The development of a cell-based diabetic wound bioassay

Chronic wounds, such as diabetic foot ulcers, result in impaired wound healing in 3-5% of the population over the age of 65. However, despite the increasing financial burden of these diabetic wounds there is, at present, no suitable diabetic chronic wound animal model. Therefore, in light of the concerns and limitations of animal models and human testing, this NC3Rs Grant application seeks to develop a simple, reproducible, in vitro diabetic wound model system. This will permit rapid, low cost testing of materials, reagents and drugs in order to reduce unnecessary animal experimentation. We have already demonstrated that venous leg ulcer derived fibroblasts are phenotypically distinct from patient-matched normal fibroblasts (NF) and are currently, as part of an ongoing project, characterising the phenotype of diabetic fibroblasts (DF). As part of this ongoing project we are immortalising both the NF and DF (by retroviral insertion of the human telomerase) to create stable, disease-specific cell lines. The aim of this NC3Rs Grant application will be initially to investigate the differences in the global, genetic profiles of these cells utilising gene microarrays. Confirmation of these alterations by other methodologies will lead to the identification of diabetic wound disease marker genes. The disease marker genes identified will be used to develop a well controlled, cell-based reporter system, enabling automated testing and pre-screening of reagents which may ameliorate the diabetic wound disease state. It is anticipated that future projects leading from the present application will develop a commercial in vitro bioassay and a high-throughput screening system, which will reduce the amount of unnecessary animal studies undertaken with respect to wound product/materials testing. We believe that the development of such an in vitro diabetic wound model will represent an important and unique resource for wound healing researchers worldwide.