化妆品眼刺激试验替代方法标准化与展望

眼刺激试验是化妆品安全评价的主要测试项目,目前正尝试建立代替传统动物实验的体外方法,这些方法处于研发、验证和认可的不同阶段。尽管还没有单一体外试验能完全替代兔眼实验,但系统分析替代方法建立的原理、科学相关性和标准化程度的不同,有助于合理选择运用体外方法指导化妆品眼刺激的标识分类、质量检验、产品开发和机制研究。     

GLP规定和安全试验

<正>一、所谓GLP GLP(Good Laboratory practices)可译为“有关安全试验的非临床试验实验规范”,“前临床试验的动物试验规范’或“关于安全试验标准。”GLP规定的目的是为了确保各种安全试验数据的可靠性,从实施     

皮肤刺激实验替代模型的研究新进展

皮肤刺激性是日常使用化妆品最常见的不良反应之一。人类健康相关产品危险性评价常做皮肤刺激性实验,皮肤刺激性试验是化妆品原料及产品安全性评价的主要项目。传统皮肤刺激试验采用实验动物进行,2013年3月11日欧盟已经禁止销售基于动物实验研发的化妆品原料及产品.随着组织工程技术和现代生物技术的发展,多种替代动物试验的体外模型被开发和应用,新的的皮肤刺激物陆续被发现。欧盟多采纳重组人表皮实验方法作为新体外皮肤实验指南(包括模型Episkin和模型Epiderm),随着体外模型重建技术的不断改善,不仅拓展了皮肤模型的临床应用范围,也必然推动新的敏感而特异的皮肤标志物的发现和应用。     

一种体外皮肤腐蚀性试验替代方法的建立

目的建立皮肤腐蚀性的体外检测方法—大鼠经皮电阻试验(TER),优化OECD指南标准TG 430大鼠经皮电阻方法结果判定标准,并评价该方法作为替代方法用于评估化妆品原料的皮肤腐蚀性检测可行性。方法参照经合组织(OECD)化学品检测指南中TG 430体外皮肤腐蚀性大鼠经皮电阻试验的流程,使用Wistar大鼠对16种参考化学物质进行皮肤腐蚀性检测,设定不同的判定标准方案,通过对比筛选出最优的判定标准方案;并在5个实验室间进行11种参考物质的结果比对。结果改良后的TER结果判定方案结论与参考化学物质的实际腐蚀性结果具一致性(Kappa值0.64),改良方案B的特异性为66.7%,敏感性为100%,效能最高;实验室间比对结论差异无显著性(P0.05),实验室间对11种参考物质的判定一致率为72.7%。结论经改良的大鼠经皮电阻试验可较好的筛选化学物质的腐蚀性,有望在我国的替代毒理学及化学物质安全性评价中发挥重要的作用。     

动物实验替代方法与21世纪毒性测试发展策略

随着新化学物质日益增多以及“3R”原则的广泛实施,传统的毒性测试面临着严峻挑战.毒性测试的发展正经历着一个关键时期,即从耗时、耗费的传统整体动物试验转向快速高通量的、含定量参数分析和机制研究的体外替代试验.实验动物替代方法不仅是出于遵行“3R”原则的考虑,也是毒理学学科发展以及社会经济发展的需要与科学要求.实验动物替代方法的发展与应用已成为21世纪毒性测试的重要方向,获得越来越广泛的支持和管理认可,具有广阔的发展前景和十分重要的应用价值.     

