1.1 本试验方法提供了一种评估纳米颗粒材料悬浮液在猪近端小管细胞（LLC-PK1）和人肝癌细胞（Hep G2）中的细胞毒性的方法，这两种细胞代表了全身给药后的潜在靶器官。 1.2 本试验方法是 体外 临床前表征级联。 1.3 该试验方法包括使用两种方法评估细胞毒性的方案，3-（4,5-二甲基噻唑基-2）-2,5-二苯基四唑溴化铵（MTT）还原和乳酸脱氢酶（LDH）释放。 1.4 以国际单位制表示的数值应视为标准值。本标准不包括其他计量单位。 1.5 本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.6 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 评估纳米材料对靶器官细胞产生细胞毒性的倾向有助于临床前开发。 5.2 材料和材料提取物的标准历史细胞毒性试验已使用成纤维细胞，并在实践中得到充分证明 F813 ，试验方法 F895 巨噬细胞和微米颗粒的使用也提供了有关细胞毒性和刺激的信息 F1903 . 5.3 该试验方法通过使用靶器官细胞添加到细胞毒性试验方案中。两种定量测定LDH漏出量和MTT减少量用于评估细胞毒性。 5.4 该测试方法可能无法预测所有类型的纳米材料应用中发生的事件，并且提醒用户考虑测试对各种类型的纳米材料及其应用的适当性。 该程序只能用于比较一系列相关纳米材料的细胞毒性。如果没有对分析基质中每个单独纳米材料的物理化学性质进行额外表征，则不可能对不相关的纳米材料进行有意义的比较。

1.1 This test method provides a methodology to assess the cytotoxicity of suspensions of nanoparticulate materials in porcine proximal tubule cells (LLC-PK1) and human hepatocarcinoma cells (Hep G2), which represent potential target organs following systemic administration. 1.2 This test method is part of an in vitro preclinical characterization cascade. 1.3 This test method consists of a protocol utilizing two methods for estimation of cytotoxicity, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) release. 1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. 1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 5.1 Assessing the propensity of a nanomaterial to cause cytotoxicity to the cells of a target organ can assist in preclinical development. 5.2 The standard historical cytotoxicity testing of materials and extracts of materials has used fibroblasts and is well documented in Practice F813 , Test Method F895 , and ISO 10993-5. The use of macrophages and micron size particles has also provided information on cytotoxicity and stimulation using Practice F1903 . 5.3 This test method adds to the cytotoxicity test protocols by using target organ cells. Two quantitative assays measuring LDH leakage and MTT reduction are used to estimate cytotoxicity. 5.4 This test method may not be predictive of events occurring in all types of nanomaterial applications, and the user is cautioned to consider the appropriateness of the test for various types of nanomaterial and their applications. This procedure should only be used to compare the cytoxicity of a series of related nanomaterials. Meaningful comparison of unrelated nanomaterials is not possible without additional characterization of physicochemical properties of each individual nanomaterial in the assay matrix.