骨水泥的遗传毒性研究

Genotoxicity Study of Bone Cement

目的检测医疗器械进口产品骨水泥的遗传毒性。材料与方法以骨水泥粉体浸提液与其液体混合液为供试液,采用细菌回复突变试验(Ames试验)和小鼠淋巴瘤细胞试验(MLA),在非代谢活化(-S9)和代谢活化(+S9)条件下检测。Ames试验采用平板掺入法,检测了25、50、100 ml/皿3个剂量组诱发鼠伤寒沙门氏菌(S.typhimurium)TA98、TA100、TA1535、TA1537及大肠杆菌(E.coli)WP2 uvrA的回变菌落数;MLAT采用微孔法检测了终浓度0.125%、0.25%、0.5%(V/V)3个浓度组,在+S9/3 h、-S9/3 h、-S9/24 h三种处理条件下诱发L5178Y细胞tk+/-基因突变频率(MF)和小集落突变百分率(SC%)。结果 Ames试验,50、100 ml/皿剂量组在加入磷酸缓冲液或S9混合液后略有沉淀产生,并对TA100、TA1535、WP2 uvrA产生不同程度的抑菌作用,无抑菌剂量诱发五株菌的回变菌落数对比DMSO溶媒(阴性)对照组,均无明显增加;MLA,各浓度组在三种处理条件下诱发的MF、SC%与其溶媒(阴性)对照组比较,无明显增加(P>0.05)。结论本实验条件下,骨水泥对测试菌株his-/trp-无明显诱变作用,对测试细胞tk+/-及染色体无明显损伤作用。

Objective To investigate the genotoxicity of bone cement. Material and Methods Test article was mixed of liquid and the powder extract of bone cement. Using bacterial reverse mutation test （Ames test） and mouse lymphoma Assay （MLA） with and without metabolic activation S9. Ames test was performed by the plate incorporation method for its ability to induce reverse mutations in three treatment dosage groups： 25 μl/plate, 50 μl/plate, 100 μl/plate using S. typhimurium TA98. TA100. TA1535 TA1537 and E. coil WP2 uvrA; In MLA, the mutation frequency （MF） and the percentage of small colony mutants （SC%） of L5178 tk＋/- cells induced by bone cement at final concentration of 0.125%, 0.25% and 0.5% （V/V） were calculated and assayed with ＋S9/3 h, -S9/3 h, -S9/24 h. Results Slight precipitations of two dosage group 50 μl/plate and 100 μl/plate were observed after phosphoric acid buffer solution or S9mix added. Growth inhibition effect exists in TA100, TA1535 and WP2 uvrA to varying degrees. Other groups did not cause obvious increases in the mean number of revertant per plate compared with negative control （DMSO）. The result of MLA indicated no significant MF or SC% increases （P〉0.05） observed comparing with negative control under all test conditions. Conclusions The bone cement did not induce reverse mutation at his-/trp- and there was no obvious damage effect to gene tk＋/- or chromosome under this study condition.