An experimental murine model to study periodontitis


Periodontal disease (PD) is a common dental disease associated with the interaction between dysbiotic oral microbiota and host immunity. It is a prevalent disease, resulting in loss of gingival tissue, periodontal ligament, cementum and alveolar bone. PD is a major form of tooth loss in the adult population. Experimental animal models have enabled the study of PD pathogenesis and are used to test new therapeutic approaches for treating the disease. The ligature-induced periodontitis model has several advantages as compared with other models, including rapid disease induction, predictable bone loss and the capacity to study periodontal tissue and alveolar bone regeneration because the model is established within the periodontal apparatus. Although mice are the most convenient and versatile animal models used in research, ligature-induced periodontitis has been more frequently used in large animals. This is mostly due to the technical challenges involved in consistently placing ligatures around murine teeth. To reduce the technical challenge associated with the traditional ligature model, we previously developed a simplified method to easily install a bacterially retentive ligature between two molars for inducing periodontitis. In this protocol, we provide detailed instructions for placement of the ligature and demonstrate how the model can be used to evaluate gingival tissue inflammation and alveolar bone loss over a period of 18 d after ligature placement. This model can also be used on germ-free mice to investigate the role of human oral bacteria in periodontitis in vivo. In conclusion, this protocol enables the mechanistic study of the pathogenesis of periodontitis in vivo.

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Fig. 1: Tools and technical procedures required to set up the simplified ligature model in mice.
Fig. 2: Time course of alveolar bone loss and osteoclast activation after insertion of ligature.
Fig. 3: Time course of gingival tissue inflammation after insertion of ligature.
Fig. 4: The expression of innate and adaptive immune genes at different time points after insertion of ligature.
Fig. 5: Human bacteria induced alveolar bone loss and gingival tissue inflammation when the ligature model was used in GF mice.


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This study was supported by grants T90DE021986 and F32DE026688 (Y.J.), an IBM Junior Faculty Development Award from the University of North Carolina at Chapel Hill and grants K01DE027087 (J.M.), 5R01DE023836 (S.O.) and 5R01DE025207 (M.H.S.). We thank W.V. Giannobile of the University of Michigan School of Dentistry for constructive suggestions and helpful discussion. We thank B. Huang and J. Ashley of the Histology Research Core Facility at UNC for histological embedding and sectioning. We thank the Microscope Core facility for the images. We thank the UNC Biomedical Research Imaging Center for microCT scanning. We thank the Germ Free & Gnotobiotic Mouse Facilities at the University of Michigan and the Gnotobiotic Rodent Core Facility at the University of North Carolina for assistance with the human bacterial infection experiment. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

Author information




Y.J. and J.M. designed the experiments. Y.J., J.M. and M.S.G. performed the in vivo experiments. L.J. performed H&E and MPO staining and analysis. M.Z.M. performed TRAP staining and analysis. M.S.G. performed qRT-PCR and analysis using the RT2 Profiler Array. S.Z. and L.S. performed the microCT analysis. Y.J., J.M., T.M. and M.S.G. developed the murine dental bed and ligature holder. J.M. and Y.J. wrote and edited the manuscript together. S.O., N.I. and M.H.S. helped to develop the model and edited the manuscript. All authors approved the final version of the manuscript.

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Correspondence to Yizu Jiao.

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1. Jiao, Y. et al. Cell Host Microbe 13, 595–601 (2013):

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Supplementary Figure 1

Typical colony morphology for S. gordonii, F. nucleatum and V. parvula on a TSA plate.

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Supplementary Text and Figures

Supplementary Figure 1

Reporting Summary

Supplementary Manual

Detailed top view of blueprints required for manufacture of the murine dental bed

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Marchesan, J., Girnary, M.S., Jing, L. et al. An experimental murine model to study periodontitis. Nat Protoc 13, 2247–2267 (2018).

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