基于皮肤模型与THP-1细胞共培养建立皮肤致敏体外检测方法的初步研究

建立了皮肤模型与THP-1细胞共培养方法,对石化产品进行了致敏性评价。不同浓度测试物与表皮模型作用15 min,去除受试物后孵育6 h,使用MTT法检测皮肤模型组织活率,利用流式细胞法检测THP-1细胞表面标志物CD86、CD54表达变化情况以及细胞活率。对上述参数进行统计学分析及是否存在剂量效应关系,判定测试物是否具有潜在致敏特性。对SLS、肉桂醛、柠檬醛的测试表明,该方法可以正确区分刺激物和致敏物;对4种未知润滑油添加剂的预测结果表明,HVI60、Cuvan826、ATM PAN为非致敏物,Irgamet30为致敏物,高浓度时可导致表皮组织和THP-1细胞活性降低及表面标志物CD86、CD54的特异性升高。

A preliminary study on in vitro skin sensitization test based on skin model and THP-1 cell co-culture

An in vitro skin sensitization test based on skin model and THP-1 cell co-culture was established, and used to evaluate the sensitizing potency of petrochemicals. Skin model was treated with different concentrations of samples for 15 min, then removing the samples and was incubated for another 6 h. Skin tissue activity was measured by MTT assay, while cell viability and cell surface markers (CD54,CD86) expression levels of THP-1 cells were detected by flow cytometry. The skin sensitization potential of test substances was determined by statistically analyzing the above parameters and reaching a dose-response relationship. The results of SLS, cinnamaldehyde and citral show that this co-culture method can correctly distinguish irritants and sensitizers. The results of 4 unknown lubricant additives show that HVI60, Cuvan826 and ATM PAN are predicted to be non-sensitizers, while Irgamet30 is predicted to be a sensitizer, which at higher concentration can decrease the skin tissue activity and THP-1 cell viability, and specifically increase the expression of surface markers.