{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T07:47:47Z","timestamp":1792655267254,"version":"4.5.0"},"reference-count":47,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,30]],"date-time":"2021-10-30T00:00:00Z","timestamp":1635552000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Background: Periodontitis is characterized by excessive osteoclastic activity, which is closely associated with inflammation. It is well established that MAPK\/NF-kB axis is a key signaling pathway engaged in osteoclast differentiation. It is stated that that biphasic calcium phosphate (BCP) and platelet-rich fibrin (PRF) have significant antiostoeclastogenic effects in chronic periodontitis. Objective: We aimed to elucidate the synergetic effect of PRF\/BCP involvement of the nuclear factor kappa\u2013light\u2013chain\u2013enhancer of activated B cells (NF-kB) and the mitogen-activated protein kinase (MAPK) signaling pathway in osteoclast differentiation in chronic periodontitis. Methods: We induced osteoclast differentiation in vitro using peripheral blood mononuclear cells (PBMCs) derived from patients with chronic periodontitis. We assessed osteoclast generation by tartrate-resistant acid phosphatase (TRAP) activity, proinflammatory cytokines were investigated by ELISA and NF-\u03baB, and IKB by immunoblot, respectively. MAPK proteins and osteoclast transcription factors were studied by Western blot analysis and osteoclast transcriptional genes were assessed by RT-PCR. Results: The results showed that the potent inhibitory effect of PRF\/BCP on osteoclastogenesis was evidenced by decreased TRAP activity and the expression of transcription factors, NFATc1, c-Fos, and the osteoclast marker genes, TRAP, MMP-9, and cathepsin-K were found to be reduced. Further, the protective effect of PRF\/BCP on inflammation-mediated osteoclastogenesis in chronic periodontitis was shown by decreased levels of proinflammatory cytokines, NF-kB, IKB, and MAPK proteins. Conclusions: PRF\/BCP may promote a synergetic combination that could be used as a strong inhibitor of inflammation-induced osteoclastogenesis in chronic periodontitis.<\/jats:p>","DOI":"10.3390\/molecules26216578","type":"journal-article","created":{"date-parts":[[2021,11,1]],"date-time":"2021-11-01T22:22:24Z","timestamp":1635805344000},"page":"6578","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["Synergistic Effect of Biphasic Calcium Phosphate and Platelet-Rich Fibrin Attenuate Markers for Inflammation and Osteoclast Differentiation by Suppressing NF-\u03baB\/MAPK Signaling Pathway in Chronic Periodontitis"],"prefix":"10.3390","volume":"26","author":[{"given":"Anil","family":"Kumar","sequence":"first","affiliation":[{"name":"Department of Periodontology, Meenakshi Ammal Dental College and Hospital, Meenakshi Academy of Higher Education and Research, Chennai 600095, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4726-3727","authenticated-orcid":false,"given":"Jaideep","family":"Mahendra","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Meenakshi Ammal Dental College and Hospital, Meenakshi Academy of Higher Education and Research, Chennai 600095, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Little","family":"Mahendra","sequence":"additional","affiliation":[{"name":"Maktoum Bin Hamdan Dental University College, Dubai 213620, United Arab Emirates"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hesham H.","family":"Abdulkarim","sequence":"additional","affiliation":[{"name":"Advanced Periodontal and Dental Implant Care, Missouri School of Dentistry and Oral Health, A. T. Still University, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8667-8987","authenticated-orcid":false,"given":"Mohammed","family":"Sayed","sequence":"additional","affiliation":[{"name":"Department of Prosthetic Dental Sciences, College of Dentistry, Jazan University, Jazan 45412, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5613-9760","authenticated-orcid":false,"given":"Maryam H.","family":"Mugri","sequence":"additional","affiliation":[{"name":"Department of Maxillofacial Surgery and Diagnostic Sciences, College of Dentistry, Jazan University, Jazan 45412, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeeshan Heera","family":"Ahmad","sequence":"additional","affiliation":[{"name":"Dental College Hospital, King Saud University Medical City, Riyadh 12372, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashok Kumar","family":"Bhati","sequence":"additional","affiliation":[{"name":"Division of Periodontics Department of Preventive Dental Sciences, College of Dentistry, Jazan University, Jazan 45412, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hadeel