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Updated: Jul 16, 2025
Protocol to Create Chronic Wounds in Diabetic Mice
Published on: September 25, 2019
Evaluation of topical methylene blue nanoemulsion for wound healing in diabetic mice
Yu Gao1, Zhounan Jiang2, Bin Xu3
1Nanjing Drum Tower Hospital Clinical College of Nanjing Medical University, Nanjing, China.
Context:
Diabetic wounds (DW) are a complication of diabetes and slow wound healing is the main manifestation. Methylene blue (MB) has been shown to exhibit therapeutic effects on diabetes-related diseases.
Objective:
To investigate the mechanisms of action of MB-nanoemulsion (NE) in the treatment of DW.
Materials And Methods:
The concentration of MB-NE used in the in vivo and in vitro experiments was 0.1 mg/mL. Streptozocin-induced diabetic mice were used as models. The mice were separated into nondiabetic, diabetic, MB-NE treated, and NE-treated groups. Intervention of high glucose-induced human umbilical vein endothelial cells using MB-NE. The mechanism by which MB-NE promotes DW healing is investigated by combining histological analysis, immunofluorescence analysis, TUNEL and ROS assays and western blotting.
Results:
In diabetic mice, the MB-NE accelerated DW healing (p < 0.05), promoted the expression of endothelial cell markers (α-SMA, CD31 and VEGF) (p < 0.05), and reduced TUNEL levels. In vitro, MB accelerated the migration rate of cells (p < 0.05); promoted the expression of CD31, VEGF, anti-apoptotic protein Bcl2 (p < 0.05) and decreased the expression of the pro-apoptotic proteins cleaved caspase-3 and Bax (p < 0.05). MB upregulated the expression of Nrf2, catalase, HO-1 and SOD2 (p < 0.05). In addition, MB reduced the immunofluorescence intensity of TUNEL and ROS in cells and reduced apoptosis. The therapeutic effect of MB was attenuated after treatment with an Nrf2 inhibitor (ML385).
Discussion And Conclusion:
This study provides a foundation for the application of MB-NE in the treatment of DW.
