{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:17:40Z","timestamp":1785421060377,"version":"3.56.0"},"reference-count":34,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,7,26]],"date-time":"2026-07-26T00:00:00Z","timestamp":1785024000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Spanish Government and the Ministry of Science and Innovation of Spain","award":["PID2022-137496OB-I00"],"award-info":[{"award-number":["PID2022-137496OB-I00"]}]},{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/01rkhv978","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Gobierno de Espa\u00f1a"},{"id":[{"id":"https:\/\/ror.org\/038jjxj40","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Guided bone regeneration (GBR) heavily relies on barrier membranes, with collagen being the clinical standard material. Human amniotic epithelial cells (hAECs) represent a promising, non-controversial stem cell source with substantial osteogenic and immunomodulatory potential. This study aimed to comparatively evaluate the structural characteristics of three commercial collagen membranes (Biocollagen\u00ae, Derma\u00ae, and VantyColl\u00ae) and their influence on the biological behavior, viability, and osteogenic differentiation of hAECs and hFOB 1.19 human fetal osteoblasts. The microarchitecture was assessed via scanning electron microscopy (SEM). Biological response was evaluated over 14 days, using MTT assays, calcium and phosphorus quantification, alkaline phosphatase (ALP) activity, and quantitative real-time PCR (qRT-PCR) for osteogenic markers (Runx2, Osterix, ALP, and OPN). SEM revealed a dense lamellar structure for Biocollagen\u00ae, a fibrillar and oriented architecture for Derma\u00ae, and a highly porous network for VantyColl\u00ae. Both cell types adhered to and proliferated on all membranes. Derma\u00ae provided the best long-term proliferative support for both lineages. Conversely, VantyColl\u00ae induced robust early osteoblastic differentiation, marked by exceptional upregulation of Osterix (24.93-fold) and Runx2 (2.64-fold), though it exhibited diminished long-term hAEC viability. Ultimately, collagen membrane microarchitecture dictates cell fate; dense fibrillar networks (Derma\u00ae) favor sustained growth and late matrix maturation (OPN), whereas high-porosity scaffolds (VantyColl\u00ae) amplify early osteoinductive cascades.<\/jats:p>","DOI":"10.3390\/ijms27156660","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T09:47:41Z","timestamp":1785145661000},"page":"6660","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Human Amniotic Epithelial Stem Cells and Osteoblast Cells Behavior on Collagen Membranes for Bone Guided Regeneration"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5660-735X","authenticated-orcid":false,"given":"Antonio","family":"P\u00e9rez-P\u00e9rez","sequence":"first","affiliation":[{"name":"Departamento de Bioqu\u00edmica M\u00e9dica y Biolog\u00eda Molecular e Inmunolog\u00eda, Hospital Universitario Virgen Macarena, Facultad de Medicina, Universidad de Sevilla, Avenida S\u00e1nchez Pizju\u00e1n 4, 41009 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6824-1412","authenticated-orcid":false,"given":"Javier","family":"Gil","sequence":"additional","affiliation":[{"name":"Biomaterials i Interf\u00edcies Orals Bioinspirats (BOBI), Departament de Ci\u00e8ncia i Enginyeria de Materials i Centre de Recerca en Ci\u00e8ncia i Enginyeria Multiescala de Barcelona (CCEM), Escola d\u2019Enginyeria de Barcelona Est (EEBE), Universitat Polit\u00e8cnica de Catalunya \u00b7 BarcelonaTech (UPC), Av. Eduard Maristany, 16, 08019 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabela","family":"Bueno-Bianchi","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Loreto","family":"Monsalve-Guil","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iv\u00e1n","family":"Ortiz-Garcia","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alvaro","family":"Jim\u00e9nez-Guerra","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrique","family":"N\u00fa\u00f1ez-M\u00e1rquez","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0228-484X","authenticated-orcid":false,"given":"Eugenio","family":"Velasco-Ortega","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7766-1462","authenticated-orcid":false,"given":"Jos\u00e9 Luis","family":"Rond\u00f3n Romero","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8638-8680","authenticated-orcid":false,"given":"Victor","family":"S\u00e1nchez-Margalet","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica M\u00e9dica y Biolog\u00eda Molecular e Inmunolog\u00eda, Hospital Universitario Virgen Macarena, Facultad de Medicina, Universidad de Sevilla, Avenida S\u00e1nchez Pizju\u00e1n 4, 41009 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0157-9643","authenticated-orcid":false,"given":"Jes\u00fas","family":"Moreno-Mu\u00f1oz","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Universidad de Sevilla, 41004 Sevilla, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bubalo, M., Dugonjic, S., Dubovina, D., Stojanovic, Z., Gardasevic, M., Mijatovic, J., Milovanovic, B., Stevic, M., Stepovic, M., and Jeremic, D. (2026). Bone substitutes in alveolar ridge augmentation: A narrative literature review. J. Funct. Biomater., 17.","DOI":"10.3390\/jfb17040176"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e3681","DOI":"10.31661\/gmj.v13iSP1.3681","article-title":"The role of guided bone regeneration in enhancing dental implant success: Guided bone regeneration in dental implant","volume":"13","author":"Taghizadeh","year":"2024","journal-title":"Galen. Med. J."