{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T18:00:30Z","timestamp":1791223230246,"version":"4.1.0"},"reference-count":40,"publisher":"Georg Thieme Verlag KG","issue":"04","license":[{"start":{"date-parts":[[2025,3,12]],"date-time":"2025-03-12T00:00:00Z","timestamp":1741737600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Eur J Dent"],"published-print":{"date-parts":[[2025,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Regenerative periodontal surgical approaches require scaffolds in a form that can fill narrow and irregular defects. Each scaffold must be specially designed to conform to the shape of the specific defect. The aim of this study was to fabricate nanohydroxyapatite chitosan-gelatin (nHA\/KG) pastes with different composition percentages and to analyze the differences in physical, chemical, and biological characteristics in response to periodontal tissue regeneration in vitro.<\/jats:p><jats:p>The nHA\/KG paste was prepared at three different concentrations of inorganic and organic contents (70\/30; 75\/25; and 80\/20) by mixing nHA powder, chitosan flakes, and gelatin powder. The ratio of chitosan and gelatin on all nHA\/KG pastes is 1:1. The three nHA\/KG pastes were tested for the following rheology and bioactivity properties in simulated body fluid (SBF): pH value, swelling, degradability, surface morphology, and cell attachment by scanning electron microscopy and chemical structure by Fourier transform infrared (FTIR). Osteoblasts and fibroblasts were analyzed for proliferation using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and for cell proliferation by reverse transcription quantitative real-time polymerase chain reaction of COL1, alkaline phosphatase (ALP), osteocalcin (OCN), and RUNX2.<\/jats:p><jats:p>Analysis of variance followed by Tukey's post hoc, Kruskal\u2013Wallis, Wilcoxon, and paired sample t-tests were performed according to each data type.<\/jats:p><jats:p>The nHA\/KG paste showed gel-like physical characteristics. The pH value after SBF immersion was stable at pH\u2009\u00b1\u20097.0, although the pH of the nHA\/KG 80\/20 paste decreased to pH 6.3 on day 14. The three paste preparations showed significant differences in swelling (p\u2009&lt;\u20090.05) on days 1 and 14 and in the degradability ratio on days 1, 2, and 7 (p\u2009&lt;\u20090.05). The three-dimensional scaffold surface morphology differed depending on the immersion time. The FTIR test showed the presence of PO4\n                  3-, CO3\n                  2-, -OH, amide I, and amide II functional groups in all paste variants. The nHA\/KG 75\/25 paste had the most stable structure during the immersion period. Biological tests showed a viability ratio of osteoblasts and fibroblasts \u2265 70%. The paste could stimulate the messenger ribonucleic acid expression of the COL1, ALP, OCN, and RUNX2.<\/jats:p><jats:p>The nHA\/KG bone graft paste showed good potential as an injectable scaffold, with the nHA\/KG 75\/25 paste being the best of the three pastes tested here.<\/jats:p>","DOI":"10.1055\/s-0044-1800826","type":"journal-article","created":{"date-parts":[[2025,3,12]],"date-time":"2025-03-12T20:09:46Z","timestamp":1741810186000},"page":"1035-1045","source":"Crossref","is-referenced-by-count":2,"title":["Bone Graft Paste Nanohydroxyapatite Chitosan-Gelatin (nHA\/KG) for Periodontal Regeneration: Study on Three-Dimensional Cell Culture"],"prefix":"10.1055","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3586-8788","authenticated-orcid":false,"given":"Nadhia Anindhita","family":"Harsas","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Endang Winiati","family":"Bachtiar","sequence":"additional","affiliation":[{"name":"Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"},{"name":"Oral Science Research Center, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa Rinanda","family":"Amir","sequence":"additional","affiliation":[{"name":"Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"},{"name":"Oral Science Research Center, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachmat","family":"Mauludin","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, School of Pharmacy, Bandung Institute of Technology, Bandung, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0035-4821","authenticated-orcid":false,"given":"Sunarso","family":"Sunarso","sequence":"additional","affiliation":[{"name":"Department of Dental Material, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viona","family":"Yosefa","sequence":"additional","affiliation":[{"name":"Periodontology