BMP-2-encapsulated AM were fabricated following Lee et al. [15 (link)]: A solution of 1.8% (w/v) sodium alginate from Büchi Labortechnik AG (Flawil, Switzerland) was diluted to 1.2% (w/v) by adding distilled water. Next, BMP-2 (Novosis, CGBio, Gyeonggi-do, Korea) was added to the 1.2% (w/v) sodium alginate solution at a protein/polymer ratio of 3.47 μg/mg. BMP-2-encapsulated AM were produced by a vibrational nozzle technique through a Büchi Encapsulator B-395 pro (Dottikon, Switzerland) under the following optimized parameters [16 (link)]: 23-gauge needle; 120 μm nozzle size; 1000 Hz frequency; 23 mL/min flow rate; and 1000 V electrode potential (Figure 1 A). The mixture droplets of alginate and BMP-2 were gelled in a 250 mL 100 mM calcium chloride solution for 30 min. Finally, BMP-2-encapsulated AM were purified through a 40 m cell strainer and twice washed with phosphate-buffered saline (PBS).
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