Property and biological effects of the cuttlebone derived calcium phosphate particles, a potential bioactive bone substitute material

Cuttlebone (CB) is a marine waste-derived biomaterial and a rich source of calcium carbonate for the biosynthesis of the calcium phosphate (CaP) particles. The current study aimed to synthesize CB derived biphasic calcium phosphate (CB-BCP) and investigate biological activity of the CB-CaP: hydroxyapatite (CB-HA), beta-tricalcium phosphate (CB-b-TCP) and biphasic 60:40 (w/w) HA/b-TCP (CB-BCP) with the human dental pulp stem cells (hDPSCs). The particles were synthesized using solid state reactions under mild condition and properties of the particles were compared with a commercial BCP as a reference material. Morphology, particle size, physicochemical properties, mineral contents, and the ion released patterns of the particles were examined. Then the particle/cell interaction, cell cytotoxicity and osteogenic property of the particles were investigated in the direct and indirect cell culture models. It was found that an average particles size of the CB-HA was 304.73 ± 4.19 nm, CB-b-TCP, 503.17 ± 23.06 nm and CB-BCP, 1394.67 ± 168.19 nm. The physicochemical characteristics of the CB-CaP were consistent with the HA, b-TCP and BCP. The highest level of calcium (Ca) was found in the mineral contents and the preincubated medium of the CB-BCP and traces of fluoride, magnesium, strontium, and zinc were identified in the CB-CaP. The cell cytotoxicity and osteogenic property of the particles were dose dependent. The particles adhered on cell surface and were internalized into the cell cytoplasm. The CB-BCP and CB-HA indirectly and directly promote osteoblastic differentiations of the hDPSCs in stronger levels than other groups. The CB-BCP and CB-HA were potential bioactive bone substitute materials.