Specialist Program, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Savvyana","sequence":"additional","affiliation":[{"name":"Periodontology Specialist Program, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8767-0941","authenticated-orcid":false,"given":"Fatimah Maria","family":"Tadjoedin","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4660-3045","authenticated-orcid":false,"given":"Yuniarti","family":"Soeroso","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"194","published-online":{"date-parts":[[2025,3,12]]},"reference":[{"issue":"01","key":"ref1","first-page":"135","article-title":"The role of scaffolds in periodontal regeneration","volume":"45","author":"M Shalini","year":"2017","journal-title":"Int J Pharm Sci Rev Res"},{"issue":"01","key":"ref2","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.cden.2021.06.004","article-title":"Scaffolds in periodontal regenerative treatment","volume":"66","author":"S Yamada","year":"2022","journal-title":"Dent Clin North Am"},{"key":"ref3","doi-asserted-by":"crossref","first-page":"111158","DOI":"10.1016\/j.msec.2020.111158","article-title":"Injectable hydrogel-loaded nano-hydroxyapatite that improves bone regeneration and alveolar ridge promotion","volume":"116","author":"Y Pan","year":"2020","journal-title":"Mater Sci Eng C"},{"issue":"02","key":"ref4","doi-asserted-by":"crossref","first-page":"435","DOI":"10.3390\/ijms20020435","article-title":"Current trends in fabrication of biomaterials for bone and cartilage regeneration: materials modifications and biophysical stimulations","volume":"20","author":"A Przekora","year":"2019","journal-title":"Int J Mol Sci"},{"issue":"04","key":"ref5","doi-asserted-by":"crossref","first-page":"423","DOI":"10.4103\/0972-124X.184036","article-title":"Comparative evaluation of the efficacy of synthetic nanocrystalline hydroxyapatite bone graft (Ostim\u00ae) and synthetic microcrystalline hydroxyapatite bone graft (Osteogen\u00ae) in the treatment of human periodontal intrabony defects: a clinical and denta scan study","volume":"20","author":"M Kamboj","year":"2016","journal-title":"J Indian Soc Periodontol"},{"issue":"06","key":"ref6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3390\/ma10060602","article-title":"Chitosan biomaterials for current and potential dental applications","volume":"10","author":"S Husain","year":"2017","journal-title":"Materials (Basel)"},{"issue":"02","key":"ref7","first-page":"107","article-title":"Evaluation of regenerative therapy using cell sheet through cementum protein-1 expression on macaca nemestrina","volume":"9","author":"S W Astuty","year":"2017","journal-title":"Int J Appl Pharmaceut"},{"issue":"02","key":"ref8","doi-asserted-by":"crossref","first-page":"13","DOI":"10.30880\/ijie.2022.14.02.003","article-title":"The characteristics, swelling ratio and water content percentage of chitosan-gelatin\/limestone-based carbonate hydroxyapatite composite scaffold","volume":"14","author":"D Rianti","year":"2022","journal-title":"Int J Integ Eng"},{"key":"ref9","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1016\/j.msec.2016.03.060","article-title":"Fabrication and characterization of novel nano-biocomposite scaffold of chitosan-gelatin-alginate-hydroxyapatite for bone tissue engineering","volume":"64","author":"C Sharma","year":"2016","journal-title":"Mater Sci Eng C"},{"issue":"02","key":"ref10","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.jare.2013.02.004","article-title":"In vitro study of nano-hydroxyapatite\/chitosan-gelatin composites for bio-applications","volume":"5","author":"K R Mohamed","year":"2014","journal-title":"J Adv Res"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.22203\/eCM.v032a01","article-title":"Hydroxyapatite nanoparticle injectable hydrogel scaffold to support osteogenic differentiation of human mesenchymal stem cells","volume":"32","author":"A A Thorpe","year":"2016","journal-title":"Eur Cell Mater"},{"issue":"03","key":"ref12","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S1532-3382(12)70021-0","article-title":"Minimally invasive surgical techniques in periodontal regeneration","volume":"12","author":"P Cortellini","year":"2012","journal-title":"J Evid Based Dent Pract"},{"key":"ref13","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.jmbbm.2011.12.007","article-title":"Mechanical properties and in vitro bioactivity of injectable and self-setting calcium sulfate\/nano-HA\/collagen bone graft substitute","volume":"12","author":"N M Hu","year":"2012","journal-title":"J Mech Behav Biomed