Enhancement of the Cytokine Adsorption Capacity of Octacalcium Phosphate by Interaction-Controlled Glycosaminoglycan Modification to Promote Osteoblastic Differentiation.

This study was designed to investigate how the strength of the interaction between octacalcium phosphate (OCP) and modified chondroitin-A sulfate (CS-A), a glycosaminoglycan, regulates the adsorption-desorption of cytokines and subsequently affects the osteoblastic differentiation of mesenchymal stem cells (MSCs) in vitro. The utilization of cytokines produced by cells, such as macrophages, stimulated by the hydrolysis of OCP, is expected to enhance the bone regeneration capacity of the OCP. CS-Na was used to modify CS-A on the OCP immobilized with the amino group through electrostatic interactions. On the other hand, -hydroxysuccinimide (NHS)-esterified CS-A was used to form the covalent bond between CS-A and the amino group on the surface of OCP. X-ray diffraction and Raman spectroscopy indicated that the CS-A-modified OCP maintained its structure, regardless of the modification process. The remaining ratio of the modified CS-A in the buffer suggests that increasing the immobilized density of the amino group and the modification using NHS ester may enhance interaction strength between OCP and CS-A. The adsorption amount and retention rate of recombinant human bone morphological protein-2 (rhBMP-2, an endogenous cytokine model) increased onto CS-A-modified OCP under physiological conditions when the interaction strength between OCP and the protein was stronger. The higher interaction strength between the OCP and CS-A could be associated with the enhanced adsorption affinity for the lower-molecular-weight basic protein. The alkaline phosphatase activity of MSCs increased depending on the remaining rate of rhBMP-2 adsorption on the CS-A-modified-OCP. Fourier transform infrared spectroscopy and chemical analysis indicated that the hydrolysis of the OCP progressed regardless of CS-A modification after MSC incubation. The present study suggests that stronger interactions between the OCP and glycosaminoglycans could contribute to the capture and retention of endogenous cytokines on the surface to further promote osteoblastic differentiation under the chemical environment induced by the hydrolysis of the OCP.