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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

This protocol demonstrates step-by-step details of how to extract the mandibular first molar in the mouse. It provides an alternative method for researchers focusing on jawbone healing and regeneration.

Abstract

This study introduces the development of a molar extraction model in the murine mandible to provide a practicable model for studying alveolar bone regeneration and intramembranous ossification. C57/J6 mice were used to extract the mandibular first molar to establish this model. They were euthanized, and the bilateral mandibles harvested, at 1 week and 4 weeks post-surgery, respectively. Subsequent serial stereoscopic harvest, histological assessment, and immunofluorescence staining were performed to demonstrate successful surgery. Immediately after surgery, the stereoscopic images displayed an empty extraction socket. The hematoxylin and eosin (H&E) at 1 week and Masson staining at 4 weeks post-surgery showed that the area of the original root was partially and fully filled with bone trabeculae, respectively. The immunofluorescence staining showed that, compared with the homeostasis side, the Sp7 expression increased at 1 week post-surgery, suggesting vigorous osteogenesis in the alveolar fossa. All these results demonstrated a practicable murine tooth extraction socket healing model. Upcoming studies revealing the mechanisms of jawbone defect healing or socket healing could adopt this method.

Introduction

Socket healing after tooth extraction is a common clinical scenario, which can result in unbecoming complications such as socket hemorrhage, dry socket, or even jaw osteomyelitis under undesirable healing1,2,3. These comorbidities may impair the quality of life of patients and, even worse, vitally challenge prosthetic rehabilitation due to massive bone loss4. Though the socket healing stages have been elucidated, they are inadequate to direct clinical care post-tooth extraction surgery when encountering various prognosis challenges4

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-2017-041). Adult C57BL/6 mice, obtained from a commercial source (see Table of Materials), were used for the present study.

1. Presurgical preparation

  1. Instrument preparation
    1. Prepare various needles of disposable syringes (26G, 25G, 23G; see Table of Materials) to .......

Representative Results

To elucidate the practical use of this method, the right mandibular first molar of two healthy C57BL/6 mice (3 months old, both female) were extracted and followed for 1 week and 4 weeks, respectively. The undamaged left mandibles were used as healthy controls. Figure 1A shows the specific features of the surgical appliance, including 26-23 G needles and a toothed ophthalmic tweezer. The 26 G needle is pinpoint-removed and bent. The 25 G needle is bent at approximately 25° at the pinpoi.......

Discussion

The murine socket healing model is an important method for unraveling the underlying mechanisms in bone healing and regeneration, ultimately solving clinical challenges. Existing studies have demonstrated the possibility of the incisor extraction model and the maxillary molar extraction model, whereas studies have not used the mandibular first molar model13,17,18. However, incisors are crucial for rodent living, and their impair.......

Acknowledgements

This work is supported by National Natural Science Foundation of China 81825005 (L.Y.), 82201045 (F.Y.), and 82100982 (F.L.), and by Sichuan Province Science and Technology Program 2021JDRC0144 (F.L.), 2022JDRC0130 (F.Y.).

....

Materials

NameCompanyCatalog NumberComments
23/25/26 G needleChengdu Xinjin Shifeng Medical Apparatus & Instruments Co. LTD.SB1-074(IV)
C57/B6J Gempharmatech Experimental Animals Company, Chengdu, ChinaC57/B6J
DAPI Staining Solution Beyotime Cat#C1005
Embedding CassettesCITOTEST Scientific80106-1100-16
Hematoxylin and Eosin Stain KitBiosharpBL700B
IsofluraneRWD Life Science Co.,LtdR510-22-10
Masson’s Trichrome Stain KitSolarbioG1340
Microtome LeicaRM2235
Pentobarbital SodiumHuaxia Chemical Reagent Co., Ltd2018042001
Rabbit polyclonal anti-Sp7 Abcam Cat# ab22552
TweezersChengdu Xinjin Shifeng Medical Apparatus & Instruments Co. LTD.SB2-115

References

  1. Mamoun, J. Dry socket etiology, diagnosis, and clinical treatment techniques. Journal of the Korean Association of Oral and Maxillofacial Surgeons. 44 (2), 52-58 (2018).
  2. Laraki, M., Chbicheb, S., El Wady, W. Alveolitis: review of the literature. ....

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Murine Mandibular Molar ExtractionBone RegenerationSocket Healing ModelAlveolar BoneTooth ExtractionClot FormationTrabecular BoneSp7 ExpressionAnesthesiaAnalgesia

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