Mater"},{"issue":"18","key":"ref14","doi-asserted-by":"crossref","first-page":"3413","DOI":"10.1016\/j.biomaterials.2006.01.039","article-title":"Biodegradable and bioactive porous polymer\/inorganic composite scaffolds for bone tissue engineering","volume":"27","author":"K Rezwan","year":"2006","journal-title":"Biomaterials"},{"issue":"01","key":"ref15","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.mseb.2009.11.012","article-title":"Study of nanobiomaterial hydroxyapatite in simulated body fluid: formation and growth of apatite","volume":"168","author":"P N Chavan","year":"2010","journal-title":"Mater Sci Eng B"},{"issue":"04","key":"ref16","first-page":"1","article-title":"The use of simulated body fluid (SBF) for assessing materials bioactivity in the context of tissue engineering: Review and challenges","volume":"5","author":"F Baino","year":"2020","journal-title":"Biomimetics (Basel)"},{"issue":"01","key":"ref17","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1111\/prd.12079","article-title":"In vitro models for evaluation of periodontal wound healing\/regeneration","volume":"68","author":"M Weinreb","year":"2015","journal-title":"Periodontol 2000"},{"issue":"01","key":"ref18","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1186\/s12903-020-01124-4","article-title":"Development and application of a 3D periodontal in vitro model for the evaluation of fibrillar biomaterials","volume":"20","author":"F Koch","year":"2020","journal-title":"BMC Oral Health"},{"issue":"04","key":"ref19","doi-asserted-by":"crossref","first-page":"176","DOI":"10.20473\/j.djmkg.v43.i4.p176-180","article-title":"Biocompatibility and osteoconductivity of injectable bone xenograft, hydroxyapatite and hydroxyapatite-chitosan on osteoblast culture","volume":"43","author":"E W Bachtiar","year":"2010","journal-title":"Dent J (Majalah Kedokteran Gigi)"},{"issue":"12","key":"ref20","first-page":"ZF01","article-title":"Potency of injectable hydroxyapatite chitosan scaffold for bone regeneration","volume":"11","author":"E W Bachtiar","year":"2017","journal-title":"J Clin Diagn Res"},{"key":"ref21","first-page":"905","article-title":"Physicochemical characterization of nanohydroxyapatite powder in simulated body fluid immersion: a pilot study","volume":"16","author":"N A Harsas","year":"2023","journal-title":"Int J Nanoelectron Mater"},{"issue":"05","key":"ref22","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1002\/jbm.a.36620","article-title":"Simulated body fluid and the novel bioactive materials derived from it","volume":"107","author":"T Kokubo","year":"2019","journal-title":"J Biomed Mater Res A"},{"issue":"06","key":"ref23","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.3892\/ijmm.2013.1516","article-title":"Effects of apatite particle size in two apatite\/collagen composites on the osteogenic differentiation profile of osteoblastic cells","volume":"32","author":"W Hatakeyama","year":"2013","journal-title":"Int J Mol Med"},{"issue":"04","key":"ref24","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1089\/ast.2012.0922","article-title":"Studies on culture and osteogenic induction of human mesenchymal stem cells under CO2-independent conditions","volume":"13","author":"J Chen","year":"2013","journal-title":"Astrobiology"},{"issue":"12","key":"ref25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3390\/biomedicines8120616","article-title":"Enhancing mechanical properties and biological performances of injectable bioactive glass by gelatin and chitosan for bone small defect repair","volume":"8","author":"M Sohrabi","year":"2020","journal-title":"Biomedicines"},{"issue":"06","key":"ref26","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1016\/j.cocis.2014.09.002","article-title":"The contribution of rheology for designing hydroxyapatite biomaterials","volume":"19","author":"M Maas","year":"2014","journal-title":"Curr Opin Colloid Interface Sci"},{"issue":"04","key":"ref27","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1515\/cti-2022-0010","article-title":"Basic principle and good practices of rheology for polymers for teachers and beginners","volume":"4","author":"H Ramli","year":"2022","journal-title":"Chem Teach Int"},{"issue":"01","key":"ref28","doi-asserted-by":"crossref","first-page":"15957","DOI":"10.1038\/s41598-019-52042-7","article-title":"Nanohydroxyapatite reinforced chitosan composite hydrogel with tunable mechanical and biological properties for cartilage regeneration","volume":"9","author":"B YS Kumar","year":"2019","journal-title":"Sci