{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T21:48:02Z","timestamp":1775166482583,"version":"3.50.1"},"reference-count":58,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2020,6,9]],"date-time":"2020-06-09T00:00:00Z","timestamp":1591660800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antioxidants"],"abstract":"<jats:p>Bisphosphonates (BPs) are classified into two groups, according to their side chain structures, as nitrogen-containing BPs (NBPs) and non-nitrogen-containing BPs (non-NBPs). In this study, we examined the effects of NBPs and non-NBPs on inflammatory responses, by quantifying the inflammatory mediators, prostaglandin E2 (PGE2) and nitric oxide (NO), in cultured neonatal mouse calvaria. All examined NBPs (pamidronate, alendronate, incadronate, risedronate, zoledronate) stimulated lipopolysaccharide (LPS)-induced PGE2 and NO production by upregulating COX-2 and iNOS mRNA expression, whereas non-NBPs (etidronate, clodronate, tiludronate) suppressed PGE2 and NO production, by downregulating gene expression. Additionally, [4-(methylthio) phenylthio] methane bisphosphonate (MPMBP), a novel non-NBP with an antioxidant methylthio phenylthio group in its side chain, exhibited the most potent anti-inflammatory activity among non-NBPs. Furthermore, results of immunohistochemistry showed that the nuclear translocation of NF-\u03baB\/p65 and tyrosine nitration of cytoplasmic protein were stimulated by zoledronate, while MPMBP inhibited these phenomena, by acting as a superoxide anion (O2\u2212) scavenger. These findings indicate that MPMBP can act as an efficacious agent that causes fewer adverse effects in patients with inflammatory bone diseases, including periodontitis and rheumatoid arthritis.<\/jats:p>","DOI":"10.3390\/antiox9060503","type":"journal-article","created":{"date-parts":[[2020,6,9]],"date-time":"2020-06-09T08:49:11Z","timestamp":1591692551000},"page":"503","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Structure-Dependent Effects of Bisphosphonates on Inflammatory Responses in Cultured Neonatal Mouse Calvaria"],"prefix":"10.3390","volume":"9","author":[{"given":"Keiko","family":"Suzuki","sequence":"first","affiliation":[{"name":"Department of Dental Pharmacology, School of Dentistry, Showa University, Tokyo 142-8555, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sadaaki","family":"Takeyama","sequence":"additional","affiliation":[{"name":"Department of Molecular Cell Pharmacology, Graduate School of Dental Medicine, Hokkaido University, Sapporo 060-8586, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shinobu","family":"Murakami","sequence":"additional","affiliation":[{"name":"Center for Environmental Dentistry, Graduate School of Dentistry, Tohoku University, Sendai 980-8575, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Nagaoka","sequence":"additional","affiliation":[{"name":"Department of Dental Pharmacology, School of Dentistry, Ohu University, Fukushima 963-8611, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mirei","family":"Chiba","sequence":"additional","affiliation":[{"name":"Division of Oral Physiology, Tohoku University Graduate School of Dentistry, Sendai 980-8575, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0017-357X","authenticated-orcid":false,"given":"Kaoru","family":"Igarashi","sequence":"additional","affiliation":[{"name":"Division of Craniofacial Anomalies, Tohoku University Graduate School of Dentistry, Sendai 980-8575, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hisashi","family":"Shinoda","sequence":"additional","affiliation":[{"name":"Center for Environmental Dentistry, Graduate School of Dentistry, Tohoku University, Sendai 980-8575, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,6,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.bone.2011.04.022","article-title":"Bisphosphonates: The first 40 years","volume":"49","author":"Russell","year":"2011","journal-title":"Bone"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1652","DOI":"10.1002\/(SICI)1097-0142(19971015)80:8+<1652::AID-CNCR15>3.0.CO;2-Z","article-title":"Overview of bisphosphonates","volume":"80","author":"Rogers","year":"1997","journal-title":"Cancer"},{"key":"ref_3","unstructured":"Fleisch, H. (2000). Bisphosphonates in Bone Disease. From the Laboratory to the Patient, Academic Press. [4th ed.]."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1016\/j.bone.2003.07.007","article-title":"Differentiating the mechanisms of antiresorptive action of nitrogen containing bisphosphonates","volume":"33","author":"Cohen","year":"2003","journal-title":"Bone"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2961","DOI":"10.1002\/1097-0142(20000615)88:12+<2961::AID-CNCR12>3.0.CO;2-L","article-title":"Cellular and molecular mechanisms of action of bisphosphonates","volume":"88","author":"Rogers","year":"2000","journal-title":"Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1359\/jbmr.0707onj","article-title":"Bisphosphonate-associated osteonecrosis of the jaw: Report of a task force of the American society for bone and mineral research","volume":"22","author":"Khosla","year":"2007","journal-title":"J. Bone Miner. