nach oben

Erschienen in:

06.11.2019 | Original Article

Orthodontic cell stress modifies proinflammatory cytokine expression in human PDL cells and induces immunomodulatory effects via TLR-4 signaling in vitro

verfasst von: Jana Marciniak, Stefan Lossdörfer, Isabel Knaup, Asisa Bastian, Rogerio B. Craveiro, Andreas Jäger, Michael Wolf

Erschienen in: Clinical Oral Investigations | Ausgabe 4/2020

Einloggen, um Zugang zu erhalten

Abstract

Objective

Biomechanical orthodontics loading of the periodontium initiates a cascade of inflammatory signaling events that induce periodontal remodeling and finally facilitate orthodontic tooth movement. Pattern recognition receptors such as toll-like receptors (TLRs) have been well characterized for their ability to induce the activation of inflammatory, immunomodulatory cytokines. Here, we examined whether the cellular response of human periodontal ligament (hPDL) cells to mechanical stress involves TLR-4 signaling in vitro.

Materials and methods

Confluent hPDL cells were cultured in the presence of 5 μg/ml TLR-4 antibody (TLR-4ab) for 1 h prior to the induction of compressive forces by the use of round glass plates for 24 h. At harvest, interleukin-6 and interleukin-8 (IL-6, IL-8) mRNA and protein expression were analyzed by real-time PCR and ELISA. The immunomodulatory role of mechanical cell stress and TLR-4 signaling was addressed in co-culture experiments of hPDL and THP-1 cells targeting monocyte adhesion and by culturing osteoclastic precursors (RAW 264.7) in the presence of the conditioned medium of hPDL cells that had been mechanically loaded before.

Results

Basal expression of IL-6 and IL-8 was not affected by TLR-4ab, but increased significantly upon mechanical loading of hPDL cells. When cells were mechanically stressed in the presence of TLR-4ab, the effect seen for loading alone was markedly reduced. Likewise, monocyte adhesion and osteoclastic differentiation were enhanced significantly by mechanical stress of hPDL cells and this effect was partially inhibited by TLR-4ab.

Conclusions

The results of the present study indicate a proinflammatory and immunomodulatory influence of mechanical loading on hPDL cells. Intracellular signaling involves a TLR-4-dependent pathway.

Clinical relevance

These findings hold out the prospect of interfering with the cellular response to mechanical cell stress in order to minimize undesired side effects of orthodontic tooth movement.

Nakao K, Goto T, Gunjigake KK, Konoo T, Kobayashi S, Yamaguchi K (2007) Intermittent force induces high RANKL expression in human periodontal ligament cells. J Dent Res 86:623–628PubMedCrossRef

Pinkerton MN, Wescott DC, Gaffey BJ, Beggs KT, Milne TJ, Meikle MC (2008) Cultured human periodontal ligament cells constitutively express multiple osteotropic cytokines and growth factors, several of which are responsive to mechanical deformation. J Periodontal Res 43:343–351PubMedCrossRef

Wolf M, Lossdorfer S, Craveiro R et al (2014) High-mobility group box protein-1 released by human-periodontal ligament cells modulates macrophage migration and activity in vitro. Innate Immun 20:688–696PubMedCrossRef

Wolf M, Lossdorfer S, Kupper K, Jager A (2014) Regulation of high mobility group box protein 1 expression following mechanical loading by orthodontic forces in vitro and in vivo. Eur J Orthod 36:624–631PubMedCrossRef

Jäger A, Radlanski RJ, Götz W (1993) Demonstration of cells of the mononuclear phagocyte lineage in the periodontium following experimental tooth movement in the rat. An immunohistochemical study using monoclonal antibodies ED1 und ED2 on paraffin-embedded tissues. Histochemistry 100:161–166PubMedCrossRef

Kim YS, Lee YM, Park JS et al (2010) SIRT1 modulates high-mobility group box 1-induced osteoclastogenic cytokines in human periodontal ligament cells. J Cell Biochem 111:1310–1320PubMedCrossRef

Wolf M, Lossdorfer S, Romer P et al (2016) Short-term heat pre-treatment modulates the release of HMGB1 and pro-inflammatory cytokines in hPDL cells following mechanical loading and affects monocyte behavior. Clin Oral Investig 20:923–931PubMedCrossRef

Medzhitov R (2007) Recognition of microorganisms and activation of the immune response. Nature 449:819–826PubMed

Sun Y, Shu R, Zhang MZ et al (2008) Toll-like receptor 4 signaling plays a role in triggering periodontal infection. FEMS Immunol Med Microbiol 52:362–369PubMedCrossRef

10.

Takeuchi O, Akira S (2010) Pattern recognition receptors and inflammation. Cell 140:805–820PubMedCrossRef

11.

Wang PL, Ohura K, Fujii T, Oido-Mori M, Kowashi Y, Kikuchi M, Suetsugu Y, Tanaka J (2003) DNA microarray analysis of human gingival fibroblasts from healthy and inflammatory gingival tissues. Biochem Biophys Res Commun 305:970–973PubMedCrossRef

12.

