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In vitro antibacterial activity of green tea–loaded chitosan nanoparticles on caries-related microorganisms and dentin after Er:YAG laser caries removal

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Abstract

This study aimed to determine the inhibitory effects of green tea (Gt), EGCG, and nanoformulations containing chitosan (Nchi) and chitosan+green tea (Nchi+Gt) against Streptococcus mutans and Lactobacillus casei. In addition, the antibacterial effect of nanoformulations was evaluated directly on dentin after the selective removal of carious lesion. At first, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against S. mutans and L. casei isolates were investigated. In parallel, dentin specimens were exposed to S. mutans to induce carious lesions. Soft dentin was selectively removed by Er:YAG laser (n=33) or bur (n=33). Remaining dentin was biomodified with Nchi (n=11) or Gt+Nchi (n=11). Control group (n=11) did not receive any treatment. Dentin scraps were collected at three time points. Microbiological analyses were conducted and evaluated by agar plate counts. Gt at 1:32 dilution inhibited S. mutans growth while 1:16 was efficient against L. casei. EGCG at 1:4 dilution completely inhibited S. mutans and L. casei growth. Independently of the association with Gt, Nchi completely inhibited S. mutans at 1:4 dilution. For L. casei, different concentrations of Nchi (1:32) and Nchi+Gt (1:8) were required to inhibit cell growth. After selective carious removal, viability of S. mutans decreased (p<0.001), without difference between bur and Er:YAG laser (p>0.05). Treatment with Nchi and Nchi+Gt did not influence the microbial load of S. mutans on dentin (p>0.05). Although variations in concentrations were noticed, all compounds showed antibacterial activity against S. mutans and L. casei. Both bur and Er:YAG laser have effectively removed soft dentin and reduced S. mutans counts. Nanoformulations did not promote any additional antibacterial effect in the remaining dentin.

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Data availability

The data supporting the findings of this study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Ana Paula Macedo for assistance with statistical data analysis.

Funding

This study received financial support from the São Paulo Research Foundation (FAPESP), grant nos. #2017/11582-1, #2017/00720-4, #2018/23862-1, #2019/04807-2, thematic project #2013/50181-1 and FINEP project 01.10.0758.01) and from the National Council for Scientific and Technological Development (CNPq), grant nos. PRONON-SIPAR project #25000.077093/2015-86), CNPq-SCTIE-Decit-DGITIS-CGCIS/CNPq no. 26/2020 – Innovative Platforms in Advanced Therapies, # 441673/2020-1 and CNPq-UN # 404416/2021-7, #2021-3/130399.

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Authors and Affiliations

  1. Department of Restorative Dentistry, School of Dentistry of Ribeirao Preto, University of Sao Paulo (USP), Cafe Avenue, s/n 14040-904, Sao Paulo, Ribeirao Preto, Brazil

    Fabiana Almeida Curylofo-Zotti, Analu Rodriguez Marchesin & Silmara Aparecida Milori Corona

  2. Department of Dental Materials and Prosthodontics, School of Dentistry of Ribeirao Preto, University of Sao Paulo (USP), Ribeirao Preto, Sao Paulo, SP, Brazil

    Viviane De Cássia Oliveira

  3. Department of Chemistry, Center of Nanotechnology and Tissue Engineering - Photobiology and Photomedicine Research Group, Faculty of Philosophy, Sciences and Letters of Ribeirao Preto, University of Sao Paulo (USP), São Paulo, Brazil

    Hiago Salge Borges & Antonio Claudio Tedesco

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  1. Fabiana Almeida Curylofo-Zotti
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Fabiana Almeida Curylofo Zotti, Viviane de Cássia Oliveira, Analu Rodriguez Marquesin, and Hiago Salge Borges. The first draft of the manuscript was written by Fabiana Almeida Curylofo Zotti and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Fabiana Almeida Curylofo-Zotti.

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Ethical approval

All procedures performed in the study were in accordance with the Research Ethics Committee of the University of São Paulo (Institutional Review Board protocol CAAE 69600217.4.0000.5419 and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants who agree to donate their extracted teeth to be included in the study.

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Curylofo-Zotti, F.A., Oliveira, V.D.C., Marchesin, A.R. et al. In vitro antibacterial activity of green tea–loaded chitosan nanoparticles on caries-related microorganisms and dentin after Er:YAG laser caries removal. Lasers Med Sci 38, 50 (2023). https://doi.org/10.1007/s10103-023-03707-3

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