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Objective:Through the observation of expansion、purification and biology characteristic of rat BMSCs,Study the effect of the Chinese herbal on BMSCs’ proliferation and differentiation into cartilage,make sure the best function concentration of NG promoting BMSCs to proliferate and inducementing differentiation to the cartilage,And compared With the Classical Induction Group,Collaborative Group,study the possible function mechanism of repair to the bone and the joint,then provides theories for the prevention and treatment of OA.Methods:1 Make use of the method of the whole marrowses stick to the wall,separated,culture the BMSCs,and to identify their biological characteristics;2 With MTT method,to survey the effect that NG-intervention liquid on proliferation of rat BMSCs.make sure which concentration is the best one,then assurance the dose-effect relationship between NG and BMSCs proliferation.3 Using the Toluidine blue staining and typeⅡcollagen immunofluorescence technique,the two specific composition of cartilage-matrix proteoglycan and collagen typeⅡwere identified,to confirm whether rats BMSCs induced differentiation into cartilage by NG intervention liquid,and make sure of its best function concentration,we make a quantitativing analysis of the glycosaminoglycan,which is the cartilage-specific components.4 With different concentrations of NG intervention liquid United classical fluid-induced induced rats BMSCs differentiation into cartilage(Collaborative Group),and simultaneously, with the Classical Group、Blank Control Group for comparison,by dimethyl methylene blue staining,measure the quantitative of the cartilage-specific cells composition—Glycosaminoglycan,study NG intervention on inducing BMSCs differentiation into cartilage. Results:1 Being isolated,BMSCs adherent the base of the culture flask in 24h,and about 9～11days reach fusion,after three generations,as it was carried on inducemented into cartilage、osteoplast and lipocyte,BMSCs successful show this three kinds of cell phenotype.2 With the function of different concentrations NG intervention liquid,the MTT result shows: different concentrations of NG intervention liquid makes different effects on BMSCs’ proliferation,low-concentration group boost proliferation,the most obviously group is the 1μg/ml group,compared with the Blank Group,P=0.043＜0.05,there is statistical significance,then the other groups compared with Blank Group,P＞0.05,no statistically significant;NG intervention liquid of different concentrations and cell proliferation quantity changes disproportionately.3 The identification by toluidine blue staining and typeⅡcollagen immunofluorescence technique shows:Blank Group and different concentrations of pure NG intervention liquid can not induced the rat BMSCs differentiation into chondrocytes,cartilage cells’positive expression can be seen in the classic group.4 After 10 days and 15 days Inducing rat BMSCs differentiation into chondrocytes by different cooperative groups,the methyl-methylene blue staining shows:the content of glycosaminoglycan in each groups of the two compared with the Classical Group,the Collaborative Groups＞the Classical Group,two groups have significant differences(P＜0.05),the 0.1μg/ml、0.2μg/ml two Collaborative Groups separately compared with the Classical Group,the Cooperative Groups＜the Classic Group,there are not significant difference(P＞0.05).Conclusions:1 Make use of the method of the whole marrowses stick to the wall,can obtain good uniformity of vigorous growth rats BMSCs,establish a stable system of vitro culture;2 Different concentrations of NG intervention liquid on rat BMSCs’ proliferation are different, 1μg/ml group is the most obviously group;3 Different concentrations of pure NG interfere liquid could not induced rat BMSCs differentiation into chondrocytes;1μg/ml,2μg/ml NG intervention liquid have synergy with the Classic Group,they can enhance the effect of induction,however,the 0.1μg/ml,and 0.2μg/ml groups have not significant synergy with the Classic Group.
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