体外重建人皮肤与THP-1共培养在化妆品皮肤增敏中的应用。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters Pub Date : 2025-09-30 DOI:10.1016/j.toxlet.2025.111738

Bo Wang , Hongyu Ma , Yuying Cheng , Dangdang Cheng , Feifei Wang

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引用次数: 0

摘要

背景：化妆品配方的敏感性测试主要涉及人体斑贴试验。细胞模型，包括KeratinoSensTM， ARE-Nrf2荧光素酶LuSens和h-CLAT，可以根据OECD 442E标准预测化学致敏。目的：本研究的目的是利用皮肤模型与THP-1细胞共培养，确定一种评估化妆品致敏潜力的潜在方法。方法：采用流式细胞术和酶联免疫吸附法（elisa）鉴定THP-1细胞表面标志物CD54和CD86；检测IL-18、MTT、7AAD的相对分泌；并评估组织和细胞的活力。用人斑贴试验分析不良反应。结果：暴露于肉桂醛、2,4-二硝基氯苯、没食子酸丙酯和香豆素后IL-18的相对分泌水平均小于0.79。CD54的相对荧光强度大于150，CD86的相对荧光强度大于200。以RFI CD54≥150、RFI CD86≥200或RS IL-18≤0.79作为潜在致敏标准，18项化妆品配方引起致敏。此外，在人体贴片试验中，一种含有23种成分的化妆品配方引起了致敏。体外试验对该产品的敏感性为56.52%，对清洁产品的敏感性为75%。结论：RFI CD54/CD86和IL-18可作为评价化妆品潜在致敏性的新方法。体外皮肤细胞和THP-1细胞共培养结合人斑贴试验可以更好地预测潜在的化妆品致敏性。

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Application of an in vitro reconstructed human skin coculture with THP-1 on cosmetics in skin sensitization

Background

Sensitivity testing for cosmetic formulations mainly involves human patch tests. Cell models, including KeratinoSens^TM, ARE-Nrf2 luciferase LuSens and h-CLAT, can predict chemical sensitization according to the OECD 442E standard.

Objective

The aim of this study was to identify a potential approach for the assessment of the sensitization potential of cosmetics using skin models cocultured with THP-1 cells.

Methods

We identified the surface markers CD54 and CD86 in THP-1 cells using flow cytometry and enzyme-linked immunosorbent assays (ELISAs); detected the relative secretion of IL-18, MTT and 7AAD; and evaluated tissue and cell viability. Adverse reactions were analysed using the human patch test.

Results

The relative level of IL-18 secretion after exposure to cinnamyl aldehyde, 2,4-dinitrochlorobenzene, propyl gallate and coumarin was less than 0.79. The relative fluorescence intensity of CD54 was greater than 150, and that for CD86 was greater than 200. Using RFI CD54 ≥ 150, RFI CD86 ≥ 200 or RS IL-18 ≤ 0.79 as the potential sensitization standard, an 18-item cosmetic formulation caused sensitization. Moreover, a 23-item cosmetic formulation caused sensitization in the human patch test. The sensitivity to the products in the in vitro test reached 56.52 %, compared with 75 % for the cleaning products.

Conclusion

RFI CD54/CD86 and IL-18 may serve as a new approach to evaluate potential sensitization to cosmetics. In vitro SkinEthic and THP-1 cell coculture combined with the human patch test may better predict potential sensitization to cosmetics.

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