{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T03:30:29Z","timestamp":1771039829881,"version":"3.50.1"},"reference-count":56,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,10,19]],"date-time":"2025-10-19T00:00:00Z","timestamp":1760832000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JFB"],"abstract":"<jats:p>This study characterized the biological response of MG-63 cells to synthetic, hydroxyapatite scaffolds (HAsint) fabricated via fused filament fabrication. Scaffolds were compared to 2D plate-adherent cultures using six assays: cell morphology and distribution with scanning electron microscopy and confocal laser scanning microscopy; cell proliferation and cytotoxicity via WST-1 tetrazolium assay; relative osteogenic gene expression through reverse-transcription\u2013quantitative polymerase chain reaction, and protein synthesis via multiplex immunoassay. Data were analyzed using one-way ANOVA. Results confirmed high cell viability and uniform distribution on HAsint scaffolds. Proliferation increased significantly over 7 days, though direct cytotoxicity also increased, likely due to the static conditions of the experiment and, subsequently, the high ion reprecipitation from scaffold degradation. Importantly, HAsint scaffolds significantly enhanced osteogenic marker expression of phosphatase alkaline (ALPL), osteopontin (OPN), and osteocalcin (OCN) genes, and elevated concentrations of interleukins (IL)-6, IL-8 and matrix metalloproteinase 1 compared to plate-adherent controls. It can be concluded that 3D-printed HAsint scaffolds support robust osteogenic differentiation and proliferation despite inducing a transient cytotoxic response in vitro. The marked upregulation of key osteogenic genes and proteins confirms the scaffolds\u2019 bioactivity and highlights their potential for bone tissue engineering applications.<\/jats:p>","DOI":"10.3390\/jfb16100392","type":"journal-article","created":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T10:44:10Z","timestamp":1760957050000},"page":"392","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Biological Characterization of 3D-Printed, Sintered Hydroxyapatite Scaffolds Obtained by Fused Filament Fabrication: An In Vitro Study"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8655-5290","authenticated-orcid":false,"given":"Eddy","family":"Shan","sequence":"first","affiliation":[{"name":"Etiology and Therapy of Periodontal and Peri-Implant Diseases (ETEP) Research Group, Complutense University of Madrid, 28040 Madrid, Spain"},{"name":"Doctorate Programme in Dental Sciences, Complutense University of Madrid, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristina","family":"Chamorro","sequence":"additional","affiliation":[{"name":"Etiology and Therapy of Periodontal and Peri-Implant Diseases (ETEP) Research Group, Complutense University of Madrid, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3399-1069","authenticated-orcid":false,"given":"Ana","family":"Ferr\u00e1ndez-Montero","sequence":"additional","affiliation":[{"name":"Institute of Ceramics and Glass (ICV), Spanish National Research Council (CSIC), 28049 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-1139-1907","authenticated-orcid":false,"given":"Rosa M.","family":"Martin-Rodriguez","sequence":"additional","affiliation":[{"name":"Doctorate Programme in Dental Sciences, Complutense University of Madrid, 28040 Madrid, Spain"},{"name":"Institute of Ceramics and Glass (ICV), Spanish National Research Council (CSIC), 28049 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3376-5232","authenticated-orcid":false,"given":"Leire","family":"Virto","sequence":"additional","affiliation":[{"name":"Etiology and Therapy of Periodontal and Peri-Implant Diseases (ETEP) Research Group, Complutense University of Madrid, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda Jos\u00e9","family":"Mar\u00edn","sequence":"additional","affiliation":[{"name":"Etiology and Therapy of Periodontal and Peri-Implant Diseases (ETEP) Research Group, Complutense University of Madrid, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3377-6844","authenticated-orcid":false,"given":"Bego\u00f1a","family":"Ferrari","sequence":"additional","affiliation":[{"name":"Institute of Ceramics and Glass (ICV), Spanish National Research Council (CSIC), 28049 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2874-6640","authenticated-orcid":false,"given":"Antonio Javier","family":"Sanchez-Herencia","sequence":"additional","affiliation":[{"name":"Institute of Ceramics and Glass (ICV), Spanish National Research Council (CSIC), 28049 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3129-1416","authenticated-orcid":false,"given":"Elena","family":"Figuero","sequence":"additional","affiliation":[{"name":"Etiology and Therapy of Periodontal and Peri-Implant Diseases (ETEP) Research Group, Complutense University of Madrid, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6293-5755","authenticated-orcid":false,"given":"Mariano","family":"Sanz","sequence":"additional","affiliation":[{"name":"Etiology and Therapy of Periodontal and Peri-Implant Diseases (ETEP) Research Group, Complutense University of Madrid, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.actbio.2017.08.030","article-title":"Regenerating bone with bioactive glass scaffolds: A review of in vivo studies in bone defect models","volume":"62","author":"Roether","year":"2017","journal-title":"Acta Biomater."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1111\/jcpe.13058","article-title":"Bone grafts: Which is the ideal biomaterial?","volume":"46","author":"Haugen","year":"2019","journal-title":"J. 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