Effects of highly leukotoxic Aggregatibacter actinomycetemcomitans associated with ligature-induced periodontitis on oral and gut tissues in male rats

Authors

  • Gabriela Furian Trama Ferraz Universidade Federal do Rio de Janeiro, Faculdade de Odontologia, Departamento de Clínica Odontológica
  • Camila Philipe de Araujo Camilo Universidade Federal do Rio de Janeiro, Faculdade de Odontologia, Departamento de Clínica Odontológica
  • Ana Luisa Palhares de Miranda Universidade Federal do Rio de Janeiro, Faculdade de Farmácia, Departamento de Biotecnologia Farmacêutica, Laboratório de Estudos em Farmacologia Experimental (LEFEx)
  • Mariana Alves Soares Universidade Federal do Rio de Janeiro, Faculdade de Farmácia, Departamento de Biotecnologia Farmacêutica, Laboratório de Estudos em Farmacologia Experimental (LEFEx)
  • Carmelo Sansone Universidade Federal do Rio de Janeiro, Faculdade de Odontologia, Departamento de Clínica Odontológica, Rio de Janeiro
  • Ricardo Tadeu Lopes Universidade Federal do Rio de Janeiro, Instituto Alberto Luiz de Coimbra de Pós-graduação e Pesquisa de Engenharia, Programa de Engenharia Nuclear, Laboratório de Instrumentação Nuclear
  • Aline Correa Abrahão Universidade Federal do Rio de Janeiro, Faculdade de Odontologia, Departamento de Patologia Oral
  • Ana Paula Vieira Colombo Universidade Federal do Rio de Janeiro, Instituto de Microbiologia
  • Carina Maciel Silva-Boghossian Universidade Federal do Rio de Janeiro, Faculdade de Odontologia, Departamento de Clínica Odontológica

DOI:

https://doi.org/10.1590/1678-7765-2025-0731

Abstract

Objective  To evaluate the effects of gastric administration of the highly leukotoxic Aggregatibacter actinomycetemcomitans JP2 strain (Aa JP2), with or without ligature-induced periodontitis, on periodontal and small intestinal tissues in a rat model. Methodology  Forty-eight-week-old male Wistar rats were divided into four groups (n=10/group): Control (CG), Periodontitis (PG), Periodontitis and Aa JP2 (PAaG), and Aa JP2 only (AaG). Ligatures were placed in PG and PAaG on day one and maintained for 42 days. After six weeks of twice-weekly gastric Aa JP2 inoculation, mandibles and small intestines were analyzed using stereomicroscopy, micro-computed tomography (CT), and histopathology to assess alveolar bone loss, trabecular architecture, inflammatory infiltrate, vascular congestion, goblet cell density, and villus morphology. Statistical analyses included the Kruskal–Wallis, Mann–Whitney, and Chi-square tests for group comparisons (p<0.05), as well as Spearman’s correlation test. Results  PG and PAaG exhibited significantly greater bone loss compared to CG and AaG (p<0.05). Micro-CT analysis revealed reduced trabecular thickness in AaG (342.4±23.1µm) compared to PG (378.2±28.1µm) and PAaG (385.5±45.1µm) (p<0.05). PAaG and AaG presented higher inflammatory infiltrate scores (>751 inflammatory cells) compared to CG and/or PG (p<0.05). Elevated vascular congestion and/or goblet cell hyperplasia in the jejunum and ileum were observed in PAaG and AaG compared to CG and PG (p<0.05). Villus height (duodenum/ileum) and villus width (jejunum/ileum) differed significantly among groups (p<0.0001). Significant correlations were identified: the alveolar bone volume/total volume ratio was positively associated with duodenal vascular congestion (rho=0.738; p=0.037); the periodontal inflammatory infiltrate was positively associated with jejunal goblet cell counts (rho=0.704; p=0.007); alveolar bone loss was positively associated with both duodenal and jejunal villus height (rho≥0.762; p<0.05); and bone volume was negatively associated with both jejunal vascular congestion (rho=-0.696; p=0.003) and jejunal goblet cell counts (rho=-0.617; p=0.011). Conclusions  Gastric inoculation with Aa JP2 exacerbates ligature-induced periodontitis and independently induces intestinal morphological changes.

