The main purpose of this project is to develop a skin equivalent model that incorporates microvasculature, which will then be used to address several important questions in the area of skin ageing. Researchers have for several decades used translational assays to mimic different aspects of human skin in a 3-D culture system. These skin equivalent models replicate the dermis and epidermis, providing a way to investigate signalling mechanisms between these cell types. However, a significant drawback is that they lack a blood supply. Thus, they are unable to examine inflammatory changes occurring within the skin during natural processes such as ageing, or in pathology such as wound healing.
The project will first incorporate endothelial cells into tissue engineered vessels, which will subsequently be integrated into skin equivalents to address questions on skin ageing. We are aiming for a skin equivalent culture system that can be maintained by perfusion via the vessels, which we expect take 12-18 months. Effectiveness of the assay will be demonstrated by histological assessment for skin viability (e.g. H&E morphology; IHC for proliferation and apoptosis), but also assessing vessel integrity and the extent angiogenesis that occurs (e.g. CD31, aSMA).
The assay will then be used to address important biological questions related to skin health and ageing.
By passing inflammatory cells such as neutrophils through the vasculature their migration through the skin can be followed, to examine how this influences skin behaviour. Intrinsic skin ageing is associated with a chronic elevation of inflammatory markers, so the expression of cytokines (e.g. TNFa, IL6) in the skin will be examined. Differences occurring when inflammatory cells with an aged profile are added will help us understand skin ageing more clearly.
Key techniques
Electrospinning, tissue culture, histology and immunohistochemistry, image analysis, RT-qPCR.
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