Institute for Clinical & Experimental Surgery, Saarland University, Homburg/Saar, Germany.
INM-Leibniz Institute for New Materials, Saarbrücken, Germany.
Exp Dermatol. 2019 Aug;28(8):940-947. doi: 10.1111/exd.13983. Epub 2019 Jul 4.
In this study, we introduce the mouse dorsal skinfold chamber model as a valuable approach for the in vivo evaluation of topical formulations. For this purpose, dorsal skinfold chambers were implanted into BALB/c mice. Tumor necrosis factor (TNF)-α was administered to the chamber tissue for the local induction of inflammation followed by the application of diclofenac-containing or diclofenac-free (control) gel onto the skin of the chamber backside. Intravital fluorescence microscopy was repetitively performed throughout an observation period of 24 hours to study macromolecular leakage, leucocyte-endothelial cell interactions and microhaemodynamic parameters. In addition, infiltration of the inflamed tissue with different immune cell subtypes was assessed by immunohistochemistry. In a second set of experiments, the effect of dermal application of a diclofenac-containing gel on photochemically induced thrombus formation was analysed. It was observed that macromolecular leakage, numbers of adherent leucocytes and tissue infiltrating myeloperoxidase (MPO)-positive neutrophilic granulocytes and CD68-positive macrophages were significantly reduced in dorsal skinfold chambers treated with diclofenac-containing gel when compared to controls. Moreover, the diclofenac-containing gel exerted an anti-thrombotic activity, as indicated by a significantly prolonged complete vessel occlusion time. These findings demonstrate that the mouse dorsal skinfold chamber represents a valid and versatile tool to evaluate the effects of topical formulations in vivo.
在这项研究中，我们引入了小鼠背部皮褶室模型，作为评估局部制剂的体内评估的一种有价值的方法。为此，将背部皮褶室植入 BALB/c 小鼠体内。向室组织中给予肿瘤坏死因子 (TNF)-α，以局部诱导炎症，然后将含有双氯芬酸或不含双氯芬酸的（对照）凝胶（游离型）应用于室背面的皮肤。通过活体荧光显微镜在 24 小时的观察期内重复进行，以研究大分子渗漏、白细胞-内皮细胞相互作用和微血管血流动力学参数。此外，通过免疫组织化学评估炎症组织中不同免疫细胞亚型的浸润情况。在第二组实验中，分析了局部应用含有双氯芬酸的凝胶对光化学诱导血栓形成的影响。观察到与对照组相比，用含有双氯芬酸的凝胶处理的背部皮褶室中的大分子渗漏、黏附的白细胞数量以及组织浸润的髓过氧化物酶 (MPO)阳性中性粒细胞和 CD68 阳性巨噬细胞明显减少。此外，含有双氯芬酸的凝胶具有抗血栓形成活性，因为完全血管闭塞时间显著延长。这些发现表明，小鼠背部皮褶室是一种有效的、多功能的工具，可用于体内评估局部制剂的效果。
