CC BY 4.0 · Eur J Dent
DOI: 10.1055/s-0044-1779426
Original Article

Alveolar Bone Loss in a Ligature-Induced Periodontitis Model in Rat Using Different Ligature Sizes

1   Department of Periodontology, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
,
1   Department of Periodontology, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
,
2   Center of Excellence in Periodontal Disease and Dental Implant, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
3   Immunology Research Center, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
,
4   Biomaterial Testing Center, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
,
1   Department of Periodontology, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
2   Center of Excellence in Periodontal Disease and Dental Implant, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
3   Immunology Research Center, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
,
Wichaya Wisitrasameewong
1   Department of Periodontology, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
2   Center of Excellence in Periodontal Disease and Dental Implant, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
3   Immunology Research Center, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
› Author Affiliations
Funding This study was supported by Thailand Science Research and Innovation (DIG6280002), Thailand Science Research and Innovation Fund Chulalongkorn University (CU_FRB65_hea(3)_009_32_04), Chulalongkorn Academic Advancement into Its 2nd Century Project and Grants for Development of New Faculty Staff, Ratchadaphiseksomphot Endowment Fund.

Abstract

Objectives Ligature-induced periodontitis model has been widely used as a preclinical stage for investigating new treatment modalities. However, the effect of different ligature sizes on alveolar bone loss has never been studied. Therefore, we examined alveolar bone loss in this rat model using different sizes of silk ligatures, as well as healing after ligature removal.

Materials and Methods Left maxillary second molars of Sprague-Dawley rats were ligated with 3-0, 4-0, or 5-0 silk ligatures (n = 4–5/group) for 14 days before harvested maxillae and gingival tissues. For subsequent experiment, animals were ligated for 14 days using the ligature size that induced the most alveolar bone loss before ligature removal and sacrificed at 0, 7 and 14 days (n = 5–6/group). All maxillae and gingival tissues were harvested to evaluate alveolar bone level, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) levels.

Statistical Analysis Data was analyzed using SPSS Statistics 23.0 software (SPSS Inc., Chicago, Illinois, United States). Data from all experiments were tested for normality using Shapiro–Wilk test. Data between ligatured and nonligatured teeth were compared using Student's t-test or Wilcoxon signed-rank test. Differences among different ligature sizes were analyzed by analysis of variance followed by multiple comparisons with post-hoc test. A p-value less than 0.05 was considered statistically significant.

Results The alveolar bone loss of ligated teeth was substantially higher than that of control after 14 days of ligation. While 3-0 and 4-0 resulted in significantly greater bone loss than 5-0 silk, the 3-0 group had the lowest rate of ligature loss. Therefore, alveolar bone healing postligature removal was investigated further using 3-0 silk. The results showed no significant bone level change at 2 weeks after ligature removal. In term of IL-1β and TNF-α levels, there was no statistically significant difference in IL-1β level between groups at any time point, while TNF-α was undetectable.

Conclusion These data showed that 3-0 silk was the most effective ligature size in promoting alveolar bone loss comparing with 4-0 and 5-0 silk. During the 2-week period following ligature removal, spontaneous bone healing was not observed.