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Yang, Z., Wu, C., Shi, H., Luo, X., Sun, H., Wang, Q., and Zhang, D. (2022). Advances in barrier membranes for guided bone regeneration techniques. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.921576"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Sbricoli, L., Guazzo, R., Annunziata, M., Gobbato, L., Bressan, E., and Nastri, L. (2020). Selection of collagen membranes for bone regeneration: A literature review. Materials, 13.","DOI":"10.3390\/ma13030786"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5457","DOI":"10.1021\/acsbiomaterials.3c00690","article-title":"Guided bone regeneration using barrier membrane in dental applications","volume":"9","author":"Kim","year":"2023","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/j.actbio.2015.10.005","article-title":"Guided bone regeneration using resorbable membrane and different bone substitutes: Early histological and molecular events","volume":"29","author":"Elgali","year":"2016","journal-title":"Acta Biomater."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1007\/s13770-025-00704-1","article-title":"Implantable dental barrier membranes as regenerative medicine in dentistry: A comprehensive review","volume":"22","author":"Chen","year":"2025","journal-title":"Tissue Eng. Regen. Med."},{"key":"ref_8","first-page":"1","article-title":"Functional requirements for guided bone regeneration\/guided tissue regeneration membrane design: Progress and challenges","volume":"12","author":"Zhan","year":"2025","journal-title":"Periodontology 2000"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"032003","DOI":"10.1088\/2057-1976\/ad1e75","article-title":"Advances in guided bone regeneration membranes: A comprehensive review of materials and techniques","volume":"10","author":"Ali","year":"2024","journal-title":"Biomed. Phys. Eng. Express"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1177\/0963689717742819","article-title":"The immunomodulatory properties of amniotic cells: The two sides of the coin","volume":"27","author":"Magatti","year":"2018","journal-title":"Cell Transplant."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1007\/978-1-0716-3609-1_13","article-title":"Amniotic membrane and amniotic epithelial cell culture","volume":"Volume 2749","author":"Canciello","year":"2024","journal-title":"Epithelial Cell Culture"},{"key":"ref_12","first-page":"5717","article-title":"Differentiation of human amniotic epithelial cells into osteoblasts is induced by mechanical stretch via the Wnt\/\u03b2-catenin signalling pathway","volume":"18","author":"Luan","year":"2018","journal-title":"Mol. Med. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Riedel, R., P\u00e9rez-Amodio, S., Cabo-Zabala, L., Velasco-Ortega, E., Maym\u00f3, J., Gil, J., Monsalve-Guil, L., Ortiz-Garcia, I., P\u00e9rez-P\u00e9rez, A., and S\u00e1nchez-Margalet, V. (2024). Influence of the surface topography of titanium dental implants on the behavior of human amniotic stem cells. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25137416"},{"key":"ref_14","first-page":"685","article-title":"Alveolar ridge preservation procedures performed with freeze-dried bone allograft: Clinical and histologic outcomes in a case series: Part ii","volume":"44","author":"Barone","year":"2024","journal-title":"Int. J. Periodontics Restor. Dent."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Bertran Faus, A., Cordero Bayo, J., Velasco-Ortega, E., Torrejon-Moya, A., Fern\u00e1ndez-Velilla, F., Garc\u00eda, F., and L\u00f3pez-L\u00f3pez, J. (2022). Customized titanium mesh for guided bone regeneration with autologous bone and xenograft. Materials, 15.","DOI":"10.3390\/ma15186271"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2157","DOI":"10.1002\/jbm.b.35060","article-title":"Asymmetric resorbable-based dental barrier membrane for periodontal guided tissue regeneration and guided bone regeneration: A review","volume":"110","author":"Bee","year":"2022","journal-title":"J. Biomed. Mater. Res. B"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1111\/prd.12518","article-title":"Bone regeneration in implant dentistry: Which are the factors affecting the clinical outcome?","volume":"93","author":"Donos","year":"2023","journal-title":"Periodontology 2000"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1545","DOI":"10.1002\/term.2670","article-title":"Using biomimetically mineralized collagen membranes with different surface stiffness to guide regeneration of bone defects","volume":"12","author":"Wang","year":"2018","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"101607","DOI":"10.1016\/j.jormas.2023.101607","article-title":"Surface topography of resorbable porcine collagen membranes, and their effect on early osteogenesis: An In Vitro study","volume":"124","author":"Marques","year":"2023","journal-title":"J. Stomatol. Oral. Maxillofac. Surg."