Untersuchungen zur Regulation sekretorischer Aspartatproteinasen in einem oralen Candidose-Modell und in vivo. [Investigations on the regulation of secreted aspartyl proteases in a model of oral candidiasis in vivo]

By means of RT-PCR and specific primers the expression of SAP1-6 and SAP8 was investigated with respect to the time course in an in vitro candidosis model based on reconstituted human mucosal epithelium. Corresponding morphological alterations of the epithelium were documented by light microscopy. The detection of Sap was performed immunoelectron microscopically using a monoclonal antibody. In the oral candidosis model SAP1 and SAP3 transcripts were detected 42 h after inoculation corresponding to first histopathological changes. Additional SAP6 expression was observed six hours later concomitantly with germ-tube formation. Later on SAP2 and SAP8 transcripts were found after 60 h. On protein level it was possible to demonstrate Sap antigens within Candida and markedly deteriorated epithelial cells. Initial experiments with proteinase mutants and proteinase inhibitors showed reduction of histological damage. In a clinical specimen obtained from a twenty nine-year-old female patient suffering from acute oral candidosis SAP1, 3 and 6 could be demonstrated corresponding to the findings in vitro after 48 h. Investigating a clinical specimen obtained from a lesion of chronic oral candidosis in an HIV-infected patient also showed SAP2 expression. On the basis of our results a relationship between the expression on of particular SAP genes and the turn up of lesions looks as probable as a relevant contribution to the in vivo infection

Influence of massage and occlusion on the ex vivo skin penetration of rigid liposomes and invasomes

Liposomes are frequently described as drug delivery systems for dermal and transdermal applications. Recently, it has been shown that particulate substances penetrate effectively into hair follicles and that the follicular penetration depth can be increased by massaging the skin, which simulates the in vivo movement of hairs in the hair follicles. In the present study, massage was applied to skin mounted to Franz diffusion cells. By means of confocal laser scanning microscopy, the influence of massage and occlusion on the follicular penetration depths of rigid and flexible liposomes loaded with a hydrophilic and lipophilic dye was investigated. The application of massage increased follicular penetration significantly. Occlusion resulted in an increased follicular penetration depth only for rigid liposomes, whereas invasomes did not penetrate more effectively if occlusion was applied. The results confirm that massage is an important tool for increasing follicular penetration in ex vivo studies using Franz diffusion cells. Occlusion may reduce the efficacy of follicular penetration depending on the specific liposomal preparation. Rigidity in particular appears to be a relevant paramete