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1056\/NEJM200507073530120","article-title":"Osteonecrosis of the jaw and bisphosphonates","volume":"353","author":"Durie","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_8","first-page":"8580","article-title":"Osteonecrosis of the jaw in cancer after treatment with bisphosphonates: Incidence and risk factors","volume":"34","author":"Bamias","year":"2006","journal-title":"J. Clin. Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"826","DOI":"10.1359\/jbmr.080205","article-title":"Frequency and risk factors associated with osteonecrosis of the jaw in cancer patients treated with intravenous bisphosphonates","volume":"23","author":"Hoff","year":"2008","journal-title":"J. Bone Miner. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1111\/j.1365-2249.2005.02665.x","article-title":"The bisphosphonate acute phase response: Rapid and copious production of proinflammatory cytokines by peripheral blood \u03b3\u03c6 T cells in response to aminobisphosphonates is inhibited by statins","volume":"139","author":"Hewitt","year":"2005","journal-title":"Clin. Exp. Immunol."},{"key":"ref_11","first-page":"147","article-title":"Nitrogen containing bisphosphonates associated osteonecrosis of the jaws: A review for past 10 year literature","volume":"11","author":"Kumar","year":"2014","journal-title":"Dent. Res. J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.jphs.2016.04.011","article-title":"The novel bisphosphonate disodium dihydrogen-4-[(methylthio) phenylthio] methane bisphosphonate increases bone mass in post-ovariectomy rats","volume":"131","author":"Takizawa","year":"2016","journal-title":"J. Pharmacol. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1159\/000028189","article-title":"TRK-530 inhibits accumulation of superoxide anions derived from human polymorphonuclear leukocytes and bone resorption induced by activated osteoclasts","volume":"56","author":"Tanahashi","year":"1998","journal-title":"Pharmacology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1248\/bpb.20.1147","article-title":"The effect of TRK-530 on experimental arthritis in mice","volume":"20","author":"Takaoka","year":"1997","journal-title":"Biol. Pharm. Bull."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1159\/000028203","article-title":"Inhibitory effect of TRK-530 on inflammatory cytokines in bone marrow of rats with adjuvant arthritis","volume":"56","author":"Tanahashi","year":"1998","journal-title":"Pharmacology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1159\/000028204","article-title":"Inhibitory effects of TRK-530 on rat adjuvant arthritis","volume":"56","author":"Tanahashi","year":"1998","journal-title":"Pharmacology"},{"key":"ref_17","first-page":"120","article-title":"Role of reactive nitrogen species and antioxidant defense systems in the pathogenesis of rheumatoid arthritis","volume":"22","author":"Attia","year":"2015","journal-title":"Wulfenia"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1254\/jphs.FM0070272","article-title":"Pharmacological topics of bone metabolism: A novel bisphosphonate for the treatment of periodontitis","volume":"106","author":"Shinoda","year":"2008","journal-title":"J. Pharmacol. Sci."},{"key":"ref_19","unstructured":"Suzuki, K., Takeshita, F., Yamamoto, K., Yamada, S., Shinoda, H., and Ochiya, T. (2010, January 15\u201319). In vivo imaging of osteoclast precursor recruitment to the inflammatory site where extensive bone destruction occurs. Proceedings of the ASBMR 2010 Annual Meeting, Toronto, ON, Canada. MO0263."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1038\/nature01320","article-title":"Points of control in inflammation","volume":"420","author":"Nathan","year":"2002","journal-title":"Nature"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/j.intimp.2007.11.003","article-title":"Inhibition of LPS-induced NO and PGE2 production by asiatic acid via NF-kappa B inactivation in RAW 264.7 macrophages: Possible involvement of the IKK and MAPK pathways","volume":"8","author":"Lee","year":"2008","journal-title":"Int. Immunopharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1525","DOI":"10.1369\/jhc.5A6630.2005","article-title":"Osteoclast responses to lipopolysaccharide, parathyroid hormone and bisphosphonates in neonatal murine calvaria analyzed by laser scanning confocal microscopy","volume":"53","author":"Suzuki","year":"2005","journal-title":"J. Histochem. Cytochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1038\/nature02924","article-title":"NF-kappaB functions as a tumour promoter in inflammation-associated