van de Sandt J J, Rutten A A, Koëter H B
TNO Toxicology and Nutrition Institute, Department of Biological Toxicology, PO Box 360, 3700 AJ Zeist, The Netherlands.
Toxicol In Vitro. 1993 Jan;7(1):81-6. doi: 10.1016/0887-2333(93)90115-l.
Rabbit full thickness skin can be maintained viable for 7 days using a two-compartment organ culture model. We report that this model is useful for assessing cutaneous toxicity. 11 chemicals were each applied topically and then cytotoxicity was determined using two different assays. Mitochondrial activity was assessed by measuring reduction of the tetrazolium salt MTT, while uptake of the vital dye neutral red was used as a parameter of membrane damage. Conversion of MTT was inhibited in a dose-dependent way by all of the chemicals tested except dimethyl sulphoxide. Furthermore, when the skin explants were kept in culture after exposure to the test agents, both repair of initial damage and delayed toxicity could be observed to some extent. Uptake of neutral red was affected dose-dependently by six out of the 11 chemicals tested. The data indicate that a strong correlation exists between the cytotoxicity of chemicals and their effect on the conversion of MTT in skin organ culture. The MTT assay reported here offers the possibility of studying both acute and delayed cytotoxicity, and to some extent, recovery from initial damage.
使用双室器官培养模型，兔全层皮肤可保持存活7天。我们报告该模型可用于评估皮肤毒性。分别局部应用11种化学物质，然后使用两种不同的检测方法测定细胞毒性。通过测量四唑盐MTT的还原评估线粒体活性，而将活性染料中性红的摄取用作膜损伤的参数。除二甲基亚砜外，所有测试的化学物质均以剂量依赖性方式抑制MTT的转化。此外，当皮肤外植体在接触测试剂后保持在培养中时，在一定程度上可以观察到初始损伤的修复和延迟毒性。11种测试化学物质中有6种对中性红的摄取有剂量依赖性影响。数据表明，化学物质的细胞毒性与其对皮肤器官培养中MTT转化的影响之间存在很强的相关性。本文报道的MTT检测方法提供了研究急性和延迟细胞毒性以及在一定程度上研究从初始损伤中恢复的可能性。
