Geranylgeraniol Reverses the Toxicity Induced by Clinical Doses of Zoledronic Acid on Gingival Epithelial Cells and Gingival Fibroblasts

Filip Koneski; David Tipton; Jegdish Babu; Danica Popovic-Monevska; Vladimir Popovski; Alberto Benedetti; Suzana Dvojakovska-Bozovic; Aleksandar Iliev; Goran Panchevski; Antonio Kirkov; Aleksandar Grchev; Aleksandar Stamatoski; Franklin Garcia-Godoy

doi:10.3889/oamjms.2021.5592

Authors

Filip Koneski Department of Bioscience Research, College of Dentistry, University of Tennessee, Memphis, Tennessee, USA; University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
David Tipton Department of Bioscience Research, College of Dentistry, University of Tennessee, Memphis, Tennessee, USA
Jegdish Babu Department of Bioscience Research, College of Dentistry, University of Tennessee, Memphis, Tennessee, USA
Danica Popovic-Monevska University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Vladimir Popovski University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Alberto Benedetti University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Suzana Dvojakovska-Bozovic University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Aleksandar Iliev University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Goran Panchevski University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Antonio Kirkov University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Aleksandar Grchev University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Aleksandar Stamatoski University Clinic for Maxillofacial Surgery in Skopje, Faculty of Dentistry, Ss. Cyril and Methodius University in Skopje, Macedonia
Franklin Garcia-Godoy Department of Bioscience Research, College of Dentistry, University of Tennessee, Memphis, Tennessee, USA

DOI:

https://doi.org/10.3889/oamjms.2021.5592

Keywords:

medication-related osteonecrosis of the jaw, geranylgeraniol, gingival epithelial cells, gingival fibroblasts, wound healing

Abstract

BACKGROUND: Medication-related osteonecrosis of the jaw (MRONJ) is of considerable concern among clinicians and researchers, with no clear pathology mechanism, preventive, or treatment protocols.

AIM: This study aimed to assess the effects of geranylgeraniol (GGOH) on the toxicity induced by clinical doses of zoledronic acid (ZOL) on gingival epithelial cells and gingival fibroblasts in vitro.

METHODS: Human gingival fibroblasts and gingival epithelial cells were treated with 5, 25, or 50 μM ZOL ± 50 μM GGOH for 3 days. Viability of the cells was determined using the 3-[4, 5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay. Calculation of percentage of the control group, analysis of variance and Tukey post-hoc comparisons were performed to test the significance between groups, which was set at p = 0.05. Cell morphology was evaluated using light microscopy.

RESULTS: ZOL significantly reduced the viability of both epithelial cells and fibroblasts at all concentrations (p < 0.05), with the exception of fibroblasts at concentration of 5 μM (p = 0.44). GGOH had positive effects on the viability of the cells treated with ZOL at all concentrations. However, statistically significant improvement was obtained only in epithelial cells at 5 and 25 μM ZOL. The cell morphology of both types of cells was improved after addition of GGOH.

CONCLUSION: GGOH reverses the toxic effects of clinical doses of ZOL on gingival epithelial cells and has slightly positive, but not significant effects on gingival fibroblasts. This study suggests that GGOH may be effective in the prevention and treatment of MRONJ.

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References

Ruggiero SL, Dodson TB, Fantasia J, Goodday R, Aghaloo T, Mehrotra B. American association of oral and maxillofacial surgeons position paper on medication-related osteonecrosis of the jaw-2014 update. J Oral Maxillofac Surg. 2014;72(10):1938- 56. https://doi.org/10.1016/j.joms.2014.04.031 PMid:25234529

Otto S, Pautke C, Opelz C, Westphal I, Drosse I, Schwager J, et al. Osteonecrosis of the jaw: Effect of bisphosphonate type, local concentration, and acidic milieu on the pathomechanism. J Oral Maxillofac Surg. 2010;68(11):2837-45. https://doi.org/10.1016/j.joms.2010.07.017 PMid:20971371

Landesberg R, Cozin M, Cremers S, Woo V, Kousteni S, Sinha S, et al. Inhibition of oral mucosal cell wound healing by bisphosphonates. J Oral Maxillofac Surg. 2008;66:839-47. https://doi.org/10.1016/j.joms.2008.01.026 PMid:18423269

Scheper MA, Badros A, Chaisuparat R, Cullen KJ, Meiller TF. Effect of zoledronic acid on oral fibroblasts and epithelial cells: A potential mechanism of bisphosphonate-associated osteonecrosis. Br J Haematol. 2009;144(5):667-76. https://doi.org/10.1111/j.1365-2141.2008.07504.x PMid:19036117

