發(fā)布時(shí)間：2018-05-03 04:19

  本文選題：表皮模型 + HaCaT　； 參考：《中國(guó)修復(fù)重建外科雜志》2017年10期

【摘要】：目的采用人永生化上皮角質(zhì)形成細(xì)胞系HaCaT構(gòu)建表皮模型,探討其作為皮膚刺激性替代實(shí)驗(yàn)的可行性。方法取HaCaT細(xì)胞采用氣-液面培養(yǎng)的方式構(gòu)建表皮模型,同法以采用人原代上皮角質(zhì)形成細(xì)胞構(gòu)建的表皮模型EpiKutis~?作為對(duì)照。HE染色觀察兩種模型的組織形態(tài);采用快速滲透實(shí)驗(yàn)方法檢測(cè)兩種模型屏障功能(ET_(50)值);取20種化學(xué)標(biāo)準(zhǔn)品分別于兩種模型表面受試30 min,孵育42 h后MTT法測(cè)定組織相對(duì)活力百分比,判斷化學(xué)品刺激性,并與聯(lián)合國(guó)-化學(xué)品分類及標(biāo)記全球協(xié)調(diào)制度(UN GHS)體內(nèi)測(cè)試結(jié)果分類和經(jīng)濟(jì)合作與發(fā)展組織(OECD)TG439(2013版)中采用的體外替代方法(validated reference method,VRM)體外測(cè)試結(jié)果分類進(jìn)行比較。結(jié)果 HE染色示,構(gòu)建的HaCaT模型無(wú)完整復(fù)層化結(jié)構(gòu);屏障功能檢測(cè)結(jié)果示,ET_(50)值為0.99 h;通過(guò)20種化學(xué)品檢測(cè),HaCaT模型的靈敏度為100%,特異性為50%,準(zhǔn)確率為75%;EpiKutis~?模型靈敏度為100%,特異性為70%,準(zhǔn)確率為85%。結(jié)論 HaCaT細(xì)胞構(gòu)建的表皮模型不具備完整的表皮皮膚生理結(jié)構(gòu)特征,屏障功能較低,化學(xué)品體外皮膚刺激性測(cè)試結(jié)果不能完全滿足OECD關(guān)于刺激物的正確判定標(biāo)準(zhǔn),不適宜作為化學(xué)品皮膚刺激性體外替代檢測(cè)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.

【作者單位】： 武警后勤學(xué)院附屬醫(yī)院皮膚科;廣東博溪生物科技有限公司;
【基金】：天津市自然科學(xué)基金面上項(xiàng)目(14JCYBJC29200)~~
【分類號(hào)】：R99;TQ658
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本文編號(hào)：1836942