In vitro evaluation of the genotoxicity of a naturally occurring crosslinking agent (genipin) for biologic tissue fixation.
In vitro evaluation of the genotoxicity of a naturally occurring crosslinking agent (genipin) for biologic tissue fixation.
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DOI：
10.1002/1097-4636(200010)52:1
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发表时间：
2000-10
期刊：
影响因子：
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通讯作者：
C. Tsai;R. Huang;H. Sung;H. Liang
中科院分区：
文献类型：
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作者：
C. Tsai;R. Huang;H. Sung;H. Liang
本研究的目的是使用中国仓鼠卵巢（CHO-K1）细胞在体外评价京尼平（一种天然交联剂）的遗传毒性。戊二醛是生物组织固定最常用的交联剂，用作参比化学品。本评价的选定程序为微核（MN）和姐妹染色单体交换（SCE）试验，添加或不添加代谢活化系统（S9混合物）。在开始遗传毒性试验之前，使用MTT试验确定了戊二醛和京尼平的最大非细胞毒性量。MTT法结果显示京尼平的细胞毒性显著低于戊二醛（有或无S9混合物）。在存在或不存在S9混合物的情况下，在使用不同浓度戊二醛或京尼平给药的病例中观察到的MN频率与阴性对照（空白）中观察到的MN频率无统计学差异。然而，注意到戊二醛显著抑制细胞周期进程，而京尼平给药的细胞没有导致细胞周期延迟。在SCE试验中，在使用不同浓度戊二醛给药的情况下观察到的每个细胞的SCE数量显著大于在使用或不使用S9混合物的阴性对照中观察到的SCE数量。然而，与用作阳性对照品的强诱变剂诱导的SCE数量相比，这些数量仍然较低。这表明戊二醛可能在CHO-K1细胞中产生弱致染色体断裂反应。相比之下，在京尼平给药的情况下获得的SCE/细胞数与在阴性对照中观察到的SCE/细胞数相当，但在最高剂量（50 ppm）给药的情况下除外。这表明京尼平在CHO-K1细胞中不引起致染色体断裂反应，只要其浓度低于50 ppm。总之，就细胞毒性和遗传毒性而言，京尼平是一种有前途的生物组织固定交联剂。
The objective of the present study was to evaluate in vitro, using Chinese hamster ovary (CHO-K1) cells, the genotoxicity of genipin, a naturally occurring crosslinking agent. Glutaraldehyde, the most commonly used crosslinking agent for biologic tissue fixation, was employed as a reference chemical. The selected procedures for this evaluation were the micronucleus (MN) and sister chromatid exchange (SCE) assays with or without the addition of a metabolic activation system (S9 mix). Before starting the genotoxicity assays, the maximum noncytotoxic amounts of glutaraldehyde and genipin were determined using the MTT assay. The results obtained in the MTT assay revealed that the cytotoxicity of genipin was significantly lower than that of glutaraldehyde with or without S9 mix. The frequencies of MN observed in the cases drugged with varying concentrations of glutaraldehyde or genipin were not statistically different from those seen in the negative controls (blank) in the presence or absence of S9 mix. However, it was noted that glutaraldehyde significantly inhibited the cell-cycle progression while the cells drugged with genipin did not result in cell-cycle delay. In the SCE assay, the numbers of SCE per cell observed in the cases drugged with varying concentrations of glutaraldehyde were significantly greater than those found in the negative controls with or without S9 mix. Nevertheless, these numbers were still low compared to the numbers of SCE induced by the strong mutagens used as our positive control substances. This suggests that glutaraldehyde may produce a weakly clastogenic response in CHO-K1 cells. In contrast, the numbers of SCE per cell obtained in the cases drugged with genipin were comparable to those observed in the negative controls in those that were except drugged with the highest dose (50 ppm). This suggests that genipin does not cause clastogenic response in CHO-K1 cells provided its concentration is lower than 50 ppm. In conclusion, as far as cytotoxicity and genotoxicity are concerned, genipin is a promising crosslinking agent for biologic tissue fixation.