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01-06-2016 | Original Article

Altered macrophagic THP-1 cell phagocytosis and migration in bisphosphonate-related osteonecrosis of the jaw (BRONJ)

Authors: Sebastian Hoefert, Claudia Sade Hoefert, Adelheid Munz, Hinnak Northoff, Anna Yuan, Kathrin Reichenmiller, Siegmar Reinert, Martin Grimm

Published in: Clinical Oral Investigations | Issue 5/2016

Abstract

Objectives

Local immune dysfunction via macrophages is a proposed aetiology of bisphosphonate-related osteonecrosis of the jaw (BRONJ). This study aimed to clarify the effects of various bisphosphonates on macrophage function using a THP-1 monocytic model to examine migration, phagocytosis, and fibrin structure.

Materials and methods

THP-1 cell migration was measured in the presence and absence of zoledronate, ibandronate, risedronate, alendronate, pamidronate (0.5, 5 and 50 μM) and clodronate (125, 250 and 500 μM) using the real-time xCELLigence system. Phagocytosis and actin fibre assays were performed after 72 h with zoledronate, ibandronate, alendronate and clodronate.

Results

Time to maximum migration for THP-1 cells was significantly reduced (p < 0.05) for high dosages of zoledronate, ibandronate and alendronate compared to controls. All dosages of clodronate and a low dose of zoledronate exhibited prolonged migrations. Phagocytic capacity was significantly reduced in high dosages of all bisphosphonates and for 5 μM zoledronate and ibandronate (p < 0.05). Low bisphosphonate exposure was accompanied by overcharged phagosoms. Altered appearance in F-actin fibrin structure was observed in bisphosphonate-exposed cells.

Conclusions

All bisphosphonates altered the migration of THP-1 cells dose-dependently. Low doses also prolonged migration and altered cell morphology. These findings support the idea of a disturbed local immune function of macrophages even in jaw bone exposed to low concentrations of bisphosphonate.

Clinical relevance

These are the first real-time results for disrupted migration and function of macrophagic THP-1 cells in high doses. Low dosages also demonstrated altered macrophage phagocytosis and cell morphology, suggesting a disturbed local immune function in BRONJ pathogenesis.

Ruggiero SL, Dodson TB, Fantasia J, Goodday R, Aghaloo T, Mehrotra B, O’Ryan F (2014) American Association of Oral and Maxillofacial Surgeons position paper on medication-related osteonecrosis of the jaw-2014 update. J Oral Maxillofac Surg 72(10):1938–1956. doi:10.1016/j.joms.2014.04.031 CrossRefPubMed

Russell RG (2011) Bisphosphonates: the first 40 years. Bone 49(1):2–19. doi:10.1016/j.bone.2011.04.022 CrossRefPubMed

Fantasia JE (2009) Bisphosphonates—what the dentist needs to know: practical considerations. J Oral Maxillofac Surg 67(5 Suppl):53–60. doi:10.1016/j.joms.2009.01.011 CrossRefPubMed

Stopeck AT, Lipton A, Body JJ, Steger GG, Tonkin K, de Boer RH, Lichinitser M, Fujiwara Y, Yardley DA, Viniegra M, Fan M, Jiang Q, Dansey R, Jun S, Braun A (2010) Denosumab compared with zoledronic acid for the treatment of bone metastases in patients with advanced breast cancer: a randomized, double-blind study. J Clin Oncol 28(35):5132–5139. doi:10.1200/JCO.2010.29.7101 CrossRefPubMed

Saad F, Brown JE, Van Poznak C, Ibrahim T, Stemmer SM, Stopeck AT, Diel IJ, Takahashi S, Shore N, Henry DH, Barrios CH, Facon T, Senecal F, Fizazi K, Zhou L, Daniels A, Carriere P, Dansey R (2012) Incidence, risk factors, and outcomes of osteonecrosis of the jaw: integrated analysis from three blinded active-controlled phase III trials in cancer patients with bone metastases. Ann Oncol 23(5):1341–1347. doi:10.1093/annonc/mdr435 CrossRefPubMed

Russell RG, Watts NB, Ebetino FH, Rogers MJ (2008) Mechanisms of action of bisphosphonates: similarities and differences and their potential influence on clinical efficacy. Osteoporos Int 19(6):733–759. doi:10.1007/s00198-007-0540-8 CrossRefPubMed

Frith JC, Rogers MJ (2003) Antagonistic effects of different classes of bisphosphonates in osteoclasts and macrophages in vitro. J Bone Miner Res 18(2):204–212. doi:10.1359/jbmr.2003.18.2.204 CrossRefPubMed

