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Histomorphometric Changes of the Masseter Muscle of Rats After a Single Injection of Botulinum Toxin Type A

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  • Basic Science/Experimental
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Abstract

Background

It has been reported that botulinum toxin type A (BoNT-A) produces structural changes in masticatory muscles. However, not all histomorphometric parameters affected by BoNT-A parameters have been assessed. This study investigated the histomorphometric changes in the masseter muscle of rats after a single injection of BoNT-A.

Methods

Forty-four adult animals were randomly divided into control group (n = 22) and BoNT-A group (n = 22). Controls received a single dose of 0.14 mL/kg of saline in masseter muscles, and the BoNT-A group received a 7 U/Kg of BoNT-A. The groups received the same volume of injected substances. Animals were sacrificed on 7th (n = 5), 14th (n = 5), 21st (n = 5), 28th (n = 4) and 90th (n = 3) days post-treatment. Histological masseter tissue slides were obtained from hematoxylin–eosin treatment and analyzed in optical microscopy regarding muscle cross-sectional area, amount of connective tissue and quantity and diameter of myocytes. For statistical analysis, generalized linear models were used to compare the data (ANOVA). In all test, the significance level of 5% was set.

Results

BoNT-A values of cross-sectional area of the masseter muscle were significantly lower than controls (p < 0.01) throughout the study. Regarding myocytes quantity, BoNT-A subgroups presented higher values than controls (p < 0.0001) since the 14th day until the end of the study; however, the diameter of myocytes was smaller in all BoNT-A subgroups (p < 0.0001) in all assessment points. The amount of connective tissue was higher in BoNT-A subgroups (p < 0.0001) throughout the study.

Conclusion

A single injection of BoNT-A altered the structure of masseter muscle of rats, regarding its histomorphometric parameters.

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Data Availability

Datasets related to this article will be available upon request to the corresponding author.

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Acknowledgments

The authors acknowledge the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES) for the PhD scholarship of D.M.R.—grant code 001 and the Sao Paulo Research Foundation—FAPESP for the post-doctoral scholarship of G. D. T. C. (grant 2017/21674-0).

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Authors and Affiliations

  1. Department of Oral Biology, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil

    Douglas Massoni Ramos, Raira de Brito Silva, Nádia Cristina Fávaro Moreira & Célia Marisa Rizzatti-Barbosa

  2. Egas Moniz Center for Interdisciplinary Research (CiiEM), Egas Moniz School of Health & Science, Caparica, Almada, Portugal

    Giancarlo De la Torre Canales

  3. Araras Dental School, University Center Herminio Ometto, Araras, SP, Brazil

    Marcelo Augusto Marretto Esquisatto

  4. Department of Dental Medicine, Karolinska Institutet, and the Scandinavian Network for Orofacial Neurosciences (SCON), Huddinge, Sweden

    Giancarlo De la Torre Canales & Malin Ernberg

  5. Ingá University Center Uningá, Department of Dentistry, Maringá, PR, Brazil

    Giancarlo De la Torre Canales, Luciana Resende & Célia Marisa Rizzatti-Barbosa

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  1. Douglas Massoni Ramos
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Correspondence to Giancarlo De la Torre Canales.

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The authors declare that they have no conflicts of interest to disclose.

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For this type of study, informed consent is not required.

Institutional Review Board Statement

The present study was approved by the Ethics Committee in Animals Research of the State University of Campinas (CEUA/UNICAMP), Sao Paulo, Brazil under protocol no. 4554-1/2017 and followed the National Council for Control of Animal Experimentation (CONCEA). All applicable institutional and/or national guidelines for the care and use of animals were followed.

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Ramos, D.M., de Brito Silva, R., De la Torre Canales, G. et al. Histomorphometric Changes of the Masseter Muscle of Rats After a Single Injection of Botulinum Toxin Type A. Aesth Plast Surg 48, 1037–1044 (2024). https://doi.org/10.1007/s00266-023-03572-z

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