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In This Article

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

Summary

This article outlines a step-by-step procedure for establishing a mice model with an oronasal fistula. The oronasal fistula was created by employing heated ophthalmologic cautery to damage the midline portion of the hard palate, resulting in the formation of an opening between the oral and nasal cavities.

Abstract

This study presents a method utilizing heated ophthalmologic cautery to develop a viable model for investigating oronasal fistulas. C57BL/6 mice were used to establish the oronasal fistula (ONF) model. To create the ONF, the mice were anesthetized, immobilized, and their hard palates were exposed. During the surgical procedure, a 2.0 x 1.5 mm full-thickness mucosal injury was induced in the midline of the hard palate using ophthalmologic cautery. It was crucial to control the size of the ONF and minimize bleeding in order to ensure the success of the experiment. Verification of the ONF model's effectiveness was conducted on the 7th-day post-operation, encompassing both anatomical and functional assessments. The presence of the nasal septum within the oral cavity and the outflow of sterile water from the nostrils upon injection into the oral cavity confirmed the successful establishment of the ONF model. The model demonstrated a practical and successful oronasal fistula, characterized by a low mortality rate, significant weight changes, and minimal variation in ONF size. Future studies may consider adopting this methodology to elucidate the mechanisms of palate wound healing and explore novel treatments for oronasal fistulas.

Introduction

Oronasal fistula (ONF), an abnormal opening between the oral and nasal cavities, clinically manifests as a defect in a structural area from the alveolar process to the uvula, which commonly occurs as a complication following cleft palate repair1. Patients with ONF experience food reflux, articulation disorders, and impaired velopharyngeal function, significantly impacting their quality of life2,3,4. The rate of post-operative ONF ranges from 2.4% to 55% due to factors such as cleft width, Veau type, and surgical method5,

Protocol

All animal procedures in this study were reviewed and approved by the Ethical Committee of the West China School of Stomatology, Sichuan University. Adult C57BL/6 mice (female) were used for the present study.

1. Surgical preparation

  1. Gather the necessary surgical instruments for the procedure: germinator, ophthalmologic cautery, microsurgical scissors, microsurgical tweezers, syringes, and needles (26 g x  0.63 inch) (Figure 1A

Representative Results

To assess the feasibility and stability of this experimental method, the same procedure was performed on ten mice, and observations were made regarding mortality, changes in wound size, body weight, and histologic analysis. The mice were euthanized on day 7.

The procedure exhibited a low mortality rate. The ophthalmologic cautery and germinator, depicted in Figure 1A-C, were the key instruments utilized in this experiment. The ONF.......

Discussion

Researchers have explored various materials, drugs, and novel techniques for treating ONF10,11,12,13,14,15,16,17. With advancements in surgical procedures, the incidence and recurrence of ONF have been reduced. However, due to the unique characteristics of .......

Acknowledgements

This work was supported by the Research and Development Program, West China Hospital of Stomatology, Sichuan University (RD-02-202107), Sichuan Province Science and Technology Support Program (2022NSFSC0743), and Sichuan Postdoctoral Science Foundation (TB2022005) grant to H. Huang.

....

Materials

NameCompanyCatalog NumberComments
GerminatorElectron Microscopy Sciences 66118-20Heating and disinfection equipment
Latex glovesAllmedor similar
LightsOlympusA1813
MeloxicamMedChemExpressHY-B0261crushed; 5 mg/kg
Microsurgical instruments (scissors and tweezers)Jiangsu Tonghui Medical Devices Co.M-Y-0087Surgical instrument
Ophthalmologic cauterySuqian Wenchong Medical Equipment Co.1.00234E+13Surgical instrument
Sterile cotton,Yancheng Begu Technology Co.or similar
Sterile metal foilBiosharpor similar
Sterile sheets3MXH003801129or similar
Surgical drapesYancheng Begu Technology Co.or similar
SyringesYancheng Begu Technology Co.S-015301or similar
TapeBkmamlabor similar
Temperature maintenance deviceHarvard Apparatus LE-13-2104
Zoletil50Virbac80 mg/kg

References

  1. Alonso, V., et al. Three-layered repair with a collagen membrane and a mucosal rotational flap reinforced with fibrine for palatal fistula closure in children. International Journal of Pediatric Otorhinolaryngology. 127, 109679 (2019).
  2. Garg, R., Shah, S., Uppal, S., Mittal, R. K.

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