Hussain","family":"Faqehi","sequence":"additional","affiliation":[{"name":"Dental School, Jazan University, Jazan 45412, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Waleed Omar","family":"Algregri","sequence":"additional","affiliation":[{"name":"King Fahad Armed Forces Hospital, Jeddah 23311, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7051-1143","authenticated-orcid":false,"given":"Saranya","family":"Varadarajan","sequence":"additional","affiliation":[{"name":"Department of Oral Pathology and Microbiology, Sri Venkateswara Dental College and Hospital, Chennai 600130, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thodur Madapusi","family":"Balaji","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Tagore Dental College and Hospital, Chennai 600127, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hosam Ali","family":"Baeshen","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, College of Dentistry, King Abdulaziz University, Jeddah 22254, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7246-5497","authenticated-orcid":false,"given":"Shankargouda","family":"Patil","sequence":"additional","affiliation":[{"name":"Department of Maxillofacial Surgery and Diagnostic Sciences, Division of Oral Pathology, College of Dentistry, Jazan University, Jazan 45412, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1038\/nri3785","article-title":"Periodontitis: From microbial immune subversion to systemic inflammation","volume":"15","author":"Hajishengallis","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/s41577-020-00488-6","article-title":"Local and systemic mechanisms linking periodontal disease and inflammatory comorbidities","volume":"21","author":"Hajishengallis","year":"2021","journal-title":"Nat. Rev. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"627","DOI":"10.14202\/vetworld.2018.627-635","article-title":"The crucial roles of inflammatory mediators in inflammation: A review","volume":"11","author":"Abdulkhaleq","year":"2018","journal-title":"Vet. World"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Braicu, C., Buse, M., Busuioc, C., Drula, R., Gulei, D., Raduly, L., Rusu, A., Irimie, A., Atanasov, A.G., and Slaby, O. (2019). A Comprehensive Review on MAPK: A Promising Therapeutic Target in Cancer. Cancers, 11.","DOI":"10.3390\/cancers11101618"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"705","DOI":"10.3389\/fimmu.2019.00705","article-title":"NF-\u03baB Signaling in Macrophages: Dynamics, Crosstalk, and Signal Integration","volume":"10","author":"Dorrington","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"a008151","DOI":"10.1101\/cshperspect.a008151","article-title":"Signaling in Cell Differentiation and Morphogenesis","volume":"4","author":"Basson","year":"2012","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"55684","DOI":"10.18632\/oncotarget.18264","article-title":"The p38 pathway, a major pleiotropic cascade that transduces stress and metastatic signals in endothelial cells","volume":"8","author":"Corre","year":"2017","journal-title":"Oncotarget"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"682","DOI":"10.18632\/genesandcancer.155","article-title":"JNK-signaling: A multiplexing hub in programmed cell death","volume":"8","author":"Dhanasekaran","year":"2017","journal-title":"Genes Cancer"},{"key":"ref_9","first-page":"2266","article-title":"Correlation of ERK\/MAPK signaling pathway with proliferation and apoptosis of colon cancer cells","volume":"17","author":"Zhou","year":"2019","journal-title":"Oncol. Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"706","DOI":"10.14348\/molcells.2017.0225","article-title":"Current Understanding of RANK Signaling in osteoclast differentiation and Maturation","volume":"40","author":"Park","year":"2017","journal-title":"Mol. Cells"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"233","DOI":"10.11005\/jbm.2014.21.4.233","article-title":"Regulation of NFATc1 in osteoclast differentiation","volume":"21","author":"Kim","year":"2014","journal-title":"J. Bone Metab."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ausenda, F., Rasperini, G., Acunzo, R., Gorbunkova, A., and Pagni, G. (2019). New Perspectives in the Use of Biomaterials for Periodontal Regeneration. Materials, 12.","DOI":"10.3390\/ma12132197"},{"key":"ref_13","first-page":"2277","article-title":"Clinical effect of platelet-rich fibrin on the preservation of the alveolar ridge following tooth extraction","volume":"15","author":"Zhang","year":"2018","journal-title":"Exp. Ther. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s13036-018-0133-4","article-title":"Tissue engineering strategies for the induction of angiogenesis using biomaterials","volume":"12","author":"Saberianpour","year":"2018","journal-title":"J. Biol. Eng."