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References

1- Papapanou PN, Sanz M, Buduneli N, Dietrich T, Feres M, Fine DH, et al. Periodontitis: Consensus report of workgroup 2 of the 2017 World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions. J Periodontol. 2018;89 Suppl 1:S173-S82. doi: 10.1002/JPER.17-0721

» https://doi.org/10.1002/JPER.17-0721

2- Colombo AP, Tanner AC. The role of bacterial biofilms in dental caries and periodontal and peri-implant diseases: a historical perspective. J Dent Res. 2019;98(4):373-85. doi: 10.1177/0022034519830686

» https://doi.org/10.1177/0022034519830686

3- Belibasakis GN, Maula T, Bao K, Lindholm M, Bostanci N, Orsarsson J, et al. Virulence and pathogenicity properties of Aggregatibacter actinomycetemcomitan s. Pathogens. 2019;8(4):222. doi: 10.3390/pathogens8040222

» https://doi.org/10.3390/pathogens8040222

4- Haubek D. The highly leukotoxic JP2 clone of Aggregatibacter actinomycetemcomitans: evolutionary aspects, epidemiology and etiological role in aggressive periodontitis. APMIS Suppl. 2010(130):1-53. doi: 10.1111/j.1600-0463.2010.02665.x

» https://doi.org/10.1111/j.1600-0463.2010.02665.x

5- Talapko J, Juzbasic M, Mestrovic T, Matijevic T, Mesaric D, Katalinic D, et al. Aggregatibacter actinomycetemcomitans: from the oral cavity to the heart valves. Microorganisms. 2024;12(7):1491. doi: 10.3390/microorganisms12071451

» https://doi.org/10.3390/microorganisms12071451

6- Vega BA, Schober LT, Kim T, Belinka BA Jr, Kachlany SC. Aggregatibacter actinomycetemcomitans leukotoxin (LtxA) requires death receptor Fas, in addition to LFA-1, to trigger cell death in T lymphocytes. Infect Immun. 2019;87(8):e00309-19. doi: 10.1128/IAI.00309-19

» https://doi.org/10.1128/IAI.00309-19

7- Sohn J, Sun Y, Genco RJ, Kirkwood KL. The periodontal microenvironment: a potential reservoir for intestinal pathobionts in Crohn's disease. Current Oral Health Reports. 2020;7:37-44. doi: 10.1007/s40496-020-00251-9

» https://doi.org/10.1007/s40496-020-00251-9

8- Kato T, Yamazaki K, Nakajima M, Date Y, Kikuchi J, Hase K, et al. Oral administration of Porphyromonas gingivalis alters the gut microbiome and serum metabolome. mSphere. 2018;3(5):e00460-18. doi: 10.1128/mSphere.00460-18

» https://doi.org/10.1128/mSphere.00460-18

9- Kitamoto S, Nagao-Kitamoto H, Jiao Y, Gillilland 3rd MG, Hayashi A, Imai J, et al. The intermucosal connection between the mouth and gut in commensal pathobiont-driven colitis. Cell. 2020;182(2):447-62 e14. doi: 10.1016/j.cell.2020.05.048

» https://doi.org/10.1016/j.cell.2020.05.048

10- Zilberstein NF, Engen PA, Swanson GR, Naqib A, Post Z, Alutto J, et al. The bidirectional effects of periodontal disease and oral dysbiosis on gut inflammation in inflammatory bowel disease. J Crohns Colitis. 2025;19(4):jjae162. doi: 10.1093/ecco-jcc/jjae162

» https://doi.org/10.1093/ecco-jcc/jjae162

11- Fine DH, Velliyagounder K, Furgang D, Kaplan JB. The Actinobacillus actinomycetemcomitans autotransporter adhesin Aae exhibits specificity for buccal epithelial cells from humans and old world primates. Infect Immun. 2005;73(4):1947-53. doi: 10.1128/IAI.73.4.1947-1953.2005

» https://doi.org/10.1128/IAI.73.4.1947-1953.2005

12- Yue G, Kaplan JB, Furgang D, Mansfield KG, Fine DH. A second Aggregatibacter actinomycetemcomitans autotransporter adhesin exhibits specificity for buccal epithelial cells in humans and Old World primates. Infect Immun. 2007;75(9):4440-8. doi: 10.1128/IAI.02020-06

» https://doi.org/10.1128/IAI.02020-06

13- Weiss EI, Shaniztki B, Dotan M, Ganeshkumar N, Kolenbrander PE, Metzger Z. Attachment of Fusobacterium nucleatum PK1594 to mammalian cells and its coaggregation with periodontopathogenic bacteria are mediated by the same galactose-binding adhesin. Oral Microbiol Immunol. 2000;15(6):371-7. doi: 10.1034/j.1399-302x.2000.150606.x

14- Ebersole JL, Graves CL, Gonzalez OA, Dawson D 3rd, Morford LA, Huja PE, et al. Aging, inflammation, immunity and periodontal disease. Periodontol 2000. 2016;72(1):54-75. doi: 10.1111/prd.12135

» https://doi.org/10.1111/prd.12135

15- Oliveira AM, Lourenco TG, Colombo AP. Impact of systemic probiotics as adjuncts to subgingival instrumentation on the oral-gut microbiota associated with periodontitis: a randomized controlled clinical trial. J Periodontol. 2022;93(1):31-44. doi: 10.1002/jper.21-0078

