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Bacteria-mediated resistance of neutrophil extracellular traps to enzymatic degradation drives the formation of dental calculi

Nature Biomedical Engineering (2024)Cite this article

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Abstract

Dental calculi can cause gingival bleeding and periodontitis, yet the mechanism underlying the formation of such mineral build-ups, and in particular the role of the local microenvironment, are unclear. Here we show that the formation of dental calculi involves bacteria in local mature biofilms converting the DNA in neutrophil extracellular traps (NETs) from being degradable by the enzyme DNase I to being degradation resistant, promoting the nucleation and growth of apatite. DNase I inhibited NET-induced mineralization in vitro and ex vivo, yet plasma DNases were ineffective at inhibiting ectopic mineralization in the oral cavity in rodents. The topical application of the DNA-intercalating agent chloroquine in rodents fed with a dental calculogenic diet reverted NET DNA to its degradable form, inhibiting the formation of calculi. Our findings may motivate therapeutic strategies for the reduction of the prevalence of the deposition of bacteria-driven calculi in the oral cavity.

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Fig. 1: Evidence of neutrophil extracellular traps in human body stones.
Fig. 2: NETs adhered on calculus and induced further mineralization ex vivo.
Fig. 3: NETs induced CaP mineralization in vitro.
Fig. 4: Molecular dynamics simulation of NET-induced mineralization.
Fig. 5: The elimination of NET disrupted ectopic mineralization ex vivo and in vitro, but not in vivo.
Fig. 6: DNase I could not inhibit NET-induced mineralization with the involvement of mature biofilm in vitro.
Fig. 7: Mature bacterial biofilm induced NET B-DNA to become NET Z-DNA in vitro and in vivo.
Fig. 8: Inhibition of ectopic mineralization in vivo via the regulation of NET-DNA configuration.

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The main data supporting the results of this study are available within the paper and its Supplementary Information. The raw and analysed datasets generated during the study are available for research purposes from the corresponding author on reasonable request.

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Acknowledgements

This work was supported by the National Clinical Research Center for Oral Diseases (LCA202004), National Natural Science Foundation of China (no. 82325012 to Li-na Niu and no. 82170978 to K.J.), the Shaanxi Key Scientific and Technological Innovation Team (no. 2020TD-033 to Li-na Niu). Parts of the figures were drawn using pictures from Servier Medical Art. Servier Medical Art by Servier is licenced under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licences/by/3.0).

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Author notes

  1. These authors contributed equally: Mei-chen Wan, Kai Jiao, Yi-na Zhu, Qian-qian Wan.

Authors and Affiliations

  1. State Key Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University, Xi’an, Shaanxi, P.R. China

    Mei-chen Wan, Yi-na Zhu, Qian-qian Wan, Yi-peng Zhang, Long-zhang Niu, Chen Lei, Jing-han Song, Wei-cheng Lu, Hua-jie Liu, Zhao-yang Ren & Li-na Niu

  2. Department of Stomatology, Tangdu hospital; State Key Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Stomatology, School of Stomatology, The Fourth Military Medical University, Xi’an, Shaanxi, P.R. China

    Kai Jiao

  3. The Dental College of Georgia, Augusta University, Augusta, GA, USA

    Franklin Tay

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Contributions

M.W., K.J., Yi-na Zhu and Q.W. contributed to the study design, data acquisition and interpretation. Yi-peng Zhang contributed to the DNA manipulation of Escherichia coli. Long-zhang Niu, C.L. and J.S. contributed to data analysis and interpretation. W.L. and Z.R. performed the animal experiments. H.L. provided the clinical samples. F.T. refined the paper. Li-na Niu developed the concept and supervised experiments. All authors contributed to writing the paper.

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Correspondence to Li-na Niu.

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Wan, Mc., Jiao, K., Zhu, Yn. et al. Bacteria-mediated resistance of neutrophil extracellular traps to enzymatic degradation drives the formation of dental calculi. Nat. Biomed. Eng (2024). https://doi.org/10.1038/s41551-024-01186-7

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