Preparation and detection of calcium alginate/bone powder hybrid microbeads for in vitro culture of ADSCs.

Calcium alginate microbeads have been widely used in tissue engineering application, due to their excellent biocompatibility, biodegradability, enhanced mechanical strength and toughness. Bone powder containing abundant hydroxylapatite, type I collagen and growth factors such as BMP2 and BMP4, possesses good osteoinductive activity. Herein, a hybrid calcium alginate/bone powder microbead was therefore prepared. Afterwards, different seeding density of adipose-derived stem cells (ADSCs) in these hybrid microbeads was discussed systematically for further in vitro expansion. Optimised microbeads suitable for in vitro expansion and differentiation of ADSCs were prepared using the droplet method under overall considering suitable concentrations of calcium alginate and calcium chloride as well as the density of bone powder through an orthogonal experiment. The results showed that the concentration of sodium alginate had the most influence on inside mass transfer and mechanical strength of the hybrid microbeads, secondly the calcium chloride, then the density of bone powder. The hybrid microbeads could be optimally performed while the concentrations of sodium alginate and calcium chloride were 2.5% and 4.5%, as well as 5.0 mg/mL bone powder, respectively. Live/Dead assay showed that the expanded ADSCs differentiated well with an initial embedding density of 5 × 10(6) cells/mL.