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

Objectives: 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: 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: 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.