{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T10:07:04Z","timestamp":1779444424778,"version":"3.53.1"},"reference-count":33,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,5,17]],"date-time":"2026-05-17T00:00:00Z","timestamp":1778976000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation","doi-asserted-by":"publisher","award":["2019\/04468-3"],"award-info":[{"award-number":["2019\/04468-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"name":"S.I.N. Implant System"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JFB"],"abstract":"<jats:p>Poor bone quality in osteoporotic patients remains a major challenge for achieving predictable osseointegration. This study serves as a mechanistic complement to previously reported structural data, aiming to investigate the molecular pathways underlying the synergy between nanostructured surfaces and autologous blood concentrates in compromised bone. Ninety-six Wistar rats were divided into healthy (SHAM) and osteoporotic (OVX) groups. Implants with nanostructured hydroxyapatite (NanoHA) or dual acid-etched (DAE) surfaces were installed in the tibiae, associated or not with leukocyte- and platelet-rich fibrin (L-PRF). Gene expression (RT-qPCR) for Runx2, Alpl, Bglap, Spp1, Tnfrsf11, and Tnfrsf11b was assessed at 7 and 30 days. In compromised systemic conditions (OVX), the NanoHA + L-PRF association promoted a robust \u201cmolecular rescue\u201d of bone metabolism. At 30 days, this synergistic group exhibited a significant upregulation of Alpl (mean: 11.69 \u00b1 1.65) and Runx2 (mean: 4.49 \u00b1 0.82) compared to DAE controls (p &lt; 0.05). Crucially, the therapy orchestrated a protective remodeling environment by significantly inducing Tnfrsf11b expression (5.50 \u00b1 0.88), effectively balancing the Tnfrsf11\/Tnfrsf11b ratio. Late-stage maturation markers (Bglap and Spp1) were also significantly elevated, effectively mimicking healthy physiological levels observed in the SHAM group. NanoHA biofunctionalization, synergistically with L-PRF, triggers a transcriptional reprogramming of the peri-implant microenvironment, mitigating the catabolic effects of estrogen deficiency. These findings provide a biological foundation for enhanced clinical predictability in high-risk patients, suggesting that local interfacial modifications can overcome systemic bone compromise.<\/jats:p>","DOI":"10.3390\/jfb17050250","type":"journal-article","created":{"date-parts":[[2026,5,18]],"date-time":"2026-05-18T10:25:16Z","timestamp":1779099916000},"page":"250","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Molecular Insights into the Synergistic Effect of Nano-Hydroxyapatite and L-PRF on Osteoporotic Osseointegration: An In Vivo Gene Expression Study"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8711-6791","authenticated-orcid":false,"given":"Ana Carolina Loyola","family":"Barbosa","sequence":"first","affiliation":[{"name":"School of Dentistry of Ribeir\u00e3o Preto, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 14040-904, SP, Brazil"},{"name":"Research & Development Department, S.I.N. Implant System, S\u00e3o Paulo 03033-021, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Augusto Gabarra","family":"J\u00fanior","sequence":"additional","affiliation":[{"name":"School of Dentistry of Ribeir\u00e3o Preto, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 14040-904, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1673-264X","authenticated-orcid":false,"given":"Lilian Eslaine Costa Mendes","family":"da Silva","sequence":"additional","affiliation":[{"name":"Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 14049-900, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0121-0996","authenticated-orcid":false,"given":"Fernando","family":"N\u00f3brega","sequence":"additional","affiliation":[{"name":"School of Dentistry of Ribeir\u00e3o Preto, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 14040-904, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edmara Tatiely Pedroso","family":"Bergamo","sequence":"additional","affiliation":[{"name":"Department of Biomaterials and Biomimetics, New York University, New York, NY 10010, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3693-3035","authenticated-orcid":false,"given":"Bruna","family":"Ghiraldini","sequence":"additional","affiliation":[{"name":"Research & Development Department, S.I.N. Implant System, S\u00e3o Paulo 03033-021, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1763-3691","authenticated-orcid":false,"given":"Roberto Sales","family":"e Pessoa","sequence":"additional","affiliation":[{"name":"Research & Development Department, S.I.N. Implant System, S\u00e3o Paulo 03033-021, SP, Brazil"},{"name":"Department of Periodontology and Implantology, School of Dentistry, University Centre of Triangulo-UNITRI, Uberl\u00e2ndia 38411-106, MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8485-9645","authenticated-orcid":false,"given":"Michel Reis","family":"Messora","sequence":"additional","affiliation":[{"name":"School of Dentistry of Ribeir\u00e3o Preto, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 