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In This Article

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

Summary

Here, we present a protocol for studying orthodontic tooth movement (OTM), serving as a suitable model for investigating the mechanisms of bone adaptation, root resorption, and the response of bone cells to mechanical stimuli. This comprehensive guide provides detailed information on the OTM model, micro-computed tomography acquisition, and subsequent analysis.

Abstract

Orthodontic tooth movement (OTM) represents a dynamic process in which the alveolar bone undergoes resorption at compression sites and deposition at tension sites, orchestrated by osteoclasts and osteoblasts, respectively. This mechanism serves as a valuable model for studying various aspects of bone adaptation, including root resorption and the cellular response to mechanical force stimuli. The protocol outlined here offers a straightforward approach to investigate OTM, establishing 0.35 N as the optimal force in a mouse model employing a nickel-titanium (NiTi) coil spring. Utilizing micro-computed tomography analysis, we quantified OTM by assessing the discrepancy in the linear distance at the cement-enamel junction. The evaluation also included an analysis of orthodontic-induced inflammatory root resorption, assessing parameters such as root mineral density and the percentage of root volume per total volume. This comprehensive protocol contributes to advancing our understanding of bone remodeling processes and enhancing the ability to develop effective orthodontic treatment strategies.

Introduction

Bone remodeling is an ongoing process orchestrated by osteoclasts, osteoblasts, bone lining cells, and osteocytes, essential for maintaining the integrity of the adult skeleton1,2. Primarily driven by the differentiation and activity of osteoclasts and osteoblasts, this dynamic process involves the resorption and deposition of bone, triggered by mechanical stress and loading3,4,5.

Animal experiments play a pivotal role in elucidating the intricate biological and cellular mechanisms underpin....

Protocol

All procedures strictly adhered to the ethical standards established by the Universidade Federal de Minas Gerais Ethics Committee (No. 166/2022). Before each experiment, a sample size calculation is mandatory. Use 8-10-week-old male C57BL6/J wild-type mice weighing approximately 20-30 g. The mice must be housed in a cage within a room maintained at 25 °C, adhering to a 12 h light/12 h dark cycle. Following coil attachment, the animal should be fed with a soft diet. Daily monitoring should include assessments of body.......

Representative Results

This protocol enables the investigation of an OTM mouse model using a NiTi coil spring. With a force of 0.35 N applied, the mean CEJ distance on the control side between the first and second molars was 243.69 µm (Figure 1A, line A), whereas on the OTM side was measured at 284.66 µm (Figure 1A, line B). The difference between the OTM and control sides was 40.97 µm (Figure 1B). The linear distance betwe.......

Discussion

Here, we describe a standardized protocol designed to elucidate the cellular and molecular mechanisms underlying bone remodeling during OTM. A thorough understanding of these mechanisms in mice requires a meticulously planned protocol to ensure accuracy and reliability7,11. Studies conducted by our research group have shown that this protocol effectively reduces operator variability by incorporating a tension gauge and a specially designed apparatus, establishing.......

Acknowledgements

We wish to express our sincere appreciation to Miss Beatriz M. Szawka for her contribution to the schematic diagram and to Mrs. Ilma Marçal de Souza for her technical support. J.A.A.A. is the recipient of a fellowship granted by Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ, E-26/200.331/2024), Brazil. This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (406928/2023-1), Fundação de Amparo a Pesquisa do Estado de Minas Gerais and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Finance code 001), Brazil. The authors ....

Materials

NameCompanyCatalog NumberComments
AcetoneSigma-Aldrich67-64-1
Distal cut pliersQuinelatoQO.700.00
DynamometerSHIMPOFGE-5XY
Fiber Optic IlluminatorCole-ParmerN/A
ketamineSyntec100477-72-3
NiTi open-coil spring 0.25 x 0.76Lancer Orthodontics
Ø 0.20 mm round chrome-nickel (CrNi)Morelli55.01.208
Round CrNi Hard Elastic Orthodontic Wire Ø0.50 mm (.020 inch)Morelli55.01.050
Round CrNi Tie Wire Ø0.20 mm (.008 inch)Morelli55.01.208
StereomicroscopeQuimisQ7740SZ
Transbond Plus Self Etching Primer3MLE-Q100-1004-7
Weingart PlierQuinelatoQO.120.00
XylazineSyntec23076-35-9
MicroCT Analysis
Skyscan 1174v2Bruker1174v2
Software
NReconSkyscanN/A
DataViewerSkyscanN/A
CTAnSkyscanN/A
MimicsMaterialiseN/A

References

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Orthodontic Tooth MovementOTMAlveolar BoneResorptionDepositionOsteoclastsOsteoblastsBone AdaptationRoot ResorptionMechanical Force StimuliNickel titanium Coil SpringMicro computed TomographyInflammatory Root ResorptionRoot Mineral DensityBone RemodelingOrthodontic Treatment Strategies

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