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1002\/jbm.b.34478","article-title":"The effect of platelet-rich fibrin exudate addition to porous poly(lactic-co-glycolic acid) scaffold in bone healing: An in vivo study","volume":"108","author":"Witek","year":"2020","journal-title":"J. Biomed. Mater. Res. B Appl. Biomater."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1007\/s00784-019-03156-9","article-title":"Effect of platelet-rich fibrin on cell proliferation, migration, differentiation, inflammation, and osteoclastogenesis: A systematic review of in vitro studies","volume":"24","author":"Strauss","year":"2020","journal-title":"Clin. Oral Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1002\/JPER.19-0109","article-title":"Platelet-rich fibrin suppresses in vitro osteoclastogenesis","volume":"91","author":"Kargarpour","year":"2020","journal-title":"J. Periodontol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1002\/jbm.a.10544","article-title":"Effect of biphasic calcium phosphate on human macrophage functions in vitro","volume":"65","author":"Silva","year":"2003","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1238","DOI":"10.1111\/cid.12384","article-title":"Addition of a Synthetically Fabricated Osteoinductive Biphasic Calcium Phosphate Bone Graft to BMP2 Improves New Bone Formation","volume":"18","author":"Zhang","year":"2016","journal-title":"Clin. Implant Dent. Relat. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1089\/ten.tec.2010.0387","article-title":"Consistent osteoblastic differentiation of human mesenchymal stem cells with bone morphogenetic protein 4 and low serum","volume":"17","author":"Cordonnier","year":"2011","journal-title":"Tissue Eng. Part C Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/s40824-018-0149-3","article-title":"Bioactive calcium phosphate materials and applications in bone regeneration","volume":"23","author":"Jeong","year":"2019","journal-title":"Biomater. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3495","DOI":"10.1089\/ten.tea.2010.0227","article-title":"Biphasic calcium phosphate microparticles for bone formation: Benefits of combination with blood clot","volume":"16","author":"Balaguer","year":"2010","journal-title":"Tissue Eng. Part A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1002\/JPER.17-0306","article-title":"Platelet-rich fibrin\/biphasic calcium phosphate impairs osteoclast differentiation and promotes apoptosis by the intrinsic mitochondrial pathway in chronic periodontitis","volume":"90","author":"Kumar","year":"2019","journal-title":"J. Periodontol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"190","DOI":"10.4014\/jmb.1707.07042","article-title":"Inhibitory Effects of Panduratin A on Periodontitis-Induced Inflammation and Osteoclastogenesis through Inhibition of MAPK Pathways In Vitro","volume":"28","author":"Kim","year":"2018","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.actbio.2017.01.076","article-title":"Biphasic calcium phosphate ceramics for bone reconstruction: A review of biological response","volume":"53","author":"Bouler","year":"2017","journal-title":"Acta Biomater."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1902\/annals.1999.4.1.1","article-title":"Development of a classification system for periodontal diseases and conditions","volume":"4","author":"Armitage","year":"1999","journal-title":"Ann. Periodontol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e00780","DOI":"10.1016\/j.heliyon.2018.e00780","article-title":"Peripheral blood monocytes show increased osteoclast differentiation potential compared to bone marrow monocytes","volume":"4","author":"Nakamura","year":"2018","journal-title":"Heliyon"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kany, S., Vollrath, J.T., and Relja, B. (2019). Cytokines in Inflammatory Disease. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20236008"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1111\/j.1601-0825.1998.tb00255.x","article-title":"The role of IL-6 in the pathogenesis of periodontal disease","volume":"4","author":"Irwin","year":"1998","journal-title":"Oral Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"104997","DOI":"10.1016\/j.archoralbio.2020.104997","article-title":"Identification of the different salivary Interleukin-6 profiles in patients with periodontitis: A cross-sectional study","volume":"122","author":"Isola","year":"2021","journal-title":"Arch. Oral Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"403","DOI":"10.4049\/jimmunol.160.1.403","article-title":"IL-1 and TNF antagonists inhibit the inflammatory response and bone loss in experimental periodontitis","volume":"160","author":"Assuma","year":"1998","journal-title":"J. