Nanotechnologies such as nanoparticles are established components of new medical devices and pharmaceuticals. The use and distribution of these materials increases the requirement for standardized evaluation of possible adverse effects, starting with a general cytotoxicity screening. The Horizon 2020 project “Regulatory Science Framework for Nano(bio)material-based Medical Products and Devices (REFINE)” identified in vitro cytotoxicity quantification as a central task and first step for risk assessment and development for medical nanocarriers. We have performed an interlaboratory comparison on a cell-assay matrix including a kinetic lactate dehydrogenase (LDH) release cell death and WST-8 cell viability assay adapted for testing organic nanocarriers in four well-characterized cell lines of different organ origins. Identical experiments were performed by three laboratories, namely the Biomedical Technology Center (BMTZ) of the University of Münster, SINTEF Materials and Chemistry (SINTEF), and the National Institute for Public Health and the Environment (RIVM) of the Netherlands according to new standard operating procedures (SOPs). The experiments confirmed that LipImage™ 815 lipidots^® are non-cytotoxic up to a concentration of 128 µg/mL and poly(alkyl cyanoacrylate) (PACA) nanoparticles for drug delivery of cytostatic agents caused dose-dependent cytotoxic effects on the cell lines starting from 8 µg/mL. PACA nanoparticles loaded with the active pharmaceutical ingredient (API) cabazitaxel showed a less pronounced dose-dependent effect with the lowest concentration of 2 µg/mL causing cytotoxic effects. The mean within laboratory standard deviation was 4.9% for the WST-8 cell viability assay and 4.0% for the LDH release cell death assay, while the between laboratory standard deviation was 7.3% and 7.8% for the two assays, respectively. Here, we demonstrated the suitability and reproducibility of a cytotoxicity matrix consisting of two endpoints performed with four cell lines across three partner laboratories. The experimental procedures described here can facilitate a robust cytotoxicity screening for the development of organic nanomaterials used in medicine.

Graphical abstract

中文翻译：

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用于评估有机纳米载体细胞毒性的体外测定基质的标准化：实验室间试验性比较

纳米技术（如纳米粒子）是新型医疗设备和药物的成熟组成部分。这些材料的使用和分布增加了对可能的不良反应进行标准化评估的要求，从一般的细胞毒性筛选开始。Horizo​​n 2020 项目“基于纳米（生物）材料的医疗产品和设备（REFINE）的监管科学框架”将体外细胞毒性量化确定为医疗纳米载体风险评估和开发的核心任务和第一步。我们对细胞测定基质进行了实验室间比较，包括动态乳酸脱氢酶 (LDH) 释放细胞死亡和 WST-8 细胞活力测定，适用于测试四种不同器官来源的良好表征的细胞系中的有机纳米载体。三个实验室根据新标准进行了相同的实验，即明斯特大学生物医学技术中心（BMTZ）、SINTEF材料与化学（SINTEF）和荷兰国家公共卫生与环境研究所（RIVM）操作程序（SOP）。实验证实 LipImage™ 815 脂质体^®浓度高达 128 µg/mL 时无细胞毒性，用于细胞抑制剂药物递送的聚（氰基丙烯酸烷基酯）（PACA）纳米颗粒从 8 µg/mL 开始对细胞系产生剂量依赖性细胞毒性作用。载有活性药物成分 (API) 卡巴他赛的 PACA 纳米颗粒显示出不太明显的剂量依赖性效应，最低浓度为 2 µg/mL，引起细胞毒性效应。WST-8 细胞活力测定的实验室标准偏差平均值为 4.9%，LDH 释放细胞死亡测定为 4.0%，而两种测定的实验室标准偏差分别为 7.3% 和 7.8%。在这里，我们展示了一个细胞毒性矩阵的适用性和可重复性，该矩阵由三个合作伙伴实验室的四个细胞系组成的两个端点组成。

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