Assessment of Safety and Efficacy of Cellular Therapy on Foot Ulcer in Diabetic Animal: A Preclinical Investigation

Aim and Objectives

Around 20% of diabetic patients are at increased risk of chronic foot ulcers. Current treatments mainly focus on glycemic control and supportive care, with limited effectiveness. Mononuclear Cell (MNC) therapy, containing hematopoietic, mesenchymal, and progenitor cells, is emerging as a novel and effective regenerative approach for Diabetic Foot Ulcers (DFU). The objective of the study is to evaluate the safety profile and healing efficacy of MNC-based therapy in a diabetic rat model of non-healing wounds.

Materials and Methods

36- male Wistar rats (150–200 g) were randomized into different groups (n=6): Normal Healthy Control, Diabetic Control (STZ + Ulcer control), Mupirocin, Insulin + Mupirocin, MNCs (~10 × 10⁶ cells), and MNCs (~10 × 10⁶ cells) + Insulin. A diabetic model was induced in experimental rats using STZ (50 mg/kg, i.p.). From each diabetic rat, 1.5 mL of blood was collected to isolate MNCs, which were resuspended in 0.5 mL of autologous plasma and injected once at the ulcer site. Insulin groups received 4 IU/day (s.c.). The end points of our study evaluated various parameters, including wound closure, serum insulin, blood glucose, lymphocyte/NLR profiles, inflammatory markers (TNF-α, IL-6), and histology.

Results

The combination of insulin and MNC-based therapy was well tolerated and resulted in a significant reduction in hyperglycemia, reduced inflammation, and enhanced the wound closure rate. This combinatorial approach facilitated tissue homeostasis and aided in the remodeling of cells for functional restoration. MNC therapy combined with insulin resulted in a marked glycemic reduction (p<0.001) and pro-inflammatory cytokines TNF-α and IL-6 levels were significantly attenuated (p<0.001) compared to diabetic controls. The combination therapy enhanced wound closure (~95% by day 14) relative to MNCs alone (~75%) and insulin + mupirocin (~80%) (p<0.01). Lymphocyte sets and the NL ratio were also restored, indicating immune stabilization. Histological analysis revealed increased collagen deposition, angiogenesis, and re-epithelialization in the MNC + insulin group.

Conclusion

MNC-based therapy can serve as a promising candidate for treating diabetic patients with non-healing ulcers.