HaCaT表皮模型作为皮肤刺激性体外替代实验的可行性研究

发布时间：2018-05-03 04:19

  本文选题：表皮模型 + HaCaT　； 参考：《中国修复重建外科杂志》2017年10期

【摘要】：目的采用人永生化上皮角质形成细胞系HaCaT构建表皮模型,探讨其作为皮肤刺激性替代实验的可行性。方法取HaCaT细胞采用气-液面培养的方式构建表皮模型,同法以采用人原代上皮角质形成细胞构建的表皮模型EpiKutis~?作为对照。HE染色观察两种模型的组织形态;采用快速渗透实验方法检测两种模型屏障功能(ET_(50)值);取20种化学标准品分别于两种模型表面受试30 min,孵育42 h后MTT法测定组织相对活力百分比,判断化学品刺激性,并与联合国-化学品分类及标记全球协调制度(UN GHS)体内测试结果分类和经济合作与发展组织(OECD)TG439(2013版)中采用的体外替代方法(validated reference method,VRM)体外测试结果分类进行比较。结果 HE染色示,构建的HaCaT模型无完整复层化结构;屏障功能检测结果示,ET_(50)值为0.99 h;通过20种化学品检测,HaCaT模型的灵敏度为100%,特异性为50%,准确率为75%;EpiKutis~?模型灵敏度为100%,特异性为70%,准确率为85%。结论 HaCaT细胞构建的表皮模型不具备完整的表皮皮肤生理结构特征,屏障功能较低,化学品体外皮肤刺激性测试结果不能完全满足OECD关于刺激物的正确判定标准,不适宜作为化学品皮肤刺激性体外替代检测的工具。
[Abstract]:Objective to construct the epidermis model by human immortalized epithelial keratinocyte line HaCaT and explore its feasibility as a skin irritant substitute experiment. Methods the epidermal model was constructed by using gas liquid surface culture of HaCaT cells, and the epidermal model EpiKutis~ constructed by human primary epithelial keratinocytes was used as a control.HE dye. Color observation of the tissue morphology of the two models; detection of two models of barrier function (ET_ (50)) by rapid penetration test; 20 chemical standard products were 30 min on the two model surface and 42 h after incubating for 42 h to determine the percentage of relative vigor of the tissue, to judge the irritation of chemical products, and to classify and mark the world with the United Nations chemicals. The classification of in vivo test results of UN GHS and the in vitro substitution method (validated reference method, VRM) used in the organization for economic cooperation and development (OECD) TG439 (2013 Edition) was compared. Results HE staining showed that the constructed HaCaT model had no complete complex layer structure; the barrier function test results showed that ET_ (50) was 0 .99 h, the sensitivity of the HaCaT model was 100%, the sensitivity of the HaCaT model was 50%, the accuracy was 75%, the sensitivity of the model was 100%, the specificity was 70%, the accuracy was 85%. conclusion that the epidermal model constructed by HaCaT cells did not have the complete physiological structure of skin skin, the barrier function was low and the skin irritation of the chemicals in vitro. The test results can not fully meet the OECD's criteria for determining irritants and are not suitable for chemical skin irritation in vitro.

【作者单位】： 武警后勤学院附属医院皮肤科;广东博溪生物科技有限公司;
【基金】：天津市自然科学基金面上项目(14JCYBJC29200)~~
【分类号】：R99;TQ658
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本文编号：1836942