cancer","volume":"431","author":"Pikarsky","year":"2004","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1042\/bj2810419","article-title":"Production of hydroxyl radicals from the simultaneous generation of superoxide and nitric oxide","volume":"281","author":"Hogg","year":"1992","journal-title":"Biochem. J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"8526","DOI":"10.1074\/jbc.C300511200","article-title":"Structural basis for bisphosphonate-mediated inhibition of isoprenoid biosynthesis","volume":"279","author":"Hosfield","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1084\/jem.20030015","article-title":"An essential role of cytosolic phospholipase A2alpha in prostaglandin E2-mediated bone resorption associated with inflammation","volume":"197","author":"Miyaura","year":"2003","journal-title":"J. Exp. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1006\/bbrc.1999.1606","article-title":"Transcriptional regulation of the cyclooxygenase-2 gene by diverse ligands in murine osteoblasts","volume":"264","author":"Wadleigh","year":"1999","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1042\/bj3420021","article-title":"S-adenosylmethionine attenuates the lipopolysaccharide-induced expression of the gene for the tumour necrosis factor-\u03b1","volume":"342","author":"Watson","year":"1999","journal-title":"Biochem. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/S0002-9343(00)00480-0","article-title":"Nitric oxide and intestinal inflammation","volume":"109","author":"Kubes","year":"2000","journal-title":"Am. J. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.jot.2017.05.002","article-title":"Macrophages and bone inflammation","volume":"10","author":"Gu","year":"2017","journal-title":"J. Orthop. Translat."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"106085","DOI":"10.1016\/j.intimp.2019.106085","article-title":"MPMBP down-regulates Toll-like receptor (TLR) 2 ligand-induced proinflammatory cytokine production by inhibiting NF-\u03baB but not AP-1 activation","volume":"79","author":"Tamai","year":"2020","journal-title":"Int. Immunopharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1006\/abbi.1998.0755","article-title":"Biological tyrosine nitration: A pathophysiological function of nitric oxide and reactive oxygen species","volume":"356","author":"Ischiropoulos","year":"1998","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_33","first-page":"1607","article-title":"NF-\u03baB activation by peroxynitrite through I\u03baB\u03b1-dependent phosphorylation versus nitration in colon cancer cells","volume":"31","author":"Gochman","year":"2011","journal-title":"Anticancer Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1165\/ajrcmb.20.5.3452","article-title":"Cooperativity between oxidants and tumor necrosis factor in the activation of nuclear factor (NF)-kappaB: Requirement of Ras\/mitogen-activated protein kinases in the activation of NF-kappaB by oxidants","volume":"20","author":"Macara","year":"1999","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"11671","DOI":"10.1021\/bi701107z","article-title":"Tyrosine nitration of I\u03baB\u03b1: A novel mechanism for NF-\u03baB activation","volume":"46","author":"Yakovlev","year":"2007","journal-title":"Biochemistry"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Nagaoka, M., Maeda, T., Chatani, M., Handa, K., Yamakawa, T., Kiyohara, S., Negishi-Koga, T., Kato, Y., Takami, M., and Niida, S. (2019). A delphinidin-enriched maqui berry extract improves bone metabolism and protects against bone loss in osteopenic mouse models. Antioxidants, 8.","DOI":"10.3390\/antiox8090386"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1097\/SHK.0b013e3181e7e9ba","article-title":"Pathophysiological roles of peroxynitrite in circulatory shock","volume":"34","year":"2010","journal-title":"Shock"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3465","DOI":"10.1074\/jbc.272.6.3465","article-title":"Metabolic fate of peroxynitrite in aqueous solution: Reaction with nitric oxide and pH-dependent decomposition to nitrite and oxygen in a 2:1 stoichiometry","volume":"272","author":"Pfeiffer","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1523\/JNEUROSCI.18-03-00923.1998","article-title":"Nitric oxide and superoxide contribute to motor neuron apoptosis induced by trophic factor deprivation","volume":"18","author":"Spear","year":"1998","journal-title":"J. Neurosci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"14243","DOI":"10.1073\/pnas.94.26.14243","article-title":"Peroxynitrite rapidly permeates phospholipid membranes","volume":"94","author":"Marla","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","first-page":"1","article-title":"S-demethylation by microsomal enzymes","volume":"143","author":"Mazel","year":"1964","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"313","DOI":"10.3109\/00498258809041667","article-title":"In