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Kiara Boodhoo, Mare Vlok, David L Tabb, Kathryn H Myburgh, and Mari van de Vyver

Chronic wounds are a serious and debilitating complication of diabetes. A better understanding of the

dysregulated healing responses following injury will provide insight into the optimal time frame for

therapeutic intervention. In this study a direct comparison was done between the healing dynamics and

the proteome of acute and obese diabetic wounds on day 2 and day 7 following injury. Full thickness

excisional wounds were induced on obese diabetic (B6.Cg-lepob/J, ob/ob, n=14) (blood glucose

423,25±127,92 mg/dL) and wild-type control (C57BL/6J, n=14) (blood glucose 186,67±24,5 mg/dL)

mice. Histological analysis showed no signs of healing in obese DM wounds whereas complete wound

closure/ re-epithelization, the formation of granulation tissue and signs of re-vascularization was evident

in acute wounds on day 7. In obese DM wounds, substance P deficiency and increased MMP-9 activity

on day 2 coincided with increased cytokine/chemokine levels within wound fluid. LC-MS/MS identified

906 proteins, of which 23 (Actn3, Itga6, Epb41, Sncg, Nefm, Rsp18, Rsp19, Rpl22, Macroh2a1, Rpn1,

Ppib, Snrnp70, Ddx5, Eif3g, Tpt1, FABP5, Cavin1, Stfa1, Stfa3, Cycs, Tkt, Mb, Chmp2a) were

differentially expressed in wounded tissue on day 2 (p<0.05; >2 fold) and 6 (Cfd, Ptms, Hp, Hmga1,

Cbx3, Syap1) (p<0.05; >2 fold) on day 7. A large number of dysregulated proteins on day 2 was

associated with an inability to progress into the proliferative stage of healing and suggest that early

intervention might be pivotal for effective healing outcomes. The proteomic approach highlighted the

complexity of obese DM wounds in which the dysregulation involves multiple regulatory pathways and

biological processes.