Chessboard-like Burn Wound Healing Model of Mice Based on Digital Heating Device

Summary

A streamlined protocol is presented for establishing a burn wound healing model in mice using a digital heating device. The chessboard-like experimental sites created on the skin facilitate further functional analysis for the wound healing assay.

Abstract

Severe burn injuries are among the most traumatic and physically debilitating conditions, impacting nearly every organ system and resulting in considerable morbidity and mortality. Given their complexity and the involvement of multiple organs, various animal models have been created to replicate different facets of burn injury. Methods used to produce burned surfaces vary among experimental animal models. This study describes a simple, cost-effective, and user-friendly mouse burn model for creating consistent full-thickness burns using a digital heating device. The tip of this device was applied to the dorsum of mice for 10 s at 97 °C to establish a chessboard-like burn and examine wound healing under the treatment of an experimental dressing. Skin samples were collected for histological analysis, including Hematoxylin and Eosin (H&E) staining and Masson's staining. Wound healing was assessed through analysis of the wound area and microscopic examination of inflammatory infiltration, re-epithelialization, and granulation tissue formation. The mouse burn injury model can serve as a fundamental tool in studying the pathophysiology of thermal injuries and evaluating therapeutic interventions.

Introduction

Burns are considered one of the critical injuries to the skin, caused by heat exposure, electricity, chemical materials, and radiation exposure^1,2. It can be classified into four degrees depending on the depth of the injury, ranging from the epidermis to the full thickness of the skin, and even the muscles and bones. Small burns can lead to scar formation and increase the risk of infection. A large area of burn injury not only causes local damage, but also stimulates disorders of the body's heart, kidneys, and other organs or systems through severe and long-term inflammation and immun....

Protocol

All animal procedures in this study were reviewed and approved by the Ethical Committee of the West China School of Stomatology, Sichuan University (WCHSIRB-D-2024-499). Twenty-four eight-week-old C57BL/6 mice (female, body weight 25-30 g) were used for the present study. The details of the reagents and the equipment used are listed in the Table of Materials.

1. Preparing equipment and mouse before burn injury

1. Acquire the animals and house them in .......

Discussion

For burn studies, in vitro models typically focus on the inhibitory effects of local antimicrobial agents or antibiotics on bacteria associated with burns, such as Staphylococcus aureus and Pseudomonas aeruginosa¹⁹, as well as the impact of various biomaterials (like elastin, silk, and hydrogel dressings^14,20) on post-burn inflammatory cells (such as neutrophils, macrophages) or stem cells (like mesenchymal stem .......

Materials

Name	Company	Catalog Number	Comments
1 mL pipette tip	KIRGEN,USA	KG1333	Used to locate burn wound sites
3 M Tegaderm film	3M,USA	1624WCN 6 cm x 7 cm	For the wound cover after burn induction
4% paraformaldehyde(PFA)	Biosharp,China	BL539A	Used to fix the skin samples
Buprenorphine	Sigma-Aldrich,USA	PHR8955-50MG	For the pain management of the mice
C57BL/6 mice	Chengdu Dashuo experimental animal company,China	none	For the establishment of burn model
Depilatory cream	Veet,China	—	For the dorsum hair removal of the mice
Digital Heating Device	Shenzhen Kapper Technology Company,China	No.936D	For the burn induction of the mice
Electric shaver	AUX,China	AUX-A5	For the dorsum hair removal of the mice
Filter paper	—	—	Used to unfold of the skin samples
GraphPad software	—	GraphPad prism 9.5.0	For the analysis of burn wound area
Heat-resistant gloves	—	—	Used to hold the digital heating device tip
Hematoxylin and Eosin Stain kit	Solarbio,China	G1120	For the histological analysis of the slides
ImageJ software	—	ImageJ 1.54f	For the analysis of burn wound area
Isoflurane	RWD,China	R510-22-10	For the anesthesia of the mice
Masson's Trichrome Stain Kit	Solarbio,China	G1340	For the histological analysis of the slides
Microscope	Olympus,Japan	VS200 ASW	Used to scan the H&E and Masson stained slides
Tissue cassette	CITOTEST LABWARE MANUFACTURING Co., LTD,China	31050102W	For tissue paraffin dehydration and paraffin embedding