14040-904, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6199-7348","authenticated-orcid":false,"given":"Sergio Scombatti de","family":"Souza","sequence":"additional","affiliation":[{"name":"School of Dentistry of Ribeir\u00e3o Preto, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 14040-904, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3859","DOI":"10.7150\/thno.96516","article-title":"Osteoporotic Osseointegration: Therapeutic Hallmarks and Engineering Strategies","volume":"14","author":"Chen","year":"2024","journal-title":"Theranostics"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/j.actbio.2016.06.035","article-title":"The Effects of Implant Topography on Osseointegration under Estrogen Deficiency Induced Osteoporotic Conditions: Histomorphometric, Transcriptional and Ultrastructural Analysis","volume":"42","author":"Du","year":"2016","journal-title":"Acta Biomater."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1111\/j.1600-0501.2010.01989.x","article-title":"The Effect of Oestrogen and Alendronate Therapies on Postmenopausal Bone Loss around Osseointegrated Titanium Implants","volume":"22","author":"Giro","year":"2011","journal-title":"Clin. Oral. Implant. Res."},{"key":"ref_4","first-page":"638043","article-title":"Platelet-Rich Fibrin Promotes Periodontal Regeneration and Enhances Alveolar Bone Augmentation","volume":"2013","author":"Li","year":"2013","journal-title":"Biomed. Res. Int."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"10203","DOI":"10.1038\/s41598-020-67167-3","article-title":"TGF\u03b2 Activity Released from Platelet-Rich Fibrin Adsorbs to Titanium Surface and Collagen Membranes","volume":"10","author":"Kargarpour","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"9507342","DOI":"10.1155\/2016\/9507342","article-title":"The Synergistic Effect of Leukocyte Platelet-Rich Fibrin and Micrometer\/Nanometer Surface Texturing on Bone Healing around Immediately Placed Implants: An Experimental Study in Dogs","volume":"2016","author":"Neiva","year":"2016","journal-title":"BioMed Res. Int."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"597","DOI":"10.11607\/jomi.5887","article-title":"Evaluation of a Titanium Surface Treated with Hydroxyapatite Nanocrystals on Osteoblastic Cell Behavior: An In Vitro Study","volume":"33","author":"Martinez","year":"2018","journal-title":"Int. J. Oral Maxillofac. Implant."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.5005\/jp-journals-10024-3786","article-title":"Evaluation of Effectiveness of Nanocrystalline Hydroxyapatite and Demineralized Bone Matrix Combined with Titanium-Platelet Rich Fibrin for Ridge Preservation: A Randomized Controlled Clinical Trial","volume":"25","author":"Bajaj","year":"2025","journal-title":"J. Contemp. Dent. Pract."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2846","DOI":"10.1016\/j.msec.2013.03.004","article-title":"Effect of Grain Size on Mechanical, Surface and Biological Properties of Microwave Sintered Hydroxyapatite","volume":"33","author":"Dasgupta","year":"2013","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Almeida, D., Sartoretto, S.C., Calasans-Maia, J.D.A., Ghiraldini, B., Bezerra, F.J.B., Granjeiro, J.M., and Calasans-Maia, M.D. (2023). In Vivo Osseointegration Evaluation of Implants Coated with Nanostructured Hydroxyapatite in Low Density Bone. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0282067"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Gabarra J\u00fanior, J.A., N\u00f3brega, F., Oliveira, P.G., Bergamo, E.T., Cadore, U., Gomes, M.Z.D.V., Kjellin, P., Chaushu, L., Bezerra, F., and Ghiraldini, B. (2023). Evaluation of Implant Surface Modification with Nanohydroxyapatite Associated with the Use of L-PRF: In Vivo Study in Rats. J. Funct. Biomater., 14.","DOI":"10.3390\/jfb14070370"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1002\/jbm.a.37052","article-title":"Influence of Nano-hydroxyapatite Coating Implants on Gene Expression of Osteogenic Markers and micro-CT Parameters. An in Vivo Study in Diabetic Rats","volume":"109","author":"Taba","year":"2021","journal-title":"J. Biomed. Mater. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.biomaterials.2010.09.011","article-title":"The Correlation between Gene Expression of Proinflammatory Markers and Bone Formation during Osseointegration with Titanium Implants","volume":"32","author":"Omar","year":"2011","journal-title":"Biomaterials"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1111\/j.1600-0501.2012.02496.x","article-title":"Correlation between Molecular Signals and Bone Bonding to Titanium Implants","volume":"24","author":"Monjo","year":"2013","journal-title":"Clin. Oral Implant. