Allogenic Vertebral Body Adherent Mesenchymal Stromal Cells Promote Muscle Recovery in Diabetic Mouse Model of Limb Ischemia

Mackenzie K Madison; Theresa S Doiron; Jennifer Stashevsky; Nancy Zhang; Marlee Yancey; Chang-Hyun Gil; Hanaa Dakour Aridi; Erik J Woods; Michael P Murphy; Steven J Miller

doi:10.1016/j.avsg.2024.08.004

Allogenic Vertebral Body Adherent Mesenchymal Stromal Cells Promote Muscle Recovery in Diabetic Mouse Model of Limb Ischemia

Ann Vasc Surg. 2024 Sep 27;110(Pt A):522-533. doi: 10.1016/j.avsg.2024.08.004. Online ahead of print.

Affiliations

¹ Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA. Electronic address: madisonm@iupui.edu.
² Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA.
³ Ossium Health Inc., Indianapolis, IN, USA.

PMID: 39343376
DOI: 10.1016/j.avsg.2024.08.004

Abstract

Background: Chronic limb-threatening ischemia (CLTI) carries a significant risk for amputation, especially in diabetic patients with poor options for revascularization. Phase I trials have demonstrated efficacy of allogeneic mesenchymal stromal cells (MSC) in treating diabetic CLTI. Vertebral bone-adherent mesenchymal stromal cells (vBA-MSC) are derived from vertebral bodies of deceased organ donors, which offer the distinct advantage of providing a 1,000x greater yield compared to that of living donor bone aspiration. This study describes the effects of intramuscular injection of allogenic vBA-MSC in promoting limb perfusion and muscle recovery in a diabetic CLTI mouse model.

Methods: A CLTI mouse model was created through unilateral ligation of the femoral artery in male polygenic diabetic TALLYHO mice. The treated mice were injected with vBA-MSC into the gracilis muscle of the ischemic limb 7 days post ligation. Gastrocnemius or tibialis muscle was assessed postmortem for fibrosis by collagen staining, capillary density via immunohistochemistry, and mRNA by quantitative real-time polymerase chain reaction (PCR). Laser Doppler perfusion imaging and plantar flexion muscle testing (MT) were performed to quantify changes in limb perfusion and muscle function.

Results: Compared to vehicle (Veh) control, treated mice demonstrated indicators of muscle recovery, including decreased fibrosis, increased perfusion, muscle torque, and angiogenesis. PCR analysis of muscle obtained 7 and 30 days post vBA-MSC injection showed an upregulation in the expression of MyoD1 (P = 0.03) and MyH3 (P = 0.008) mRNA, representing muscle regeneration, vascular endothelial growth factor A (VEGF-A) (P = 0.002; P = 0.004) signifying angiogenesis as well as interleukin (IL-10) (P < 0.001), T regulatory cell marker Foxp3 (P = 0.04), and M2-biased macrophage marker Mrc1 (CD206) (P = 0.02).

Conclusions: These findings indicate human allogeneic vBA-MSC ameliorate ischemic muscle damage and rescue muscle function. These results in a murine model will enable further studies to develop potential therapies for diabetic CLTI patients.