Three-dimensional human skin model infected with Staphylococcus aureus as a tool for evaluation of bioactivity and biocompatibility of antiseptics

Abstract

In the light of pandemic spreads of multi-drug-resistant micro-organisms, alternative antimicrobial strategies to the use of antibiotics are the focus of research attention. As a prerequisite for medical application, the aim of this study was to develop a three-dimensional full skin infection model to evaluate the bioactivity and biocompatibility of antiseptics in application-relevant concentrations. A three-dimensional (3D) full skin model consisting of collagen-embedded fibroblasts as dermis and a fully differentiated epidermis built from keratinocytes was infected with Staphylococcus aureus. Infected skin models were treated for 24 h with the antiseptics polihexanide, octenidine dihydrochloride, chlorhexidine digluconate and povidone-iodine. Infection resulted in detrimental effects, a strong immune response with increased secretion of lactate dehydrogenase and pro-inflammatory cytokines, and increased gene expression of pro-inflammatory cytokines and antimicrobial peptides after 24 h. Application of antiseptics protected the skin models from damage due to S. aureus infection while demonstrating good biocompatibility. The best ratio of bioactivity to biocompatibility was observed for polihexanide. Polihexanide also enhanced the innate immune response by increasing the gene expression levels of antimicrobial peptides such as human β-defensin 2, human β-defensin 3, psoriasin and ribonuclease 7. The developed model provides an excellent tool to investigate the response of human cells to microbial infections in a complex 3D structure. Furthermore, the infection model is appropriate for evaluation of bioactivity and biocompatibility of antiseptics. As such, the model presented in this study is a promising approach to evaluate the mechanisms and effectiveness of new antimicrobial strategies.