3D bioprinted complex skin models to investigate in vitro human simplex virus infection and enable high-throughput therapeutic screenings

Authors

• K Kim
• S Haller
• M Brotman
• Z Laky

Abstract

Herpes simplex virus (HSV) is a highly infectious virus that causes painful sores and affects 3.7billion people. Current treatments manage symptoms but do not cure the disease and have been shown to have adverse effects. There is a critical need for the development of predictive in vitro models of skin that can help investigate the infection pathway of HSV-1 and enable testing of therapeutic interventions. Skin is a complex tissue made up of the dermis, a layer of fibroblasts and extracellular matrix, and the epidermis, an epithelium composed of stratified keratinocytes. To accurately recapitulate skin, we have developed a 3D-bioprinting technique to layer human fibroblasts and keratinocytes in a 96-multi-well plate. The bioprinted tissues are provided various media formulations and exposed to air liquid interface to stratify the epidermis, forming a skin tissue equivalent. We infect the constructs with HSV-1 in two methods: by adding the virus directly to the epidermis or adding the virus to the media. The infection pathway of the HSV-1 in these engineered skin tissues is assessed by confocal imaging and histology. The fibroblasts are transfected with RFP signal and the HSV-1 expresses GFP signal when a cell is infected, allowing the healthy and infected cells to be visualized, and differentiation between fibroblast and keratinocyte infection. Leveraging the fabricated skin, the multi-well plate format, and the fluorescent markers, we are able to test over 700 drugs for potential treatments for HSV-1, determine level of efficacy and cytotoxicity, and observe the virus’ reaction to various drug mechanisms.

Scientific Focus Area: Biomedical Engineering and Biophysics

This page was last updated on Monday, September 25, 2023