一种肠镜润滑消泡剂的细胞毒性和遗传毒性生物学评价

Cytotoxicity and Genotoxicity Biological Evaluation of a Colonoscopy Lubrication Defoamer

目的对一种以二甲基硅油、甘油、羧甲基纤维素钠为成分的肠镜润滑消泡剂进行细胞毒性和遗传毒性评价,为这种肠镜润滑消泡剂在临床使用提供生物安全依据。方法选择琼脂扩散试验对肠镜润滑消泡剂进行体外细胞毒性检测,选择细菌回复突变试验(Ames试验)、用小鼠淋巴瘤细胞进行的体外哺乳动物细胞基因突变试验(MLA试验)、体外哺乳动物细胞染色体畸变试验进行遗传毒性检测。细胞毒试验,将样品放置在固化的琼脂层上与L929细胞间接接触24h,用中性红对细胞进行染色后显微镜下观察细胞毒性。Ames试验选用TA97a、TA98、TA100、TA102、TA1535五种组氨酸缺陷型鼠伤寒沙门氏菌株,观察肠镜润滑消泡剂对细菌回复突变率的影响;MLA试验采用小鼠淋巴瘤细胞与样品溶液接触3h和24h,通过计算其平板效率和大小集落形成数目计算突变频率,判断受试样品对小鼠淋巴瘤细胞突变率的影响;体外哺乳动物细胞染色体畸变试验,使样品溶液与中华地鼠肺细胞(CHL细胞)接触6h和24h,通过对处于有丝分裂中期的CHL细胞的染色体畸变情况进行分析,评价肠镜润滑消泡剂对CHL细胞潜在的致突变性。结果在细胞毒试验中,受试样品与阴性对照相比,细胞数量相对较少且受试样品显微镜下观察有少量畸形和退化的细胞,为轻微的细胞毒性;Ames试验样品组回变菌落数均未比阴性对照回变菌落数增加1倍或者1倍以上且无重复性;MLA试验样品组MF值与阴性对照相比无超过126×10^-6的增长;染色体畸变试验受试样品组染色体结构畸变率与阴性对照相比,差异无统计学意义(P>0.05)。结论以二甲基硅油、甘油、羧甲基纤维素钠为成分的肠镜润滑消泡剂无潜在的细胞毒性和遗传毒性,可进一步用于临床研究。

Objective To evaluate the cytotoxicity and genotoxicity of a colonoscopy lubricant defoamer based on dimethyl silicone oil,glycerin and sodium carboxymethyl cellulose,and provide clinical use for this colonoscopy lubrication defoamer and biosafety basis.Methods The agar diffusion test was used to detect the in vitro cytotoxicity of the colon gel lubrication defoamer.The bacterial back mutation test(Ames test),the in vitro mammalian cell gene mutation test(MLA test)using mouse lymphoma cells,and the in vitro breastfeeding were selected.Genotoxicity testing for animal cell chromosome aberration test.In the cytotoxicity test,the sample was placed on the solidified agar layer for indirect contact with L929 cells for 24 h,and the cells were stained with neutral red,and the cytotoxicity was observed under a microscope.Ames test used TA97a,TA98,TA100,TA102,TA1535 five histidine-deficient Salmonella typhimurium strains to observe the effect of colonoscopy lubrication defoamer on bacterial reversion rate;MLA test using mouse lymphoma cells and sample solution After 3 h and 24 h of exposure,the mutation frequency was calculated by calculating the plate efficiency and the number of colony formation,and the effect of the test sample on the mutation rate of mouse lymphoma cells was determined.In vitro mammalian cell chromosome aberration test,the sample solution and the Chinese hamster lung cells(CHL cells)were exposed for 6 h and 24 h.The chromosomal aberrations of CHL cells in the middle stage of mitosis were analyzed to evaluate the potential mutagenicity of colon gel lubrication defoamers on CHL cells.Results In the cytotoxicity test,the number of cells in the test sample was relatively small compared with the negative control,and a small amount of abnormal and degraded cells were observed under the microscope of the test sample,which was mild cytotoxicity;The number of colonies did not increase by 1 time or more than the number of colonies of the negative control and there was no repeatability;the MF value of the MLA test sample group did not exceed 126×10-6 growth compared with the negative control;the chromosome aberration test was subjected to the sample.There was no significant difference in the chromosome structural aberration rate between the group and the negative control(P>0.05).Conclusion The colonoscopy lubricant defoamer with dimethyl silicone oil,glycerin and sodium carboxymethyl cellulose has no potential cytotoxicity and genotoxicity and can be further used in clinical research.