An evaluation of a cultured human corneal epithelial tissue model for the determination of the ocular irritation potential of pharmaceutical process materials.
An evaluation of a cultured human corneal epithelial tissue model for the determination of the ocular irritation potential of pharmaceutical process materials.
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DOI：
10.1016/j.tiv.2010.03.004
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发表时间：
2010-09
期刊：
影响因子：
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通讯作者：
Christopher W Seaman;A. Whittingham;R. Guest;N. Warren;M. J. Olson;F. Guerriero;E. Adriaens;B. De
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文献类型：
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作者：
Christopher W Seaman;A. Whittingham;R. Guest;N. Warren;M. J. Olson;F. Guerriero;E. Adriaens;B. De
接触眼睛后的刺激和其他形式的局部毒性是职业接触化学品引起的潜在严重问题。传统上，评价新化学品的潜在刺激性依赖于使用家兔进行的体内研究。对人体危害的体内方法的预测潜力的关注和对经济和快速筛选化学品的需求刺激了大量的工作，以研究体外替代品，以评估眼部刺激潜力。该出版物描述了一项筛选研究，以评估重建的角膜上皮培养系统，作为药物工艺材料眼刺激试验的替代方法，扩展了该系统已测试的化学领域。将总共21种试验化学品应用于市售的重建人角膜上皮（HCE）培养物，并评估对组织活力（MTT还原试验）、组织组织学和IL-α表达的影响。阳性对照（0.5%和1% SDS）对组织显示出与剂量和时间相关的不良反应，与已知的刺激作用一致。阴性对照未显示组织学变化，并在整个实验时间过程中保持高活力。当将活力（MTT降低）与供试品眼部刺激的现有EU分类（基于体内评价结果的分类）进行比较时，一致性极佳，每个采样时间点的准确度均超过80%。通过MTT减少估计的组织活力似乎是最有用的，作为评估化学品刺激潜力的主要手段。组织学检查结果与MTT法基本一致。然而，需要进一步考虑使用细胞因子分析，因为该参数的结果显示与已知的刺激潜力无关。这些结果表明，HCE培养，单独或作为一个分层的危险筛选计划的一部分，有希望用于减少对活的受试者测试的依赖，并有助于生成一个适当的危险分类和标签的建议。
Irritation and other forms of local toxicity following contact with eyes is a potentially serious problem arising from occupational exposure to chemicals. Traditionally, evaluation of the irritant potential of novel chemicals has relied on the use of in vivo studies with rabbits. Concerns about the predictive potential of in vivo methods for human hazard and demand for economical and rapid screening of chemicals has stimulated a great deal of work to investigate in vitro alternatives for evaluating ocular irritation potential. This publication describes a screening study to assess a reconstituted corneal epithelial culture system, as an alternative for testing for ocular irritation with pharmaceutical process materials, extending the chemical domain with which this system has been tested. A total of 21 test chemicals were applied to commercially supplied reconstituted human corneal epithelial (HCE) cultures and effects on tissue viability (MTT reduction assay), tissue histology and IL-alpha expression were assessed. Positive controls (0.5% and 1% SDS) showed dose- and time-related adverse effects on tissues, consistent with known irritant effects. Negative controls showed no histological changes and retained high viability throughout the time-course of the experiment. Concordance was excellent with accuracy at each sampling time point of over 80% when viability (MTT reduction) was compared with existing EU classification of the test articles for ocular irritation (classification based on results of in vivo evaluation). Tissue viability as estimated by MTT reduction appears most useful as the primary means of assessing the irritation potential of the chemicals. Histopathological examination generally agreed with the results of the MTT assay. However, the use of cytokine analysis will need further consideration as results for this parameter showed no relationship with known irritation potential. These results infer that HCE cultures, alone or as a part of a tiered hazard screening programme, have promise for use in reducing reliance on live subject tests and contribute to generation of an appropriate hazard classification and label advice.