Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres

In this study, the behavior of bone marrow stromal cells cultured on calcium titanium phosphate (CTP) microspheres was analyzed. Cell adhesion and proliferation were estimated by the neutral red assay and by total DNA quantification. Morphology and deposition of extracellular matrix were assessed by confocal laser scanning microscopy and/or scanning electron microscopy. The expression of the osteoblastic phenotype was evaluated by monitoring alkaline phosphatase activity and osteocalcin secretion. Results revealed that cells were able to attach and spread on the surface of CTP microspheres, and gradually grow into nearly confluent monolayers. Moreover, cells were able to bridge adjacent microspheres forming microsphere-cell clusters. Cells produced an abundant amount of fibrillar extracellular matrix that covered the substrate surface. Alkaline phosphatase activity peaked around days 7-14 and then decreased until day 21. Cells secreted osteocalcin, with higher levels being detected at day 14 than at day 21. Taken together, these results suggest that CTP microspheres are appropriate scaffolds for the growth and differentiation of cells along the osteoblastic lineage.