KeratinoSens试验在医疗器械致敏性检测中的应用研究

doi:10.3969/j.issn.1004-616x.2021.06.010

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (6): 455-460.doi: 10.3969/j.issn.1004-616x.2021.06.010

KeratinoSens试验在医疗器械致敏性检测中的应用研究

陈虹, 黄元礼, 王涵, 连环, 韩倩倩, 王春仁

中国食品药品检定研究院, 北京 102629

收稿日期:2020-11-05 修回日期:2021-07-01 出版日期:2021-11-30 发布日期:2021-12-04
通讯作者: 韩倩倩,E-mail:hanqianqian2005@163.com;王春仁,E-mail:chunrenwang@263.net E-mail:hanqianqian2005@163.com;chunrenwang@263.net
作者简介:陈虹,E-mail:cherry118000@163.com。
基金资助:
科技部“十三五”重点研发计划（2016YFC1103202）；中国科学院“器官重建与制造”战略性先导专项（XDA16040602）

Research on KeratinoSens assay applied to medical devices

CHEN Hong, HUANG Yuanli, WANG Han, LIAN Huan, HAN Qianqian, WANG Chunren

National Institute for Food and Drug Control, Beijing 102629, China

Received:2020-11-05 Revised:2021-07-01 Online:2021-11-30 Published:2021-12-04

摘要/Abstract

摘要： 目的： 介绍一种迟发型皮肤超敏反应的新型体外人源细胞系替代评价方法──KeratinoSens试验，并探讨其在两种医疗器械致敏性检测中的应用。方法： 构建质粒pGL4.17-AKR1C2-ARE-SV40转染人类永生化角质细胞系HaCaT，用G418筛选得到稳定转染细胞系KeratinoSens（采用此细胞系检测致敏性的方法称为KeratinoSens试验）。采用KeratinoSens试验对已知非致敏物和致敏物进行分析，通过计算阴性对照组的变异度，测试细胞株的稳定性；将试验结果和已知的致敏性进行对比，得出KeratinoSens试验的致敏预测性。采用KeratinoSens试验测试临床上致敏报道较多的医疗器械医用天然乳胶手套和含镍金属的致敏性。结果： 阴性对照组的变异度为18.6%，符合稳定性要求，KeratinoSens试验的致敏预测性良好；两种医疗器械的KeratinoSens试验结果均呈阴性。结论： KeratinoSens细胞系构建成功，将其初步应用于医疗器械的致敏性检测，可为该方法转化为医疗器械检测方法提供经验数据。

关键词: 致敏性检测, 体外替代, KeratinoSens试验, 医疗器械

Abstract: OBJECTIVE: This article introduces a new in vitro human-derived cell line as an alternative method (KeratinoSens assay) of evaluation for delayed-type hypersensitivity to,KeratinoSens assay,and discusses its application in the detection of two medical devices. METHODS: Plasmid pGL4.17-AKR1C2-ARE-SV40 was constructed to transfect the human immortalized keratinocytes HaCaT,and the stablye transfected cell line KeratinoSens was obtained by screening with G418. TheUsing KeratinoSens assaywas used to evaluateprocess sensitizers with different sensitization ranks and non-sensitizers. By calculating the Variabilities of gene activity induction variability ofin the negative control were used to identify cell stability. The data were,compared toing the test results with the sensitization properties of the compounds in the literature,and were used to indicateget the predictability of the sensitization of the cell line. After the successful cell transfection is identified,the KeratinoSens assay wasis used to test the medical devices with more clinical sensitization reports,medical natural latex gloves and nickel-containing metals. RESULTS: The negative control hads a variation of 18.6% which meets the stability requirements,and the synthesized cell hadve good sensitization predictability. KeratinoSens assay rResults fromof the two medical devices were are negative. CONCLUSION: The KeratinoSens was successfully constructed and used to initially applied to medical device testing to provide empirical data for itsthe method to be transformationed into a medical device testing method.