vitro metabolism of the methylthio group of 2-methylthiobenzothiazole by rat liver","volume":"18","author":"Larsen","year":"1988","journal-title":"Xenobiotica"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1161\/01.CIR.100.12.1330","article-title":"Inhibition of NF-\u03baB activation by pyrrolidine dithiocarbamate prevents in vivo expression of proinflammatory genes","volume":"100","author":"Liu","year":"1999","journal-title":"Circulation"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3976","DOI":"10.4049\/jimmunol.159.8.3976","article-title":"In vivo inhibition of nuclear factor-kappa B activation prevents inducible nitric oxide synthase expression and systemic hypotension in a rat model of septic shock","volume":"159","author":"Liu","year":"1997","journal-title":"J. Immunol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1210\/jc.2013-3648","article-title":"Nitrogen-bisphosphonate therapy is linked to compromised coenzyme Q10 and vitamin E status in postmenopausal women","volume":"99","author":"Kalyan","year":"2014","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2332","DOI":"10.1073\/pnas.68.10.2332","article-title":"Deficiency of coenzyme Q 10 in gingival tissue from patients with periodontal disease","volume":"68","author":"Littarru","year":"1971","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"334","DOI":"10.4103\/0253-7613.71884","article-title":"Role of coenzyme Q (10) as an antioxidant and bioenergizer in periodontal diseases","volume":"42","author":"Prakash","year":"2010","journal-title":"Indian J. Pharmacol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/S0891-5849(00)00307-5","article-title":"Regulation of ubiquinone metabolism","volume":"29","author":"Dallner","year":"2000","journal-title":"Free Radic. Biol. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1097\/01.crd.0000154790.42283.a1","article-title":"HMG-CoA Reductase inhibitors and coenzyme Q10","volume":"13","author":"Nawarskas","year":"2005","journal-title":"Cardiol. Rev."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1432-1033.1987.tb13379.x","article-title":"In vitro and in vivo synthesis of dolichol and other main mevalonate products in various organs of the rat","volume":"168","author":"Elmberger","year":"1987","journal-title":"Eur. J. Biochem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1016\/S0278-2391(03)00720-1","article-title":"Pamidronate (Aredia) and zoledronate (Zometa) induced avascular necrosis of the jaws: A growing epidemic","volume":"9","author":"Marx","year":"2003","journal-title":"J. Oral Maxillofac. Surg."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1016\/j.bone.2009.07.011","article-title":"Bisphosphonate-associated osteonecrosis of the jaw: A key role of inflammation?","volume":"45","author":"Lesclous","year":"2009","journal-title":"Bone"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1097\/MJT.0b013e318247148f","article-title":"Bisphosphonates: Focus on inflammation and bone loss","volume":"19","author":"Iannitti","year":"2012","journal-title":"Am. J. Ther."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"2272","DOI":"10.1210\/jc.2012-1027","article-title":"Benefits and risks of bisphosphonate therapy for osteoporosis","volume":"97","author":"Khosla","year":"2012","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.bone.2011.01.014","article-title":"Pharmacology of bisphosphonates","volume":"49","author":"Cremers","year":"2011","journal-title":"Bone"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1508","DOI":"10.1126\/science.289.5484.1508","article-title":"Therapeutic approaches to bone diseases","volume":"289","author":"Rodan","year":"2000","journal-title":"Science"},{"key":"ref_57","first-page":"1173","article-title":"Distribution of protein nitrotyrosine in synovial tissues of patients with rheumatoid arthritis and osteoarthritis","volume":"30","author":"Sandhu","year":"2003","journal-title":"J. Rheumatol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.bone.2019.03.027","article-title":"Rescue bisphosphonate treatment of alveolar bone improves extraction socket healing and reduces osteonecrosis in zoledronate-treated mice","volume":"123","author":"Hokugo","year":"2019","journal-title":"Bone"}],"container-title":["Antioxidants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3921\/9\/6\/503\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:37:01Z","timestamp":1760175421000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3921\/9\/6\/503"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,6,9]]},"references-count":58,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2020,6]]}},"alternative-id":["antiox9060503"],"URL":"https:\/\/doi.org\/10.3390\/antiox9060503","relation":{},"ISSN":["2076-3921"],"issn-type":[{"value":"2076-3921","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,6,9]]}}}