The Experiments Of MMP-2,MMP-13Expression In Human Gingival Fibroblasts Influence By PFM Alloy

ObjectivePFM has both the metal crowns the intensity and the has the beautifuleffect of porcelain, in the clinical application were accepted by doctors andpatients. But in recent years, along with ceramometal crown restoration iswidely used, and also gradually exposed some problems, patients withrestoration wear after a period of time will appear gingival recession, gingivalbleeding, Gum margin dye ash and so on. The aim of this study is to explorethe nickel chrome, cobalt-chromium alloy, pure titanium, low content gold alloysoaks raises the in vitro culture of liquid gum fibroblasts secretion MMP-2,MMP-13influence. Thus, the analysis matrix metalloproteinases to gumfibroblasts mechanism of action, for clinical porcelain crowns base alloy choiceto provide the evidence.Methods1. With improved tissue mass method to original cultivate the human gingivalfibroblasts, with the third generation gum fibroblasts to identify the source ofthe cells in immunofluorescence.2. Preparating diameter of5.5mm, high2.4mm cylinder alloy,after processingsoaked in DMEM medium, after14days, collecting the base metals extract.3. Take the fifth generation cells grow well made3x104/ml singlecelled levitation liquid, each hole300ul uniform vaccination in48orifice plateto stick wall,discarded original DMEM medium.the experimental group to joinbase metals extract, the control group to join contain10%FBS of DMEM medium processing. Respectively in1h,6h,12h,24h collect cells of theliquid, with ELISA kit to detect.human gingival fibroblasts secretion of MMP-2,MMP-13.4. take the fifth generation cells made into3x104/ml cell Levitation liquidvaccinated cover glass in24hole plate, wait for after the stick wall discardedoriginal DMEM medium. the experimental group to join base metals extract,the control group to join contain10%FBS of DMEM medium.After12hours tocollectors gingival fibroblasts climb piece, with immunofluorescence testMMP-2, MMP-13human gingival fibroblasts in the expression.Results1. The modified tissue mass method has successfully cultured human gingivalfibroblasts in vitro, immunofluorescence and drawing the growth curveconfirmed cells derived from mesoderm with fibroblasts characteristics.2. The elisa test results show: nickel chrome and cobalt chromium alloy groupcompared with controls are statistically significant differences; Pure titaniumand rhotanium group compared with controls the difference was no statisticallysignificant; Nickel chrome, cobalt chromium alloy groups and pure titanium andrhotanium comparing statistical significance; Nickel chrome group and cobaltchromium alloy is not statistically significant differences between the groups.3. The cellular immune fluorescence method to observe human gingivalfibroblasts in processing of base metals extract,nickel chrome and cobaltchromium alloy group MMP-2, MMP-13express hyperchromatic, comparedwith controls obviously deepened; Pure titanium and rhotanium groupcompared with controls no obvious distinction.Conclusion1. The success of in vitro cultured human gingival fibroblasts, modified tissuemass method compared with pure tissue mass method is more advantage.2. Base metals extract of Nickel chrome and cobalt chromium can significantlyimprove human gingival fibroblasts MMP-2, MMP-13expression level. 3. Base metals extract of pure titanium and rhotanium processing of humangingival fibroblasts MMP-2, MMP-13expression effect is not obvious.4.Base metals extract influence human gingival fibroblasts matrixmetalloproteinases expression of mechanism research providede experimentbasis for clinical porcelain crowns base alloy selection.