Wheeler AP, Wells CM, Smith SD, Vega FM, Henderson RB, Tybulewicz VL, Ridley AJ (2006) Rac1 and Rac2 regulate macrophage morphology but are not essential for migration. J Cell Sci 119(Pt 13):2749–2757. doi:10.1242/jcs.03024 CrossRefPubMed

Rougerie P, Miskolci V, Cox D (2013) Generation of membrane structures during phagocytosis and chemotaxis of macrophages: role and regulation of the actin cytoskeleton. Immunol Rev 256(1):222–239. doi:10.1111/imr.12118 CrossRefPubMed

10.

Pixley FJ (2012) Macrophage migration and its regulation by CSF-1. Int J Cell Biol 2012:501962. doi:10.1155/2012/501962 CrossRefPubMedPubMedCentral

11.

Guasch RM, Scambler P, Jones GE, Ridley AJ (1998) RhoE regulates actin cytoskeleton organization and cell migration. Mol Cell Biol 18(8):4761–4771CrossRefPubMedPubMedCentral

12.

Hackam DJ, Rotstein OD, Schreiber A, Zhang W, Grinstein S (1997) Rho is required for the initiation of calcium signaling and phagocytosis by Fcgamma receptors in macrophages. J Exp Med 186(6):955–966CrossRefPubMedPubMedCentral

13.

Russell RG, Xia Z, Dunford JE, Oppermann U, Kwaasi A, Hulley PA, Kavanagh KL, Triffitt JT, Lundy MW, Phipps RJ, Barnett BL, Coxon FP, Rogers MJ, Watts NB, Ebetino FH (2007) Bisphosphonates: an update on mechanisms of action and how these relate to clinical efficacy. Ann N Y Acad Sci 1117:209–257. doi:10.1196/annals.1402.089 CrossRefPubMed

14.

Hoefert S, Schmitz I, Tannapfel A, Eufinger H (2010) Importance of microcracks in etiology of bisphosphonate-related osteonecrosis of the jaw: a possible pathogenetic model of symptomatic and non-symptomatic osteonecrosis of the jaw based on scanning electron microscopy findings. Clin Oral Investig 14(3):271–284. doi:10.1007/s00784-009-0300-6 CrossRefPubMed

15.

Otto S, Hafner S, Mast G, Tischer T, Volkmer E, Schieker M, Sturzenbaum SR, von Tresckow E, Kolk A, Ehrenfeld M, Pautke C (2010) Bisphosphonate-related osteonecrosis of the jaw: is pH the missing part in the pathogenesis puzzle? J Oral Maxillofac Surg 68(5):1158–1161. doi:10.1016/j.joms.2009.07.079 CrossRefPubMed

16.

Sato M, Grasser W (1990) Effects of bisphosphonates on isolated rat osteoclasts as examined by reflected light microscopy. J Bone Miner Res 5(1):31–40. doi:10.1002/jbmr.5650050107 CrossRefPubMed

17.

Hoefert S, Hoefert CS, Albert M, Munz A, Grimm M, Northoff H, Reinert S, Alexander D (2014) Zoledronate but not denosumab suppresses macrophagic differentiation of THP-1 cells. An aetiologic model of bisphosphonate-related osteonecrosis of the jaw (BRONJ). Clin Oral Investig. doi:10.1007/s00784-014-1358-3

18.

Miyagawa F, Tanaka Y, Yamashita S, Minato N (2001) Essential requirement of antigen presentation by monocyte lineage cells for the activation of primary human gamma delta T cells by aminobisphosphonate antigen. J Immunol 166(9):5508–5514CrossRefPubMed

19.

Langermans JA, Hazenbos WL, van Furth R (1994) Antimicrobial functions of mononuclear phagocytes. J Immunol Methods 174(1–2):185–194CrossRefPubMed

20.

Jeganathan S, Fiorino C, Naik U, Sun HS, Harrison RE (2014) Modulation of osteoclastogenesis with macrophage M1- and M2-inducing stimuli. PLoS One 9(8), e104498. doi:10.1371/journal.pone.0104498 CrossRefPubMedPubMedCentral

21.

Chambers TJ (1980) Diphosphonates inhibit bone resorption by macrophages in vitro. J Pathol 132(3):255–262. doi:10.1002/path.1711320307 CrossRefPubMed

22.

