Optimizing Chitosan/Collagen Type I/Nanohydroxyapatite Cross-linked Porous Scaffolds for Bone Tissue Engineering

Bio-composite scaffolds mimicking the natural microenvironment of bone tissue offer striking advantages in material-guided bone regeneration. The combination of biodegradable natural polymers and bioactive ceramics that leverage potent bio-mimicking cues has been an active strategy to achieve success in bone tissue engineering. Herein, a competitive approach was followed to point out an optimized bio-composite scaffold in terms of scaffold properties and stimulation of osteoblast differentiation. The scaffolds, composed of chitosan/collagen type I/nanohydroxyapatite (Chi/Coll/nHA) as the most attractive components in bone tissue engineering, were analyzed. The scaffolds were prepared by freeze-drying method and cross-linked using different types of cross-linkers. Based on the physicochemical and mechanical characterization, the scaffolds were eliminated comparatively. All types of scaffolds displayed highly porous structures. The cross-linker type and collagen content had prominent effects on mechanical strength. Glyoxal cross-linked structures displayed optimum mechanical and structural properties. The MC3T3-E1 proliferation, osteogenic-related gene expression, and matrix mineralization were better pronounced in collagen presence and triggered as collagen type I amount was increased. The results highlighted that glyoxal cross-linked scaffolds containing equal amounts of Chi and Coll by mass and 1% (w/v) nHA are the best candidates for osteoblast differentiation and matrix mineralization.