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Antonacci, D., Padula, R., Gaudelli, F., Catalano, I., and Mastrangelo, F. (2025). In Vitro evaluation and comparative analysis of resorbable membranes for guided bone regeneration. Medicina, 61.","DOI":"10.3390\/medicina61091720"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1080\/03008207.2017.1384471","article-title":"Contact guidance and collective migration in the advancing epithelial monolayer","volume":"59","author":"Lee","year":"2018","journal-title":"Connect. Tissue Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1038\/s41536-019-0076-5","article-title":"Synthetic scaffolds for musculoskeletal tissue engineering: Cellular responses to fiber parameters","volume":"4","author":"Jenkins","year":"2019","journal-title":"npj Regen. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"111787","DOI":"10.1016\/j.msec.2020.111787","article-title":"The effects of alignment and diameter of electrospun fibers on the cellular behaviors and osteogenesis of BMSCs","volume":"120","author":"Xie","year":"2021","journal-title":"Mater. Sci. Eng. C Mater. Biol. Appl."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"21066","DOI":"10.1039\/D6RA00640J","article-title":"A novel PCL fiber membrane with a gradient structure for guided bone regeneration","volume":"16","author":"Jin","year":"2026","journal-title":"RSC Adv."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"11142","DOI":"10.1039\/D4TB00962B","article-title":"Scalable fabrication of porous membrane incorporating human extracellular matrix-like collagen for guided bone regeneration","volume":"12","author":"Wang","year":"2024","journal-title":"J. Mater. Chem. B"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ashoka Sreeja, H., Couso-Queiruga, E., Raabe, C., Chappuis, V., and Asparuhova, M.B. (2025). Biofunctionalization of collagen barrier membranes with bone-conditioned medium, as a natural source of growth factors, enhances osteoblastic cell behavior. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26041610"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1007\/s11033-026-11925-5","article-title":"Autologous guided bone regeneration: A comprehensive narrative review","volume":"53","author":"Yu","year":"2026","journal-title":"Mol. Biol. Rep."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ren, Y., Fan, L., Alkildani, S., Liu, L., Emmert, S., Najman, S., Rimashevskiy, D., Schnettler, R., Jung, O., and Xiong, X. (2022). Barrier membranes for guided bone regeneration (GBR): A focus on recent advances in collagen membranes. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232314987"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Luo, Q., Gao, K., Li, Y., Zhang, Z., Chen, S., and Zhou, J. (2025). Osteogenesis activity and porosity effect of biodegradable MG-GA alloys barrier membrane for guided bone regeneration: An in vitro and in vivo study in rabbits. Biomedicines, 13.","DOI":"10.3390\/biomedicines13081940"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1504","DOI":"10.5966\/sctm.2014-0118","article-title":"Osteogenic differentiation of human amniotic epithelial cells and its application in alveolar defect restoration","volume":"3","author":"Jiawen","year":"2014","journal-title":"Stem Cells Transl. Med."},{"key":"ref_31","first-page":"791","article-title":"Human amniotic epithelial cells regulate osteoblast differentiation through the secretion of TGF\u03b21 and microRNA-34a-5p","volume":"41","author":"Wang","year":"2018","journal-title":"Int. J. Mol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1007\/978-1-4419-1050-9_5","article-title":"Regulation of osteoblast differentiation by Runx2","volume":"658","author":"Komori","year":"2010","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"39322","DOI":"10.1038\/srep39322","article-title":"The mechanism of a chitosan-collagen composite film used as biomaterial support for MC3T3-E1 cell differentiation","volume":"6","author":"Wang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Romasco, T., Tumedei, M., Inchingolo, F., Pignatelli, P., Montesani, L., Iezzi, G., Petrini, M., Piattelli, A., and Di Pietro, N. (2022). A Narrative review on the effectiveness of bone regeneration procedures with Osteobiol\u00ae collagenated porcine grafts: The translational research experience over 20 years. J. Funct. Biomater., 13.","DOI":"10.3390\/jfb13030121"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6660\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T04:21:19Z","timestamp":1785212479000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6660"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,26]]},"references-count":34,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27156660"],"URL":"https:\/\/doi.org\/10.3390\/ijms27156660","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,26]]}}}