Experimental Study On The Treatment Of The Rabbits' Aseptic Necrosis Of The Femoral Head With Adipose-derived Mesenchymal Stem Cells (ADMSCs) Combining Nano-hap/Collagen(NHAC)

PARTⅠ: The Experimental Study on Inducing Rabbit Adipose-derived Mesenchymal Stem Cells into Osteoblast in VitroObjective: To study the bone tissue osteogenesis ability of adipose mesenchymal stem cells (ADMSCs) in vitro in order to offer the ideal seed cells for tissue engineering.Methods: In vitro for the third generation of rabbit adipose-derived mesenchymal stem cells,and then the biologic characters of the ADMSCs was observed and test.Results: Osteoblast can be induced from ADMSCs by osteoblast conditioned medium,moreover can secret ALP and format the calcific nodules.Conclusions: ADMSCs can be induced successfully to osteoblast in vitro and can be used as ideal seed cells for bone tissue engineering.PARTⅡ: Chondrogenic Induction of Rabbit Adipose Mesenchymal Stem Cells in VitroObjective: To study the bone tissue chondrogenic ability of adipose mesenchymal stem cells (ADMSCs) in vitro in order to offer the experimental statements for tissue engineering.Methods: The third generation of rabbit adipose-derived mesenchymal stem cells which taking in vitro,inducing to Chondrocytes and then the biologic characters of the chondrocyte were observed and tested.Results: ADMSCs because round or ovoid from spindle shape exposure to TGF-β1 and vitamin C under cartilage culture fluid,then turned into chondrogenic nudoles.The test of Alcian blue staining,safrarine"O"are positive,and immunohistochemistry staining for typeⅡcollagen show positive.Conclusions: Rabbit ADMSCs may be induced into chondrocytes under conditional culture media in vitro.PART III: The study on the biocompatibility of the nano-hap/collagen with adipose mesenchymal stem cellsObjective: To observe the biocompatibility of ADMSCs and NHAC by coculturing them in vitro.Methods: ADMSCs and NHAC were cocultured in vitro .After 1,3,5 day,we examined the ADMSCs and NHAC by inverted microscope , laser scanning confocal microscope observation.Results: The ADMSCs can adhere to, grow and proliferate normally and perfectly on the serface and holes of NHAC.Conclusions: NHAC has good biocompatibility and can be used as ideal biomaterial in bone tissue engineering.PARTⅣ: Experimental study on repairing the rabbits bone perthes'disease with ADMSCs combining NHACObjective: To explore the impact of NHAC seeded with ADMSCs repairing the rabbits perthes disease.Methods: The adipose-derived mesenchymal stem cells combined with the new nano-materials implanted perthes disease animal models, in both simple system modules, modeling after the implantation of pure nano-bone group comparison, respectively, 2,4,6-week observation femoral head shape recovery, X-ray examination, MRI examination, histopathological examination, viewing vascular growth by FⅧ-Ag by immunohistochemistry, Observing the marked ADMSCs distribution and proliferation in vivo by BrdU immunohistochemistry of inspection.Results: The observation group are able to repair the femoral head, but the experimental group, the femoral head shape restoration and recovery time are better than the control group.Conclusions: The adipose-derived mesenchymal stem cells combined with the new nano-materials can effectively shorten the rabbit perthes disease course and speed up the recovery rate and extent of the femoral head.