Development of an In Vitro Diabetic Keratinocyte Wound-Relevant Model Incorporating a Mechanical Microenvironment

Abstract Cells in living organisms respond to chemical and mechanical cues from their microenvironment to maintain healthy function. In diabetes, these cues are disrupted, impairing signaling pathways and contributing to complications such as nonhealing skin wounds. Conventional in vitro models using plastic tissue culture plates (TCP) fail to replicate the mechanical interface of human skin, particularly the altered substrate rigidity associated with diabetic skin, and typically expose cells to supraphysiological glucose levels, limiting their physiological relevance. To address these gaps, we developed a diabetic-like wound-relevant model integrating polyacrylamide hydrogels engineered to mimic skin-relevant substrate stiffness with human keratinocytes acclimatized to physiological glucose (NG) and subsequently exposed to diabetic glucose conditions (DG). This tunable cell–material interface enabled assessment of how substrate mechanics and glucose milieu jointly govern keratinocyte viability, proliferation, and migration. The model’s utility was further validated using the standard antidiabetic drug metformin (MET) and the phytochemical curcumin (CUR) to evaluate therapeutic effects on diabetic cell dynamics. Beyond its application in drug evaluation, this work offers design insights for engineering hydrogel substrates with optimized mechanical properties, contributing to materials strategies for improving diabetic wound healing outcomes.