Microengineered diabetic wound-on-a-chip model for emulating chronic wound dynamics.

Abstract

Diabetic wounds, which affect 19-34% of individuals with diabetes mellitus, remain a serious complication due to their chronic, non-healing nature and high risk of limb amputation. Traditional 2D <i>in vitro</i> systems and animal models often fail to replicate the intricate cellular interactions and microenvironmental complexity of human diabetic wounds. To address this gap, a humanized 3D diabetic wound-on-a-chip (DWOC) model was developed to simulate key aspects of diabetic wound pathology under physiologically relevant conditions. This four-channel microfluidic platform integrates human dermal fibroblasts and macrophages within a collagen I matrix to mimic the dermis, alongside endothelial cells embedded in Matrigel to represent the vascular compartment. The system was subjected to hyperglycemic conditions with added advanced glycation end-products (AGEs) and lipopolysaccharide (LPS), alongside normoglycemic controls. Cellular viability, extracellular matrix (ECM) remodeling, myofibroblast differentiation, angiogenesis, and intercellular signaling were assessed using immunofluorescence markers (CD68, α-SMA, CD31, VE-cadherin, SLUG). Cytokine profiling (ELISA, multiplex assays) evaluated inflammatory responses. The DWOC effectively replicated hallmarks of diabetic wound pathology, including impaired ECM remodeling, disrupted dermal-vascular cell crosstalk, defective angiogenesis, and signs of endothelial-to-mesenchymal transition (EndMT) in endothelial cells under diabetic stress. Elevated pro-inflammatory markers (IL-1β, TNF-α, MMP9) and reduced anti-inflammatory/angiogenic factors (IL-10, VEGF-A) reflected the chronic inflammatory and angiogenic imbalance characteristic of non-healing diabetic ulcers. This advanced DWOC platform offers a physiologically relevant, human-specific model for studying diabetic wound healing, highlighting endothelial-to-mesenchymal transition as a critical pathological feature and enabling preclinical evaluation of targeted therapies.