Kinane DF, Shiba H, Stathopoulou PG, Zhao H, Lappin DF, Singh A, Eskan MA, Beckers S, Waigel S, Alpert B, Knudsen TB (2006) Gingival epithelial cells heterozygous for toll-like receptor 4 polymorphisms Asp299Gly and Thr399ile are hypo-responsive to Porphyromonas gingivalis. Genes Immun 7:190–200PubMedCrossRef

13.

Hatakeyama J, Tamai R, Sugiyama A et al (2003) Contrasting responses of human gingival and periodontal ligament fibroblasts to bacterial cell-surface components through the CD14/toll-like receptor system. Oral Microbiol Immunol 18:14–23PubMedCrossRef

14.

Tang L, Zhou XD, Wang Q, Zhang L, Wang Y, Li XY, Huang DM (2011) Expression of TRAF6 and pro-inflammatory cytokines through activation of TLR2, TLR4, NOD1, and NOD2 in human periodontal ligament fibroblasts. Arch Oral Biol 56:1064–1072PubMedCrossRef

15.

Hoshino K, Takeuchi O, Kawai T et al (1999) Cutting edge: toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J Immunol 162:3749–3752PubMed

16.

Gordon S (2002) Pattern recognition receptors: doubling up for the innate immune response. Cell 111:927–930PubMedCrossRef

17.

Lee SI, Park KH, Kim SJ, Kang YG, Lee YM, Kim EC (2012) Mechanical stress-activated immune response genes via Sirtuin 1 expression in human periodontal ligament cells. Clin Exp Immunol 168:113–124PubMedPubMedCentralCrossRef

18.

Yamaba S, Yamada S, Kajikawa T, Awata T, Sakashita H, Tsushima K, Fujihara C, Yanagita M, Murakami S (2015) PLAP-1/Asporin regulates TLR2- and TLR4-induced inflammatory responses. J Dent Res 94:1706–1714PubMedCrossRef

19.

Chen GY, Nunez G (2010) Sterile inflammation: sensing and reacting to damage. Nat Rev Immunol 10:826–837PubMedPubMedCentralCrossRef

20.

Nogueira AV, Nokhbehsaim M, Eick S, Bourauel C, Jäger A, Jepsen S, Cirelli JA, Deschner J (2014) Regulation of visfatin by microbial and biomechanical signals in PDL cells. Clin Oral Investig 18:171–178PubMedCrossRef

21.

Boas Nogueira AV, Chaves de Souza JA, Kim YJ, Damião de Sousa-Neto M, Chan Cirelli C, Cirelli JA (2013) Orthodontic force increases interleukin-1beta and tumor necrosis factor-alpha expression and alveolar bone loss in periodontitis. J Periodontol 84:1319–1326PubMedCrossRef

22.

Kirschneck C, Maurer M, Wolf M, Reicheneder C, Proff P (2017) Regular nicotine intake increased tooth movement velocity, osteoclastogenesis and orthodontically induced dental root resorptions in a rat model. Int J Oral Sci 9:174–184PubMedPubMedCentralCrossRef

23.

Kirschneck C, Fanghanel J, Wahlmann U et al (2017) Interactive effects of periodontitis and orthodontic tooth movement on dental root resorption, tooth movement velocity and alveolar bone loss in a rat model. Ann Anat 210:32–43PubMedCrossRef

24.

Lossdörfer S, Götz W, Jäger A (2011) PTH(1-34)-induced changes in RANKL and OPG expression by human PDL cells modify osteoclast biology in a co-culture model with RAW 264.7 cells. Clin Oral Investig 15:941–952PubMedCrossRef

25.

Wolf M, Lossdörfer S, Römer P et al (2014) Anabolic properties of high mobility group box protein-1 in human periodontal ligament cells in vitro. Mediat Inflamm 2014:347585CrossRef

26.

Samelko L, Landgraeber S, McAllister K et al (2016) Cobalt alloy implant debris induces inflammation and bone loss primarily through danger signaling, not TLR4 activation: implications for DAMP-ening implant related inflammation. PLoS One 11:e0160141PubMedPubMedCentralCrossRef

27.

Kanzaki H, Chiba M, Shimizu Y, Mitani H (2002) Periodontal ligament cells under mechanical stress induce osteoclastogenesis by receptor activator of nuclear factor kappaB ligand up-regulation via prostaglandin E2 synthesis. J Bone Miner Res 17:210–220PubMedCrossRef

28.

Konermann A, Stabenow D, Knolle PA, Held SA, Deschner J, Jäger A (2012) Regulatory role of periodontal ligament fibroblasts for innate immune cell function and differentiation. Innate Immun 18:745–752PubMedCrossRef

29.

Romer P, Wolf M, Fanghanel J et al (2014) Cellular response to orthodontically-induced short-term hypoxia in dental pulp cells. Cell Tissue Res 355:173–180PubMedCrossRef

30.

Kraus D, Deschner J, Jager A et al (2012) Human beta-defensins differently affect proliferation, differentiation, and mineralization of osteoblast-like MG63 cells. J Cell Physiol 227:994–1003PubMedCrossRef

31.

