Effect of fluoride on the expression of CTGF in two-dimensional and three-dimensional cultured fibroblasts

Objective To explore the effects of fluoride on the expression of CTGF in two-dimensional(2D)and three-dimensional(3D)cultured fibroblsats.Methods 2D and 3D cultured fibroblsat were exposed to different concentrations of fluoride(0,0.000 1,0.001,0.1,1,10 and 20 mg/L).The levels of CTGF protein at time of 2,4,24,48 and 72h were measured by using Elisa method.The expression of CTGF mRNA was tested with RT-PCR method.By using the method of IHC,we detected the expression of the masculine protein in cytoplasm of FB.Results By using the method of ELISA,either in 2D or 3D cultured system,Compared with the group of 0 mg F-/L the content of CTGF protein was increased obviously in certain groups.In 2D cultured system,compared with control(1.02±0.03),the level of CTGF protein at time of 4 h in 0.001 mg F-/L group(1.50±0.40) increased obviously(P0.01),and 0.1 mg F-/L group(1.42±0.20) increased significantly(P0.05),and compared with control(1.59±0.40) that at time of 72 h in 20 mg F-/L group(1.59±0.40) increased obviously(P0.01).In 3D cultured system,at time of 48 h,compared with control(1.21±0.05),the level of CTGF protein in 20 mg F-/L group(1.45±0.17) increased significantly(P0.05).At time of 72h,compared with control(1.17±0.09),the level of CTGF protein in 10 mg F-/L group(1.43±0.22) increased significantly(P0.05).At time of 96h,compared with control(1.17±0.17),that in 0.001,10 and 20 mg F-/L groups(1.39±0.04,1.52±0.08,1.45±0.03) increased obviously(P0.01).The result showed that the expression of CTGF mRNA at 48h had been enhanced in the group of higher fluoride dosage in 2D and 3D cultured system.It,however,did not rich the apparent degree in statistics.We found that fluoride can stimulate the masculine protein of CTGF by using the mothod of IHC.Conclusion Above-mentioned result shows that the expression of CTGF in FB has been stimulated by fluoride in 2D and 3D cultured system.CTGF stimulated the function of osteogenesis and it may have an important usage in osteoplastic potency of FB.Fluoride increased the expression of CTGF in three-dimensional cultured fibroblasts,nevertheless,the effect was fairly slow,the degree was relatively feeble.