细胞培养法评价自制镍铬烤瓷合金的细胞毒性(英文)

Cytotoxicity evaluation of self-made nickel-chromium ceramic alloy using cell culture method

背景:具有良好抗腐蚀性能的镍铬铸造烤瓷合金正在临床广泛应用,但不容忽视的是其释放出的镍离子、铍离子具有明显的毒副作用。因此,改进合金的组成成分,提高其理化性能和生物相容性是目前研究的热点。目的:检测自制镍铬烤瓷合金的体外细胞毒性,并与口腔科常用的镍铬合金、纯钛金属的细胞毒性进行比较。设计、时间及地点:对比观察实验,于2006-05/12在中国医科大学中心实验室完成。材料:细胞选用对数生长期的小鼠成纤维细胞(L-929细胞)。自制镍铬烤瓷合金由中国科学院金属研究所提供,纯钛由日本松风公司提供,镍铬合金由贺利氏古莎齿科有限公司提供。方法:实验分为自制镍铬烤瓷合金、纯钛组、镍铬合金组,阳性对照组,阴性对照组。取96孔培养板,每板选出5组孔,每组重复孔12个,每孔中均加入浓度为1.0×108L-1的细胞悬液100μL。在自制镍铬烤瓷合金、纯钛组、镍铬合金组孔中分别加入各种材料浸提液100μL,阳性对照组孔中加入铅浸提液100μL,使材料浸提液浓度最终为50%,在阴性对照组孔内加入RPMI1640培养液100μL。主要观察指标:分别于培养1,2,3,4,5d时采用四唑盐比色法测定各组吸光度值,计算相对增殖率。结果:3种金属材料组与阴性对照组之间的吸光度值差异均无显著性意义(P>0.05),与阳性对照组之间的吸光度值差异有显著性意义(P<0.05或P<0.01)。纯钛组细胞增殖率>100%,细胞毒性为0级,自制镍铬烤瓷合金组和镍铬合金组的细胞增殖率为86.36%～99.49%,细胞毒性为1级,阳性对照组在培养3d后细胞毒性增长至2级。结论:自制镍铬烤瓷合金仅具有极微弱的细胞毒性,符合口腔材料临床应用的要求。

BACKGROUND： Nickel-chromium ceramic alloy with well anti-causticity has been widely used in clinic; however, it should be paid much attention to the toxic and side effects of the released nickel and glucinium ions. Therefore, studies about modifying alloy component, enhancing physical and chemical performance, and improving biocompatibility are hot topics recently. OBJECTIVE： To measure the in vitro cytotoxicity of self-made nickel-chromium ceramic alloy comparing with nickel-chromium alloy and pure titanium metal. DESIGN, TIME AND SETTING; A contrast observational experiment was performed at Central Laboratory of China Medical University from May to December 2006. MATERIALS： L-929 cells at logarithmic phase were used in this study. Self-made nickel-chromium ceramic alloy was provided by Metal Institute of Chinese Academy of Science, pure titanium was provided by Matsukaze Japan Co., Ltd., and nickel-chromium alloy was provided by Heraeus Kulzer Dental Co., Ltd. METHODS： L-929 cells were assigned into nickel-chromium ceramic alloy, pure titanium, nickel-chromium alloy, positive control, and negative control groups. The cells were cultured in 96-well culture plate. Every five wells were randomly selected from each group, and 12 wells were used for the duplication. Cell suspension （100 μL） at concentration of 1.0×10^8/L was added in each well. Leaching liquor （100 μ L） was added in self-made nickel-chromium ceramic alloy, pure titanium alloy, and nickel-chromium alloy groups, respectively, lead leaching liquor （100 μ L） was added in positive control group to make the final concentration at 50%, and RPMI 1640 culture solution （100μL）was added in the negative control group. MAIN OUTCOME MEASURES： Absorbance was measured at day 1, 2, 3, 4 and 5 using MTT colorimetric assay to calculate relative growth rate. RESULTS： There were no significant differences in absorbance between three metal groups and negative control group （P 〉 0.05）, but there were significant differences in absorbance between three metal groups and positive control group （P 〈 0.05 or P 〈 0.01）. Cell growth rate was 〉 100% and cytotoxicity was grade 0 in the pure titanium group; while, cell growth rate ranged from 86.36% to 99.49% and cytotoxicity was grade 1 in both self-made nickel-chromium ceramic alloy and nickel-chromium alloy groups. At day 3 after culture, cytotoxicity was grade 2 in the positive control group. CONCLUSION： Self-made nickel-chromium ceramic alloy has a little cytotoxicity and requires for the clinical application.