Express Of Conditions In Synovial-derived Mesenchymal Stem Cells Differentiating Into Hyaline Cartilage

Objective: To explore the effects of TGFβ1、BMP-2、bFGF、IGF、BMP-7、 ASA、Dex on the differentiation of synovial-derivedmesenchymal stem cells intohyaline cartilage.Methods:The synovial tissue of a rabbit knee was obtained under sterile condition and the synovial-derived mesenchymal stem cells were acquired by enzymatic digestion method and limited dilution method.. The synovial-derivedmesenchymal stem cells of the third generation were used for osteogenic/adipogenic/chondrogenic differentiation. References were checked to find relevant conditions of inducing the differentiation of synovial mesenchymal stem cells into hyaline cartilage and TGFβ1、BMP-2、bFGF、IGF、BMP-7、AsA、Dex were included in the orthogonal experiment.L8 (27) orthogonal experiment and table heading were designed by SPSS22.0statistical software while 2 level conditions were defined. The CD151+/CD44+ cell number was detected by a flow cytometer and the ratio of high expressed Col Ⅱ cells detected by immunohistochemistry stain and the total cartilage cell was regarded ashyaline cartilage conversion rate(N). Rate equaled to the result of CD151+/CD44+cell number multiplying by N, which was set as the test index. The result was further validated by immunohistochemistry stain and toluidine blue stain. And the experiment result was analyzed through direct observation as well as covariance analysis method.Results:Homogeneous cells were obtained from synovial tissue after adherent culture and purification. They were in spindle or stellate shapes, looking approximately like fibroblast. These cells gave positive reacts in oil red o staining,alkaline phosphatase staining and toluidine blue stain. TGF-β1 was found to have the strongest effect while BMP-7 was inferior to it in the orthogonal experiment.Covariance analysis showed that TGFβ1、bFGF、IGF、BMP-7、AsA had significant influences on the differentiation of SMSCs into hyaline cartilage and they were important regulating factors of this process which were in accord with the result of direct observation.Conclusions: SMSCs could achieve increment and amplification stably with a strong multidirectional differentiation ability so they were seemed as ideal seed cells.TGFβ1、bFGF、IGF、BMP-7、AsA had significant influences on thedifferentiation of SMSCs into hyaline cartilage while TGF(31 showed the strongest effect. More explorations on the factors and levels of inducing the differentiation of synovial mesenchymal stem cells into hyaline cartilage will contribute to the search of a more reasonable and precise combination in tissue engineering for the repair of cartilage defects.