Rep"},{"issue":"06","key":"ref29","first-page":"266","article-title":"Injectable bone substitute paste based on hydroxyapatite, gelatin and streptomycin for spinal tuberculosis","volume":"04","author":"H N Maulida","year":"2015","journal-title":"J Spine"},{"issue":"04","key":"ref30","doi-asserted-by":"crossref","first-page":"266","DOI":"10.3390\/ma9040226","article-title":"Bioactive glasses with low Ca\/P ratio and enhanced bioactivity","volume":"9","author":"M Ara\u00fajo","year":"2016","journal-title":"Materials (Basel)"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"e27664","DOI":"10.4161\/biom.27664","article-title":"In vitro and in vivo bioactivity assessment of a polylactic acid\/hydroxyapatite composite for bone regeneration","volume":"4","author":"C B Danoux","year":"2014","journal-title":"Biomatter"},{"key":"ref32","first-page":"17","article-title":"Increased osteoblast viability at alkaline pH in vitro provides a new perspective on bone regeneration","volume":"10","author":"A M Galow","year":"2017","journal-title":"Biochem Biophys Rep"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ijbiomac.2020.01.035","article-title":"Fabrication and in-vitro biocompatibility of freeze-dried CTS-nHA and CTS-nBG scaffolds for bone regeneration applications","volume":"149","author":"P Kumar","year":"2020","journal-title":"Int J Biol Macromol"},{"key":"ref34","doi-asserted-by":"crossref","first-page":"117765","DOI":"10.1016\/j.carbpol.2021.117765","article-title":"Two-step strategy for constructing hierarchical pore structured chitosan-hydroxyapatite composite scaffolds for bone tissue engineering","volume":"260","author":"T T Li","year":"2021","journal-title":"Carbohydr Polym"},{"issue":"12","key":"ref36","doi-asserted-by":"crossref","first-page":"3171","DOI":"10.1016\/j.eurpolymj.2006.08.009","article-title":"A study on the bioactivity of chitosan\/nano-hydroxyapatite composite scaffolds for bone tissue engineering","volume":"42","author":"L Kong","year":"2006","journal-title":"Eur Polym J"},{"issue":"03","key":"ref37","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1007\/s00784-011-0570-7","article-title":"Human periodontal ligament fibroblasts stimulated by nanocrystalline hydroxyapatite paste or enamel matrix derivative. An in vitro assessment of PDL attachment, migration, and proliferation","volume":"16","author":"A Kasaj","year":"2012","journal-title":"Clin Oral Investig"},{"issue":"16","key":"ref38","doi-asserted-by":"crossref","first-page":"1190","DOI":"10.1080\/09205063.2015.1082809","article-title":"Development of gelatin-chitosan-hydroxyapatite based bioactive bone scaffold with controlled pore size and mechanical strength","volume":"26","author":"K Maji","year":"2015","journal-title":"J Biomater Sci Polym Ed"},{"key":"ref39","doi-asserted-by":"crossref","first-page":"213280","DOI":"10.1016\/j.bioadv.2023.213280","article-title":"Full physicochemical and biocompatibility characterization of a supercritical CO2 sterilized nano-hydroxyapatite\/chitosan biodegradable scaffold for periodontal bone regeneration","volume":"146","author":"M Souto-Lopes","year":"2023","journal-title":"Biomater Adv"},{"key":"ref40","doi-asserted-by":"crossref","first-page":"103259","DOI":"10.1016\/j.jdent.2019.103259","article-title":"Stem cells in the periodontal ligament differentiated into osteogenic, fibrogenic and cementogenic lineages for the regeneration of the periodontal complex","volume":"92","author":"J Liu","year":"2020","journal-title":"J Dent"},{"key":"ref41","doi-asserted-by":"crossref","first-page":"651945","DOI":"10.1155\/2013\/651945","article-title":"Modification of decellularized goat-lung scaffold with chitosan\/nanohydroxyapatite composite for bone tissue engineering applications","volume":"2013","author":"S K Gupta","year":"2013","journal-title":"BioMed Res Int"}],"container-title":["European Journal of Dentistry"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.1055\/s-0044-1800826.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T22:34:13Z","timestamp":1759530853000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/s-0044-1800826"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,12]]},"references-count":40,"journal-issue":{"issue":"04","published-online":{"date-parts":[[2025,10,3]]},"published-print":{"date-parts":[[2025,10]]}},"URL":"https:\/\/doi.org\/10.1055\/s-0044-1800826","archive":["Portico","CLOCKSS"],"relation":{},"ISSN":["1305-7456","1305-7464"],"issn-type":[{"value":"1305-7456","type":"print"},{"value":"1305-7464","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,12]]}}}