Key words: sensitization test, alternative in vitro, KeratinoSens assay, medical device

中图分类号:

R331

陈虹, 黄元礼, 王涵, 连环, 韩倩倩, 王春仁. KeratinoSens试验在医疗器械致敏性检测中的应用研究[J]. 癌变·畸变·突变, 2021, 33(6): 455-460.

CHEN Hong, HUANG Yuanli, WANG Han, LIAN Huan, HAN Qianqian, WANG Chunren. Research on KeratinoSens assay applied to medical devices[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(6): 455-460.

参考文献

[1] CUMBERBATCH M, DEARMAN R J, GRIFFITHS C E, et al. Epidermal Langerhans cell migration and sensitisation to chemical allergens[J]. APMIS, 2003, 111(7/8):797-804.
[2] VAN OCH F M, VAN LOVEREN H, VAN WOLFSWINKEL J C, et al. Assessment of potency of allergenic activity of low molecular weight compounds based on IL-1alpha and IL-18 production by a murine and human keratinocyte cell line[J]. Toxicology, 2005, 210(2/3):95-109.
[3] ANTONOPOULOS C, CUMBERBATCH M, MEE J B, et al. IL-18 is a key proximal mediator of contact hypersensitivity and allergen-induced Langerhans cell migration in murine epidermis[J]. J Leukoc Biol, 2008, 83(2):361-367.
[4] BERGSTRÖM M A, LUTHMAN K, NILSSON J L, et al. Conjugated dienes as prohaptens in contact allergy:in vivo and in vitro studies of structure-activity relationships, sensitizing capacity, and metabolic activation[J]. Chem Res Toxicol, 2006, 19(6):760-769.
[5] ANDREAS N, CAROLINE B, LESLIE F, et al. The intra-and inter-laboratory reproducibility and predictivity of the KeratinoSens assay to predict skin sensitizers in vitro:results of a ring-study in five laboratories[J]. Toxicol in Vitro, 2011, 25(3):733-744.
[6] NATSCH A. The Nrf2-Keap1-ARE toxicity pathway as a cellular sensor for skin sensitizers-functional relevance and a hypothesis on innate reactions to skin sensitizers[J]. Toxicol Sci, 2010, 113(2):284-292.
[7] EMTER R, ELLIS G, NATSCH A. Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro[J]. Toxicol Appl Pharmacol, 2010, 245(3):281-290.
[8] 冷建梅, 黄沛力. 医用乳胶手套中水溶性蛋白质的含量分析[J]. 分析科学学报, 2009, 25(1):114-116.
[9] KIMBER I, BASKETTER D A, BUTLER M, et al. Classification of contact allergens according to potency:proposals[J]. Food Chem Toxicol, 2003, 41(12):1799-1809.
[10] SOMMER S, WILKINSON S M, BECK M H, et al. Type IV hypersensitivity reactions to natural rubber latex:results of a multicentre study[J]. Br J Dermatol, 2002, 146(1):114-117.
[11] KOH D, NG V, LEOW Y H, et al. A study of natural rubber latex allergens in gloves used by healthcare workers in Singapore[J]. Br J Dermatol, 2005, 153(5):954-959.
[12] ARTIK S, VON VULTéE C, GLEICHMANN E, et al. Nickel allergy in mice:enhanced sensitization capacity of nickel at higher oxidation states[J]. J Immunol, 1999, 163(3):1143-1152.
[13] BAUCH C, KOLLE S N, FABIAN E, et al. Intralaboratory validation of four in vitro assays for the prediction of the skin sensitizing potential of chemicals[J]. Toxicol Vitro, 2011, 25(6):1162-1168.
[14] URBISCH D, MEHLING A, GUTH K, et al. Assessing skin sensitization hazard in mice and men using non-animal test methods[J]. Regul Toxicol Pharmacol, 2015, 71(2):337-351.

KeratinoSens试验在医疗器械致敏性检测中的应用研究

Research on KeratinoSens assay applied to medical devices

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