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Expression and localisation of amelotin, laminin and protein secreted by follicular dendritic cells after ligature-induced experimental periodontitis in rats

British Dental Journal (2025)Cite this article

Abstract

Objective The junctional epithelium (JE) is composed of tightly interconnected layers of squamous epithelial cells that play a crucial role as a defence mechanism. Recently, several enamel-derived proteins have been shown to play a role in the adhesion of JE cells to the mineralised tooth surface. This study aims to explore the in vivo protein expression levels of amelotin (AMTN), laminin (LAM332) and the protein secreted by follicular dendritic cells (FDC-SP) within the JE, both in the presence and absence of experimental periodontitis (EP) in rats.

Materials and methods In total, 16 rats were randomly divided into two groups: a control group without EP and a group with induced periodontitis. EP was established by placing cotton ligatures around the cervical region of the lower first molars. Fifteen days after EP induction, all animals were euthanised and mandibles were harvested. Micro-computed tomography, histomorphometric, and immunohistochemistry analyses were employed to assess volumetric bone alterations, architectural bone parameters and number of osteoclasts. The presence of inflammatory cells and enamel protein expression were evaluated by immunofluorescence.

Results The results demonstrated that the EP group showed a significant increase in alveolar bone loss, an elevated number of tartrate-resistant acid phosphatase-positive cells, and enhanced inflammatory processes compared to the control group. Immunofluorescence staining revealed a significant increase in the expression of AMTN in the EP group. However, there were no significant differences between the expression of FDC-SP and LAM332. Correlation analysis of AMTN, LAM332 and FDC-SP with the intensity of inflammatory markers (CD45+, CD66b+ and CD8+, CD163+ and CD80+) showed no significant differences for the EP group.

Conclusion Our data suggest that the expression of AMTN increased after inflammatory stimuli and that AMTN may be associated with the onset and progression of EP.

Key points

  • It is essential to investigate the role of junctional epithelium proteins during periodontitis progression to understand their function under local inflammatory conditions. In this context, our study assessed the expression levels of enamel matrix proteins between healthy and diseased sites in the periodontium.

  • We showed that the localisation and expression of amelotin were significantly increased after inducing experimental periodontitis at the junctional epithelium. FDC-SP and LAM332 protein levels were not overexpressed during periodontitis progression. These findings suggest a potential role for amelotin in the inflammatory events that occur during the initiation and progression of periodontitis.

  • Increased protein expression of amelotin under inflammatory conditions underscores the role of amelotin as a diagnostic marker for disease onset/progression with direct clinical implications and paves the way for new approaches to managing periodontitis.

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Data availability

The datasets used or analysed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors express their special thanks to the staff at the Faculty of Dentistry, University of Toronto, for the extensive help during immunofluorescence processing.

Funding

Rafael Scaf de Molon is currently supported by The Sao Paulo Research Foundation (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo) FAPESP grant #2023/15750-7. The authors acknowledge the financial support provided by the Brazilian funding agency coordination for the improvement of higher education (CAPES), financial code 01 in the scope of CAPES/print - funding code: 001 (process: 88887.194785/2018-00).

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Authors and Affiliations

  1. Department of Diagnostic and Surgery, School of Dentistry, São Paulo State University (UNESP), Araçatuba, São Paulo, Brazil

    Natália Amanda Gomes, Gabriel Mulinari-Santos, Tiago Esgalha da Rocha, Rafael Scaf de Molon & Letícia Helena Theodoro

  2. Department of Diagnostic and Surgery, School of Dentistry, São Paulo State University (UNESP), Araçatuba, São Paulo, Brazil; Latin American Institute of Dental Research and Teaching (ILAPEO), Curitiba, PR, Brazil

    Valdir Gouveia Garcia

  3. Department of Basic Sciences, School of Dentistry, São Paulo State University (UNESP), Araçatuba, São Paulo, Brazil

    Edilson Ervolino

  4. Faculty of Dentistry, University of Toronto, Toronto, ON, Canada

    Bernhard Ganss, Aiman Ali & Marco Magalhães

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Contributions

Conceptualisation: VGG, RSdM, LHT; methodology: NAG, VGG, GMS, E E, TER, BG, AA, MM, RSdM, LHT; validation: NAG, VGG, GMS, E E, TER, BG, AA, MM, RSdM, LHT; formal analysis: NAG, VGG, GMS, E E, TER, BG, AA, MM, RSdM, LHT; investigation: NAG, VGG, GMS, RSdM, LHT; data curation: NAG, VGG, GMS, E E, TER, BG, AA, MM, RSdM, LHT; writing - original draft preparation, RSdM; writing - review and editing: NAG, VGG, GMS, E E, TER, BG, AA, MM, RSdM, LHT; visualisation: NAG, VGG, GMS, E E, TER, BG, AA, MM, RSdM, LHT; supervision: VGG and LHT. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Rafael Scaf de Molon.

Ethics declarations

The authors declare no conflicts of interest. This study was performed in line with the principles of the ARRIVE guidelines. The approval for the study was granted by the Animal Use Ethics Committee (#00608-2020) from the Sao Paulo State University, UNESP, School of Dentistry at Aracatuba.

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Gomes, N., Garcia, V., Mulinari-Santos, G. et al. Expression and localisation of amelotin, laminin and protein secreted by follicular dendritic cells after ligature-induced experimental periodontitis in rats. Br Dent J (2025). https://doi.org/10.1038/s41415-025-8508-7

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