Effects of platelet derived growth factor BB and basic fibroblast growth factor on phenotype of tenocytes and the mRNA expression of tenocyte phenotypic and differentiation markers

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Objective To evaluate the the changes of tenocyte phenotype and the expression of the phenotypic markers by adding platelet derived growth factor BB (PDGFBB) and basic fibroblast growth factor(bFGF) to human tenocyte in vitro culture.Methods Human tenocytes were cultured in α-MEM medium by adding FBS at the concentration of 1％ and supplementing either/both PDGFBB and bFGF.Sirius red staining was employed to evaluate the characteristics of the tenocytes cultured.Real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) technique was used to detect the mRNA expression of the tenocyte phenotypic and differentiation markers.Results The tenocytes cultured for 14 days with 1％ FBS,50 μg/L PDGFBB + 50 μg/L bFGF showed less prominent collagen synthesis in comparison with those cultured in 10％ FBS.The tenocytes cultured in the treated group also showed down-regulated mRNA expression of those markers [Scleraxis (Scx) 0.33,t =374.055 ; Tenomodulin (Tnmd) 0.003,t =28 199.418; Collagen type Ⅰ (Col Ⅰ) 0.57,t =164.195 ; Decorin (Den) 0.26,t =177.52).Conclusion These findings suggest that human tenocytes could not only maintain their phenotype in the culture media with low concentration of FBS,but also keep the cells at less differentiated state,having this approach a suitable one for tendon tissue engineering. Key words: Tissue engineering; Tenocyte; Basic fibroblast growth factor; Differentiation

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