Effect of umbilical cord mesenchymal stem cell-derived mitochondrial transplantation on ischemia-reperfusion injury in a rat model

Abstract

Background: Despite advancements in reconstructive procedures, ischemia-reperfusion (I/R) injury remains a significant challenge in reconstructive surgery, with mitochondrial dysfunction playing a pivotal role. Mitochondrial transplantation has emerged as a promising therapeutic strategy to address this issue. This study aims to evaluate the impact of umbilical cord mesenchymal stem cell-derived mitochondrial transplantation on skin flap I/R models in rats.

Material and methods: Twenty male rats underwent I/R injury on skin flaps, with or without mitochondrial transplantation administered via intravenous or subcutaneous routes. Analysis encompassed histopathology, inflammatory, apoptotic, oxidative stress, and hypoxia markers.

Results: Results revealed a reduction in inflammation, apoptosis, oxidative stress, and hypoxia in the transplantation group compared to controls.

Conclusion: The findings suggest that umbilical cord mesenchymal stem cell-derived mitochondrial transplantation shows promise in enhancing flap viability and attenuating I/R injury, offering valuable insights for improved outcomes in reconstructive surgery. However, further exploration in larger animal models and refinement of delivery methods and dosage are warranted to fully elucidate its clinical translatability.

Keywords: ischemia reperfusion injury; mitochondria; umbilical cord mesenchymal stem cells.

FIGURE 1

Left inferior epigastric flap as a rat I/R model. (A) Preoperative skin marking photograph, (B) Postoperative photograph, (C) Schematic illustration of the experimental model.

FIGURE 2

Study design.

FIGURE 3

Histological analysis of samples stained with Hematoxylin and Eosin. This figure shows the infiltration of inflammatory cells (indicated by asterisks) in the samples. The samples are from the Control group on post‐surgery day 1 (C1), the Control group on post‐surgery day 7 (C7), the Transplantation group on post‐surgery day 1 (T1), and the Transplantation group on post‐surgery day 7 (T7).

FIGURE 4

TNF‐alpha immunohistochemical staining. This figure shows fewer brown‐colored TNF‐alpha positive cells (indicated by asterisks) in the transplantation group compared to the control group, indicating a reduction in inflammation in the transplanted tissue.

FIGURE 5

RT‐qPCR analysis of inflammatory markers. This figure shows the results of an RT‐qPCR analysis, which revealed a statistically significant decrease in the expression of TNF‐alpha, IL‐1β, and IL‐6 within the transplantation group ("I/V before", "I/V after", "S/C before", "S/C after") compared to the control group.

FIGURE 6

Casp3 immunohistochemical staining. A noticeable decrease in the number of positively stained cells (indicated by asterisks) was observed in the transplantation group compared to the control group.

FIGURE 7

RT‐qPCR analysis of Casp3 expression. This figure revealed a significant reduction in Casp3 expression across all transplantation subgroups ("I/V before", "I/V after", "S/C before", "S/C after") compared to the control group.

FIGURE 8

RT‐qPCR analysis of Bcl2 expression. This figure reveals a statistically significant increase in Bcl2 expression across all transplantation groups, as compared to the control group.

FIGURE 9

SOD1 immunohistochemical staining. This figure shows a higher count of brown‐colored positive cells (indicated by asterisks) in the transplantation group compared to the control group.

FIGURE 10

RT‐qPCR analysis of SOD1 expression. This figure demonstrates that the transplantation groups (“I/V before”, “I/V after”, “S/C before”, “S/C after”) exhibited significantly increased SOD1 expression levels compared to the control group.

FIGURE 11

HIF‐1‐alpha immunohistochemical staining: This figure shows a lower count of positive cells in the transplantation group compared to the control group.

FIGURE 12

RT‐qPCR analysis of HIF‐1‐alpha expression. This figure demonstrates a significant decrease in HIF‐1‐alpha levels in all transplantation subgroups (“I/V before”, “I/V after”, “S/C before”, “S/C after”) compared to the control group.