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 the light of the concerns and limitations of animal models and human testing, this NC3Rs Grant application will develop a stable, 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 (CWF) and diabetic foot ulcer fibroblasts (DF) are phenotypically distinct from patient-matched normal fibroblasts (NF). We have also immortalised both the NF and DF (by retroviral insertion of the human telomerase) to create stable, disease-specific cell lines and within these cells have identified disease specific marker genes by microarray analysis. The aim of this NC3Rs grant application will be to extend these initial studies/findings and, using a virus integration approach, stably transduce our immortalised disease cells with fluorescent disease marker gene reporter constructs. This will give us a robust, cell-based reporter system, enabling automated testing and pre-screening of reagents which may ameliorate the diabetic wound disease state. It is anticipated that at the end of the funding we will be close to commercialising the bioassay and developing 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.
Caley M et al. (2018). Development and Characterisation of a Human Chronic Skin Wound Cell Line-Towards an Alternative for Animal Experimentation. International Journal of Molecular Sciences 19(4). doi: 10.3390/ijms19041001
Roper JA et al. (2015). Ultrasonic Stimulation of Mouse Skin Reverses the Healing Delays in Diabetes and Aging by Activation of Rac1. The Journal of Investigative Dermatology 135(11):2842-51. doi: 10.1038/jid.2015.224
Peake MA et al. (2014). Identification of a transcriptional signature for the wound healing continuum. Wound Repair and Regeneration 22(3):399-405. doi: 10.1111/wrr.12170
Stephens P (2010). Development of a cell-based diabetic wound assay. Alternatives to Laboratory Animals 38(supplement 1):45-48. doi: 10.1177/026119291003801S15
Principal investigatorProfessor Phil Stephens
Co-InvestigatorProfessor David Kipling
Professor David William Thomas