Kasten F H, Felder S M, Gettleman L, Alchediak T
In Vitro. 1982 Jul;18(7):650-60. doi: 10.1007/BF02796398.
A model experimental culture system and protocol are described to screen polymerized dental materials for diffusible toxic products. The system employs cultures of human gingival fibroblasts grown in plates containing immobilized samples of polymerized resins. Comparative cytoxicity is evaluated by counting viable cells with the aid of phase optics at several time periods up to 48 h. To achieve adequate statistical sampling, multiple counts are made in four different zones at 90 degrees angles from each sample and at three distances from the centers of samples. The most significant data were generated during a 24 to 48 h test period in climate. This cytotoxicity test measured cell death as a function of time of exposure and distance from the sample (24 h, 0 to 3 mm; 48 h, 3 to 6 mm) and permitted a calculation of the relative cytoxicity for each material, which is termed the viability index (VI). This can be expressed as a percentage related to the control, which is called the time-distance cytotoxicity index (TDCI). This method is simple to carry out because it used basic laboratory equipment, is rapid, and has a sound scientific basis. It focuses on times and distances when or where, or both, the greatest cellular changes are taking place. Some data illustrated are based on the screening of eight different restorative resins. The literature of cell culture testing of dental materials is reviewed. It is concluded that biotoxicity studies ideally should employ diploid human target cells from the oral cavity because the cells retain specialized features. Secondary cultures or strains of human diploid gingival fibroblasts, which are relatively easy to obtain and maintain, are recommended as cells of choice for screening dental restorative materials in vitro.
本文描述了一种用于筛选聚合牙科材料中可扩散有毒产物的模型实验培养系统及方案。该系统采用在含有聚合树脂固定样品的平板中培养的人牙龈成纤维细胞。通过在长达48小时的多个时间段借助相差光学显微镜对活细胞进行计数来评估相对细胞毒性。为了获得足够的统计样本，在与每个样品呈90度角的四个不同区域以及距样品中心三个不同距离处进行多次计数。在气候条件下的24至48小时测试期内产生了最显著的数据。这种细胞毒性测试测量了细胞死亡与暴露时间和距样品距离的函数关系（24小时，0至3毫米；48小时，3至6毫米），并允许计算每种材料的相对细胞毒性，即活力指数（VI）。这可以表示为与对照相关的百分比，称为时间 - 距离细胞毒性指数（TDCI）。该方法易于实施，因为它使用基本的实验室设备，速度快且有坚实的科学基础。它关注细胞变化最大的时间和距离，无论是何时或何地，或两者皆是。所展示的一些数据基于对八种不同修复树脂的筛选。对牙科材料细胞培养测试的文献进行了综述。得出的结论是，生物毒性研究理想情况下应使用来自口腔的二倍体人类靶细胞，因为这些细胞保留了特殊特征。建议使用相对容易获得和维持的人二倍体牙龈成纤维细胞的传代培养物或菌株作为体外筛选牙科修复材料的首选细胞。
