Chatterjee Abhijit, Babu R Jayachandra, Klausner M, Singh Mandip
College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL 32307, USA.
Toxicol Lett. 2006 Dec 1;167(2):85-94. doi: 10.1016/j.toxlet.2006.08.017. Epub 2006 Sep 10.
The purpose of this study was to evaluate an in vitro EpiDerm human skin model (EPI-200) to study the irritation potential of jet fuels (JP-8 and JP-8+100). Parallel in vivo studies on hairless rats on the dermal irritancy of jet fuels were also conducted. Cytokines are an important part of an irritation and inflammatory cascade, which are expressed in upon dermal exposures of irritant chemicals even when there are no obvious visible marks of irritation on the skin. We have chosen two primary cytokines (IL-1alpha and TNF-1alpha) as markers of irritation response of jet fuels. Initially, the EPI-200 was treated with different quantities of JP-8 and JP-8+100 to determine quantities which did not cause significant cytotoxicity, as monitored using the MTT assay and paraffin embedded histological cross-sections. Volumes of 2.5-50 microl/tissue (approximately 4.0-78 microl/cm2) of JP-8 and JP-8+100 showed a dose dependent loss of tissue viability and morphological alterations of the tissue. At a quantity of 1.25 microl/tissue (approximately 2.0 microl/cm2), no significant change in tissue viability or morphology was observed for exposure time extending to 48 h. Nonetheless, this dose induced significant increase in IL-1alpha and TNF-alpha release versus non-treated controls after 24 and 48 h. In addition, IL-1alpha release for JP-8+100 was significantly higher than that observed for JP-8, but TNF-alpha release after 48 h exposure to these two jet fuels was the same. These findings parallel in vivo studies on hairless rats, which indicated higher irritation levels due to JP-8+100 versus JP-8. In vivo, transepidermal water loss (TEWL) and IL-1alpha expression levels followed the order JP-8+100 > JP-8 > control. Further, in vivo TNF-alpha levels for JP-8 and JP-8+100 were also elevated but not significantly different from one another. In aggregate, these findings indicate that EPI-200 tissue model can be utilized as an alternative to the use of animals in evaluating dermal irritation.
本研究的目的是评估一种体外表皮人皮肤模型（EPI - 200），以研究喷气燃料（JP - 8和JP - 8 + 100）的刺激潜力。同时也对无毛大鼠进行了关于喷气燃料皮肤刺激性的体内平行研究。细胞因子是刺激和炎症级联反应的重要组成部分，即使在皮肤上没有明显可见的刺激痕迹时，在皮肤接触刺激性化学物质后也会表达。我们选择了两种主要细胞因子（IL - 1α和TNF - 1α）作为喷气燃料刺激反应的标志物。最初，用不同量的JP - 8和JP - 8 + 100处理EPI - 200，以确定不会引起明显细胞毒性的量，这通过MTT法和石蜡包埋组织学横截面进行监测。2.5 - 50微升/组织（约4.0 - 78微升/平方厘米）的JP - 8和JP - 8 + 100显示出剂量依赖性的组织活力丧失和组织形态改变。在1.25微升/组织（约2.0微升/平方厘米）的量时，暴露时间延长至48小时，未观察到组织活力或形态有显著变化。尽管如此，与未处理的对照相比，该剂量在24小时和48小时后诱导IL - 1α和TNF - α释放显著增加。此外，JP - 8 + 100的IL - 1α释放显著高于JP - 8，但在暴露于这两种喷气燃料48小时后，TNF - α释放相同。这些发现与无毛大鼠的体内研究结果一致，表明JP - 8 + 100比JP - 8具有更高的刺激水平。在体内，经表皮水分流失（TEWL）和IL - 1α表达水平遵循JP - 8 + 100 > JP - 8 >对照的顺序。此外，JP - 8和JP - 8 + 100的体内TNF - α水平也升高，但彼此之间无显著差异。总体而言，这些发现表明EPI - 200组织模型可作为评估皮肤刺激性时替代动物使用的方法。