Walter C, Klein MO, Pabst A, Al-Nawas B, Duschner H, Ziebart T. Influence of bisphosphonates on endothelial cells, fibroblasts, and osteogenic cells. Clin Oral Investig. 2010;14(1):35-41. https://doi.org/10.1007/s00784-009-0266-4 PMid:19294435

Cozin M, Pinker BM, Solemani K, Zuniga JM, Dadaian SC, Cremers S, et al. Novel therapy to reverse the cellular effects of bisphosphonates on primary human oral fibroblasts. J Oral Maxillofac Surg. 2011;69(10):2564-78. https://doi.org/10.1016/j.joms.2011.03.005 PMid:21807448

Ravosa MJ, Ning J, Liu Y, Stack MS. Bisphosphonate effects on the behaviour of oral epithelial cells and oral fibroblasts. Arch Oral Biol. 2011;56(5):491-8. https://doi.org/10.1016/j.archoralbio.2010.11.003 PMid:21146154

Tipton DA, Seshul BA, Dabbous MK. Effect of bisphosphonates on human gingival fibroblast production of mediators of osteoclastogenesis: RANKL, osteoprotegerin and interleukin-6. J Periodontal Res. 2011;46(1):39-47. https://doi.org/10.1111/j.1600-0765.2010.01306.x PMid:20701670

Açil Y, Möller B, Niehoff P, Rachko K, Gassling V, Wiltfang J, et al. The cytotoxic effects of three different bisphosphonates in vitro on human gingival fibroblasts, osteoblasts and osteogenic sarcoma cells. J Craniomaxillofac Surg. 2012;40(8):e229-35. https://doi.org/10.1016/j.jcms.2011.10.024 PMid:22082730

Basso FG, Pansani TN, de Oliveira CF, Turrioni AP, Soares DG, Hebling J, et al. Cytotoxic effects of zoledronic acid on human epithelial cells and gingival fibroblasts. Braz Dent J. 2013;24(6):551-8. https://doi.org/10.1590/0103-6440201302229 PMid:24474348

Walter C, Pabst A, Ziebart T, Klein MO, Al-Nawas B. Bisphosphonates affect migration ability and cell viability of HUVEC, fibroblasts and osteoblasts in vitro. Oral Dis. 2017;17(2):194-9. https://doi.org/10.1111/j.1601-0825.2010.01720.x PMid:20796232

Açil Y, Arndt ML, Gülses A, Wieker H, Naujokat H, Ayna M, et al. Cytotoxic and inflammatory effects of alendronate and zolendronate on human osteoblasts, gingival fibroblasts and osteosarcoma cells. J Craniomaxillofac Surg. 2018;46(4):538- 46. https://doi.org/10.1016/j.jcms.2017.12.015 PMid:29454604

Otto S, Pautke C, Jurado OM, Nehrbass D, Stoddart MJ, Ehrenfeld M, et al. Further development of the MRONJ minipig

large animal model. J Craniomaxillofac Surg. 2017;45(9):1503- 14. https://doi.org/10.1016/j.jcms.2017.07.002 PMid:28803745

Soundia A, Hadaya D, Esfandi N, Gkouveris I, Christensen R, Dry SM, et al. Zoledronate impairs socket healing after extraction of teeth with experimental periodontitis. J Dent Res. 2018;97(3):312-20. https://doi.org/10.1177/0022034517732770 PMid:28954199

Zandi M, Dehghan A, Janbaz P, Malekzadeh H, Amini P. The starting point for bisphosphonate-related osteonecrosis of the jaw: Alveolar bone or oral mucosa? A randomized, controlled experimental study. J Craniomaxillofac Surg. 2017;45(1):157- 61. https://doi.org/10.1016/j.jcms.2016.10.015 PMid:27919595

Popovik-Monevska D, Muratovska I, Popovska M, Dvojakovska S, Koneski F. Osteoradionecrosis of the mandible: A case report. Balkan J Dent Med. 2015;20:54-8. https://doi.org/10.1515/bjdm-2016-0009

Luckman SP, Hughes DE, Coxon FP, Russell RG, Rogers MJ. Nitrogen-containing bisphosphonates inhibit the mevalonate pathway and prevent post-translational prenylation of GTP-binding proteins, including Ras. J Bone Miner Res. 1998;13(4):581-9. https://doi.org/10.1359/jbmr.1998.13.4.581 PMid:9556058