GLP认证对动物实验管理的要求

依据我国GLP法规和GLP检查办法,结合本单位申请GLP认证的实践,对GLP认证中与实验动物管理有关的人员、动物饲养设施设备、相关的标准操作规程（SOP）的制定和配备、动物实验的管理以及相关原始资料的归档保存等作了论述。     

转基因动物在毒理学中的应用

转基因动物作为研究代谢调节、分化、发育中有关基因功能的工具已在各领域发挥了巨大的作用。在新药开发和研究中,因转基因动物的准确、经济、实验次数少和显著缩短实验时间等优点,现已成为一种进行"快速筛选"和药物非临床前安全评价的的手段。本文将对毒理学研究方面常用的转基因小鼠和大鼠进行阐述,旨在更全面的了解其在毒理学研究中的重要作用,为利用转基因技术技术培育应用于毒理学研究方面的转基因小鼠和大鼠的制备提供参考性的依据。     

斑马鱼模型在化妆品功效评价研究中的应用进展

斑马鱼具有个体小、易饲养、胚胎透明、易观察等优势,是一种广泛应用于医学和毒理学研究的模式生物。因其具有可高通量筛选、测试周期短、实验结果可信度高等优势,近年,斑马鱼逐步用于化妆品功效评价,且成为化妆品行业认可的评价方式,也是学者们研究的热点。目前,化妆品功效种类繁多,而斑马鱼模型评价标准正逐步完善,本文阐述了斑马鱼模型在化妆品美白、抗炎、抗氧化等功效评价中的应用进展。     

GLP条件下非实验因素对实验用豚鼠生理状态的影响

目的研究GLP条件下非实验因素（包括实验操作、不同笼养方式、对照组因素以及维生素C的添加等）对全身过敏实验中豚鼠质量及生理状况的影响情况,为探讨在GLP条件下如何有效的提高实验动物的质量以及规范过敏性反应的评价提供实验依据。方法实验采用计算机软件进行随机分组,雌雄各半。实验在GLP实验室进行,按要求对豚鼠饲养的环境温度、湿度和噪音进行严格的控制。实验按照SFDA颁布的指导原则推荐的全身过敏实验的程序和方案进行。结果空白对照组和溶剂对照组（生理盐水对照组）在致敏期,体重变化差异明显（P〈0．05）;溶剂对照组与阳性对照组相比较,豚鼠的体重增长在实验的各阶段无明显差异（P〉0．05）;对比托盘笼架饲养,大塑料盒饲养在检疫期更有利于雌性豚鼠体重的增长（P〈0．05）;不添加维生素C组豚鼠在致敏期体重减轻明显（P〈0．05）,并出现明显的脱毛症状。结论在GLP条件下,虽然环境条件得到了有效的改善和控制,但是一些容易忽视的非实验因素明显影响了豚鼠的质量及生理状况。     

体外模型的发展及其在鲸豚研究中的应用

体外补充替代模型“细胞系”为生命科学研究提供了新的平台,在一定程度上突破了科学研究中伦理、法律、动物福利和动物保护等的限制,从细胞和分子视角更深层次地揭示复杂生命体的生物效应和 调控机制。尤其对于濒危动物,细胞系的建立与超低温冷冻技术相结合,既可以保存濒危动物具有生物表达活性的遗传种质,又可以提供体外保育研究的新平台（如动物毒理学实验）,对动物保护意义 重大。目前细胞培养体系已作为多功能平台被应用于鲸豚类细胞遗传学、病毒学和毒理学的相关研究中,但从物种和组织来源以及细胞类型来看,能长期稳定传代的鲸豚类细胞系仍较单一。优化细胞培 养条件,运用鲸豚类体外细胞揭示更多的生命机制问题,仍是当前鲸豚类体外细胞模型研究所面临的挑战。本文对动物体外模型及其在鲸豚研究中的应用进行了概述,以期推动该技术在鲸豚保育研究中 的创新和发展。     

Integrated data acquisition system for medical device testing and physiology research in compliance with good laboratory practices

