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ORIGINAL RESEARCH
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Gelatin/chitosan delivery system improves stability and regenerative potential of Ca(OH)2 in an open dental pulp model

Endytiastuti Endytiastuti1 ,  Retno Ardhani2* ,  Bidhari Pidhatika3 ,  Yogi A. Swasono4 ,  Reza P. Rudianto5 ,  Juni Handajani6 ,  Ghadah A. Al-qatta7 ,  Iwa S. R. Sudarso1 ,  Fauzi Mh Busra8
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1 Department of Pediatric Dentistry, Faculty of Dentistry, Universitas Gadjah Mada, Sleman, Yogyakarta, Indonesia
2 Department of Dental Biomedical Sciences, Faculty of Dentistry, Universitas Gadjah Mada, Sleman, Yogyakarta, Indonesia
3 Polytechnic Institute of Nuclear Technology, National Research and Innovation Agency, Sleman, Yogyakarta, Indonesia
4 Research Centre for Food Technology and Processing, National Research and Innovation Agency, Gunung Kidul, Yogyakarta, Indonesia
5 Research Centre for Polymer Technology, National Research and Innovation Agency, Tangerang Selatan, Banten, Indonesia
6 Department of Oral Biology, Faculty of Dentistry, Universitas Gadjah Mada, Sleman, Yogyakarta, Indonesia
7 Faculty of Dentistry, Universitas Gadjah Mada, Sleman, Yogyakarta, Indonesia
8 Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, Wilayah Persekutuan Kuala Lumpur, Malaysia
BMT, 00059 https://doi.org/10.12336/bmt.25.00059
Submitted: 8 June 2025 | Revised: 21 August 2025 | Accepted: 22 August 2025 | Published: 2 October 2025
Copyright © 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution–NonCommercial–ShareAlike 4.0 License.
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Abstract

Calcium hydroxide (Ca(OH)2) has a long history as an agent to induce hard tissue regeneration in teeth. However, its high solubility requires inefficient repeated applications. Its alkalinity has two-sided effects: antibacterial property, but simultaneously compromises cell viability. This study prepared a composite of gelatin/chitosan to deliver Ca(OH)2 using tetraethyl orthosilicate (TEOS) as the crosslinker. The chemical and physical properties of the composite were compared with unmodified Ca(OH)2 aloneusing Fourier-transform infrared spectroscopy, X-ray diffraction, and scanning electron microscopy with energy-dispersive X-ray spectroscopy, followed by an investigation into the release or dissolution of Ca2+ from both materials. A total of 16 Wistar rats were allocated to receive either the composite or Ca(OH)2 after dental pulp exposure. Regenerative potential was assessed after 7 and 14 days by histological evaluation of odontoblast-like cell numbers and transforming growth factor β1 (TGF-β1) expression, with statistical analysis performed at a 95% confidence level. The gelatin/chitosan/Ca(OH)2/TEOS composite was successfully synthesized and exhibited controlled Ca2+ release. The results demonstrated a higher odontoblast-like cell proliferation and stronger TGF-β1 expression in the composite-treated group after 7 days of application, indicating a more intensive regeneration than the Ca(OH)2 control. After 14 days, the number of odontoblast-like cells in both groups did not differ significantly. However, TGF-β1 expression was significantly more pronounced. In conclusion, the incorporation of Ca(OH)2 into a gelatin/chitosan matrix using TEOS as a crosslinker successfully decreases its solubility without impairing the ability to induce dental pulp regeneration. 

Keywords
Polymer
Pulp capping
Regenerative endodontic
Tetraethyl orthosilicate
SDG 3
Funding
This study was funded by the Regular Fundamental Research Scheme of The Directorate of Research, Technology, and Community Services, Directorate General of Higher Education, Research, and Technology, Ministry of Education, Culture, Research, and Technology, Republic of Indonesia 122/ E5/PG/02.00PL/2023 with contract number 3135/UN1/DITLIT/Dit- Lit/PT.01.03/2023 to Retno Ardhani. Part of the animal model work was also funded by the Dana Masyarakat Research Funding from the Faculty of Dentistry, Universitas Gadjah Mada, with contract number 3857/UN1/FKG/ Set.KG1/LT/2023 to Juni Handajani.
Conflict of interest
The authors declare no conflicts of interest.
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