Li X, Liu L, Tupper JC, Bannerman DD, Winn RK, Sebti SM, et al. Inhibition of protein geranylgeranylation and RhoA/RhoA kinase pathway induces apoptosis in human endothelial cells. J Biol Chem. 2002;277(18):15309-16. https://doi.org/10.1074/jbc.m201253200 PMid:11839765

Hagelauer N, Ziebart T, Pabst AM, Walter C. Bisphosphonates inhibit cell functions of HUVECs, fibroblasts and osteogenic cells via inhibition of protein geranylgeranylation. Clin Oral Investig. 2015;19:1079-91. https://doi.org/10.1007/s00784-014-1320-4 PMid:25261400

Ziebart T, Koch F, Klein MO, Guth J, Adler J, Pabst A, et al. Geranylgeraniol-a new potential therapeutic approach to bisphosphonate associated osteonecrosis of the jaw. Oral Oncol. 2011;47(3):195-201. https://doi.org/10.1016/j.oraloncology.2010.12.003 PMid:21247791

Zafar S, Coates DE, Cullinan MP, Drummond BK, Milne T, Seymour GJ. Zoledronic acid and geranylgeraniol regulate cellular behaviour and angiogenic gene expression in human gingival fibroblasts. J Oral Pathol Med. 2014;43(9):711-21. https://doi.org/10.1111/jop.12181 PMid:24762323

Zafar S, Coates DE, Cullinan MP, Drummond BK, Milne T, Seymour GJ. Effects of zoledronic acid and geranylgeraniol on the cellular behaviour and gene expression of primary human alveolar osteoblasts. Clin Oral Investig. 2016;20(8):2023-35. https://doi.org/10.1007/s00784-015-1706-y PMid:26795621

Pabst AM, Krüger M, Ziebart T, Jacobs C, Sagheb K, Walter C. The influence of geranylgeraniol on human oral keratinocytes after bisphosphonate treatment: An in vitro study. J Craniomaxillofac Surg. 2015;43(5):688-95. https://doi.org/10.1016/j.jcms.2015.03.014 PMid:25913629

Pabst AM, Krüger M, Sagheb K, Ziebart T, Jacobs C, Blatt S, et al. The influence of geranylgeraniol on microvessel sprouting after bisphosphonate substitution in an in vitro 3D-angiogenesis assay. Clin Oral Investig. 2017;21(3):771-8. https://doi.org/10.1007/s00784-016-1842-z PMid:27170294

Koneski F, Popovic-Monevska D, Gjorgoski I, Krajoska J, Popovska M, Muratovska I, et al. In vivo effects of geranylgeraniol on the development of bisphosphonate-related osteonecrosis of the jaws. J Craniomaxillofac Surg. 2018;46(2):230-6. https://doi.org/10.1016/j.jcms.2017.11.007 PMid:29233701

Nagaoka Y, Kajiya H, Ozeki S, Ikebe T, Okabe K. Mevalonates restore zoledronic acid-induced osteoclastogenesis inhibition. J Dent Res. 2015;94(4):594-601. https://doi.org/10.1177/0022034514564187 PMid:25535203

Tipton DA, Pabst MJ, Dabbous MK. Interleukin-1 beta-and tumor necrosis factor-alpha-independent monocyte stimulation of fibroblast collagenase activity. J Cell Biochem. 1990;44(4):253- 64. https://doi.org/10.1002/jcb.240440407 PMid:1965653

Smulow J, Glickman I. An epithelial-like cell line in continuous culture from normal adult human gingiva. Proc Soc Exp Biol Med. 1966;121(3):1294-6. https://doi.org/10.3181/00379727-121-31032 PMid:5937747

Sonnenschein C, Smulow J. Karyotypic analysis of an epithelial-like cell line derived from normal adult human gingiva. Exp Cell Res. 1966;43(3):633-8. https://doi.org/10.1016/0014-4827(66)90034-6 PMid:5957752

Kasten FH, Seileu K, Nefert RM. Quantitative evaluation of human gingival epithelial cell attachment to implant surfaces in vitro. Int J Periodontics Restorative Dent. 1990;10(1):68-79. PMid:2394556

Tipton DA, Lyle B, Babich H, Dabbous MK. In vitro cytotoxic and anti-inflammatory effects of myrrh oil on human gingival fibroblasts and epithelial cells. Toxicol In Vitro. 2003;17(3):301- 10. https://doi.org/10.1016/s0887-2333(03)00018-3 PMid:12781209