In vitro biocompatibility analysis of novel nano-biphasic calcium phosphate scaffolds in different composition ratios

This study aimed to evaluate in vitro biocompatibility of a composite of nanoscale biphasic calcium phosphate (BCP) and collagen (C) compared to pure BCP (P) in different composition ratios of nanohydroxyapatite to nano-β-tricalcium phosphate (HA/β-TCP). Each study group comprised of three ratios of BCP (30/70, 40/60, and 50/50). For evaluation of cellular response toward each ratio, mouse osteoblast (MC3T3-E1) cell line was cultivated on the scaffolds for 19 days. Analysis of cell proliferation, cell viability, cell attachment and morphology, alkaline phosphatase (ALP) activity, and osteocalcin synthesis were done on culture days 1, 3, 7, 13, 15, and 19, appropriately. The scanning electron microscopy showed that the osteoblasts attached successfully to scaffolds surfaces in both BCP groups and in all different ratios by spreading their filopodia and expressing similar viability that was confirmed by confocal laser scanning electron microscope. BCP scaffold (P3070) showed remarkable ALP activity, whereas BCP (P5050) showed highest osteocalcin activity. Collagen coating supported high cell proliferation on culture day 1 and possessed limited benefit restricted to early phase of cell differentiation. In conclusion, the fabricated nanoscale BCP scaffolds offered high biocompatibility and supported well the cell proliferation and differentiation regardless the composition ratio. Furthermore, higher ratio of TCP supported the early phase of cell proliferation, whereas higher HA ratio influenced the later phase. Finally, BCP scaffolds P5050 and C4060 were suggested as candidates for clinical applications.