In-vitro and In-vivo Evaluation of the DTBP Crosslinked Collagen and Gelatin Coated Porous Spherical BCP Granules for Using as Granular Bone Substitutes

DTBP (dimethyl 3,3`-dithiobispropionimidate) was applied to collagen and gelatin coating on BCP granules and a crosslinking agent. The DTBP crosslinking was done for decreasing the solubility of the coating and hence increasing the stability. The nanostructure of collagen and gelatin coating surfaces were observed by SEM technique. Based on the DSC thermograms and FT-IR spectrums, the crosslinkings were confirmed between collagen molecules and gelatin molecules. The compressive strength was measured before crosslinking and after that. In-vitro study was carried out by measuring cell viability and observing cell morphology after DTBP crosslinking. Moreover, the proliferation ability of MG-63 osteoblast-like cells on the crosslinked BCP granules was evaluated by Western blot assay. The BCP granules were implanted into rabbit femur for 4 weeks and 12 weeks. The bone tissue formation was analyzed with micro-computed tomography (micro-CT) and histological analysis was also carried out by hematoxylin and eosin (H&E) staining for visualization of cells.