Accidence Study Of The Inlfuence Of The Bone Tissue From Osteonecrosis Of The Femoral Head On The Dog Bone Marrow Mesenchymal Stem Cells Proliferation And Differentiation

Objective：To study the effects of bone tissue from osteonecrosis of the femoralhead on proliferation and differentiation of the dog bone marrow mesenchymal stemcells in vitro culture．Methods：Developing a dog model of the femoral head osteonecrosis induced byliquid nitrogen freezing. BMSCs were isolated from dog ilium bone marrow bycombination of gradient centrifugation and adherent wall culture．Different bonetissue and BMSCs were contact indirectly in vitro by co-cultured in Transwell plate.Accoding to the culture media used，three groups were set up：blank group(10％FBS／DMEM), control group (10％FBS／DMEM+bone tissue from natural femoralhead), experimental group(10％FBS／DMEM+bone tissue from osteonecrosis of thefemoral head)． Cell proliferation was measured by methylthiazol tetrazolium(MTT)method．Cell differentiation was examined by alkaline phosphatase(ALP)staining and inspect its concentration, Alizarin red staining method was used to studythe calcification effects, Oil red O staining method was used to detect if there is fatemergence．Results：As compared with blank group，the proliferation in the control groupand experiment group were significnafly promoted after culture for1day、3days and5days (P<0．05)．The proliferation of experiment group was higher than controlgroup in5th day and7th day(P<0．05). After co-culture for7days ALP staining waspositive in the control group and experiment group.In7th and9th day the ALPactivity in culture fluid was in this order control group＞experiment group＞blankgroup（P＜0.05）. Alizarin red staining show control group had the most calciumnodules（12.17±2.48,P＜0.05）,the number of calcium nodules in experiment groupwas more than blank group（P＜0.05）. Oil red O staining show there was no fatemergence after21days in every group.Conclusion：Both nature and osteonecrosis bone tissue of the femoral headcould promoted proliferation of the dog BMSCs and induced them osteogenicdifferentiation but not lipo differentiation, bone tissue from osteonecrosis of the femoral head had more competence in promoting proliferation but less capability ininducing osteogenic differentiation than natural bone tissue of the femoral head.