Top

Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie

Published in:

Open Access 21-04-2022 | Shock | Original Article

Impact of phosphorylation of heat shock protein 27 on the expression profile of periodontal ligament fibroblasts during mechanical strain

Authors: PD Dr. rer. nat. Agnes Schröder, Kathrin Wagner, PD Dr. Fabian Cieplik, DDS, Dr. Gerrit Spanier, MD, DDS, Professor Dr. Dr. Peter Proff, MD, DDS, PD Dr. Dr. Christian Kirschneck, DDS

Published in: Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie | Special Issue 2/2023

Abstract

Purpose

Orthodontic tooth movement is a complex process involving the remodeling of extracellular matrix and bone as well as inflammatory processes. During orthodontic treatment, sterile inflammation and mechanical loading favor the production of receptor activator of NF-κB ligand (RANKL). Simultaneously, expression of osteoprotegerin (OPG) is inhibited. This stimulates bone resorption on the pressure side. Recently, heat shock protein 27 (HSP27) was shown to be expressed in the periodontal ligament after force application and to interfere with inflammatory processes.

Methods

We investigated the effects of phosphorylated HSP27 on collagen synthesis (COL1A2 mRNA), inflammation (IL1B mRNA, IL6 mRNA, PTGS2 protein) and bone remodeling (RANKL protein, OPG protein) in human periodontal ligament fibroblasts (PDLF) without and with transfection of a plasmid mimicking permanent phosphorylation of HSP27 using real-time quantitative polymerase chain reaction (RT-qPCR), western blot and enzyme-linked immunosorbent assays (ELISAs). Furthermore, we investigated PDLF-induced osteoclastogenesis after compressive strain in a co-culture model with human macrophages.

Results

In particular, phosphorylated HSP27 increased gene expression of COL1A2 and protein expression of PTGS2, while IL6 mRNA levels were reduced. Furthermore, we observed an increasing effect on the RANKL/OPG ratio and osteoclastogenesis mediated by PDLF.

Conclusion

Phosphorylation of HSP27 may therefore be involved in the regulation of orthodontic tooth movement by impairment of the sterile inflammation response and osteoclastogenesis.

Available only for authorised users

Adhikari AS, Sridhar Rao K, Rangaraj N et al (2004) Heat stress-induced localization of small heat shock proteins in mouse myoblasts: intranuclear lamin A/C speckles as target for alphaB-crystallin and Hsp25. Exp Cell Res 299(2):393–403. https://doi.org/10.1016/j.yexcr.2004.05.032CrossRefPubMed

Alderson TR, Roche J, Gastall HY et al (2019) Local unfolding of the HSP27 monomer regulates chaperone activity. Nat Commun 10(1):1–16. https://doi.org/10.1038/s41467-019-08557-8CrossRef

Bozkaya E, Canigur Bavbek N, Isler SC et al (2021) Evaluation of heat shock protein 70 and toll-like receptor 4 expression in gingival crevicular fluid in response to orthodontic forces. Clin Oral Investig. https://doi.org/10.1007/s00784-021-04014-3CrossRefPubMed

Bruey JM, Paul C, Fromentin A et al (2000) Differential regulation of HSP27 oligomerization in tumor cells grown in vitro and in vivo. Oncogene 19(42):4855–4863. https://doi.org/10.1038/sj.onc.1203850CrossRefPubMed

Charette SJ, Landry J (2000) The interaction of HSP27 with Daxx identifies a potential regulatory role of HSP27 in Fas-induced apoptosis. Ann N Y Acad Sci 926:126–131CrossRefPubMed

Deng H, Gao X, Peng H et al (2018) Effect of liposome-mediated HSP27 transfection on collagen synthesis in alveolar type II epithelial cells. Mol Med Rep 17(5):7319–7324PubMed

Giuliano F, Warner TD (2002) Origins of prostaglandin E2: involvements of cyclooxygenase (COX)-1 and COX‑2 in human and rat systems. J Pharmacol Exp Ther 303(3):1001–1006CrossRefPubMed

Hansen RK, Parra I, Lemieux P et al (1999) Hsp27 overexpression inhibits doxorubicin-induced apoptosis in human breast cancer cells. Breast Cancer Res Treat 56(2):187–196CrossRefPubMed

Hirano S, Shelden EA, Gilmont RR (2004) HSP27 regulates fibroblast adhesion, motility, and matrix contraction. Cell Stress Chaperones 9(1):29–37CrossRefPubMedPubMedCentral

10.

