Abstract
OBJECTIVEHere we report a new method to increase therapeutic potential of mesenchymal stem/stromal cells (MSCs) for ischemic wound healing. We tested biologic effects of MSCs modified with E-selectin, a cell-adhesion molecule capable of inducing post-natal neovascularization, on a translational murine model. SUMMARY BACKGROUND DATATissue loss significantly worsens the risk of extremity amputation for patients with chronic limb-threatening ischemia (CLTI). MSCs-based therapeutics hold major promise for wound healing and therapeutic angiogenesis, but unmodified MSCs demonstrate only modest benefits. METHODSBone marrow cells harvested from FVB/ROSA26SormTmG donor mice were transduced with E-selectin-GFP/AAV-DJ or GFP/AAV-DJ (control). Ischemic wounds were created via a 4 mm punch biopsy in the ipsilateral limb after femoral artery ligation in recipient FVB mice and subsequently injected with PBS or 1×10^6 donor MSCGFP or MSCE-selectin-GFP. Wound closure was monitored daily for 7 postoperative days (POD), and tissues were harvested for molecular and histological analysis and immunofluorescence. Whole-body DiI perfusion and confocal microscopy was utilized to evaluate wound angiogenesis. RESULTSUnmodified MSCs do not express E-selectin, and MSCE-selectin-GFP gain stronger MSC phenotype yet maintain trilineage differentiation and colony-forming capability. MSCE-selectin-GFP therapy accelerates wound healing compared to MSCGFP and PBS treatment. Engrafted MSCE-selectin-GFP manifest stronger survival and viability in wounds at POD 7. Ischemic wounds treated with MSCE-selectin-GFP exhibit more abundant collagen deposition, and enhanced angiogenic response. CONCLUSIONWe establish a novel method to potentiate regenerative and pro-angiogenic capability of MSCs by modification with E-selectin/AAV. This innovative therapy carries potential as a platform worthy of future clinical studies.
