Immobilization of a bone and cartilage stimulating peptide to a synthetic bone graft
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A synthetic peptide fragment of human collagen type I (BCSP™-1) was linked to the surface of a commercially available ceramic in an effort to improve the properties of the bone graft substitute to accelerate local healing. BCSP™-1 was covalently immobilized on the surface of the ceramic via the linkers 3-aminopropyl-triethoxysilane (APTES) and suberic acid bis-N-hydroxysuccinimide ester (DSS). The chosen chemistry was non-cytotoxic. A rat calvaria cell assay using alkaline phosphatase (ALP) as an osteoblast differentiation marker, showed that modifying the surface of the ceramic was enough to enhance ALP activity, although the total cell population on the surface decreased. A significant increase in ALP activity/cell was noted with serum albumin bound to the surface, however, the BCSP™-1 bound surface exhibited an even greater ALP activity that showed a surface concentration dependent trend. An optimal BCSP™-1 surface density in the range of 0.87–2.24 nmol/cm2 elicited the maximum ALP activity/cell at day 6 of culture. The peptide bound ceramic generated an ALP activity/cell that was roughly 3-fold higher than the non-modified ceramic and 2-fold higher than the APTES-grafted ceramic.
KeywordsHigh Performance Liquid Chromatography Osteoprogenitor Cell High Performance Liquid Chromatography Assay Synthetic Bone Graft Disuccinimidyl Suberate
This project was funded by Materials and Manufacturing Ontario, and the Ontario Premier’s Research Excellence Award. The assistance of Dennis Sindrey of Millenium Biologix with the rat calvaria model is gratefully acknowledged.
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