Jeon HH, Teixeira H, Tsai A (2021) Mechanistic insight into orthodontic tooth movement based on animal studies: a critical review. J Clin Med 10(8):1733. https://doi.org/10.3390/jcm10081733CrossRefPubMedPubMedCentral

11.

Jovcevski B, Kelly MA, Rote AP et al (2015) Phosphomimics destabilize Hsp27 oligomeric assemblies and enhance chaperone activity. Chem Biol 22(2):186–195. https://doi.org/10.1016/j.chembiol.2015.01.001CrossRefPubMed

12.

Kampinga HH, Brunsting JF, Stege GJ et al (1994) Cells overexpressing Hsp27 show accelerated recovery from heat-induced nuclear protein aggregation. Biochem Biophys Res Commun 204(3):1170–1177. https://doi.org/10.1006/bbrc.1994.2586CrossRefPubMed

13.

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(1):14751. https://doi.org/10.1038/s41598-017-15281-0CrossRefPubMedPubMedCentral

14.

Klein Y, Fleissig O, Polak D et al (2020) Immunorthodontics: in vivo gene expression of orthodontic tooth movement. Sci Rep 10(1):8172. https://doi.org/10.1038/s41598-020-65089-8CrossRefPubMedPubMedCentral

15.

Kostenko S, Moens U (2009) Heat shock protein 27 phosphorylation: kinases, phosphatases, functions and pathology. Cell Mol Life Sci 66(20):3289–3307. https://doi.org/10.1007/s00018-009-0086-3CrossRefPubMed

16.

Li S, Zhang H, Li S et al (2015) Connexin 43 and ERK regulate tension-induced signal transduction in human periodontal ligament fibroblasts. J Orthop Res 33(7):1008–1014. https://doi.org/10.1002/jor.22830CrossRefPubMed

17.

Li X, Han L, Nookaew I et al (2019) Stimulation of Piezo1 by mechanical signals promotes bone anabolism. Elife. https://doi.org/10.7554/eLife.49631CrossRefPubMedPubMedCentral

18.

Liu X‑H, Kirschenbaum A, Yao S et al (2006) Interactive effect of interleukin‑6 and prostaglandin E2 on osteoclastogenesis via the OPG/RANKL/RANK system. Ann N Y Acad Sci 1068:225–233. https://doi.org/10.1196/annals.1346.047CrossRefPubMed

19.

Liu Z, Xi D, Kang M et al (2012) Molecular cloning and characterization of Hsp27.6: the first reported small heat shock protein from Apis cerana cerana. Cell Stress Chaperones 17(5):539–551. https://doi.org/10.1007/s12192-012-0330-xCrossRefPubMedPubMedCentral

20.

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(4):402–408. https://doi.org/10.1006/meth.2001.1262CrossRefPubMed

21.

Marciniak J, Lossdörfer S, Kirschneck C et al (2019) Heat shock protein 70 dampens the inflammatory response of human PDL cells to mechanical loading in vitro. J Periodontal Res 54(5):481–488. https://doi.org/10.1111/jre.12648CrossRefPubMed

22.

Meikle MC (2006) The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod 28(3):221–240. https://doi.org/10.1093/ejo/cjl001CrossRefPubMed

23.

Muraoka R, Nakano K, Kurihara S et al (2010) Immunohistochemical expression of heat shock proteins in the mouse periodontal tissues due to orthodontic mechanical stress. Eur J Med Res 15(11):475–482. https://doi.org/10.1186/2047-783x-15-11-475CrossRefPubMedPubMedCentral

24.

Muraoka R, Nakano K, Tsujigiwa H et al (2019) Involvement of heat-shock proteins during periodontal ligament remodeling. In: Manakil J (ed) Periodontology and dental implantology. IntechOpen,

25.

Pan J, Wang T, Wang L et al (2014) Cyclic strain-induced cytoskeletal rearrangement of human periodontal ligament cells via the Rho signaling pathway. PLoS One 9(3):e91580. https://doi.org/10.1371/journal.pone.0091580CrossRefPubMedPubMedCentral

26.

