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However, N-BPs have various side effects such as acute influenza-like reactions after their initial administration and osteonecrosis of the jawbones after repeated administration. The mechanisms underlying such effects remain unclear. To overcome these problems, it is important to profile the inflammatory nature of N-BPs. Here, we analyzed the inflammatory reactions induced in mouse ear pinnae by the N-BPs alendronate (Ale) and zoledronate (Zol). We found the following: (i) Ale and Zol each induced two phases of inflammation (early weak and late strong ear swelling); (ii) both phases were augmented by lipopolysaccharides (LPSs; cell-surface constituent of gram-negative bacteria, including oral bacteria), but prevented by inhibitors of the phosphate transporters of solute carrier 20\/34 (SLC20\/SLC34); (iii) macrophages and neutrophils were involved in both phases of Ale+LPS\u2013induced ear-swelling; (iv) Ale increased or tended to increase various cytokines, and LPS augmented these effects, especially that on interleukin 1\u03b2 (IL-1\u03b2); (v) adenosine triphosphate (ATP) was involved in both phases, and Ale alone or Ale+LPS increased ATP in ear pinnae; (vi) the augmented late-phase swelling induced by Ale+LPS depended on both IL-1 and neutrophil extracellular traps (NETs; neutrophil-derived net-like complexes); (vii) neutrophils, together with macrophages and dendritic cells, also functioned as IL-1\u03b2\u2013producing cells, and upon stimulation with IL-1\u03b2, neutrophils produced NETs; (viii) stimulation of the purinergic 2X7 (P2X7) receptors by ATP induced IL-1\u03b2 in ear pinnae; (ix) NET formation by Ale+LPS was confirmed in gingiva, too. These results suggest that (i) N-BPs induce both early-phase and late-phase inflammation via ATP-production and P2X7 receptor stimulation; (ii) N-BPs and LPS induce mutually augmenting responses both early and late phases via ATP-mediated IL-1\u03b2 production by neutrophils, macrophages, and\/or dendritic cells; and (iii) NET production by IL-1\u03b2\u2013stimulated neutrophils may mediate the late phase, leading to prolonged inflammation. These results are discussed in relation to the side effects seen in patients treated with N-BPs. \u00a9 2021 American Society for Bone and Mineral Research (ASBMR).<\/jats:p>","DOI":"10.1002\/jbmr.4384","type":"journal-article","created":{"date-parts":[[2021,6,2]],"date-time":"2021-06-02T05:33:25Z","timestamp":1622612005000},"page":"1866-1878","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Nitrogen-containing bisphosphonates and lipopolysaccharide mutually augment inflammation via adenosine triphosphate (ATP)-mediated and interleukin 1\u03b2 (IL-1\u03b2)-mediated production of neutrophil extracellular traps (NETs)"],"prefix":"10.1093","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2818-2454","authenticated-orcid":false,"given":"Kanan","family":"Bando","sequence":"first","affiliation":[{"name":"Division of Orthodontics and Dentofacial Orthopedics, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshinobu","family":"Kuroishi","sequence":"additional","affiliation":[{"name":"Division of Oral Immunology, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroyuki","family":"Tada","sequence":"additional","affiliation":[{"name":"Division of Oral Immunology, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takefumi","family":"Oizumi","sequence":"additional","affiliation":[{"name":"Department of Dentistry and Oral Surgery National Hospital Organization Sendai Medical Center Sendai Japan"},{"name":"Department of Oral and Maxillofacial Surgery, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukinori","family":"Tanaka","sequence":"additional","affiliation":[{"name":"Department of Dento-oral Anesthesiology, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tetsu","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Department of Oral and Maxillofacial Surgery, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Itaru","family":"Mizoguchi","sequence":"additional","affiliation":[{"name":"Division of Orthodontics and Dentofacial Orthopedics, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shunji","family":"Sugawara","sequence":"additional","affiliation":[{"name":"Division of Oral Immunology, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuo","family":"Endo","sequence":"additional","affiliation":[{"name":"Department of Oral and Maxillofacial Surgery, Graduate School of Dentistry Tohoku University Sendai Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,6,2]]},"reference":[{"key":"2024012713453414100_jbmr4384-bib-0001","first-page":"265","volume-title":"Bone and Mineral Research","author":"Geddes","year":"1994"},{"key":"2024012713453414100_jbmr4384-bib-0002","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.bone.2010.11.008","article-title":"Biochemical and molecular mechanisms of action of bisphosphonates","volume":"49","author":"Rogers","year":"2011","journal-title":"Bone"},{"key":"2024012713453414100_jbmr4384-bib-0003","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1677\/erc.1.01094","article-title":"Emerging anti-cancer molecular mechanisms of aminobisphosphonates","volume":"13","author":"Caraglia","year":"2006","journal-title":"Endocr Relat 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