CAJ | 학술논문

背景:采用不同方法评价材料的细胞毒性可能会得出不同的实验结果.目的:采用3种比色法评价镍铬合金、钴铬合金、3铬13及纯钛等牙科金属材料对小鼠成纤维细胞(L-929细胞)的细胞毒性.方法:以镍铬合金、钴铬合金、3铬13及纯钛4种牙科金属材料的浸提液分别作用于体外培养的 L-929细胞24,72 h.以体积分数10%小牛血清+高糖 DMEM 培养液培养的 L-929细胞为阴性对照组,以0.7%丙烯酰胺+体积分数10%小牛血清+高糖 DMEM 培养液培养的 L-929细胞为阳性对照组,分别采用 MTT、CCK-8及结晶紫3种比色法检测上述材料的细胞毒性.结果与结论:①4种材料浸提液中培养的细胞形态正常,胞内结构清晰,随着培养时间延长细胞大量增殖,与阴性对照组细胞形态无明显差异.阳性对照组细胞数量明显减少,形态完整性受破坏,形成大量细胞碎片.②培养24 h 时,CCK-8比色法检测中钴铬合金组的细胞相对增殖率低于阴性对照组(P <0.05),MTT及结晶紫比色法检测中钴铬合金组的细胞相对增殖率与阴性对照组比较差异无显著性意义(P >0.05);培养72 h 时,MTT 比色法检测中4种牙科金属材料组细胞相对增殖率低于阴性对照组(P <0.01),CCK-8及结晶紫比色法检测中4组的细胞相对增殖率与阴性对照组比较差异无显著性意义(P >0.05),但材料细胞毒性均为0-1级.表明上述4种牙科金属材料细胞毒性均在临床应用的允许范围内,具有良好的生物安全性.
배경:채용불동방법평개재료적세포독성가능회득출불동적실험결과.목적:채용3충비색법평개얼락합금、고락합금、3락13급순태등아과금속재료대소서성섬유세포(L-929세포)적세포독성.방법:이얼락합금、고락합금、3락13급순태4충아과금속재료적침제액분별작용우체외배양적 L-929세포24,72 h.이체적분수10%소우혈청+고당 DMEM 배양액배양적 L-929세포위음성대조조,이0.7%병희선알+체적분수10%소우혈청+고당 DMEM 배양액배양적 L-929세포위양성대조조,분별채용 MTT、CCK-8급결정자3충비색법검측상술재료적세포독성.결과여결론:①4충재료침제액중배양적세포형태정상,포내결구청석,수착배양시간연장세포대량증식,여음성대조조세포형태무명현차이.양성대조조세포수량명현감소,형태완정성수파배,형성대량세포쇄편.②배양24 h 시,CCK-8비색법검측중고락합금조적세포상대증식솔저우음성대조조(P <0.05),MTT급결정자비색법검측중고락합금조적세포상대증식솔여음성대조조비교차이무현저성의의(P >0.05);배양72 h 시,MTT 비색법검측중4충아과금속재료조세포상대증식솔저우음성대조조(P <0.01),CCK-8급결정자비색법검측중4조적세포상대증식솔여음성대조조비교차이무현저성의의(P >0.05),단재료세포독성균위0-1급.표명상술4충아과금속재료세포독성균재림상응용적윤허범위내,구유량호적생물안전성.
BACKGROUND: Different methods to evaluate cytotoxicity of the same material may produce different results. OBJECTIVE: To evaluate the cytotoxicity of nickel-chromium alloy, cobalt-chromium alloy, 3Cr13 stainless steel and pure titanium toward L-929 cells by using three assay systems. METHODS: The L-929 cells were cultivated in vitro in the extracts of four different dental metal materials (nickel-chromium alloy, cobalt-chromium alloy, 3Cr13 stainless steel and pure titanium) at 24 hours and 72 hours, separately. The L-929 cells, cultured in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum, served as the negative control group. And cells, cultured in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum and 0.7% acrylamide, served as the positive control group. The detection methods for the cytotoxicity included MTT, cel counting kit-8 and crystal violet cel proliferation assay system. RESULTS AND CONCLUSION: (1) Microscopy showed that the L-929 cells, cultivated in the extracts of four different dental materials, were normal and their intracel ular structure was clear. They proliferated wel with extension of incubation time. There were no significant differences between the test and negative control groups at al times. However, the number of cells in the positive control group significantly reduced as compared with the negative control group. The morphological integrity destroyed and a lot of cel debris formed. (2) After culture for 24 hours, the cel counting kit-8 assay revealed that the relative growth rate of the cobalt-chromium alloy group was significantly lower than that of the negative group (P < 0.05), while the MTT and crystal violet assay revealed that there were no significant differences in cel viability between the cobalt-chromium alloy and negative control group (P > 0.05). After culture for 72 hours, the MTT assay revealed that the relative growth rate of four dental metal materials groups was significantly lower than that of the negative group (P < 0.01), while the cel counting kit-8 and crystal violet assay revealed that there were no significant differences in cel viability between the four dental metal materials and negative control group (P > 0.05). The cytotoxicity of the tested dental metal materials was grade 0-1, which meets the safety standard. These indicate that these four dental metal materials have good biological safety.