Two-Dimensional Cellular and Three-Dimensional Bio-Printed Skin Models to Screen Topical-Use Compounds for Irritation Potential
Two-Dimensional Cellular and Three-Dimensional Bio-Printed Skin Models to Screen Topical-Use Compounds for Irritation Potential
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DOI：
10.3389/fbioe.2020.00109
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发表时间：
2020-02
影响因子：
5.7
通讯作者：
Zhengxi Wei;Xue Liu;Masato Ooka;Li Zhang;Min Jae Song;Ruili Huang;N. Kleinstreuer;A. Simeonov;M. Xia;M. Ferrer
中科院分区：
文献类型：
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作者：
Zhengxi Wei;Xue Liu;Masato Ooka;Li Zhang;Min Jae Song;Ruili Huang;N. Kleinstreuer;A. Simeonov;M. Xia;M. Ferrer
评估皮肤刺激潜力对于局部药物和其他消费品（如化妆品）的安全性评价至关重要。在消费品和成分的安全性评估中，使用先进的细胞模型作为替代动物试验的替代方法，已经被欧盟（EU）和其他国家的法律所强制要求。然而，目前还没有一个大规模的比较局部使用的化合物在不同的细胞皮肤模型的影响。本研究评估了局部使用化合物在与高通量筛选（HTS）平台兼容的不同皮肤细胞模型中的刺激潜力。首先使用原代新生儿角质形成细胞和永生化人角质形成细胞的二维（2D）单层模型测试了一组451种局部使用化合物的细胞毒性作用。从单层培养系统的初始筛选中识别出的46种毒性化合物在重建的人表皮（RhE）和全层皮肤（FTS）三维（3D）组织模型构建体上进一步测试了皮肤刺激潜力。通过测量组织活力、跨上皮电阻（TEER）以及细胞因子白细胞介素1 α（IL-1α）和白细胞介素18（IL-18）的分泌来评估化合物的皮肤刺激潜力。在已知的刺激物中，高浓度甲基紫和甲基玫瑰苯胺降低了RhE和FTS模型的活力，降低了TEER，并增加了IL-1α分泌，与刺激特性一致。然而，在低浓度下，这两种化合物在RhE或FTS模型中增加IL-18分泌而不影响分泌的IL-1α水平，并且不降低组织活力和TEER。该结果表明，在低浓度下，甲基紫和甲基玫瑰苯胺具有过敏潜力，但不会引起刺激。使用HTS兼容的2D细胞和3D组织皮肤模型，以及刺激相关活性终点，我们获得了数据，以帮助评估局部使用化合物的刺激作用并识别潜在的皮肤危害。
Assessing skin irritation potential is critical for the safety evaluation of topical drugs and other consumer products such as cosmetics. The use of advanced cellular models, as an alternative to replace animal testing in the safety evaluation for both consumer products and ingredients, is already mandated by law in the European Union (EU) and other countries. However, there has not yet been a large-scale comparison of the effects of topical-use compounds in different cellular skin models. This study assesses the irritation potential of topical-use compounds in different cellular models of the skin that are compatible with high throughput screening (HTS) platforms. A set of 451 topical-use compounds were first tested for cytotoxic effects using two-dimensional (2D) monolayer models of primary neonatal keratinocytes and immortalized human keratinocytes. Forty-six toxic compounds identified from the initial screen with the monolayer culture systems were further tested for skin irritation potential on reconstructed human epidermis (RhE) and full thickness skin (FTS) three-dimensional (3D) tissue model constructs. Skin irritation potential of the compounds was assessed by measuring tissue viability, trans-epithelial electrical resistance (TEER), and secretion of cytokines interleukin 1 alpha (IL-1α) and interleukin 18 (IL-18). Among known irritants, high concentrations of methyl violet and methylrosaniline decreased viability, lowered TEER, and increased IL-1α secretion in both RhE and FTS models, consistent with irritant properties. However, at low concentrations, these two compounds increased IL-18 secretion without affecting levels of secreted IL-1α, and did not reduce tissue viability and TEER, in either RhE or FTS models. This result suggests that at low concentrations, methyl violet and methylrosaniline have an allergic potential without causing irritation. Using both HTS-compatible 2D cellular and 3D tissue skin models, together with irritation relevant activity endpoints, we obtained data to help assess the irritation effects of topical-use compounds and identify potential dermal hazards.