Van Tienhoven E A E, Korbee D, Schipper L, Verharen H W, De Jong W H
Centre for Biological Medicines and Medical Technology, National Institute for Public Health and the Environment, P.O. Box 1, 3720 BA Bilthoven, the Netherlands.
J Biomed Mater Res A. 2006 Jul;78(1):175-82. doi: 10.1002/jbm.a.30679.
The choice for a biomaterial is partly based on the outcome of (cyto)toxicity assays. The rationales behind the selection of certain parameters, such as cell lines, controls, and animals, were evaluated using a positive and a negative control, and one experimental sample designed to induce intermediate toxicity. Extraction and direct contact assays were performed using human epidermal keratinocytes and mouse fibroblasts and mouse epithelial cells. Cell survival was measured with the tetrazolium salt (MTT) reduction assay. In addition, local implantation studies were performed in mice and rats. The positive control induced a high degree of toxicity in all in vitro tests performed, indicating that the toxicity observed in the direct contact assay was due to in situ extraction of toxic components. In the direct contact assay the negative control tested on the mouse fibroblasts resulted in a significant reduction of cell survival. No decrease in cell survival was found using the experimental sample. Subcutaneous implantation studies in mice showed that the positive control material induced a severe degeneration in mice. However, in rats just minimal alterations were noted. The experimental material induced moderate responses only in mice. Our results indicate that the direct contact assay provides limited additional information on the cytotoxicity of materials if certain limitations are not taken into account. For the in vivo implantation assay mice were superior to rats in detecting local toxic responses.
生物材料的选择部分基于（细胞）毒性试验的结果。使用阳性和阴性对照以及一个旨在诱导中度毒性的实验样品，评估了选择某些参数（如细胞系、对照和动物）背后的原理。使用人表皮角质形成细胞、小鼠成纤维细胞和小鼠上皮细胞进行了提取和直接接触试验。用四唑盐（MTT）还原试验测量细胞存活率。此外，还在小鼠和大鼠中进行了局部植入研究。阳性对照在所有进行的体外试验中均诱导了高度毒性，表明在直接接触试验中观察到的毒性是由于有毒成分的原位提取。在直接接触试验中，对小鼠成纤维细胞进行测试的阴性对照导致细胞存活率显著降低。使用实验样品未发现细胞存活率下降。小鼠皮下植入研究表明，阳性对照材料在小鼠中诱导了严重变性。然而，在大鼠中仅观察到微小变化。实验材料仅在小鼠中诱导了中度反应。我们的结果表明，如果不考虑某些局限性，直接接触试验提供的关于材料细胞毒性的额外信息有限。对于体内植入试验，小鼠在检测局部毒性反应方面优于大鼠。