Ren Y, Hazemeijer H, de Haan B et al (2007) Cytokine profiles in crevicular fluid during orthodontic tooth movement of short and long durations. J Periodontol 78(3):453–458. https://doi.org/10.1902/jop.2007.060261CrossRefPubMed

27.

Rocchi P, So A, Kojima S et al (2004) Heat shock protein 27 increases after androgen ablation and plays a cytoprotective role in hormone-refractory prostate cancer. Cancer Res 64(18):6595–6602. https://doi.org/10.1158/0008-5472.CAN-03-3998CrossRefPubMed

28.

Rogalla T, Ehrnsperger M, Preville X et al (1999) Regulation of Hsp27 oligomerization, chaperone function, and protective activity against oxidative stress/tumor necrosis factor alpha by phosphorylation. J Biol Chem 274(27):18947–18956. https://doi.org/10.1074/jbc.274.27.18947CrossRefPubMed

29.

Samali A, Cotter TG (1996) Heat shock proteins increase resistance to apoptosis. Exp Cell Res 223(1):163–170. https://doi.org/10.1006/excr.1996.0070CrossRefPubMed

30.

Santos A, Bakker AD, Zandieh-Doulabi B et al (2010) Early activation of the beta-catenin pathway in osteocytes is mediated by nitric oxide, phosphatidyl inositol‑3 kinase/Akt, and focal adhesion kinase. Biochem Biophys Res Commun 391(1):364–369. https://doi.org/10.1016/j.bbrc.2009.11.064CrossRefPubMed

31.

Schröder A (2018) Expression kinetics of human periodontal ligament fibroblasts in the early phases of orthodontic tooth movement. J Orofac Orthop 79(5):337–351. https://doi.org/10.1007/s00056-018-0145-1CrossRefPubMed

32.

Shen Y, Pan Y, Guo S et al (2020) The roles of mechanosensitive ion channels and associated downstream MAPK signaling pathways in PDLC mechanotransduction. Mol Med Rep 21(5):2113–2122. https://doi.org/10.3892/mmr.2020.11006CrossRefPubMedPubMedCentral

33.

Shoji-Matsunaga A, Ono T, Hayashi M et al (2017) Osteocyte regulation of orthodontic force-mediated tooth movement via RANKL expression. Sci Rep 7(1):8753. https://doi.org/10.1038/s41598-017-09326-7CrossRefPubMedPubMedCentral

34.

Stetler RA, Gao Y, Signore AP et al (2009) HSP27: mechanisms of cellular protection against neuronal injury. Curr Mol Med 9(7):863–872. https://doi.org/10.2174/156652409789105561CrossRefPubMedPubMedCentral

35.

Stope MB, Weiss M, Preuss M et al (2014) Immediate and transient phosphorylation of the heat shock protein 27 initiates chemoresistance in prostate cancer cells. Oncol Rep 32(6):2380–2386. https://doi.org/10.3892/or.2014.3492CrossRefPubMed

36.

Sur R, Lyte PA, Southall MD (2008) Hsp27 regulates pro-inflammatory mediator release in keratinocytes by modulating NF-kappaB signaling. J Invest Dermatol 128(5):1116–1122. https://doi.org/10.1038/sj.jid.5701157CrossRefPubMed

37.

Tomoda M, Nakano K, Muraoka R et al (2012) Immunohistochemical changes of heat shock protein 27 expression in the mouse periodontal tissues exposed to orthodontic mechanical stress. J Hard Tissue Biol 21(1):43–50. https://doi.org/10.2485/jhtb.21.43CrossRef

38.

Udagawa N, Koide M, Nakamura M et al (2021) Osteoclast differentiation by RANKL and OPG signaling pathways. J Bone Miner Metab 39(1):19–26. https://doi.org/10.1007/s00774-020-01162-6CrossRefPubMed

39.

Uematsu S, Mogi M, Deguchi T (1996) Interleukin (IL)-1 beta, IL‑6, tumor necrosis factor-alpha, epidermal growth factor, and beta 2‑microglobulin levels are elevated in gingival crevicular fluid during human orthodontic tooth movement. J Dent Res 75(1):562–567. https://doi.org/10.1177/00220345960750010801CrossRefPubMed

40.

Wang X, Su Y, Deb K et al (2004) Prostaglandin E2 is a product of induced prostaglandin-endoperoxide synthase 2 and microsomal-type prostaglandin E synthase at the implantation site of the hamster. J Biol Chem 279(29):30579–30587. https://doi.org/10.1074/jbc.M400573200CrossRefPubMed

41.

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(5):923–931. https://doi.org/10.1007/s00784-015-1580-7CrossRefPubMed

42.

Wolf M, Marciniak J, Lossdörfer S et al (2019) Role of HSP70 protein in human periodontal ligament cell function and physiology. Ann Anat 221:76–83. https://doi.org/10.1016/j.aanat.2018.09.006CrossRefPubMed

43.

Wu Q, Zhou X, Huang D et al (2017) IL‑6 enhances osteocyte-mediated osteoclastogenesis by promoting JAK2 and RANKL activity in vitro. Cell Physiol Biochem 41(4):1360–1369. https://doi.org/10.1159/000465455CrossRefPubMed

44.

Xu C, Fan Z, Shan W et al (2012) Cyclic stretch influenced expression of membrane connexin 43 in human periodontal ligament cell. Arch Oral Biol 57(12):1602–1608. https://doi.org/10.1016/j.archoralbio.2012.07.002CrossRefPubMed

45.

Yamaguchi M (2009) RANK/RANKL/OPG during orthodontic tooth movement. Orthod Craniofac Res 12(2):113–119. https://doi.org/10.1111/j.1601-6343.2009.01444.xCrossRefPubMed

46.

Yan Y, Liu F, Kou X et al (2015) T cells are required for orthodontic tooth movement. J Dent Res 94(10):1463–1470. https://doi.org/10.1177/0022034515595003CrossRefPubMed

47.

Yang C‑Y, Jeon HH, Alshabab A et al (2018) RANKL deletion in periodontal ligament and bone lining cells blocks orthodontic tooth movement. Int J Oral Sci 10(1):3. https://doi.org/10.1038/s41368-017-0004-8CrossRefPubMedPubMedCentral

48.

Young SRL, Gerard-O’Riley R, Kim J‑B et al (2009) Focal adhesion kinase is important for fluid shear stress-induced mechanotransduction in osteoblasts. J Bone Miner Res 24(3):411–424. https://doi.org/10.1359/jbmr.081102CrossRefPubMed

49.

Zhang Y, Wang X, Wang S et al (2020) Heat shock protein 27 regulates the inflammatory response of intestinal epithelial cells by the nuclear factor-κB pathway. Dig Dis Sci. https://doi.org/10.1007/s10620-020-06074-zCrossRefPubMedPubMedCentral

50.

Zininga T, Ramatsui L, Shonhai A (2018) Heat shock proteins as immunomodulants. Molecules. https://doi.org/10.3390/molecules23112846CrossRefPubMedPubMedCentral

Title: Impact of phosphorylation of heat shock protein 27 on the expression profile of periodontal ligament fibroblasts during mechanical strain
Authors: PD Dr. rer. nat. Agnes Schröder
Kathrin Wagner
PD Dr. Fabian Cieplik, DDS
Dr. Gerrit Spanier, MD, DDS
Professor Dr. Dr. Peter Proff, MD, DDS
PD Dr. Dr. Christian Kirschneck, DDS
Publication date: 21-04-2022
Publisher: Springer Medizin
Keywords: Shock
Shock
Published in: Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie / Issue Special Issue 2/2023
Print ISSN: 1434-5293
Electronic ISSN: 1615-6714
DOI: https://doi.org/10.1007/s00056-022-00391-w

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Impact of phosphorylation of heat shock protein 27 on the expression profile of periodontal ligament fibroblasts during mechanical strain

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Special Issue 2/2023

Does puberty affect oxidative stress levels and antioxidant activity of saliva in patients with fixed orthodontic appliances?

Comparative assessment of dental and basal arch dimensions of passive and active self-ligating versus conventional appliances

Relationship between craniofacial and dental arch morphology with pharyngeal airway space in adolescents

In vitro study of structural and mechanical properties of latex and non-latex intermaxillary orthodontic elastics

Periodontal health of unilateral labially vs. palatally impacted maxillary canines erupted by closed eruption technique

Effects of miniscrew-facilitated micro-osteoperforations on the rate of orthodontic tooth movement