Top

Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie

Published in:

Open Access 01-09-2020 | Original Article

Effects of sodium chloride on the gene expression profile of periodontal ligament fibroblasts during tensile strain

Authors: PD Dr. rer. nat. Agnes Schröder, Joshua Gubernator, Ute Nazet, Dr. Gerrit Spanier, MD, DDS, Jonathan Jantsch, Professor Dr. Dr. (PhD) Peter Proff, MD, DDS, PD Dr. Dr. (PhD) Christian Kirschneck, DDS

Published in: Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie | Issue 5/2020

Abstract

Purpose

During orthodontic tooth movement, pressure and tension zones develop in the periodontal ligament, and periodontal ligament fibroblasts (PDLF) become exposed to mechanical strain. Enhanced salt (NaCl) concentrations are known to modulate responses of PDLF and immune cells to different stimuli like mechanical strain. Here, we investigated the impact of tensile strain on the gene expression profile of PDLF under normal (NS) and high salt (HS) conditions.

Methods

After preincubation under NS or HS (+40 mM NaCl in medium) conditions for 24 h, PDLF were stretched 16% for 48 h using custom-made spherical cap silicone stamps using an established and published setup. After determination of cell number and cytotoxicity, we analyzed expression of genes involved in extracellular matrix reorganization, angiogenesis, bone remodeling, and inflammation by quantitative real-time polymerase chain reaction (RT-qPCR).

Results

Tensile strain did not affect the expression of genes involved in angiogenesis or extracellular matrix reorganization by PDLF, which however modulate inflammatory responses and bone remodeling in reaction to 16% static tensile strain. Salt (NaCl) treatment triggered enhanced extracellular matrix formation, expression of cyclooxygenase 2 and bone metabolism in PDLF during tensile strain.

Conclusions

Salt (NaCl) consumption may influence orthodontic tooth movement and periodontal bone loss via modulation of extracellular matrix and bone metabolism. Excessive salt intake during orthodontic therapy may cause adverse effects regarding periodontal inflammation and bone resorption.

Anastasi G, Cordasco G, Matarese G et al (2008) An immunohistochemical, histological, and electron-microscopic study of the human periodontal ligament during orthodontic treatment. Int J Mol Med 21:545–554PubMed

Bernhardt O, Krey K‑F, Daboul A et al (2019) New insights in the link between malocclusion and periodontal disease. J Clin Periodontol 46:144–159PubMedCrossRef

Binger KJ, Gebhardt M, Heinig M et al (2015) High salt reduces the activation of IL-4- and IL-13-stimulated macrophages. J Clin Invest 125:4223–4238PubMedPubMedCentralCrossRef

Birkedal-Hansen H, Moore WG, Bodden MK et al (1993) Matrix metalloproteinases: a review. Crit Rev Oral Biol Med 4:197–250PubMedCrossRef

Bolcato-Bellemin AL, Elkaim R, Abehsera A et al (2000) Expression of mRNAs encoding for alpha and beta integrin subunits, MMPs, and TIMPs in stretched human periodontal ligament and gingival fibroblasts. J Dent Res 79:1712–1716PubMedCrossRef

Deutsche Gesellschaft für Ernährung e.V. (DGE) (2017) https://www.dge.de/presse/pm/dge-aktualisiert-die-referenzwerte-fuer-natrium-chlorid-und-kalium/

Di Domenico M, D’apuzzo F, Feola A et al (2012) Cytokines and VEGF induction in orthodontic movement in animal models. J Biomed Biotechnol 2012:201689PubMedPubMedCentralCrossRef

Dutra SR, Pretti H, Martins MT et al (2018) Impact of malocclusion on the quality of life of children aged 8 to 10 years. Dental Press J Orthod 23:46–53PubMedPubMedCentralCrossRef

Favale NO, Casali CI, Lepera LG et al (2009) Hypertonic induction of COX2 expression requires TonEBP/NFAT5 in renal epithelial cells. Biochem Biophys Res Commun 381:301–305PubMedCrossRef

10.

Grundt A, Grafe IA, Liegibel U et al (2009) Direct effects of osteoprotegerin on human bone cell metabolism. Biochem Biophys Res Commun 389:550–555PubMedCrossRef

11.

Howard PS, Kucich U, Taliwal R et al (1998) Mechanical forces alter extracellular matrix synthesis by human periodontal ligament fibroblasts. J Periodontal Res 33:500–508PubMedCrossRef

12.

Jacobs C, Grimm S, Ziebart T et al (2013) Osteogenic differentiation of periodontal fibroblasts is dependent on the strength of mechanical strain. Arch Oral Biol 58:896–904PubMedCrossRef

13.

Jacobs C, Walter C, Ziebart T et al (2014) Induction of IL‑6 and MMP‑8 in human periodontal fibroblasts by static tensile strain. Clin Oral Investig 18:901–908PubMedCrossRef

14.

Jantsch J, Schatz V, Friedrich D et al (2015) Cutaneous Na+ storage strengthens the antimicrobial barrier function of the skin and boosts macrophage-driven host defense. Cell Metab 21:493–501PubMedPubMedCentralCrossRef

15.