In seeking approval from the US Food and Drug Administration (FDA) for clinical trial evaluation of an experimental medical device, a sponsor is required to submit experimental findings and support documentation to demonstrate device safety and efficacy that are in compliance with Good Laboratory Practices (GLP). The objective of this project was to develop an integrated data acquisition (DAQ) system and documentation strategy for monitoring and recording physiological data when testing medical devices in accordance with GLP guidelines mandated by the FDA. Data aquisition systems were developed as stand-alone instrumentation racks containing transducer amplifiers and signal processors, analog-to-digital converters for data storage, visual display and graphical user-interfaces, power conditioners, and test measurement devices. Engineering standard operating procedures (SOP) were developed to provide a written step-by-step process for calibrating, validating, and certifying each individual instrumentation unit and the integrated DAQ system. Engineering staff received GLP and SOP training and then completed the calibration, validation, and certification process for the individual instrumentation components and integrated DAQ system. Eight integrated DAQ systems have been successfully developed that were inspected by regulatory affairs consultants and determined to meet GLP guidelines. Two of these DAQ systems were used to support 40 of the pre-clinical animal studies evaluating the AbiCor artificial heart (ABIOMED, Danvers, MA). Based in part on these pre-clinical animal data, the AbioCor clinical trials began in July 2001. The process of developing integrated DAQ systems, SOP, and the validation and certification methods used to ensure GLP compliance are presented in this article.     

Nonhuman primates and teratological research

Nonhuman primates were first recognized as models for the study of developmental toxicity (teratology) following the thalidomide tragedy. Since that time they have played important roles in both testing of drugs for human safety and as models for studying specific malformations commonly seen in children. Although in vitro and alternative test systems using lower animal forms or simplified test systems have been incorporated into developmental toxicity studies, whole animal testing will be required for the foreseeable future because of the complex relationship of the maternal/embryofetal/placental unit. The nonhuman primate will be particularly valuable where equivocal results are experienced in other commonly used laboratory species, when the drug/chemical is likely to be used during pregnancy, and for human-derived biotechnical products which often are not bioactive in nonprimate species.     

Integrated decision-tree testing strategies for skin corrosion and irritation with respect to the requirements of the EU REACH legislation

Liverpool John Moores University and FRAME recently conducted a research project, sponsored by DEFRA, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with the REACH system. This report focuses on how to maximise the use of alternative methods (both in vitro and in silico) for skin corrosion and irritation testing within a tiered testing strategy. It considers the latest developments in in vitro testing, with particular reference to the reconstituted skin models which have now been now been successfully validated and independently endorsed as suitable for both skin corrosivity and irritancy testing within the EU.     

How omics technologies can contribute to the '3R' principles by introducing new strategies in animal testing

In Europe, in light of ethical, political and commercial pressure, every effort should be made to replace animals with alternatives (e.g. in vitro models), to reduce the number of animals used in experiments to a minimum and to refine current testing strategies in a way that ensures animals undergo minimum pain and distress. Methods currently used in toxicology for mandatory safety tests rely heavily on the dosing of animals, followed by the detection and pathological evaluation of manifested toxic lesions. Through the integration of so-called 'omics' technologies, a global analysis of treatment-related changes on the molecular level becomes feasible and therefore might provide a means for predicting toxicity before classical toxicological endpoints. This Opinion article summarizes the key features of pushing the '3R' principles in animal testing, discusses the possible impact on safety testing in toxicology and describes the potential of using omics technologies for improved toxicity prediction to meet ethical, political and commercial expectations.     

Integrated decision-tree testing strategies for acute systemic toxicity and toxicokinetics with respect to the requirements of the EU REACH legislation

Liverpool John Moores University and FRAME conducted a joint research project, sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for acute systemic toxicity and toxicokinetic testing. The paper reviews in vitro tests based on basal cytotoxicity and target organ toxicity, along with QSAR models and expert systems available for this endpoint. The use of PBPK modelling for the prediction of ADME properties is also discussed. These tests are then incorporated into a decision-tree style, integrated testing strategy, which also includes the use of refined in vivo acute toxicity tests, as a last resort. The implementation of the strategy is intended to minimise the use of animals in the testing of acute systemic toxicity and toxicokinetics, whilst satisfying the scientific and logistical demands of the EU REACH legislation.