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7033","DOI":"10.1016\/j.biomaterials.2013.05.025","article-title":"Comparison of Mesenchymal Stem Cells Derived from Gingival Tissue and Periodontal Ligament in Different Incubation Conditions","volume":"34","author":"Yang","year":"2013","journal-title":"Biomaterials"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.reprotox.2020.10.008","article-title":"The Optimal Compound Reference Genes for qRT-PCR Analysis in the Developing Rat Long Bones under Physiological Conditions and Prenatal Dexamethasone Exposure Model","volume":"98","author":"Han","year":"2020","journal-title":"Reprod. Toxicol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1016\/j.actbio.2017.07.009","article-title":"Selective Effect of Hydroxyapatite Nanoparticles on Osteoporotic and Healthy Bone Formation Correlates with Intracellular Calcium Homeostasis Regulation","volume":"59","author":"Zhao","year":"2017","journal-title":"Acta Biomater."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1111\/j.1600-0501.2009.01775.x","article-title":"Effects of Titanium Surface Topography on Bone Integration: A Systematic Review","volume":"20","author":"Wennerberg","year":"2009","journal-title":"Clin. Oral Implant. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1586\/erd.09.74","article-title":"Improving Osseointegration of Dental Implants","volume":"7","author":"Elias","year":"2010","journal-title":"Expert Rev. Med. Devices"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s13287-017-0485-6","article-title":"Differential Long Noncoding RNA\/mRNA Expression Profiling and Functional Network Analysis during Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells","volume":"8","author":"Zhang","year":"2017","journal-title":"Stem Cell Res. Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1080\/09537104.2017.1293812","article-title":"The Impact of the Centrifuge Characteristics and Centrifugation Protocols on the Cells, Growth Factors, and Fibrin Architecture of a Leukocyte- and Platelet-Rich Fibrin (L-PRF) Clot and Membrane","volume":"29","author":"Pinto","year":"2018","journal-title":"Platelets"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1080\/09537104.2017.1327654","article-title":"Leucocyte- and Platelet-Rich Fibrin (L-PRF) as a Regenerative Medicine Strategy for the Treatment of Refractory Leg Ulcers: A Prospective Cohort Study","volume":"29","author":"Pinto","year":"2018","journal-title":"Platelets"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1007\/s00784-017-2133-z","article-title":"Use of Platelet-Rich Fibrin in Regenerative Dentistry: A Systematic Review","volume":"21","author":"Miron","year":"2017","journal-title":"Clin. Oral Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1016\/S0945-053X(00)00108-6","article-title":"Osteopontin: A Versatile Regulator of Inflammation and Biomineralization","volume":"19","author":"Giachelli","year":"2000","journal-title":"Matrix Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2353","DOI":"10.1007\/s00784-016-1719-1","article-title":"Comparative Release of Growth Factors from PRP, PRF, and Advanced-PRF","volume":"20","author":"Kobayashi","year":"2016","journal-title":"Clin. Oral Investig."},{"key":"ref_27","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_28","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1111\/j.1600-051X.2011.01810.x","article-title":"The RANKL-OPG System in Clinical Periodontology","volume":"39","author":"Belibasakis","year":"2012","journal-title":"J. Clin. Periodontol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"113448","DOI":"10.1016\/j.colsurfb.2023.113448","article-title":"Effect of Osteoblasts on Osteoclast Differentiation and Activity Induced by Titanium with Nanotopography","volume":"229","author":"Ferraz","year":"2023","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1111\/clr.13575","article-title":"Titanium Implant Surface Properties Enhance Osseointegration in Ovariectomy Induced Osteoporotic Rats without Pharmacologic Intervention","volume":"31","author":"Lotz","year":"2020","journal-title":"Clin. Oral Implant. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1177\/154405910408300704","article-title":"Enhanced Bone Apposition to a Chemically Modified SLA Titanium Surface","volume":"83","author":"Buser","year":"2004","journal-title":"J. Dent. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1111\/jre.12827","article-title":"Hydrophilic Titanium Surface Modulates Early Stages of Osseointegration in Osteoporosis","volume":"56","author":"Siqueira","year":"2021","journal-title":"J. Periodontal Res."},{"key":"ref_33","unstructured":"Rezende, M.C.R.R.A., Wada, C.M., Lopes, M.R.A.N.E., Capalbo, L.C., Gon\u00e7alves, V.M., Valente, A.D.B., and de Oliveira, J.A.G. (2015). O papel da adsor\u00e7\u00e3o de prote\u00ednas na osseointegra\u00e7\u00e3o. Arch. Health Investig., 4, Available online: https:\/\/www.archhealthinvestigation.com.br\/ARCHI\/article\/view\/903."}],"container-title":["Journal of Functional Biomaterials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-4983\/17\/5\/250\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T09:47:13Z","timestamp":1779443233000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-4983\/17\/5\/250"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,17]]},"references-count":33,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["jfb17050250"],"URL":"https:\/\/doi.org\/10.3390\/jfb17050250","relation":{},"ISSN":["2079-4983"],"issn-type":[{"value":"2079-4983","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,17]]}}}