Publication History

Article published online:
05 March 2024

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  • References

  • 1 Donos N, Park JC, Vajgel A, de Carvalho Farias B, Dereka X. Description of the periodontal pocket in preclinical models: limitations and considerations. Periodontol 2000 2018; 76 (01) 16-34
  • 2 Klausen B. Microbiological and immunological aspects of experimental periodontal disease in rats: a review article. J Periodontol 1991; 62 (01) 59-73
  • 3 Kawai T, Paster BJ, Komatsuzawa H. et al. Cross-reactive adaptive immune response to oral commensal bacteria results in an induction of receptor activator of nuclear factor-kappaB ligand (RANKL)-dependent periodontal bone resorption in a mouse model. Oral Microbiol Immunol 2007; 22 (03) 208-215
  • 4 Xiao E, Mattos M, Vieira GHA. et al. Diabetes enhances IL-17 expression and alters the oral microbiome to increase its pathogenicity. Cell Host Microbe 2017; 22 (01) 120-128.e4
  • 5 de Molon RS, Mascarenhas VI, de Avila ED. et al. Long-term evaluation of oral gavage with periodontopathogens or ligature induction of experimental periodontal disease in mice. Clin Oral Investig 2016; 20 (06) 1203-1216
  • 6 Lemaitre M, Monsarrat P, Blasco-Baque V. et al. Periodontal tissue regeneration using syngeneic adipose-derived stromal cells in a mouse model. Stem Cells Transl Med 2017; 6 (02) 656-665
  • 7 Liu R, Bal HS, Desta T. et al. Diabetes enhances periodontal bone loss through enhanced resorption and diminished bone formation. J Dent Res 2006; 85 (06) 510-514
  • 8 Abe T, Hajishengallis G. Optimization of the ligature-induced periodontitis model in mice. J Immunol Methods 2013; 394 (1-2): 49-54
  • 9 Kaboosaya B, Wulansari LK, Trang Nguyen VN, Aoki K, Kasugai S. Ligation period required to induce periodontitis in mice: analysis with micro-computed tomography. J Oral Tissue Eng 2017; 15: 25-34
  • 10 Maekawa S, Katagiri S, Takeuchi Y. et al. Bone metabolic microarray analysis of ligature-induced periodontitis in streptozotocin-induced diabetic mice. J Periodontal Res 2017; 52 (02) 233-245
  • 11 Marchesan J, Girnary MS, Jing L. et al. An experimental murine model to study periodontitis. Nat Protoc 2018; 13 (10) 2247-2267
  • 12 Nemcovsky CE, Zahavi S, Moses O. et al. Effect of enamel matrix protein derivative on healing of surgical supra-infrabony periodontal defects in the rat molar: a histomorphometric study. J Periodontol 2006; 77 (06) 996-1002
  • 13 Gould TR, Melcher AH, Brunette DM. Location of progenitor cells in periodontal ligament of mouse molar stimulated by wounding. Anat Rec 1977; 188 (02) 133-141
  • 14 Lekic P, Sodek J, McCulloch CA. Osteopontin and bone sialoprotein expression in regenerating rat periodontal ligament and alveolar bone. Anat Rec 1996; 244 (01) 50-58
  • 15 Pellegrini G, Seol YJ, Gruber R, Giannobile WV. Pre-clinical models for oral and periodontal reconstructive therapies. J Dent Res 2009; 88 (12) 1065-1076
  • 16 Takeuchi T, Bizenjima T, Ishii Y. et al. Enhanced healing of surgical periodontal defects in rats following application of a self-assembling peptide nanofibre hydrogel. J Clin Periodontol 2016; 43 (03) 279-288
  • 17 Semenoff TA, Semenoff-Segundo A, Bosco AF, Nagata MJ, Garcia VG, Biasoli ER. Histometric analysis of ligature-induced periodontitis in rats: a comparison of histological section planes. J Appl Oral Sci 2008; 16 (04) 251-256
  • 18 Lee CT, Teles R, Kantarci A. et al. Resolvin E1 reverses experimental periodontitis and dysbiosis. J Immunol 2016; 197 (07) 2796-2806
  • 19 Lee HJ, Lee DR, Choi BK, Yang SH. Antiperiodontitis effects of Magnolia biondii extract on ligature-induced periodontitis in rats. Nutrients 2019; 11 (04) 934
  • 20 Hoffman MM, Schour I. Quantitative studies in the development of the rat molar. I. The growth pattern of the primary and secondary dentin (from birth to 500 days of age). Anat Rec 1940; 78: 233-251
  • 21 Liu J, Kim D, Brown L, Madsen T, Bouchard GF. Comparison of human, porcine, and rodent wound healing with new miniature swine study data. J Am Assoc Lab Anim Sci 2009; 48: 581-581
  • 22 de Molon RS, Park CH, Jin Q, Sugai J, Cirelli JA. Characterization of ligature-induced experimental periodontitis. Microsc Res Tech 2018; 81 (12) 1412-1421
  • 23 Fernandes MI, Gaio EJ, Oppermann RV, Rados PV, Rosing CK. Comparison of histometric and morphometric analyses of bone height in ligature-induced periodontitis in rats. Braz Oral Res 2007; 21 (03) 216-221
  • 24 Liu YF, Wu LA, Wang J, Wen LY, Wang XJ. Micro-computerized tomography analysis of alveolar bone loss in ligature- and nicotine-induced experimental periodontitis in rats. J Periodontal Res 2010; 45 (06) 714-719
  • 25 de Molon RS, de Avila ED, Boas Nogueira AV. et al. Evaluation of the host response in various models of induced periodontal disease in mice. J Periodontol 2014; 85 (03) 465-477
  • 26 Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012; 9 (07) 671-675
  • 27 Kwan Tat S, Padrines M, Théoleyre S, Heymann D, Fortun Y. IL-6, RANKL, TNF-alpha/IL-1: interrelations in bone resorption pathophysiology. Cytokine Growth Factor Rev 2004; 15 (01) 49-60
  • 28 Ramadan DE, Hariyani N, Indrawati R, Ridwan RD, Diyatri I. Cytokines and chemokines in periodontitis. Eur J Dent 2020; 14 (03) 483-495