Effect Of IL-1β On Inflammatory Factors Synthesis Of Human Gingival Fibroblasts Attached To Different Titaniums

OBJECTIVE:The purpose of this experiment was to compare the impact of three different titanium surface composition of human gingival fibroblasts biological behavior, and to evaluate the effects of IL-1β on inflammatory factors synthesis of human gingival fibroblasts to different titanium in vitro. Find an ideal ingredient titanium surface to be conducive to human gingival fibroblasts attached, and provides guidance for the design of the implant neck.METHODS:Partl:Human gingival fibroblasts was obtained from explants of rat normal gingival tissues by using tissue-explants technique, the cellular growth conditions were observed under inverted phase-contrast microscope. The third passage of RGFs was applied to be identified by immunochemistry of vimentin and cytokeratin. Hematocytomete was used to determine the numbers of cells and the mean of cell numbers was calculated for drawing the growth curve of human gingival fibroblasts.Part2:Round disks of titanium were placed in 24 well plates, the gingival fibroblasts seeded on three different titanium surface treatment. After 2 hours, respectively by MTT assay gingival fibroblasts attachment rate in titanium implant surfaces of different surface treatments,and After 5 days, respectively by MTT assay gingival fibroblasts proliferation rate in titanium implant surfaces of different surface treatments.Part3:The first group, round disks of titanium were placed in 24 well plates, the gingival fibroblasts seeded on three different titanium surface treatment. At the points of 2 hours, Data of the IL-8 genes expression and IL-6 genes expression in human gingival fibroblasts to different titaniums after 3 h、6 h、24 h. The second group round disks of titanium were placed in 24 well plates, the gingival fibroblasts seeded on three different titanium surface treatment. At the points of 2 hours, the gingival fibroblasts added the stimulus of interleukin-1β, Data of the IL-8 genes expression and IL-6 genes expression in human gingival fibroblasts to different titaniums after 3h、6h、24 h. The date was statistically analyzed with SPSS 13.0 software package.RESULT:1、Human gingival fibroblasts cultured with tissues-explants method started to appear after one week, most cells were fusiform with large body. The cells covered full of the floor of 25 cm2 cell culture flask in two to three weeks of primary culture. The immunochemistry study showed that vimentin was expressed in human gingival fibroblasts while cytokerate was failed of detection, which indicated that human gingival fibroblasts was originated from mesoblastma. It is a S-shape grows curve for the cultured human gingival fibroblasts. The cell number declined a little after the first 24 hours cultivation, and then it creased gradually.2、MTT method measured that treatment with titanium nitride, titanium surface compared to titanium alloy surface for cell attachment rates were significantly different after 2 hours (P<0.05), MTT method measured that treatment with titanium nitride, titanium surface compared to titanium alloy surface for cell proliferation rates were significantly different after 5 days (P<0.05).3、A significant increase in the expression of gingival fibroblasts to inflammatory factors(IL-6、IL-8) of IL-1β showed; the PCR results of the production of inflammatory factors in gingival fibroblasts with the intervention of IL-1β showed that the production of inflammatory factors (IL-6、IL-8)on titanium and titanium alloy less than that on the titanium nitride.Conclusion:1、 It can be offered high survival rate by explants culture of human gingival fibroblasts, Which can provide enough cells, the cells were in high activity and stable characterization.2、Human gingival fibroblasts attachment and proliferation were good of titanium and titanium alloy group, human gingival fibroblasts attachment and proliferation was inhibited of titanium nitride (P<0.05).3、IL-1β can stimulate high expression of inflammatory factors; the production of IL-6 and IL-8 on titanium and titanium alloy relatively less, so the implant of titanium and titanium alloy can reduce the incidence of peri-implant inflammation,and improve the successful rate of implant in clinical.