Pharmacological Preconditioning of Stem Cells with Dapagliflozin for Diabetic Wound Healing: A Physiology-Based Community KAP Study and Experimental Study in Karachi

Abstract

Background & Objective: Pakistan faces a severe epidemic of Type 2 Diabetes Mellitus, affecting approximately 30.8% of adults, with 15–25% developing diabetic foot ulcers (DFUs) and up to 60% of uncontrolled diabetic cases suffering from impaired, delayed wound healing. The metropolitan population of Karachi bears a disproportionately high burden of non healing diabetic wounds, driven by socio economic challenges, delayed presentation, and chronic metabolic dysregulation.

Methodology: This dual phase investigation integrated a community based Knowledge, Attitudes, and Practices (KAP) survey (Phase I, n = 384 diabetic patients and caregivers in Karachi) with a translationally aligned ex vivo experimental preconditioning trial (Phase II). Human adipose derived stem cells (ADSCs) isolated from diabetic donors were subjected to ex vivo pharmacological preconditioning with dapagliflozin (10 µM) under high glucose (25 mM) stress conditions. Physiological assays evaluated glucose homeostasis, reactive oxygen species (ROS) production, pro inflammatory cytokine expression (TNF α, IL 6), VEGF secretion, tube formation, and cellular migration scratch rates.

Results: Phase I survey revealed widespread gaps in community KAP: while 78.1% of respondents recognized diabetes as a chronic illness, only 28.4% demonstrated adequate knowledge regarding diabetic foot care, and 61.2% engaged in unsafe home wound management practices. Phase II experimental results demonstrated that dapagliflozin preconditioning significantly restored ADSC survival and physiological function despite high glucose oxidative stress. Dapagliflozin preconditioned diabetic ADSCs exhibited a 58.4% reduction in intracellular ROS (p < 0.001), significant down regulation of TNF α and IL 6 (p < 0.001), a 2.4 fold increase in VEGF secretion (p < 0.001), and an accelerated in vitro wound closure rate (88.6 ± 4.2% vs. 41.2 ± 5.1% in unconditioned diabetic ADSCs, p < 0.001).

Conclusion: Pharmacological preconditioning with dapagliflozin rescues diabetic stem cell exhaustion by modulating glucose toxicity, attenuating oxidative stress, suppressing inflammation, and enhancing pro angiogenic capacity. Integrating community level foot care education with novel preconditioned stem cell therapeutics offers a robust physiological strategy to address non healing diabetic ulcers in Karachi.

Keywords: Dapagliflozin; Stem Cell Preconditioning; Diabetic Foot Ulcers; Oxidative Stress; Angiogenesis; KAP Survey; Karachi; Tissue Regeneration.