Jeyraj Y, Katta AK, Vannala V et al (2015) Estimation of alkaline phosphatase in the gingival crevicular fluid during orthodontic tooth movement in premolar extraction cases to predict therapeutic progression. J Nat Sci Biol Med 6:343–346PubMedPubMedCentralCrossRef

16.

Kanzaki H, Chiba M, Shimizu Y et al (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

17.

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

18.

Kirschneck C, Maurer M, Wolf M et al (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

19.

Kirschneck C, Proff P, Fanghänel J et al (2016) Reference genes for valid gene expression studies on rat dental, periodontal and alveolar bone tissue by means of RT-qPCR with a focus on orthodontic tooth movement and periodontitis. Ann Anat 204:93–105PubMedCrossRef

20.

Kurihara N, Bertolini D, Suda T et al (1990) IL‑6 stimulates osteoclast-like multinucleated cell formation in long term human marrow cultures by inducing IL‑1 release. J Immunol 144:4226–4230PubMed

21.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(‑Delta Delta C(T)) Method. Methods 25:402–408CrossRefPubMed

22.

Machnik A, Dahlmann A, Kopp C et al (2010) Mononuclear phagocyte system depletion blocks interstitial tonicity-responsive enhancer binding protein/vascular endothelial growth factor C expression and induces salt-sensitive hypertension in rats. Hypertension 55:755–761PubMedCrossRef

23.

Machnik A, Neuhofer W, Jantsch J et al (2009) Macrophages regulate salt-dependent volume and blood pressure by a vascular endothelial growth factor-C-dependent buffering mechanism. Nat Med 15:545–552PubMedCrossRef

24.

Megat Abdul Wahab R, Md Dasor M, Senafi S et al (2013) Crevicular alkaline phosphatase activity and rate of tooth movement of female orthodontic subjects under different continuous force applications. Int J Dent 2013:245818PubMedPubMedCentralCrossRef

25.

Meikle MC (2006) The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod 28:221–240PubMedCrossRef

26.

Militi A, Cutroneo G, Favaloro A et al (2019) An immunofluorescence study on VEGF and extracellular matrix proteins in human periodontal ligament during tooth movement. Heliyon 5:e2572PubMedPubMedCentralCrossRef

27.

Miyagawa A, Chiba M, Hayashi H et al (2009) Compressive force induces VEGF production in periodontal tissues. J Dent Res 88:752–756PubMedCrossRef

28.

Müller DN, Wilck N, Haase S et al (2019) Sodium in the microenvironment regulates immune responses and tissue homeostasis. Nat Rev Immunol 19:243–254PubMedCrossRef

29.

Muluke M, Gold T, Kiefhaber K et al (2016) Diet-induced obesity and its differential impact on periodontal bone loss. J Dent Res 95:223–229PubMedPubMedCentralCrossRef

30.

Nazet U, Schröder A, Spanier G et al (2019) Simplified method for applying static isotropic tensile strain in cell culture experiments with identification of valid RT-qPCR reference genes for PDL fibroblasts. Eur J Orthod. https://doi.org/10.1093/ejo/cjz052CrossRef

31.

Nettelhoff L, Grimm S, Jacobs C et al (2016) Influence of mechanical compression on human periodontal ligament fibroblasts and osteoblasts. Clin Oral Investig 20(3):621–629PubMedCrossRef

32.

Neubert P, Weichselbaum A, Reitinger C et al (2019) HIF1A and NFAT5 coordinate Na+-boosted antibacterial defense via enhanced autophagy and autolysosomal targeting. Autophagy 15:1899–1916PubMedPubMedCentralCrossRef

33.

Neuhofer W, Woo SK, Na KY et al (2002) Regulation of TonEBP transcriptional activator in MDCK cells following changes in ambient tonicity. Am J Physiol Cell Physiol 283:C1604–C1611PubMedCrossRef

34.

Okada N, Kobayashi M, Mugikura K et al (1997) Interleukin‑6 production in human fibroblasts derived from periodontal tissues is differentially regulated by cytokines and a glucocorticoid. J Periodont Res 32:559–569CrossRef

35.

Perinetti G, Paolantonio M, D’Attilio M et al (2002) Alkaline phosphatase activity in gingival crevicular fluid during human orthodontic tooth movement. Am J Orthod Dentofacial Orthop 122:548–556PubMedCrossRef

36.

Romberger DJ (1997) Fibronectin. Int J Biochem Cell Biol 29:939–943PubMedCrossRef

37.

Sá-Pinto AC, Rego TM, Marques LS et al (2018) Association between malocclusion and dental caries in adolescents: a systematic review and meta-analysis. Eur Arch Paediatr Dent 19:73–82PubMedCrossRef

38.

Schröder A, Bauer K, Spanier G et al (2018) Expression kinetics of human periodontal ligament fibroblasts in the early phases of orthodontic tooth movement (Expression kinetics of human periodontal ligament fibroblasts in the early phases of orthodontic tooth movement). J Orofac Orthop 79:337–351. https://doi.org/10.1007/s00056-018-0145-1CrossRefPubMed

39.