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"17023","DOI":"10.1038\/sigtrans.2017.23","article-title":"NF-\u03baB signaling in inflammation","volume":"2","author":"Liu","year":"2017","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e8","DOI":"10.4110\/in.2018.18.e8","article-title":"Regulation of osteoclast differentiation by Cytokine Networks","volume":"18","author":"Amarasekara","year":"2018","journal-title":"Immune Netw."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2447","DOI":"10.1007\/s00726-015-2039-5","article-title":"Transglutaminase 2 up-regulation is associated with RANKL\/OPG pathway in cultured HPDL cells and THP-1-differentiated macrophages","volume":"47","author":"Matarese","year":"2015","journal-title":"Amino Acids"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lee, K., Seo, I., Choi, M., and Jeong, D. (2018). Roles of Mitogen-Activated Protein Kinases in Osteoclast Biology. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19103004"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1038\/bonekey.2015.80","article-title":"Focus on the p38 MAPK signaling pathway in bone development and maintenance","volume":"4","author":"Thouverey","year":"2015","journal-title":"Bonekey Rep."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"757","DOI":"10.2353\/ajpath.2006.050907","article-title":"Tumor necrosis factor-alpha mediates diabetes-enhanced apoptosis of matrix-producing cells and impairs diabetic healing","volume":"168","author":"Liu","year":"2006","journal-title":"Am. J. Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1928","DOI":"10.1016\/j.ajpath.2013.08.017","article-title":"Bacterial infection increases periodontal bone loss in diabetic rats through enhanced apoptosis","volume":"183","author":"Pacios","year":"2013","journal-title":"Am. J. Pathol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"810563","DOI":"10.1155\/2012\/810563","article-title":"Fisetin Inhibits osteoclast differentiation via Downregulation of p38 and c-Fos-NFATc1 Signaling Pathways","volume":"2012","author":"Choi","year":"2012","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.abb.2008.03.018","article-title":"Functions of RANKL\/RANK\/OPG in bone modeling and remodeling","volume":"473","author":"Boyce","year":"2008","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1454","DOI":"10.1016\/j.yexcr.2011.03.014","article-title":"Inhibition of matrix metalloproteinase-9 activity by doxycycline ameliorates RANK ligand-induced osteoclast differentiation in vitro and in vivo","volume":"317","author":"Franco","year":"2011","journal-title":"Exp. Cell Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1111\/jcmm.12125","article-title":"Calcium ions and osteoclastogenesis initiate the induction of bone formation by coral-derived macroporous constructs","volume":"17","author":"Klar","year":"2013","journal-title":"J. Cell. Mol. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"S1","DOI":"10.1186\/ar2165","article-title":"Biology of RANK, RANKL, and osteoprotegerin","volume":"9","author":"Boyce","year":"2007","journal-title":"Arthritis Res. Ther."},{"key":"ref_44","first-page":"40","article-title":"p38 MAPK Signaling in Osteoblast Differentiation","volume":"4","author":"Ventura","year":"2016","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1242\/jcs.042770","article-title":"Akt promotes BMP2-mediated osteoblast differentiation and bone development","volume":"122","author":"Mukherjee","year":"2009","journal-title":"J. Cell Sci."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Katarivas Levy, G., Birch, M.A., Brooks, R.A., Neelakantan, S., and Markaki, A.E. (2019). Stimulation of Human Osteoblast Differentiation in Magneto-Mechanically Actuated Ferromagnetic Fiber Networks. J. Clin. Med., 8.","DOI":"10.20944\/preprints201909.0149.v1"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/S1349-0079(10)80004-0","article-title":"Regulation of Osteoblast and Odontoblast Differentiation by RUNX2","volume":"52","author":"Komori","year":"2010","journal-title":"J. Oral Biosci."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/21\/6578\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:23:27Z","timestamp":1760167407000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/21\/6578"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,30]]},"references-count":47,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["molecules26216578"],"URL":"https:\/\/doi.org\/10.3390\/molecules26216578","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,30]]}}}