Behavior of macrophage and osteoblast cell lines in contact with the β-TCP biomaterial (beta-tricalcium phosphate)

Abstract

Beta-tricalcium phosphate (β-TCP) is a synthetic ceramic used for filling bone defects. It is a good alternative to autologous grafts since it is biocompatible, resorbable and osteoconductive. Previous in vivo studies have shown that macrophages are one of the first cells coming in contact with the biomaterial followed by osteoclasts and osteoblasts that will elaborate new bone packets. Studies have focused on osteoclast morphology and very few of them have investigated the role of macrophages. The aims of this study were to characterize (i) the biomaterial surface; (ii) the in vitro behavior of macrophages (J774.2 and Raw264.7 cells) using the description of cell morphology by scanning electron microscopy (SEM) at 7 and 14 days; (iii) the behavior of osteoblasts (SaOs-2 and MC3T3-E1 cells) seeded at the surface of the biomaterial 24, 48 and 72hours by SEM and confocal microscopy. Cell proliferation was analyzed by MTT assays. Viability and affinity of the macrophages for β-TCP were found significantly increased after 7 and 14d. MC3T3-E1 cells were anchored and stretched onto the β-TCP surface as early as 24h with a high proliferation rate (+190%) when compared to the surface of a well plate. SaOs-2 exhibited the same morphological profile at 72h. Proliferation became significantly higher compared to the plastic surface at only 72h (+129%). This study emphasises the importance of choice of the cell line used in exploring the osteoconductive and osteoinductive properties of a biomaterial. Additional studies are needed to analyze differentiation of macrophages into giant multinucleated cells and how the biomaterial surface influences osteoblast differentiation.