Winning S, Splettstoesser F, Fandrey J et al (2010) Acute hypoxia induces HIF-independent monocyte adhesion to endothelial cells through increased intercellular adhesion molecule-1 expression: the role of hypoxic inhibition of prolyl hydroxylase activity for the induction of NF-kappa B. J Immunol 185:1786–1793PubMedCrossRef

32.

Biswas SK, Tergaonkar V (2007) Myeloid differentiation factor 88-independent toll-like receptor pathway: sustaining inflammation or promoting tolerance? Int J Biochem Cell Biol 39:1582–1592PubMedCrossRef

33.

Medvedev AE, Lentschat A, Wahl LM et al (2002) Dysregulation of LPS-induced toll-like receptor 4-MyD88 complex formation and IL-1 receptor-associated kinase 1 activation in endotoxin-tolerant cells. J Immunol 169:5209–5216PubMedCrossRef

34.

Piao W, Song C, Chen H, Wahl LM, Fitzgerald KA, O'Neill LA, Medvedev AE (2008) Tyrosine phosphorylation of MyD88 adapter-like (Mal) is critical for signal transduction and blocked in endotoxin tolerance. J Biol Chem 283:3109–3119PubMedCrossRef

35.

Brown J, Wang H, Hajishengallis GN, Martin M (2011) TLR-signaling networks: an integration of adaptor molecules, kinases, and cross-talk. J Dent Res 90:417–427PubMedPubMedCentralCrossRef

36.

Bildt MM, Bloemen M, Kuijpers-Jagtman AM et al (2009) Matrix metalloproteinases and tissue inhibitors of metalloproteinases in gingival crevicular fluid during orthodontic tooth movement. Eur J Orthod 31:529–535PubMedCrossRef

37.

Ren Y, Vissink A (2008) Cytokines in crevicular fluid and orthodontic tooth movement. Eur J Oral Sci 116:89–97PubMedCrossRef

38.

Goto KT, Kajiya H, Nemoto T, Tsutsumi T, Tsuzuki T, Sato H, Okabe K (2011) Hyperocclusion stimulates osteoclastogenesis via CCL2 expression. J Dent Res 90:793–798PubMedCrossRef

39.

Kook SH, Jang YS (1985) Lee JC (2011) involvement of JNK-AP-1 and ERK-NF-kappaB signaling in tension-stimulated expression of type I collagen and MMP-1 in human periodontal ligament fibroblasts. J Appl Physiol 111:1575–1583CrossRef

40.

Liedert A, Kaspar D, Blakytny R, Claes L, Ignatius A (2006) Signal transduction pathways involved in mechanotransduction in bone cells. Biochem Biophys Res Commun 349:1–5PubMedCrossRef

41.

Whitmarsh AJ, Davis RJ (1996) Transcription factor AP-1 regulation by mitogen-activated protein kinase signal transduction pathways. J Mol Med (Berl) 74:589–607CrossRef

42.

Inoh H, Ishiguro N, Sawazaki S, Amma H, Miyazu M, Iwata H, Sokabe M, Naruse K (2002) Uni-axial cyclic stretch induces the activation of transcription factor nuclear factor kappaB in human fibroblast cells. FASEB J 16:405–407PubMedCrossRef

43.

Liu J, Zou L, Zheng Y, Zhao Z, Li Y, Yang P, Luo S (2007) NF-kappaB responds to mechanical strains in osteoblast-like cells, and lighter strains create an NF-kappaB response more readily. Cell Biol Int 31:1220–1224PubMedCrossRef

44.

Ning Q, Wang X (2007) Role of Rel A and IkappaB of nuclear factor kappaB in the release of interleukin-8 by cyclic mechanical strain in human alveolar type II epithelial cells A549. Respirology 12:792–798PubMedCrossRef

45.

Kirschneck C, Batschkus S, Proff P et al (2017) Valid gene expression normalization by RT-qPCR in studies on hPDL fibroblasts with focus on orthodontic tooth movement and periodontitis. Sci Rep 7:14751PubMedPubMedCentralCrossRef

Titel: Orthodontic cell stress modifies proinflammatory cytokine expression in human PDL cells and induces immunomodulatory effects via TLR-4 signaling in vitro
verfasst von: Jana Marciniak
Stefan Lossdörfer
Isabel Knaup
Asisa Bastian
Rogerio B. Craveiro
Andreas Jäger
Michael Wolf
Publikationsdatum: 06.11.2019
Verlag: Springer Berlin Heidelberg
Erschienen in: Clinical Oral Investigations / Ausgabe 4/2020
Print ISSN: 1432-6981
Elektronische ISSN: 1436-3771
DOI: https://doi.org/10.1007/s00784-019-03111-8

Bestellen Sie unseren kostenlosen Newsletter Update Zahnmedizin und bleiben Sie gut informiert – ganz bequem per eMail.

Newsletter bestellen

Springer Medizin

Orthodontic cell stress modifies proinflammatory cytokine expression in human PDL cells and induces immunomodulatory effects via TLR-4 signaling in vitro