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

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.

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).

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.

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.