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Novel Autologous Cell Based Therapy to Promote Wound Healing
Diego M. Castilla*, Zhao-Jun Liu*, Runxia Tian*, Yan Li*, Alan S. Livingstone, Omaida C. Velazquez*
University of Miami, Miami, FL

Objective(s): The development of efficacious cell based therapies (CBT) for non-healing diabetic wounds continues at the forefront of intense investigation. Our prior studies demonstrate a role for SDF-1α in wound healing, as there is a significant down regulation of this cytokine in diabetic cutaneous wounds. We now study the efficacy of a novel CBT consisting of local wound inoculation with autologous bone marrow cells (BMC) that are activated ex vivo with recombinant SDF-1α.
Methods: Unfractionated BMC from diabetic Leptin receptor-deficient (db/db) mice were incubated for 20h with SDF-1α (100ng/mL) or bovine serum albumin (control). Pre-treated BMC (1x107) were injected subcutaneously into full thickness skin wounds in db/db mice (n=8/group). Wound closure rates, capillary density and endothelial progenitor cell (EPC) numbers were assessed with serial photography, Dil perfusion, laser confocal microscopy and immunohistochemistry. Expression of pro-healing molecular targets was evaluated by PCR. Serum SDF-1α levels were measured with ELISA.
Results: SDF-1α pre-treated BMC significantly promoted wound healing (76% Vs. 42% closure by day 12; 1.8 fold difference starting at day 9; p<.0001), neovascularization (p=.0028) and EPC recruitment (p=.0059). Gene expression studies demonstrate up-regulation of EphR B4 and Plasminogen as downstream targets potentially mediating the pro-healing response. Ex vivo BMC activation did not affect systemic SDF-1α levels.
Conclusion: We report a novel CBT (autologous BMC with ex vivo SDF-1α activation) that is highly effective in promoting healing in a murine model of Type 2 Diabetes. Furthermore, we identify new molecular targets that may be important for advancing the field of wound healing.

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