Hyperresponsiveness of human gingival fibroblasts from patients with aggressive periodontitis against bacterial lipopolysaccharide

Li,Xue; Wang,Xiaoxuan; Luan,Qing-Xian

doi:10.3892/etm.2021.9861

Hyperresponsiveness of human gingival fibroblasts from patients with aggressive periodontitis against bacterial lipopolysaccharide

Authors:
- Xue Li
- Xiaoxuan Wang
- Qing-Xian Luan
View Affiliations

Affiliations: Department of Periodontology, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, P.R. China
Published online on: February 25, 2021 https://doi.org/10.3892/etm.2021.9861
Article Number: 417

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to investigate whether gingival fibroblasts (GFs) of patients with aggressive periodontitis (AgP) are more sensitive to lipopolysaccharide (LPS) stimulation than GFs of control subjects. AgP causes rapid periodontal destruction, including the production of cytokines [i.e. interleukin (IL)‑1β, IL‑6 and tumor necrosis factor (TNF)‑α] and matrix metalloproteinases (MMP)‑1, ‑3 and ‑9 in AgP GFs. LPS upregulates IL‑1β, IL‑6, TNF‑α, MMP‑1, MMP‑3, MMP‑9 and mitochondrial reactive oxygen species (mtROS). Fibroblasts are known to be associated with immune responses to bacterial virulence factors, but the precise mechanisms underlying this severe periodontal disease are unclear. In the present study, primary human GFs of four patients with AgP and four healthy subjects were challenged in vitro with LPS from Porphyromonas gingivalis (P. gingivalis). The generation of mtROS in GFs was assessed using MitoSOX Red. The expression of genes encoding inflammatory cytokines and MMPs in GFs was analyzed using reverse transcription‑quantitative polymerase chain reaction, and the expression of proteins was analyzed using ELISA and Western blotting. Human GFs of patients with AgP exhibited higher levels of mtROS, and higher mRNA and protein expression levels of proinflammatory cytokines, including IL‑1β, IL‑6, MMP‑1, MMP‑3 and MMP‑9 compared with healthy human GFs following stimulation with LPS from P. gingivalis. In the present study, it was demonstrated that GFs of patients with AgP display hyperreactivity when challenged with LPS.

