Osteogenic differentiation of human adipose-derived stem cells induced by osteoinductive calcium phosphate ceramics

Microstructure is indispensable for the osteoinduction of calcium phosphate ceramics. To study how microstructure takes its role and explore the mechanism of the osteoinduction, we evaluated attachment, proliferation, alkaline phosphatase (ALP)/DNA, protein/DNA, and mineralization of human adipose-derived stem cells cultured on two kinds of biphasic calcium phosphate (BCP) ceramic discs with the same chemistry and dimension, but different microporosity and surface area. BCP-A had been found osteoinductive in vivo while BCP-B was not. During the conventional culture, ALP/DNA and protein/DNA of the cell on BCP-A with larger surface area were significantly higher than those of the cells on BCP-B. With the adsorption of the proteins in culture medium with 50% fetal bovine serum (FBS) in advance, the increments of the ALP/DNA and protein/DNA for the BCP-A were found respectively significantly more than the increments of those for BCP-B, suggesting that the larger amount of protein adsorbed on the BCP-A was crucial. More results showed that ALP/DNA and protein/DNA of the cells on the two kinds of discs presoaked in culture medium having additional rhBMP-2 were found to be both higher than those of the cells on the discs resoaked in culture medium with 50% FBS, and that those values for BCP-A increased much more. Furthermore, larger mineral content was found on BCP-A than on BCP-B at day 7. The results indicated that by increasing microporosity and thus surface areas, osteoinductive calcium phosphate ceramics concentrate more proteins, including bone-inducing proteins, and thereafter stimulate inducible cells in soft tissues to form inductive bone.