17-P031 Gradual cell specification during formation of primitive endoderm and epiblast

wound site plays an essential role in wound healing as these can contribute directly to vasculogenesis.This process is dysregulated in diabetic wounds. It has been shown that injury-induced Hoxa3 expression is impaired in diabetic mice and this affects the recruitment of cells to the wound. Restoration of Hox transcription factor results in a decrease in inflammatory cell (CD45+) and an increase in endothelial progenitor cell (EPC) (CD34+) recruitment to the wound, which promotes quicker more efficient healing. Further analysis by our lab suggests that Hoxa3 inhibits several inflammatory cascades including the NFjB pathway, in which the expression of Myd88 and Tollip, two upstream regulators of NFjB, was repressed in response to enforced expression of Hoxa3. Therefore it is thought that reduced expression of Hoxa3 in diabetic mice contributes to the exacerbated inflammatory response in these animals. We are currently investigating whether over-activation of NFjB is associated with inhibition of EPC mobilisation, recruitment and differentiation during injury induce neovascularisation. Future data will compare the expression of members of the NFjB pathway in diabetic versus non-diabetic mice by immunohistochemistry and Western blot analysis, allowing us to assess differences in the activation of various members of the NFjB pathway. Gel shift assays will allow us to assess the activation status of NFjB in response to injury in non-diabetic and diabetic mice. We will then determine if there is an association of this pro-inflammatory pathway with the dysregulation of BMDC recruitment in the diabetic wound.