Experimental Study Of The Connective Tissue Growth Factor On Proliferation And Dose-effect Relationship Of Tendon Cells In Vitro

Objective: To study the effect of the connective tissue growthfactor of different density on proliferation oftendon cells in vitro, and toinvestigate the possible mechanism of CTGF in repairing the tendoninjury.Methods: We got the tendon from the posterior limbs of rats,tendoncells were obtained from the tendon segments after enzymatic digestion,then we cultured the cells with 10% fetal calf serum(FCS), the cells werepassaged when 80% were adherenced.we observed the morphous andgrowth regularity of the cells.The second passage celles were identifiedby immunohistochemistry to make sure that they were tendon cells. Thethird passage tendon cells of rabbit in vitro were planted in 48 wells of a96-well culture plate and every well contained 4×10~3 cells. They weredivided into 8 groups and every group contained 6 wells. According toCTGF concentration, 5ng/ml CTGF group,10ng/ml CTGF group,20ng/ml CTGF group,50ng/ml CTGF group,100ng/ml CTGF group,200ng/ml CTGF group,500ng/ml CTGF group, control group(no BFGF).When all the groups being in exponential phase of growth undermicroscope, they were dyed by MTF method and OD value of every wellwas detected by enzyme linked immunodetection meter.Results: We got the results after the MTT test by contrast to OD mean of each group.There was no difference in the proliferation of tendoncells between 5ng/ml CTGF group and control group; 5ng/ml,10ng/ml,20ng/ml,50ng/mlCTGF groups had significant difference(P＜0.05),andthe proliferation of tendon cells got faster with the higher concentrationof CTGF in this term.There were no significant difference(P＞0.05)amongthe 50ng/ml,100ng/ml,200ng/ml,500ng/ml CTGF group.Conclusions: 10ng/ml,20ng/ml,50ng/ml,100ng/ml,200ng/ml,500ng/ml,CTGF could promote proliferation of tendon cells; 5ng/mlCTGF has no significant effect on proliferation of tendon cells.theproliferation of tendon cells got faster with the higher concentration ofCTGF when CTGF concentration varying from 0ng/ml to 50ng/ml; butthe proliferation of tendon cells got a bit slower with the higherconcentration of CTGF when CTGF concentration varying from 50ng/mlto 500ng/ml.CTGF at 50ng/ml is found to be the most effectiveconcentration.