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Updated: Dec 27, 2025
In Vivo Imaging of Reactive Oxygen Species in a Murine Wound Model
Published on: November 17, 2018
Methylene blue-loaded niosome: preparation, physicochemical characterization, and in vivo wound healing assessment
Ali Farmoudeh1, Jafar Akbari2, Majid Saeedi1
1Department of Pharmaceutics, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran.
Abstract:
Following skin injury, the overproduction of reactive oxygen species (ROS) during the inflammatory phase can cause tissue damage and delay in wound healing. Methylene blue (MB) decreases mitochondrial ROS production and has antioxidant effects. The authors aimed to prepare MB-loaded niosomes using the ultra-sonication technique as a green formulation method. A Box-Behnken design was selected to optimize formulation variables. The emulsifier to cholesterol ratio, HLB of mixed surfactants (Span 60 and Tween 60), and sonication time were selected as independent variables. Vesicle size, zeta potential (ZP), and drug entrapment capacity percentage were studied as dependent variables. The optimized formulation of niosomes showed spherical shape with optimum vesicle size of 147.8 nm, ZP of - 18.0 and entrapment efficiency of 63.27%. FTIR study showed no observable interaction between MB and other ingredients. In vivo efficacy of optimized formulation was evaluated using an excision wound model in male Wistar rat. Superoxide dismutase (SOD, an endogenous antioxidant) and malondialdehyde (MDA, an end product of lipid peroxidation) levels in skin tissue samples were evaluated. After 3 days, MDA was significantly decreased in niosomal gel-treated group, whereas SOD level was increased. Histological results indicate rats that received niosomal MB were treated effectively faster than other ones. Graphical abstract.
Insights
Methylene blue-loaded niosomes were developed using green sonication methods to enhance wound healing. Optimized formulations significantly reduced lipid peroxidation and increased antioxidant levels in rats, promoting faster tissue repair.
Area of Science:
- Materials Science
- Biomedical Engineering
- Pharmacology
Background:
- Skin injury triggers reactive oxygen species (ROS) overproduction, impairing wound healing.
- Methylene blue (MB) exhibits antioxidant properties by reducing mitochondrial ROS.
- Niosomes offer a promising drug delivery system for enhanced therapeutic efficacy.
Purpose of the Study:
- To formulate and optimize methylene blue (MB)-loaded niosomes using a green ultrasonication technique.
- To evaluate the physicochemical properties and in vivo wound healing efficacy of the optimized MB-loaded niosomes.
Main Methods:
- Box-Behnken design for optimizing niosome formulation variables (emulsifier:cholesterol ratio, HLB, sonication time).
- Characterization of niosomes for vesicle size, zeta potential (ZP), and entrapment efficiency.
- Fourier-transform infrared (FTIR) spectroscopy to assess drug-excipient compatibility.
- In vivo evaluation using an excision wound model in rats, measuring malondialdehyde (MDA) and superoxide dismutase (SOD) levels, and histological analysis.
Main Results:
- Optimized niosomes exhibited a spherical shape, with a vesicle size of 147.8 nm, ZP of -18.0, and 63.27% entrapment efficiency.
- FTIR studies confirmed no interaction between MB and niosome components.
- In vivo studies showed a significant decrease in MDA and increase in SOD levels in MB-niosome treated rats after 3 days.
- Histological examination indicated accelerated wound healing in the MB-niosome group.
Conclusions:
- Green ultrasonication is an effective method for preparing MB-loaded niosomes.
- Optimized MB-niosomes demonstrate significant antioxidant and pro-healing effects in a rat wound model.
- Niosomal MB holds potential as a therapeutic agent for accelerating skin wound healing.
