP2 gene expression, Western Blot detection Cbfal, quantitative expression of BMP2 protein.[Results] By hydrocortisone hormone produced in New Zealand white rabbit femoral head necrosis in animal models.Successfully developed New Zealand white rabbit BMSCs, and has carried on the appraisal, found that the expression of specific markers CD29 and CD90.After the slow virus rLV - ZsGreen transfection BMSCs, found in the transplantation of cells with green fluorescence.By culturing and identifying good transplantation of BMSCs composite TGF beta2/geneX stent to treat femoral head necrosis in animal models, found that the slow virus rLV-ZsGreen labelled BMSCs can be distributed in experimental animals of the femoral head, present green fluorescent cells.Immunohistochemical staining results showed that Cbfal, BMP2 factor after transplantation of the enhanced its positive expression.RT-QPCR results show Cbfal, BMP2 gene expression quantity increase (P< 0.05). Western Blot results show Cbfal, BMP2 increased protein expression (P< 0.05).[Conclusion] BMSCs composite TGF-beta2/geneX by medullary core decompression of transplantation in the treatment of rabbit femoral head necrosis is better than pure treatment, its mechanism may be through raising Cbfal, the expression of BMP2 experiment, this method can be used as a new method of clinical promotion.