View Figures

View References

1	No authors listed. 1999 International International Workshop for a classification of periodontal diseases and conditions. Papers. Oak Brook, Illinois, October 30-November 2, 1999. Ann Periodontol 4: i. 1–112. 1999.PubMed/NCBI View Article : Google Scholar
2	Buckley CD, Pilling D, Lord JM, Akbar AN, Scheel-Toellner D and Salmon M: Fibroblasts regulate the switch from acute resolving to chronic persistent inflammation. Trends Immunol. 22:199–204. 2001.PubMed/NCBI View Article : Google Scholar
3	Ara T, Kurata K, Hirai K, Uchihashi T, Uematsu T, Imamura Y, Furusawa K, Kurihara S and Wang PL: Human gingival fibroblasts are critical in sustaining inflammation in periodontal disease. J Periodontal Res. 44:21–27. 2009.PubMed/NCBI View Article : Google Scholar
4	Flavell SJ, Hou TZ, Lax S, Filer AD, Salmon M and Buckley CD: Fibroblasts as novel therapeutic targets in chronic inflammation. Br J Pharmacol. 153 (Suppl 1):S241–S246. 2008.PubMed/NCBI View Article : Google Scholar
5	Dongari-Bagtzoglou AI and Ebersole JL: Increased presence of interleukin-6 (IL-6) and IL-8 secreting fibroblast subpopulations in adult periodontitis. J Periodontol. 69:899–910. 1998.PubMed/NCBI View Article : Google Scholar
6	Pathirana RD, O'Brien-Simpson NM and Reynolds EC: Host immune responses to Porphyromonas gingivalis antigens. Periodontol 2000. 52:218–237. 2010.PubMed/NCBI View Article : Google Scholar
7	Wilson M: Biological activities of lipopolysaccharides from oral bacteria and their relevance to the pathogenesis of chronic periodontitis. Sci Prog. 78:19–34. 1995.PubMed/NCBI
8	Garlet GP: Destructive and protective roles of cytokines in periodontitis: A re-appraisal from host defense and tissue destruction viewpoints. J Dent Res. 89:1349–1363. 2010.PubMed/NCBI View Article : Google Scholar
9	Liu J, Wang Y and Ouyang X: Beyond toll-like receptors: Porphyromonas gingivalis induces IL-6, IL-8, and VCAM-1 expression through NOD-mediated NF-KB and ERK signaling pathways in periodontal fibroblasts. Inflammation. 37:522–533. 2014.PubMed/NCBI View Article : Google Scholar
10	Li X, Wang X, Zheng M and Luan QX: Mitochondrial reactive oxygen species mediate the lipopolysaccharide-induced pro-inflammatory response in human gingival fibroblasts. Exp Cell Res. 347:212–221. 2016.PubMed/NCBI View Article : Google Scholar
11	Goncalves PF, Huang H, McAninley S, Alfant B, Harrison P, Aukhil I, Walker C and Shaddox LM: Periodontal treatment reduces matrix metalloproteinase levels in localized aggressive periodontitis. J Periodontol. 84:1801–1808. 2013.PubMed/NCBI View Article : Google Scholar
12	Seguier S, Gogly B, Bodineau A, Godeau G and Brousse N: Is collagen breakdown during periodontitis linked to inflammatory cells and expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gingival tissue? J Periodontol. 72:1398–1406. 2001.PubMed/NCBI View Article : Google Scholar
13	Henderson B, Poole S and Wilson M: Bacterial modulins: A novel class of virulence factors which cause host tissue pathology by inducing cytokine synthesis. Microbiol Rev. 60:316–341. 1996.PubMed/NCBI
14	Gustafsson A, Ito H, Asman B and Bergstrom K: Hyper-reactive mononuclear cells and neutrophils in chronic periodontitis. J Clin Periodontol. 33:126–129. 2006.PubMed/NCBI View Article : Google Scholar
15	Nicu EA, Van der Velden U, Everts V, Van Winkelhoff AJ, Roos D and Loos BG: Hyper-reactive PMNs in FcgammaRIIa 131 H/H genotype periodontitis patients. J Clin Periodontol. 34:938–945. 2007.PubMed/NCBI View Article : Google Scholar
16	Matthews JB, Wright HJ, Roberts A, Cooper PR and Chapple IL: Hyperactivity and reactivity of peripheral blood neutrophils in chronic periodontitis. Clin Exp Immunol. 147:255–264. 2007.PubMed/NCBI View Article : Google Scholar
17	Zhan Y, Lu R, Meng H, Wang X and Hou J: Platelet activation and platelet-leukocyte interaction in generalized aggressive periodontitis. J Leukoc Biol. 100:1155–1166. 2016.PubMed/NCBI View Article : Google Scholar
18	Nicu EA, Van der Velden U, Nieuwland R, Everts V and Loos BG: Elevated platelet and leukocyte response to oral bacteria in periodontitis. J Thromb Haemost. 7:162–170. 2009.PubMed/NCBI View Article : Google Scholar
19	Ogawa T, Ozaki A, Shimauchi H and Uchida H: Hyporesponsiveness of inflamed human gingival fibroblasts from patients with chronic periodontal diseases against cell surface components of Porphyromonas gingivalis. FEMS Immunol Med Microbiol. 18:17–30. 1997.PubMed/NCBI View Article : Google Scholar
20	Scheres N, Laine ML, Sipos PM, Bosch-Tijhof CJ, Crielaard W, de Vries TJ and Everts V: Periodontal ligament and gingival fibroblasts from periodontitis patients are more active in interaction with Porphyromonas gingivalis. J Periodontal Res. 46:407–416. 2011.PubMed/NCBI View Article : Google Scholar
21	Kang W, Hu Z and Ge S: Healthy and inflamed gingival fibroblasts differ in their inflammatory response to Porphyromonas gingivalis lipopolysaccharide. Inflammation. 39:1842–1852. 2016.PubMed/NCBI View Article : Google Scholar
22	Zdarilova A, Svobodova A, Simanek V and Ulrichova J: Prunella vulgaris extract and rosmarinic acid suppress lipopolysaccharide-induced alteration in human gingival fibroblasts. Toxicol In Vitro. 23:386–392. 2009.PubMed/NCBI View Article : Google Scholar
23	Wang PL, Ohura K, Fujii T, Oido-Mori M, Kowashi Y, Kikuchi M, Suetsugu Y and Tanaka J: DNA microarray analysis of human gingival fibroblasts from healthy and inflammatory gingival tissues. Biochem Biophys Res Commun. 305:970–973. 2003.PubMed/NCBI View Article : Google Scholar
24	Imatani T, Kato T and Okuda K: Production of inflammatory cytokines by human gingival fibroblasts stimulated by cell-surface preparations of Porphyromonas gingivalis. Oral Microbiol Immunol. 16:65–72. 2001.PubMed/NCBI View Article : Google Scholar
25	Kowaltowski AJ, de Souza-Pinto NC, Castilho RF and Vercesi AE: Mitochondria and reactive oxygen species. Free Radic Biol Med. 47:333–343. 2009.PubMed/NCBI View Article : Google Scholar
26	Yu T, Robotham JL and Yoon Y: Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology. Proc Natl Acad Sci USA. 103:2653–2658. 2006.PubMed/NCBI View Article : Google Scholar
27	Aguilera-Aguirre L, Bacsi A, Saavedra-Molina A, Kurosky A, Sur S and Boldogh I: Mitochondrial dysfunction increases allergic airway inflammation. J Immunol. 183:5379–5387. 2009.PubMed/NCBI View Article : Google Scholar
28	Vaamonde-Garcia C, Riveiro-Naveira RR, Valcarcel-Ares MN, Hermida-Carballo L, Blanco FJ and Lopez-Armada MJ: Mitochondrial dysfunction increases inflammatory responsiveness to cytokines in normal human chondrocytes. Arthritis Rheum. 64:2927–2936. 2012.PubMed/NCBI View Article : Google Scholar