The Biological Role Of Transforming Growth Factor-β₁ And Nerve Growth Factor To Proliferation And Differentiation Of Human Periodontal Ligament Cells

Objective:To explore the biological role of Transforming Growth Factor-β1 and Nerve Growth Factor and the combination of the two to human periodontal ligament cells proliferation and alkaline phosphatase activity,for providing a theoretical basis to periodontal repair after injury.Methods:Normal human periodontal ligament cells were isolated and cultured in vitro.After being identified, the fifth generation of periodontal ligament cells were cultured. In this experiment, there were the control group and the experimental group, the former added only DMEM nutrient solution containing 20% FBS, the latter added not only DMEM medium containing 2% FBS, but also different concentrations TGF-beta 1 (0.1,1.0,10.0,50.0,100.0 ng/ml) and different concentrations of NGF (0.1,1.0,10.0,50.0,100.0μg/ml). each concentration group has four holes.Cells were trained for 3 days in the standard environment, each hole was mixed with the 20μl MTT, contining to cultivate 4h. Then, culture medium in each hole was sucked. And each hole was mixed with 150μl dimethyl sulfoxide and fully oscillated 10 minutes in oscillation, enzyme immunosorbent detector measured A value of each hole under 490nm wavelength, and compute average to do statistics analysis. According to the above-mentioned methods, with the best effect of concentration measurement TGF-beta 1 and NGF, and combination of the two, cell proliferation was detected, to obtain A value. ALP activity was measured by enzyme kinetic methods.Then the values among the groups were compared by variance analysis.Results:According to the experimental results were as follows:1 Compared with the control, different concentrations of TGF-beta 1 (0.1 ng/ml,1 ng/ml,10ng/ml,100ng/ml) used alone are promoting the HPDLCs proliferation, but had different biological effects. The 10ng/ml concentration of TGF-beta 1 had the best biological effect. Compared with control group, TGF-β(0.1 ng/ml,1 ng/ml,10ng/ml,100ng/ml的TGF-β1) can increase the ALP activity, but higher than 10.0 ng/ml discrease the ALP activity,during the experiment, the effects of 10.0ng/ml was the highest(P<0.05) according to the datas of this experiment.2 Compared with the control, different concentrations of NGF (0.1μg/ml,1μg/ml,10μg/ml,100μg/ml) used alone can promote the HPDLCs proliferation and differentiation. The 10μg/ml NGF is the best concentration. ALP activities also significantly increased in 3 experiment groups (cultured with 1,10, 100μg/ml NGF-DMEM), highest in 100μg/ml NGF group(P<0.05).3 Added the best TGF-beta 1 concentration,best NGF concentration and their combination, when TGF-beta 1 used alone, testing A value was for 0.522±0.06, and added NGF alone, A value was measured for 0.304±0.020, and applied the combination of the two,A value was for 0.702±0.004.A value of the latter was higer than of the first two.4 Added the best TGF-beta 1 concentration,best NGF concentration and their combination, when TGF-beta 1 used alone, testing ALP activities A value was for 0.628±0.004, and added NGF alone, ALP activities A value was measured for 0.378±0.005, and applied the combination of the two, ALP activities A value was for 0.325±0.006. ALP activities A value of the latter was higer than of the first two.Conclusion:The best concentration of TGF-β1 and NGF can promote the proliferation and differentiation of the PDLCs, and when the combination of the two was added with a synergistic effect.