Periodontitis may induce gut microbiota dysbiosis via salivary microbiota

Abstract

The aim of this study was to identify whether periodontitis induces gut microbiota dysbiosis via invasion by salivary microbes. First, faecal and salivary samples were collected from periodontally healthy participants (PH group, n = 16) and patients with severe periodontitis (SP group, n = 21) and analysed by 16S ribosomal RNA sequencing. Significant differences were observed in both the faecal and salivary microbiota between the PH and SP groups. Notably, more saliva-sourced microbes were observed in the faecal samples of the SP group. Then, the remaining salivary microbes were transplanted into C57BL6/J mice (the C-PH group and the C-SP group), and it was found that the composition of the gut microbiota of the C-SP group was significantly different from that of the C-PH group, with Porphyromonadaceae and Fusobacterium being significantly enriched in the C-SP group. In the colon, the C-SP group showed significantly reduced crypt depth and zonula occludens-1 expression. The mRNA expression levels of pro-inflammatory cytokines, chemokines and tight junction proteins were significantly higher in the C-SP group. To further investigate whether salivary bacteria could persist in the intestine, the salivary microbiota was stained with carboxyfluorescein diacetate succinimidyl ester and transplanted into mice. We found that salivary microbes from both the PH group and the SP group could persist in the gut for at least 24 h. Thus, our data demonstrate that periodontitis may induce gut microbiota dysbiosis through the influx of salivary microbes.

References

[1] Kassebaum NJ, . Global burden of severe periodontitis in 1990-2010: a systematic review and meta-regression. J. Dent. Res., 2014, 93: 1045-1053.

[2] Kleinstein SE, Nelson KE, Freire M. Inflammatory networks linking oral microbiome with systemic health and disease. J. Dent. Res., 2020, 99: 1131-1139.

[3] Li L, . Gut microbiota may mediate the influence of periodontitis on prediabetes. J. Dent. Res., 2021, 100: 1387-1396.

[4] Huang Y, . Non-surgical periodontal treatment restored the gut microbiota and intestinal barrier in apolipoprotein E(-/-) mice with periodontitis. Front. Cell Infect. Microbiol., 2020, 10: 498.

[5] Qian J, . Periodontitis salivary microbiota Worsens colitis. J. Dent. Res., 2022, 101: 559-568.

[6] Gaffen SL, Moutsopoulos NM. Regulation of host-microbe interactions at oral mucosal barriers by type 17 immunity. Sci. Immunol., 2020, 5: eaau4594.

[7] Fine N, . Periodontal inflammation primes the systemic innate immune response. J. Dent. Res., 2021, 100: 318-325.

[8] Humphrey SP, Williamson RT. A review of saliva: normal composition, flow, and function. J. Prosthet. Dent., 2001, 85: 162-169.

[9] Slots J. Periodontitis: facts, fallacies and the future. Periodontol 2000, 2017, 75: 7-23.

[10] Zhang Y, . Human oral microbiota and its modulation for oral health. Biomed. Pharmacother., 2018, 99: 883-893.

[11] Zhang Y, . The association between periodontitis and inflammatory bowel disease: a systematic review and meta-analysis. BioMed. Res. Int., 2021, 2021: 6692420.

[12] Lourenςo TGB, Spencer SJ, Alm EJ, Colombo APV. Defining the gut microbiota in individuals with periodontal diseases: an exploratory study. J. Oral. Microbiol., 2018, 10: 1487741.

[13] Schmidt TS, . Extensive transmission of microbes along the gastrointestinal tract. Elife, 2019, 8: e42693.

[14] Saygun I, . Salivary infectious agents and periodontal disease status. J. Periodontal Res., 2011, 46: 235-239.

[15] Socransky SS, . Microbial complexes in subgingival plaque. J. Clin. Periodontol., 1998, 25: 134-144.

[16] Lourenço TG, . Microbial signature profiles of periodontally healthy and diseased patients. J. Clin. Periodontol., 2014, 41: 1027-1036.

[17] Abusleme L, Hoare A, Hong BY, Diaz PI. Microbial signatures of health, gingivitis, and periodontitis. Periodontol 2000, 2021, 86: 57-78.

[18] Willis JR, Gabaldon T. The human oral microbiome in health and disease: from sequences to ecosystems. Microorganisms, 2020, 8: 308.

[19] Belstrom D. The salivary microbiota in health and disease. J. Oral. Microbiol., 2020, 12: 1723975.

[20] Solbiati J, Frias-Lopez J. Metatranscriptome of the oral microbiome in health and disease. J. Dent. Res., 2018, 97: 492-500.

[21] Imai J, . A potential pathogenic association between periodontal disease and Crohn’s disease. JCI Insight, 2021, 6: e148543.

[22] Cai Z, . Co-pathogens in periodontitis and inflammatory bowel disease. Front. Med., 2021, 8: 723719.

[23] Tsuzuno T, . Ingestion of Porphyromonas gingivalis exacerbates colitis via intestinal epithelial barrier disruption in mice. J. Periodontal. Res., 2021, 56: 275-288.

[24] Read E, Curtis MA, Neves JF. The role of oral bacteria in inflammatory bowel disease. Nat. Rev. Gastroenterol. Hepatol., 2021, 18: 731-742.

[25] Șurlin P, . Could periodontal disease through periopathogen fusobacterium nucleatum be an aggravating factor for gastric cancer?. J. Clin. Med., 2020, 9: 3885.

[26] Engevik MA, . Fusobacteriumnucleatum adheres to Clostridioides difficile via the RadD adhesin to enhance biofilm formation in intestinal mucus. Gastroenterology, 2021, 160: 1301-1314 e1308.

