Effects Of 4 Species Of Metal Alloys On The Cell Cycle, On The Apoptosis And On The Expression Of Bcl-2 And Bax In Human Gingival Fibroblast Cells

ObjectiveMetal alloys as dental materials, application of local experience in the human body have a certain role and influence. Apoptosis mechanism of toxicity of important medical material one. This study cell biology and molecular biology from the perspective of Au-Pt alloy, titanium metal, cobalt chromium alloy and nickel-chromium alloy metal alloy of these 4 human gingival fibroblasts to explore mechanisms of apoptosis. Mainly through in vitro human gingival fibroblast cells to observe cell cycle by flow cytometry to detect 4 kinds of metal alloys extract on human gingival fibroblast cell cycle, and then discuss four kinds of metal alloy extracts of human gingival fibroblast apoptosis mechanism, and explore the biocompatibility of metallic materials of a medical evaluation of the new method.Methods1. Cell cultureCells were cultured in the clinical health of gum tissue obtained by tissue adherent to human gingival fibroblasts were cultured and passaged. By immunohistochemistry of human gingival fibroblasts were vimentin and keratin staining of the source.2. Cell cycle by flow cytometryFlow cytometry cell cycle will be four kinds of metal alloys were added to extract human gingival fibroblasts in culture medium containing 10% calf serum DMEM culture medium as a negative control. Cultured for 48 hours after PI staining and by flow cytometry cell cycle in the experimental group changed.3. Western-blot method for detection of Bax to Bcl-24 species of metal alloys were added to extract the metal alloy of human gingival fibroblasts in culture medium containing 10% calf serum DMEM culture medium as a negative control. By Western-blot method to determine the cells of Bcl-2 and Bax expression.Results1. Identification of human gingival fibroblast cells and cells from Bcl-2 and Bax expressionIdentification of human gingival fibroblast cells and cells from Bcl-2 and Bax expression in human gingival fibroblasts by immunohistochemistry, stained positive for vimentin, keratin staining was negative. The operator confirmed that fibroblast cells derived from mesoderm features. Meanwhile, Bcl-2 and Bax in human gingival fibroblast cells were positive.2. Flow cytometry detection of human gingival fibroblast cell cycleFlow cytometry of human gingival fibroblast cells in the G₀G₁ phase of cell cycle, DNA content increased percentage; in the S phase, DNA content decreased percentage; proliferation index decreased. Among them, Ni-Cr alloy group, proliferation index (20.91%) than negative control group (21.76%) small (P <0.05), suggest a significant difference. The other 3 groups the proliferation index of metal alloys in order was Au-Pt alloy group (30.22%)> Ti group (29.85%)> Co-Cr group (29.11%), compared with negative control group (21.76%) large (P <0.05), suggest a significant difference.3. Metal alloy extracts of human gingival fibroblasts in the Bcl-2 and Bax expressionBcl-2/Bax ratio of cells in each experimental group, in the control group, gold platinum alloy groups, titanium was no difference between groups was significant (P> 0.05), while the nickel-chromium alloy, cobalt-chromium alloy with control group were significant differences between the significance (P <0.05). Au-Pt alloy group and the pure titanium group, nickel-chromium alloy and cobalt-chromium alloy also no differences between groups were significant (P> 0.05).Conclusions1. Using tissue cultured in vitro adherence of human gingival fibroblasts and the successful passage, the successful establishment of human gingival fibroblasts in vitro model.2. Metal alloys can change the human gingival fibroblast cell cycle. The gold platinum alloys titanium, and cobalt-chromium on human gingival fibroblasts proliferation alloy smaller, while the nickel-chromium alloy on the proliferation of large cells.3.The metal alloy on human gingival fibroblasts in the Bcl-2 and Bax expression in different degrees. One gold platinum alloys, titanium and cobalt-chromium alloy on human gingival fibroblast cells the expression of Bcl-2 and Bax little effect on the nickel-chromium alloy on human gingival fibroblasts in the Bcl-2 and Bax expression was significantly higher than .4. For cell proliferation and on cell expression of Bcl-2 and Bax detection on metal alloys can be an indicator of mechanisms for toxicity. In turn, it may become a new evaluation of the biocompatibility for metal targets.