» https://doi.org/10.1002/jper.21-0078

16- Lourenco TG, Oliveira AM, Tsute Chen G, Colombo AP. Oral-gut bacterial profiles discriminate between periodontal health and diseases. J Periodontal Res. 2022;57(6):1227-37. doi: 10.1111/jre.13059

» https://doi.org/10.1111/jre.13059

17- Kuraji R, Ye C, Zhao C, Gao L, Martinez A, Miyashita Y, et al. Nisin lantibiotic prevents NAFLD liver steatosis and mitochondrial oxidative stress following periodontal disease by abrogating oral, gut and liver dysbiosis. NPJ Biofilms Microbiomes. 2024;10(1):3. doi: 10.1038/s41522-024-00476-x

» https://doi.org/10.1038/s41522-024-00476-x

18- Arimatsu K, Yamada H, Miyazawa H, Minagawa T, Nakajima M, Ryder MI, et al. Oral pathobiont induces systemic inflammation and metabolic changes associated with alteration of gut microbiota. Sci Rep. 2014;4:4828. doi: 10.1038/srep04828

» https://doi.org/10.1038/srep04828

19- Costa AL, Soares MA, Lourenco TG, Guimarães-Pinto K, Filardy AD, Oliveira AM, et al. Periodontal pathogen Aggregatibacter actinomycetemcomitans JP2 correlates with colonic leukocytes decrease and gut microbiome imbalance in mice. J Periodontal Res. 2024;59(5):961-73. doi: 10.1111/jre.13288

» https://doi.org/10.1111/jre.13288

20- Percie du Sert N, Hurst V, Ahluwalia A, Alam S, Avey MT, Baker M, et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLoS Biol. 2020;18(7):e3000410. doi: 10.1371/journal.pbio.3000410

» https://doi.org/10.1371/journal.pbio.3000410

21- Messora MR, Oliveira LF, Foureaux RC, Taba M Jr, Zangerônimo MG, Furlaneto FA, et al. Probiotic therapy reduces periodontal tissue destruction and improves the intestinal morphology in rats with ligature-induced periodontitis. J Periodontol. 2013;84(12):1818-26. doi: 10.1902/jop.2013.120644

» https://doi.org/10.1902/jop.2013.120644

22- Sallay K, Sanavi F, Ring I, Pham P, Behling UH, Nowotny A. Alveolar bone destruction in the immunosuppressed rat. J Periodontal Res. 1982;17(3):263-74. doi: 10.1111/j.1600-0765.1982.tb01153.x

» https://doi.org/10.1111/j.1600-0765.1982.tb01153.x

23- Kothari A, Rajagopalan P. The assembly of integrated rat intestinal-hepatocyte cultures. Bioeng Transl Med. 2020;5(1):e10146. doi: 10.1002/btm2.10146

» https://doi.org/10.1002/btm2.10146

24- Pereira JN, Murata GM, Sato FT, Marosti AR, Carvalho CR, Curi R. Small intestine remodeling in male Goto-Kakizaki rats. Physiol Rep. 2021;9(3):e14755. doi: 10.14814/phy2.14755

» https://doi.org/10.14814/phy2.14755

25- Silva VO, Lobato RV, Andrade EF, Macedo CG, Napimoga JT, Napimoga MH, et al. ß-glucans ( Saccharomyces cereviseae ) reduce glucose levels and attenuate alveolar bone loss in diabetic rats with periodontal disease. PLoS One. 2015;10(8):e0134742. doi: 10.1371/journal.pone.0134742

» https://doi.org/10.1371/journal.pone.0134742

26- Fine DH, Schreiner H, Velusamy SK. Aggregatibacter, a low abundance pathobiont that influences biogeography, microbial dysbiosis, and host defense capabilities in periodontitis: the history of a bug, and localization of disease. Pathogens. 2020;9(3):179. doi: 10.3390/pathogens9030179

» https://doi.org/10.3390/pathogens9030179

27- Frazao DR, Matos-Souza JM, Santos VR, Nazario RM, Chemelo VD, Bittencourt LO, et al. Minocycline reduces alveolar bone loss and bone damage in Wistar rats with experimental periodontitis. PLoS One. 2024;19(10):e0309390. doi: 10.1371/journal.pone.0309390

» https://doi.org/10.1371/journal.pone.0309390

28- Price U, Le HO, Powell SE, Schmid MJ, Marx DB, Zhang Y, et al. Effects of local simvastatin-alendronate conjugate in preventing periodontitis bone loss. J Periodontal Res. 2013;48(5):541-8. doi: 10.1111/jre.12036