Schröder A, Nazet U, Neubert P et al (2019) Sodium-chloride-induced effects on the expression profile of human periodontal ligament fibroblasts with focus on simulated orthodontic tooth movement. Eur J Oral Sci 127:386–395PubMedCrossRef

40.

Shimizu N, Ozawa Y, Yamaguchi M et al (1998) Induction of COX‑2 expression by mechanical tension force in human periodontal ligament cells. J Periodontol 69:670–677PubMedCrossRef

41.

Staufer K, Landmesser H (2004) Effects of crowding in the lower anterior segment—a risk evaluation depending upon the degree of crowding. J Orofac Orthop 65:13–25PubMedCrossRef

42.

Taibah SM, Al-Hummayani FM (2017) Effect of malocclusion on the self-esteem of adolescents. J Orthod Sci 6:123–128PubMedPubMedCentralCrossRef

43.

Takahashi I, Nishimura M, Onodera K et al (2003) Expression of MMP‑8 and MMP-13 genes in the periodontal ligament during tooth movement in rats. J Dent Res 82:646–651PubMedCrossRef

44.

Titze J, Rittweger J, Dietsch P et al (2004) Hypertension, sodium retention, calcium excretion and osteopenia in Dahl rats. J Hypertens 22:803–810PubMedCrossRef

45.

Titze J, Shakibaei M, Schafflhuber M et al (2004) Glycosaminoglycan polymerization may enable osmotically inactive Na+ storage in the skin. Am J Physiol Heart Circ Physiol 287:H203–H208PubMedCrossRef

46.

Tsubota M, Sasano Y, Takahashi I et al (2002) Expression of MMP‑8 and MMP-13 mRNAs in rat periodontium during tooth eruption. J Dent Res 81:673–678PubMedCrossRef

47.

Tyrovola JB, Spyropoulos MN, Makou M et al (2008) Root resorption and the OPG/RANKL/RANK system: a mini review. J Oral Sci 50:367–376PubMedCrossRef

48.

Ullrich N, Schröder A, Jantsch J et al (2019) The role of mechanotransduction versus hypoxia during simulated orthodontic compressive strain-an in vitro study of human periodontal ligament fibroblasts. Int J Oral Sci 11:33PubMedPubMedCentralCrossRef

49.

Volpi N, Maccari F, Titze J (2005) Simultaneous detection of submicrogram quantities of hyaluronic acid and dermatan sulfate on agarose-gel by sequential staining with toluidine blue and Stains-All. J Chromatogr B Analyt Technol Biomed Life Sci 820:131–135PubMedCrossRef

50.

Wescott DC, Pinkerton MN, Gaffey BJ et al (2007) Osteogenic gene expression by human periodontal ligament cells under cyclic tension. J Dent Res 86:1212–1216PubMedCrossRef

51.

WHO (2012) WHO guideling: sodium intake for adults and children. ISBN: 9789241504836

52.

Woelber JP, Bremer K, Vach K et al (2016) An oral health optimized diet can reduce gingival and periodontal inflammation in humans—a randomized controlled pilot study. BMC Oral Health 17:28PubMedPubMedCentralCrossRef

53.

Wolf M, Lossdörfer S, Römer 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

54.

Wu L, Luthringer BJC, Feyerabend F et al (2017) Increased levels of sodium chloride directly increase osteoclastic differentiation and resorption in mice and men. Osteoporos Int 28:3215–3228PubMedPubMedCentralCrossRef

55.

Yu H‑S, Kim J‑J, Kim H‑W et al (2016) Impact of mechanical stretch on the cell behaviors of bone and surrounding tissues. J Tissue Eng. https://doi.org/10.1177/2041731415618342CrossRefPubMedPubMedCentral

Title: Effects of sodium chloride on the gene expression profile of periodontal ligament fibroblasts during tensile strain
Authors: PD Dr. rer. nat. Agnes Schröder
Joshua Gubernator
Ute Nazet
Dr. Gerrit Spanier, MD, DDS
Jonathan Jantsch
Professor Dr. Dr. (PhD) Peter Proff, MD, DDS
PD Dr. Dr. (PhD) Christian Kirschneck, DDS
Publication date: 01-09-2020
Publisher: Springer Medizin
Published in: Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie / Issue 5/2020
Print ISSN: 1434-5293
Electronic ISSN: 1615-6714
DOI: https://doi.org/10.1007/s00056-020-00232-8

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Effects of sodium chloride on the gene expression profile of periodontal ligament fibroblasts during tensile strain

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 5/2020

Computerized occlusal analysis of Essix and Hawley retainers used during the retention phase: a controlled clinical trial

Evaluation of bond strength of molar orthodontic tubes subjected to reinforcement with flowable and bonding resins

Suitability of virtual plaster models superimposed with the lateral cephalogram for guided paramedian orthodontic mini-implant placement with regard to the bone support

Mitteilungen der DGKFO

Controlling incisor torque with completely customized lingual appliances

Stepwise versus single-step mandibular advancement with functional appliance in treating class II patients