Djabari Z, Bauza E, Dal Farra C, Domloge N
Vincience Research Center, Sophia Antipolis, France.
Int J Tissue React. 2002;24(4):117-21.
In the search for alternative methods to animal testing, the Hen's egg test on chorioallantoic membrane (HET-CAM) plays a central role in evaluating the innocuity of active ingredients. Therefore, in the following studies we combined the HET-CAM test with histological evaluation in order to increase the sensitivity of evaluation. Twenty active ingredients from four different categories of origin (vegetal, marine, biotechnological and chemical synthetic) were subjected to innocuity evaluation at two different concentrations (pure and 10%). We performed the HET-CAM test and histological evaluation after trypan blue and hematoxylin-eosin staining of the chorioallantoic membrane to microscopically evaluate its state of damage after application of each active ingredient. These studies showed that when the active ingredient was diluted (10%), no discrepancy was seen between the classical HET-CAM evaluation and the histological reading of the chorioallantoic membrane. The histological findings corresponded with the visual observation of the CAM. When the active ingredients were tested at pure concentration, 7 out of 20 tested products demonstrated discrepancy between the two tests. In six cases, the histological examination revealed signs of irritation, such as hyperemia, while visual HET-CAM evaluation was negative. In another case, the histological examination revealed a slight hemorrhage whereas the HET-CAM reading showed only hyperemia. Moreover, the results of trypan blue staining corroborated the histological evaluation of the CAM. These results strongly suggest that the combination of histological and visual HET-CAM tests is of interest for a more sensitive evaluation of the innocuity of cosmetic active ingredients. This additional sensitivity may help to prevent some cases of in vivo intolerance reactions.
在寻找替代动物实验的方法时，鸡胚绒毛尿囊膜试验（HET-CAM）在评估活性成分的无害性方面发挥着核心作用。因此，在接下来的研究中，我们将HET-CAM试验与组织学评估相结合，以提高评估的敏感性。对来自四类不同来源（植物、海洋、生物技术和化学合成）的20种活性成分在两种不同浓度（纯品和10%）下进行无害性评估。在对绒毛尿囊膜进行台盼蓝和苏木精-伊红染色后，我们进行了HET-CAM试验和组织学评估，以在显微镜下评估每种活性成分应用后绒毛尿囊膜的损伤状态。这些研究表明，当活性成分被稀释（10%）时，经典的HET-CAM评估与绒毛尿囊膜的组织学读数之间没有差异。组织学结果与绒毛尿囊膜的视觉观察结果一致。当以纯浓度测试活性成分时，20种受试产品中有7种在两种测试之间存在差异。在6个案例中，组织学检查显示有刺激迹象，如充血，而视觉HET-CAM评估为阴性。在另一个案例中，组织学检查显示有轻微出血，而HET-CAM读数仅显示充血。此外，台盼蓝染色的结果证实了绒毛尿囊膜的组织学评估。这些结果强烈表明，组织学和视觉HET-CAM试验相结合，对于更敏感地评估化妆品活性成分的无害性具有重要意义。这种额外的敏感性可能有助于预防一些体内不耐受反应的情况。
