Preparation and characterization of decellularized bovine bone as a bioscaffold for bone tissue engineering applications

Articles in Press

Document Type : Original Article

Authors

1 Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

2 Regenerative Medicine Research Center, Shahroud University of Medical Sciences, Shahroud, Iran

3 Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

4 Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

5 Tissue Engineering Research Group (TERG), Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

6 Department of Medical Biotechnology, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

10.22038/ijbms.2025.87816.18969

Abstract

Objective(s): This study aimed to develop and evaluate decellularized bovine bone (DBB) scaffolds and investigate their potential to promote osteogenic differentiation when combined with Crocin and Alendronate.
Materials and Methods: Bovine bone was decellularized using a combination of physical (freeze–thaw cycles, sonication), chemical (sodium dodecyl sulfate), and enzymatic (deoxyribonuclease I) treatments to preserve native bone architecture. Scaffold properties were assessed by evaluating extracellular matrix (ECM) integrity and compressive strength. Biocompatibility was confirmed through cytotoxicity and hemolysis assays. In vitro osteogenesis was analyzed using alizarin red staining and qRT-PCR (quantitative real-time polymerase chain reaction) to quantify expression of osteogenic markers RUNX2, osteocalcin, osteopontin, and osteonectin following treatment with crocin (Cr 5 mg/ml), Alendronate (ALN 1 mg/ml), and their combination (Cr/ALN 5 mg/ml).
Results: DBB scaffolds-maintained ECM structure and compressive strength (14.56 ± 0.82 MPa), comparable to native bovine bone (17.86 ± 0.14 MPa). No cytotoxic or hemolytic effects were observed. Crocin, Alendronate, and Cr/ALN treatments significantly enhanced RUNX2 expression (70%, 60%, and 65%, respectively), while Osteocalcin expression increased in Cr (50%) and Cr/ALN (25%) groups. Osteopontin and osteonectin expression also rose in Cr and Cr/ALN groups, supporting enhanced osteogenic differentiation.
Conclusion: Based on in vitro findings, DBB scaffolds demonstrate favorable mechanical and biological properties, and loading the scaffolds with crocin and Alendronate enhanced osteogenic differentiation and matrix mineralization, indicating potential for bone-regeneration applications. 

Keywords

Main Subjects

References

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Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 29 October 2025

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