Pazianas M (2011) Osteonecrosis of the jaw and the role of macrophages. J Natl Cancer Inst 103(3):232–240. doi:10.1093/jnci/djq516 CrossRefPubMed

23.

van Rooijen N, Hendrikx E (2010) Liposomes for specific depletion of macrophages from organs and tissues. Methods Mol Biol 605:189–203. doi:10.1007/978-1-60327-360-2_13 CrossRefPubMed

24.

Hoefert S, Schmitz I, Weichert F, Gaspar M, Eufinger H (2014) Macrophages and bisphosphonate-related osteonecrosis of the jaw (BRONJ): evidence of local immunosuppression of macrophages in contrast to other infectious jaw diseases. Clin Oral Investig. doi:10.1007/s00784-014-1273-7

25.

Friedl P, Zanker KS, Brocker EB (1998) Cell migration strategies in 3-D extracellular matrix: differences in morphology, cell matrix interactions, and integrin function. Microsc Res Tech 43(5):369–378. doi:10.1002/(SICI)1097-0029(19981201)43:5<369::AID-JEMT3>3.0.CO;2-6 CrossRefPubMed

26.

Boff RC, Salum FG, Figueiredo MA, Cherubini K (2014) Important aspects regarding the role of microorganisms in bisphosphonate-related osteonecrosis of the jaws. Arch Oral Biol 59(8):790–799. doi:10.1016/j.archoralbio.2014.05.002 CrossRefPubMed

27.

Ridley AJ, Hall A (1992) The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70(3):389–399CrossRefPubMed

28.

Chen WT (1981) Mechanism of retraction of the trailing edge during fibroblast movement. J Cell Biol 90(1):187–200CrossRefPubMed

29.

Herre J, Marshall AS, Caron E, Edwards AD, Williams DL, Schweighoffer E, Tybulewicz V, Reis e Sousa C, Gordon S, Brown GD (2004) Dectin-1 uses novel mechanisms for yeast phagocytosis in macrophages. Blood 104(13):4038–4045. doi:10.1182/blood-2004-03-1140 CrossRefPubMed

30.

Friedland JS, Constantin D, Shaw TC, Stylianou E (2001) Regulation of interleukin-8 gene expression after phagocytosis of zymosan by human monocytic cells. J Leukoc Biol 70(3):447–454PubMed

31.

Katsarelis H, Shah NP, Dhariwal DK, Pazianas M (2015) Infection and medication-related osteonecrosis of the jaw. J Dent Res 94(4):534–539. doi:10.1177/0022034515572021 CrossRefPubMed

32.

Lunov O, Syrovets T, Loos C, Beil J, Delacher M, Tron K, Nienhaus GU, Musyanovych A, Mailander V, Landfester K, Simmet T (2011) Differential uptake of functionalized polystyrene nanoparticles by human macrophages and a monocytic cell line. ACS Nano 5(3):1657–1669. doi:10.1021/nn2000756 CrossRefPubMed

33.

Qin Z (2012) The use of THP-1 cells as a model for mimicking the function and regulation of monocytes and macrophages in the vasculature. Atherosclerosis 221(1):2–11. doi:10.1016/j.atherosclerosis.2011.09.003 CrossRefPubMed

34.

Chanput W, Mes JJ, Wichers HJ (2014) THP-1 cell line: an in vitro cell model for immune modulation approach. Int Immunopharmacol 23(1):37–45. doi:10.1016/j.intimp.2014.08.002 CrossRefPubMed

35.

Wong B, Lumma WC, Smith AM, Sisko JT, Wright SD, Cai TQ (2001) Statins suppress THP-1 cell migration and secretion of matrix metalloproteinase 9 by inhibiting geranylgeranylation. J Leukoc Biol 69(6):959–962PubMed

36.

Pietschmann P, Stohlawetz P, Brosch S, Steiner G, Smolen JS, Peterlik M (1998) The effect of alendronate on cytokine production, adhesion molecule expression, and transendothelial migration of human peripheral blood mononuclear cells. Calcif Tissue Int 63(4):325–330CrossRefPubMed

37.

Kimachi K, Kajiya H, Nakayama S, Ikebe T, Okabe K (2011) Zoledronic acid inhibits RANK expression and migration of osteoclast precursors during osteoclastogenesis. Naunyn Schmiedeberg’s Arch Pharmacol 383(3):297–308. doi:10.1007/s00210-010-0596-4 CrossRef

38.

Stevenson PH, Stevenson JR (1986) Cytotoxic and migration inhibitory effects of bisphosphonates on macrophages. Calcif Tissue Int 38(4):227–233CrossRefPubMed

39.