[27] Strauss J, . Invasive potential of gut mucosa-derived Fusobacterium nucleatum positively correlates with IBD status of the host. Inflamm. Bowel Dis., 2011, 17: 1971-1978.

[28] Liu H, . Fusobacterium nucleatum exacerbates colitis by damaging epithelial barriers and inducing aberrant inflammation. J. Dig. Dis., 2020, 21: 385-398.

[29] Huh JW, Roh TY. Opportunistic detection of Fusobacterium nucleatum as a marker for the early gut microbial dysbiosis. BMC Microbiol., 2020, 20

[30] Cao X. Intestinal inflammation induced by oral bacteria. Science, 2017, 358: 308-309.

[31] Ohkusa T, . Commensal bacteria can enter colonic epithelial cells and induce proinflammatory cytokine secretion: a possible pathogenic mechanism of ulcerative colitis. J. Med. Microbiol., 2009, 58: 535-545.

[32] Dharmani P, . Fusobacterium nucleatum infection of colonic cells stimulates MUC2 mucin and tumor necrosis factor alpha. Infect. Immun., 2011, 79: 2597-2607.

[33] Arimatsu K, . Oral pathobiont induces systemic inflammation and metabolic changes associated with alteration of gut microbiota. Sci. Rep., 2014, 4

[34] Kitamoto S, . The intermucosal connection between the mouth and gut in commensal pathobiont-driven colitis. Cell, 2020, 182: 447-462.

[35] Garcia-Hernandez V, Quiros M, Nusrat A. Intestinal epithelial claudins: expression and regulation in homeostasis and inflammation. Ann. N. Y Acad. Sci., 2017, 1397: 66-79.

[36] Robinson K, Deng Z, Hou Y, Zhang G. Regulation of the intestinal barrier function by host defense peptides. Front. Vet. Sci., 2015, 2: 57.

[37] Koch F, . Heat stress directly impairs gut integrity and recruits distinct immune cell populations into the bovine intestine. Proc. Natl. Acad. Sci. USA, 2019, 116: 10333-10338.

[38] Obradovic M, Pasternak JA, Ng SH, Wilson HL. Use of flow cytometry and PCR analysis to detect 5-carboxyfluoroscein-stained obligate intracellular bacteria Lawsonia intracellularis invasion of McCoy cells. J. Microbiol. Methods, 2016, 126: 60-66.

[39] Kuraji R, Sekino S, Kapila Y, Numabe Y. Periodontal disease-related nonalcoholic fatty liver disease and nonalcoholic steatohepatitis: an emerging concept of oral-liver axis. Periodontol 2000, 2021, 87: 204-240.

[40] Atarashi K, . Ectopic colonization of oral bacteria in the intestine drives TH1 cell induction and inflammation. Science, 2017, 358: 359-365.

[41] Nakajima M, . Oral administration of P. gingivalis induces dysbiosis of gut microbiota and impaired barrier function leading to dissemination of Enterobacteria to the liver. PLoS ONE, 2015, 10: e0134234.

[42] Komazaki R, . Periodontal pathogenic bacteria, Aggregatibacter actinomycetemcomitans affect non-alcoholic fatty liver disease by altering gut microbiota and glucose metabolism. Sci. Rep., 2017, 7

[43] Li B, . Oral bacteria colonize and compete with gut microbiota in gnotobiotic mice. Int. J. Oral. Sci., 2019, 11: 10.

[44] Zhou D, . Total fecal microbiota transplantation alleviates high-fat diet-induced steatohepatitis in mice via beneficial regulation of gut microbiota. Sci. Rep., 2017, 7

[45] Hubálek Z. Protectants used in the cryopreservation of microorganisms. Cryobiology, 2003, 46: 205-229.

[46] Raval NC, Wadhwani CP, Jain S, Darveau RP. The interaction of implant luting cements and oral bacteria linked to peri-implant disease: an in vitro analysis of planktonic and biofilm growth–A preliminary study. Clin. implant Dent. Relat. Res., 2015, 17: 1029-1035.

[47] Amato KR, . Habitat degradation impacts black howler monkey (Alouatta pigra) gastrointestinal microbiomes. ISME J., 2013, 7: 1344-1353.

[48] Pragman AA, . The lung tissue microbiota of mild and moderate chronic obstructive pulmonary disease. Microbiome, 2018, 6

[49] Henry R, . Into the deep: evaluation of SourceTracker for assessment of faecal contamination of coastal waters. Water Res., 2016, 93: 242-253.

[50] McCarthy DT, . Source tracking using microbial community fingerprints: method comparison with hydrodynamic modelling. Water Res., 2017, 109: 253-265.

[51] Staley C, . Stable engraftment of human microbiota into mice with a single oral gavage following antibiotic conditioning. Microbiome, 2017, 5

[52] Meng Y, . Lactobacillus plantarum KLDS1.0318 ameliorates impaired intestinal immunity and metabolic disorders in cyclophosphamide-treated mice. Front. Microbiol., 2019, 10: 731.

[53] Zhou F, . Electroacupuncture attenuated anxiety and depression-like behavior via inhibition of hippocampal inflammatory response and metabolic disorders in TNBS-induced IBD rats. Oxid. Med. Cell. Longev., 2022, 2022: 8295580.

[54] Blasco-Baque V, . Periodontitis induced by Porphyromonas gingivalis drives periodontal microbiota dysbiosis and insulin resistance via an impaired adaptive immune response. Gut, 2017, 66: 872-885.

Funding

The present study was supported by the National Science Foundation of China (No. 81970939), Nanjing Clinical Research Center for Oral Diseases (No. 2019060009).