| ObjectiveMetallic alloys using in the physiological environment, as one of the oral restoration materials apploying to human body, have some invariable effect on life organism. Biocompatibility evaluation is one of the important contents of researching medical metal materials. This study estimates six metallic alloys' biocompatibility on the point of view of cytobiology and molecular biology. 1. it implys that several metallic alloys have certain effect on cell cycle and apoptosis of L929 Cell and then estimates their biocompatibility, mainly by means of cell culture and flow cytometry detection.2. it approaches mechanisms that metallic alloys produce apoptosis and provids theoral evidences for introducing new indexes of biocompatibility evaluation in the way of detecting the mRNA expression of Bcl-2 and Bax by RT-PCRMaterials and Methods1. Cells cultrueL929 cells are cultivated and passaged with RPMI 1640 cell medium containing 10% fetal calf serum in the incubator of 37℃and 5%CO2.2. Detecting cell cycle with flow cytometrysCulture L929 cell with six metallic alloys' leaching liquor and their negative controls for 24 and 48 hours, and stain them with PI, and then detect cell cycle alteration of cell in disparity experimental group by flow cytometry.3. Observing Apoptosis cell with fluorescence microscopeHave L929 cells inoculated on cultivation board of six wells and cultivate them with metallic alloy leaching liquor for 48 hours, then observe apoptosis cells with fuorescence microscope after double staining them with Annexin V-EGFP and PI.4. Detecting cell apoptosis with flow cytometry Culture L929 cell with six metallic alloys' leaching liquor or their negative controls for 24 and 48 hours, and stain them with Annexin V-EGFP and PI, and then detect apoptosis of cell with flow cytometry.5. RT-PCRCulture L929 cell with six metallic alloys' leaching liquor or their negative controls for 48 hours,and then detect the mRNA. expression of cells' Bcl-2 and Bax.Results1. The effect of Metallic alloys on the cell cycle of L929Cells in each group display different multiplication tendency after cultivation for 48 hours,in which the proportions of G2 stage in the aurum alloy group and the negative control group are the maximum, the cobalt chromium alloy group, theⅡsteel group, the argenti palladium alloy group, the nickel chromium alloy group, and theⅢsteel group one by one descent.2. The effect of Metallic alloys on the apoptosis of L929Cells in the experimental group can be observed apoptosis cells with the fluorescence microscope after Annexin V-EGFP and PI staining. The detected consequence with flow cytometry indicates that the apoptosis rates of each interclass are different. The apoptosis rate of the aurum alloy group is the minimum approaching to the one of the negative control group. While other metallic alloys' apoptosis rates are far more than the aurum alloy's, among which the one of nickel chromium alloy is the highest and much higher than the one of negative control group and the sequence is aurum alloy,cobalt chromium alloy,Ⅱsteel,Ⅲsteel,argenti palladium alloy,nickel chromium alloy descent one by one.3. The effect of metalic alloys on the mRNA expression of Bcl-2 and Bax of L929The mRNA expression of Bcl-2 and Bax of cells in each experimental group is different.The proportionalities of Bcl-2/Bax as follows: the negative control group, the aurum alloy group, the cobalt chromium alloy group, theⅡsteel group, the nickel chromium alloy group, the argenti palladium alloy group and theⅢsteel group from high to low one by one.Conclusions1. Metallic alloys are possibly to produce the alteration of the cell cycle of L929. 2. Metallic alloys can produce the alteration of the apoptosis of L929. Bcl-2 and Bax may participate in the signal conduction of the apoptosis regulation.3. Detections of the cell cycle and the apoptosis and the mRNA expression of Bcl-2 and Bax may become one of the metallic alloys' biocompatibility evaluation indexes.4. The results of six metallic alloys' biocompatibility evaluation as follows: the biocompatibility of aurum alloy and cobalt chromium alloy andⅡsteel is relativelygood comparing with the one of argenti palladium alloy and nickel chromium alloy andⅢsteel.5.Ⅱsteel as one of the new materials possesses comparatively fine biocompatibility.
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