A Modified Tooth Extraction Approach for Improving Procedural Efficiency and Consistency in a Murine Mandibular Tooth Socket Healing Model

Summary

A modified approach for extracting the first mandibular molar in mice is described and compared with the conventional approach to evaluate procedural efficiency and reproducibility. The modified approach improves procedural consistency and reduces technical challenges in establishing a murine mandibular tooth socket healing model.

Abstract

The tooth socket healing model (TSHM) is a widely used animal model for investigating the molecular mechanisms underlying bone regeneration. Although the murine mandibular TSHM is valuable for studying mandibular-specific pathological processes, establishing this model remains technically challenging because of the anatomical and biomechanical characteristics of the mandibular molar region. This protocol describes a modified approach for mandibular first molar extraction in mice and compares its performance with the conventional technique to improve procedural efficiency and reproducibility. Under controlled experimental conditions, the modified approach significantly reduced the average extraction time from 29 to 10 min. In addition, the modified group exhibited fewer extraction-related complications, including root and alveolar bone fractures, indicating improved procedural consistency. Micro-computed tomography (μCT) analysis further demonstrated improved preservation of extraction socket architecture during early healing. These findings indicate that the modified extraction approach improves procedural efficiency and reproducibility while reducing the technical challenges associated with murine mandibular tooth extraction. This protocol may facilitate more consistent establishment of murine mandibular tooth socket healing models, particularly for researchers with limited microsurgical experience.

Introduction

The tooth socket healing model (TSHM) is a classic model for studying bone regeneration¹. This model creates a challenging yet subcritical injury within the alveolar bone, enabling favorable natural healing without additional intervention¹. Mice are widely used as model organisms because of their genetic background, similarities in bone metabolism to humans, relatively low breeding costs, and ease of maintenance². Over the past few decades, rapid advances in gene-editing technology have made mice indispensable for investigating the molecular mechanisms underlying bone regeneration

Protocol

All procedures were approved by the Ethical Committee of the West China School of Stomatology, Sichuan University (WCHSIRB-D-2025-188), and were performed in accordance with institutional animal-care guidelines.

NOTE: Use 8-week-old male C57BL/6 mice weighing 22–29 g. Maintain the animals on a standard chow diet with water provided ad libitum under standard housing conditions (21°C–24°C, 40%–60% humidity, and a 12-h light-dark cycle). Use the materials and instruments listed in the Table of Materials. Use 24 mice for the comparative evaluation and presentation of failed cases. Use additional mice for operator training before the comparative....

Results

The anatomical features relevant to mandibular first molar extraction are illustrated in Figure 1, and the conventional and modified extraction approaches are shown schematically in Figure 2A,B. Representative images of the prepared needle and toothed ophthalmic tweezers, the fresh extraction sockets, and the extracted mandibular first molar are shown in Figure 2C–G. The complete procedural workflow.......

Discussion

Successful establishment of the TSHM requires complete tooth removal while preserving the surrounding alveolar architecture, as extraction-related injury may compromise subsequent bone regeneration¹⁶. Therefore, the reproducibility of the extraction procedure represents a critical determinant of model reliability¹⁷. In this study, a modified forceps-based mandibular first molar extraction approach was systematically compared with a conventional lever-based technique under s.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
25 G syringe needles	Beyotime	FS802-30pcs	Disposable syringe needles used for the mouth retractor and optional periodontal ligament pre-loosening
4% paraformaldehyde solution	Solarbio	P1110	4% paraformaldehyde solution used for specimen fixation
75% ethanol	Ouse Medical Devices Store	N/A	75% medical-grade ethanol used for surface disinfection
C57BL/6 mice	Charles River	213	8-week-old male C57BL/6 mice weighing 22–29 g
Carprofen	Solarbio	C5350	Postoperative analgesic administered subcutaneously at 5 mg/kg
Cotton balls	Ouning Medical Devices	N/A	Sterile cotton balls used for oral cleaning and hemostasis
CTAn v1.18.4.0+	SkyScan	N/A	Software used for μCT image analysis
DataViewer v1.5.6.2	SkyScan	N/A	Software used for viewing and exporting reconstructed μCT images
Finger sleeves	LeSu Office	MEKU-1/2/3B	Finger sleeves used for thumb protection during extraction
Foam board	Dongguan Lijianglong Industrial Co., Ltd.	N/A	Foam board used as the operating and fixation platform
Gel diet	ReadyDietech	J10001	Postoperative diet provided on the cage floor for 3 days, when used
GraphPad Prism v10.1.2	GraphPad	RRID: SCR_002798	Software used for statistical analysis and graph generation
Headlamp	Bazhou Pengen Protective Equipment Factory	N/A	Headlamp used to illuminate the surgical field
Heating pad	Shijiazhuang Jianuan Electrical Appliances Co., Ltd.	N/A	Heating pad preheated to approximately 38 °C for postoperative recovery
Hydroxyapatite phantom	QRM	QRM-70127	Calibration phantom used to convert CT attenuation values into bone mineral density
Magnification device	Olympus Corporation	SZX10	Device used to inspect the toothed tweezer tips and extraction socket under magnification
Mask	Senlun Medical Devices Specialty Store	N/A	Surgical mask
Micro-computed tomography scanner	Bruker SkyScan	SkyScan 1276	μCT scanner used at 80 kV, 500 μA, and 10 μm voxel resolution
Mimics Research v21.0	Materialise	RRID: SCR_015802	Software used for three-dimensional reconstruction
Pentobarbital sodium	Sigma-Aldrich	P3761	Pentobarbital sodium salt used as a 1% anesthetic solution
Phosphate-buffered saline solution	Solarbio	P1010	0.01 M phosphate-buffered saline powder, pH 7.2–7.4
Povidone-iodine antiseptic solution	Belkon Pharmacy Flagship Store	N/A	Povidone-iodine solution used for abdominal disinfection
Rubber bands	Foshan Puli Rubber Products Factory	N/A	Rubber bands used to assemble the mouth retractor
Saline solution	Thermo Fisher	BR0053G	Saline tablets used to prepare the solution for oral cleaning
Standard chow diet	Jiangsu Xietong Pharmaceutical and Biotechnology Engineering Co., Ltd.	XTC01WC-001	Standard chow used for routine housing and softened for postoperative feeding
Steam sterilizer	Institutional facility	N/A	Equipment used to sterilize the toothed tweezers and saline solution
Sterile cotton swabs	Kangbailai Medical Devices Store	N/A	Single-use sterile cotton swabs
Sterile gloves	Senlun Medical Devices Specialty Store	N/A	Sterile latex gloves
Sterile non-woven fabric	Shandong Xinhua Infection Control Supplies Hangzhou Store	N/A	Sterile non-woven fabric used to cover the operating surface and heating pad
Sterile surgical cap	Senlun Medical Devices Specialty Store	N/A	Sterile surgical cap
Surgical scrubs	Senlun Medical Devices Specialty Store	N/A	Clean surgical scrubs worn during the procedure
Tape	Thermo Fisher	15947	Adhesive tape used to secure the mouse limbs
Toothed ophthalmic tweezers	Beyotime	FS229	Toothed ophthalmic forceps used as dental forceps
Veterinary ophthalmic ointment	Dechra Veterinary Products	143-16	Ophthalmic ointment applied after anesthesia to prevent corneal drying