Adipose-Derived Mesenchymal Stem Cell Osteodifferentiation after Exposure to Beta-Tricalcium Phosphate Bioceramic Granules with 300 to 600 and 600 to 1,000 µm Sizes

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Pamela Handy Cecilia, Ida Bagus Narmada, Rini Devijanti Ridwan, Diah Savitri Ernawati, Taufan Bramantoro, Devi Rianti, Khairul Anuar Shariff, Wibi Riawan, Putri Cahaya Situmorang, Alexander Patera Nugraha

2026 European Journal of Dentistry Vol. 20 Issue 1 Article Cited by 0 Quartile

Abstract

Objective Beta-tricalcium phosphate (β-TCP) is a synthetic graft material with excellent biocompatibility, osteoconductivity, and osteoinductivity. β-TCP may induce adipose-derived mesenchymal stem cells (ADMSCs) osteodifferentiation. This study aims to investigate the osteoinductivity of 300 to 600 and 600 to 1,000μm β-TCP in ADMSCs. Materials and Methods ADMSCs were obtained from the visceral adipose tissue of young male rabbits. To determine the osteoinductive ability, bone morphogenic protein 2 (BMP-2), Osterix, runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteopontin, and osteonectin expression was examined using an immunochemical assay on ADMSCs conditioned with an osteogenic medium and a β-TCP bioceramic with granule sizes of 300 to 600 and 600 to 1,000 µm (100 ng diluted to 100 nmol as the final concentration). A 3,3′-diaminobenzidine staining kit was used for immunocytochemical staining. Anti-BMP-2, anti-Osterix, anti-Runx2, anti-ALP, anti-osteopontin, and anti-osteonectin monoclonal antibodies were employed at a 1:500 dilution. A light microscope with magnifications of 400× and 1,000× was used to manually observe and examine cultures in five different fields of view. Results BMP 2, Runx2, Osterix, and ALP expression was higher in ADMSCs þ β-TCP 300 to 600 µm compared with the control group (p < 0.05). Osteonectin and osteopontin expression was higher in ADMSCs þ 300 to 600 µm β-TCP compared with the control group (p < 0.05) and ADMSCs þ 600 to 1,000 µm β-TCP (p < 0.05). Conclusion ADMSC osteodifferentiation was influenced by β-TCP bioceramic granule size. The considerable difference in osteonectin and osteopontin expression supports the idea that 300 to 600 µm β-TCP induce ADMSCs osteodifferentiation than 600 to 1,000 µm β-TCP. © 2025. The Author(s).

Affiliations

Doctoral Program of Dental Medicine, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia; Department of Orthodontics, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia; Department of Oral Biology, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia; Department of Oral Medicine, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia; Department of Dental Public Health, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia; Department of Dental Material, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia; School of Materials and Mineral Resources Engineering, Universiti Sains Malaysia, Penang, Nibong Tebal, Malaysia; Department of Biomolecular Biochemistry, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia; Biology Study Program, Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia