| Objective:Bone tissue remodeling is the basement of Orthodontic treatment, which is the orthodontic force on the teeth. And bone tissue repair is a process of growth factor-mediated by the cell. The present study is to investigate into the proliferation,the differentiation and the mineralization of cementum cells though different concentrations of PDGF-BB. This experiment is desighed to provide the evidence in vitro experiments: whether the application of PDGF-BB has the ability of effectively promoting root repair or preventing root resorption for the clinical basis.Method:Well-cultured OCCM-30 were divided into six groups, which are one control group and five experimental groups. The experimental group were added with different concentrations of PDGF-BB(5、10、 20、 30、40ng/ml), while the control group does not add drug into the cementum cells. The following is the test:we dectect the cell proliferation under the MTT method at the different time,and request PNPP method detects the ALP activity at 48 h or 72 h. Using Alizarin Red staining cells to detect the content of mineralized nodules at day 7 and detect the expression of OPN and BSP gene by reverse transcription PCR(RT-PCR) at 72 h. All data were statistically analyzed under the software of SPSS Statistics V19.0.Results:1.MTT mehod detected proliferation of cells: The OD values of 20 ng / ml PDGF-BB is the highest at different time(24h, 48 h,72h), 5ng / ml is lower than the control group, but the difference is not statistically significant(P>0.05), which indicated that the lower concentrations of PDGF-BB has no obviously inhibition effection. The OD values of 10-40 ngml is higher than the control group, the difference is statistically significant(P<0.05), which can explain that the high concentration PDGF-BB inhibite the proliferation of OCCM-30. The OD values of72 h is higher than the other time points, which attests that the effection of PDGF-BBon the proliferation is stengthened as time going.2. Effection of alkaline phosphatase activity of cells : As the effection of different concentrations of PDGF-BB on OCCM-30 at 48 h and 72 h, the experimental group and the control group pairwise compared ALP activity of cells, the difference,which is P<0.01, is statistically significant. The promoting effect of 20 ng / ml is obviously among the different concentrations of PDGF-BB. The ALP activity of cells of 72 h is greater than that of 48 h under the pairwise testing, which approves that the ALP activity of OCCM-30 is improving as time goes by.3. Alizarin Red staining discovered content of the mineralized nodules :As the effection of different concentrations’ PDGF-BB, the OCCM-30 has obverse mineralized nodules on the fifth day. And there are verious mineralized nodules with different sizes and different shapes by staining. To compare, each groups shows the difference is obviously(P<0.05); OD570 values of 5ng / ml is lower than the control group, 10-20 ng / ml is on the growing trend, until 30 ng / ml to the peak, and 40 ng /ml declined, which implicates that the mineralization effect of 30ng/ml is the most obviously.4. OPNm RNA and BSPm RNA gene expression: The RT-PCR shows that the OPN and BSP gene expression under different concentrations of PDGF-BB effected OCCM-30 cells is higher than the control group. When compare its gray value ratio(purpose / internal) with control group, the difference is distinctly(P<0.05),suggesting that PDGF-BB has the effection of promoting the OPN and BSP gene expression of OCCM-30.Conclusion:1. PDGF-BB could promote proliferation under the differentiation of cementoblast, and the effection of 20ng/ml is the strongest. Cell proliferation and differentiation is positively correlated as time going.2. Different concentrations of PDGF-BB could promote the OCCM-30 formed mineralized nodules, and the concentration of 30ng/ml is the most obviously.3. As for the cementum cells, PDGF-BB has the ability to promote the expression of OPNm RNA and BSPm RNA. |
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