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The aim of this study was to investigate if the cysteine proteinase inhibitors cystatin C, fungal cysteine proteinase inhibitor (E-64), and N-benzyloxycarbonyl-arginyl-leucyl-valyl-glycyl-diazomethane acetate (Z-RLVG-CHN2) can inhibit LPS-induced osteoclast formation. Mouse bone marrow macrophages (BMMs) were isolated and primed with receptor activator of NF-\u03baB ligand (RANKL) for 24 h, followed by stimulation with LPS, with and without inhibitors. Adult mice were injected locally with LPS and then treated with E-64 and osteoclast formation assessed by the number of cathepsin K+ multinucleated cells. Cystatin C inhibited LPS-induced osteoclast formation time and concentration dependently (IC50 = 0.3 \u03bcM). The effect was associated with decreased mRNA and protein expression of tartrate-resistant acid phosphatase (TRAP) and cathepsin K and of the osteoclastogenic transcription factors c-Fos and NFATc1. LPS-induced osteoclast formation on bone slices was also inhibited by cystatin C, resulting in decreased pit formation and release of bone matrix proteins. Similar data were obtained with E-64 and Z-RLVG-CHN2. Cystatin C was internalized in BMMs stimulated by LPS but not in unstimulated BMMs. Osteoclast formation induced by LPS was dependent on TNF-\u03b1, and the 3 inhibitors abolished LPS-induced TNF superfamily 2 (gene encoding TNF-\u03b1; Tnfsf2) mRNA expression without affecting Il1b, Il6, or oncostatin M (Osm) expression. Formation of osteoclasts in the skull bones after local LPS stimulation was inhibited by E-64. It is concluded that cysteine proteinase inhibitors effectively inhibit LPS-induced osteoclast formation in vivo and in vitro by inhibition of TNF-\u03b1 expression. The targeting of cysteine proteinases might represent a novel treatment modality for prevention of inflammatory bone loss.<\/jats:p>","DOI":"10.1189\/jlb.3a1016-433r","type":"journal-article","created":{"date-parts":[[2017,2,15]],"date-time":"2017-02-15T01:20:13Z","timestamp":1487121613000},"page":"1233-1243","source":"Crossref","is-referenced-by-count":30,"title":["Inhibition of lipopolysaccharide-induced osteoclast formation and bone resorption in vitro and in vivo by cysteine proteinase inhibitors"],"prefix":"10.1093","volume":"101","author":[{"given":"Fredrik","family":"Str\u00e5lberg","sequence":"first","affiliation":[{"name":"Department of Molecular Periodontology, Ume\u00e5 University , Ume\u00e5 , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali","family":"Kassem","sequence":"additional","affiliation":[{"name":"Department of Molecular Periodontology, Ume\u00e5 University , Ume\u00e5 , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franciszek","family":"Kasprzykowski","sequence":"additional","affiliation":[{"name":"Institute of Chemistry, University of Gdansk , Gdansk , Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magnus","family":"Abrahamson","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Division of Clinical Chemistry and Pharmacology, Lund University , Lund , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anders","family":"Grubb","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Division of Clinical Chemistry and Pharmacology, Lund University , Lund , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catharina","family":"Lindholm","sequence":"additional","affiliation":[{"name":"Department of Rheumatology and Inflammation Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg , Gothenburg , Sweden"},{"name":"Departments of Internal Medicine and Clinical Nutrition, Centre for Bone and Arthritis Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg , Gothenburg , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulf H","family":"Lerner","sequence":"additional","affiliation":[{"name":"Department of Molecular Periodontology, Ume\u00e5 University , Ume\u00e5 , Sweden"},{"name":"Departments of Internal Medicine and Clinical Nutrition, Centre for Bone and Arthritis Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg , Gothenburg , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2017,2,14]]},"reference":[{"key":"2023011712205922200_B1","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1111\/j.1600-0757.2010.00347.x","article-title":"Mechanisms and control of pathologic bone loss in periodontitis","volume":"53","author":"Bartold","year":"2010","journal-title":"Periodontol. 2000"},{"key":"2023011712205922200_B2","doi-asserted-by":"crossref","first-page":"2354","DOI":"10.1016\/j.actbio.2014.02.003","article-title":"Contributions of human tissue analysis to understanding the mechanisms of loosening and osteolysis in total hip replacement","volume":"10","author":"Gallo","year":"2014","journal-title":"Acta Biomater."},{"issue":"Suppl 16","key":"2023011712205922200_B3","doi-asserted-by":"crossref","first-page":"S172","DOI":"10.1111\/jcpe.12346","article-title":"Risk indicators for peri-implant mucositis: a systematic literature review","volume":"42","author":"Renvert","year":"2015","journal-title":"J. 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