Cougoule C, Hoshino S, Dart A, Lim J, Caron E (2006) Dissociation of recruitment and activation of the small G-protein Rac during Fcgamma receptor-mediated phagocytosis. J Biol Chem 281(13):8756–8764. doi:10.1074/jbc.M513731200 CrossRefPubMed

40.

Small JV, Rottner K, Kaverina I (1999) Functional design in the actin cytoskeleton. Curr Opin Cell Biol 11(1):54–60CrossRefPubMed

41.

Tsuchiya S, Yamabe M, Yamaguchi Y, Kobayashi Y, Konno T, Tada K (1980) Establishment and characterization of a human acute monocytic leukemia cell line (THP-1). Int J Cancer 26(2):171–176CrossRefPubMed

42.

Imoto S, Saigo K, Bohgaki M, Matsumoto E, Goto M (2014) Effects of four bisphosphonates on macrophage phagocytosis: quantitative measurement by flow cytometry using high-fluorescence particles and human monocytic cell line THP-1. Rinsho Byori 62(1):23–30PubMed

43.

Favot CL, Forster C, Glogauer M (2013) The effect of bisphosphonate therapy on neutrophil function: a potential biomarker. Int J Oral Maxillofac Surg 42(5):619–626. doi:10.1016/j.ijom.2012.12.011 CrossRefPubMed

44.

Hagelauer N, Pabst AM, Ziebart T, Ulbrich H, Walter C (2014) In vitro effects of bisphosphonates on chemotaxis, phagocytosis, and oxidative burst of neutrophil granulocytes. Clin Oral Investig. doi:10.1007/s00784-014-1219-0

45.

Kuiper JW, Forster C, Sun C, Peel S, Glogauer M (2012) Zoledronate and pamidronate depress neutrophil functions and survival in mice. Br J Pharmacol 165(2):532–539. doi:10.1111/j.1476-5381.2011.01592.x CrossRefPubMedPubMedCentral

46.

McGrath JL, Hartwig JH, Tardy Y, Dewey CF Jr (1998) Measuring actin dynamics in endothelial cells. Microsc Res Tech 43(5):385–394. doi:10.1002/(SICI)1097-0029(19981201)43:5<385::AID-JEMT5>3.0.CO;2-Z CrossRefPubMed

47.

Ridley AJ, Hall A (1994) Signal transduction pathways regulating Rho-mediated stress fibre formation: requirement for a tyrosine kinase. EMBO J 13(11):2600–2610PubMedPubMedCentral

48.

Bailly M, Condeelis JS, Segall JE (1998) Chemoattractant-induced lamellipod extension. Microsc Res Tech 43(5):433–443. doi:10.1002/(SICI)1097-0029(19981201)43:5<433::AID-JEMT9>3.0.CO;2-2 CrossRefPubMed

49.

Murakami H, Takahashi N, Sasaki T, Udagawa N, Tanaka S, Nakamura I, Zhang D, Barbier A, Suda T (1995) A possible mechanism of the specific action of bisphosphonates on osteoclasts: tiludronate preferentially affects polarized osteoclasts having ruffled borders. Bone 17(2):137–144CrossRefPubMed

50.

Halasy-Nagy JM, Rodan GA, Reszka AA (2001) Inhibition of bone resorption by alendronate and risedronate does not require osteoclast apoptosis. Bone 29(6):553–559CrossRefPubMed

Title: Altered macrophagic THP-1 cell phagocytosis and migration in bisphosphonate-related osteonecrosis of the jaw (BRONJ)
Authors: Sebastian Hoefert
Claudia Sade Hoefert
Adelheid Munz
Hinnak Northoff
Anna Yuan
Kathrin Reichenmiller
Siegmar Reinert
Martin Grimm
Publication date: 01-06-2016
Publisher: Springer Berlin Heidelberg
Published in: Clinical Oral Investigations / Issue 5/2016
Print ISSN: 1432-6981
Electronic ISSN: 1436-3771
DOI: https://doi.org/10.1007/s00784-015-1584-3

At a glance: The STEP trials

Springer Medicine

Altered macrophagic THP-1 cell phagocytosis and migration in bisphosphonate-related osteonecrosis of the jaw (BRONJ)

Abstract

Objectives

Materials and methods

Results

Conclusions

Clinical relevance

At a glance: The STEP trials

Springer Medicine

Abstract

Objectives

Materials and methods

Results

Conclusions

Clinical relevance

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