» https://doi.org/10.1111/jre.12036

29- Pereira E Silva A, Soares JR, Mattos EB, Josetti C, Guimarães IM, Campos SM, et al. A histomorphometric classification system for normal and inflamed mouse duodenum-Quali-quantitative approach. Int J Exp Pathol. 2018;99(4):189-98. doi: 10.1111/iep.12286

» https://doi.org/10.1111/iep.12286

30- Esteves-Monteiro M, Ferreira-Duarte M, Vitorino-Oliveira C, Costa-Pires J, Oliveira S, Matafome P, et al. Oxidative stress and histomorphometric remodeling: two key intestinal features of type 2 diabetes in Goto-Kakizaki rats. Int J Mol Sci. 2024;25(22). doi: 10.3390/ijms252212115

» https://doi.org/10.3390/ijms252212115

31- Fiocchi C, Ina K, Danese S, Leite AZ, Vogel JD. Alterations of mesenchymal and endothelial cells in inflammatory bowel diseases. Adv Exp Med Biol. 2006;579:168-76. doi: 10.1007/0-387-33778-4_11

» https://doi.org/10.1007/0-387-33778-4_11

32- Corfield AP. Mucins: a biologically relevant glycan barrier in mucosal protection. Biochim Biophys Acta. 2015;1850(1):236-52. doi: 10.1016/j.bbagen.2014.05.003

» https://doi.org/10.1016/j.bbagen.2014.05.003

33- Deplancke B, Gaskins HR. Microbial modulation of innate defense: goblet cells and the intestinal mucus layer. Am J Clin Nutr. 2001;73(6):1131S-41S. doi: 10.1093/ajcn/73.6.1131S

» https://doi.org/10.1093/ajcn/73.6.1131S

34- Lauritano D, Boccalari E, Di Stasio D, Della Vella F, Carinci F, Lucchese A, et al. Prevalence of oral lesions and correlation with intestinal symptoms of inflammatory bowel disease: a systematic review. Diagnostics (Basel). 2019;9(3):77. doi: 10.3390/diagnostics9030077

» https://doi.org/10.3390/diagnostics9030077

35- Hassan GS, Helal MB, Ibrahim HF. Immunohistochemical expression of estrogen receptor alpha in the maxillary sinus, pulp, and periodontal ligament of adjacent teeth in late pregnancy in rats. Odontology. 2023;111(3):608-17. doi: 10.1007/s10266-022-00770-0

» https://doi.org/10.1007/s10266-022-00770-0

36- Rotstein I, Simon JH. Diagnosis, prognosis and decision-making in the treatment of combined periodontal-endodontic lesions. Periodontol 2000. 2004;34:165-203. doi: 10.1046/j.0906-6713.2003.003431.x

» https://doi.org/10.1046/j.0906-6713.2003.003431.x

37- Kuhr A, Popa-Wagner A, Schmoll H, Schwahn C, Kocher T. Observations on experimental marginal periodontitis in rats. J Periodontal Res 2004;39(2):101-6. doi: 10.1111/j.1600-0765.2004.00710.x

» https://doi.org/10.1111/j.1600-0765.2004.00710.x

38- Duarte PM, Tezolin KR, Figueiredo LC, Feres M, Bastos MF. Microbial profile of ligature-induced periodontitis in rats. Arch Oral Biol. 2010;55(2):142-7. doi: 10.1016/j.archoralbio.2009.10.006

» https://doi.org/10.1016/j.archoralbio.2009.10.006

39- Faber J, Fonseca LM. How sample size influences research outcomes. Dental Press J Orthod 2014;19(4):27-9. doi: 10.1590/2176-9451.19.4.027-029.ebo

» https://doi.org/10.1590/2176-9451.19.4.027-029.ebo

40- Bain JL, Lester SR, Henry WD, Bishop CM, Turnage AA, Naftel JP, et al. Comparative gender differences in local and systemic concentrations of pro-inflammatory cytokines in rats with experimental periodontitis. J Periodontal Res. 2009;44(1):133-40. doi: 10.1111/j.1600-0765.2008.01110.x

» https://doi.org/10.1111/j.1600-0765.2008.01110.x

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Published

2026-03-09

Issue

Vol. 34 (2026)

Section

Original Articles

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How to Cite

Ferraz, G. F. T., Camilo, C. P. de A., Miranda, A. L. P. de, Soares, M. A., Sansone, C., Lopes, R. T., Abrahão, A. C., Colombo, A. P. V., & Silva-Boghossian, C. M. (2026). Effects of highly leukotoxic Aggregatibacter actinomycetemcomitans associated with ligature-induced periodontitis on oral and gut tissues in male rats. Journal of Applied Oral Science, 34, e2025-0731. https://doi.org/10.1590/1678-7765-2025-0731