Potentiation effect on accelerating diabetic wound healing using 2-N,6-O-sulfated chitosan-doped PLGA scaffold

Accelerating the wound healing of diabetes-impaired cutaneous tissue is still a challenge due to the aberrant cellular behavior, poor angiogenesis, and pathological micro-environment. Activation with growth factors and modulation of the redox micro-environment of the diabetic wound are considered to be effective strategies. Herein, we have described a highly sulfated heparin-like polysaccharide 2-N, 6-O-sulfated chitosan (26SCS)-doped poly(lactic-co-glycolic acid) scaffold (S-PLGA), which can achieve controlled and sustained release of heparin-binding epidermal growth factor (HB-EGF) owing to its affinity for heparin-binding growth factors. Interestingly, the antioxidant effect of 26SCS was confirmed and it was shown to have a strong scavenging activity towards superoxide radicals, a moderate scavenging activity towards hydroxyl radicals and a lower scavenging activity towards hydrogen peroxide. It also exhibited stronger protective effects in a human keratinocyte cell line (Ha-cat) against H₂O₂-induced oxidative damage. The Ha-cat cells cultured in the presence of the S-PLGA scaffold were significantly protected against oxidative stress during proliferation. In a full thickness excisional wound model of a diabetic rat, the wound treated with the HB-EGF-loaded S-PLGA scaffold was basically healed after 28 days. Conversely, the wounds in the other diabetic groups were not closed. The migration effect of the keratinocytes was enhanced by the 26SCS-induced sustainable release of HB-EGF and the scavenging of ROS which led to rapid re-epithelialization. Furthermore, histopathological evaluation demonstrated the positive effects on wound contraction, epithelial regeneration, and collagen deposition when treated with the HB-EGF loaded S-PLGA scaffold. These findings highlight that 26SCS may serve as a promising coagent for both controlled release of growth factors and alleviation of excessive ROS production, thus leading to increased regeneration of the diabetic wounds.