Engineered cell-laden hemostatic hydrogels for all-round rapid repair of diabetic wounds

Abstract

The complex microenvironment of diabetic wounds, characterized by persistent inflammation, impaired angiogenesis, and compromised hemostasis, severely delays healing and presents a significant clinical challenge. Current therapies often lack the multi-faceted approach required to address these interwoven pathologies effectively. Herein, we present a novel cell-based therapeutic platform: precision-engineered human embryonic kidney (HEK 293T) cells encapsulated within alginate hydrogels, designed for diabetic wound repair and liver hemostasis. HEK 293T cells are stably engineered via a PiggyBac transposon system to co-express pro-angiogenic human Vascular Endothelial Growth Factor A (hVEGFA), hemostatic Factor VIII (hFVIII), and immunomodulatory Interleukin-10 (hIL10), ensuring sustained secretion of these therapeutic factors. In vitro studies confirmed hVEGFA-mediated elevation of cell migration and wound closure, hFVIII stimulated acceleration of blood clotting, and hIL10's role as a potent anti-inflammation reducing oxidative stress. A combination of hVEGFA, hFVIII, and hIL10 notably achieved superior in vitro and in vivo wound healing. In a diabetic rat full-thickness excisional wound model, administration of these engineered cell-laden hydrogels significantly accelerated wound closure within 2 weeks. Furthermore, cell-secreted hFVIII demonstrated remarkable hemostatic efficacy when mixed with human blood ex vivo and in a liver puncture model, reducing activated partial thromboplastin time, blood loss, and clotting time in vivo. On the other hand, capsules did not show any xenogeneic effect and immunogenicity in the long term. This multi-functional cell-based hydrogel system offers a comprehensive and promising platform for rapid attenuation of homeostasis for rapid diabetic wound healing.