RGD-coupled alginate (NovaMatrix FMC Biopolymer, Norway) was used as the scaffold material. Alginate was purified and partially oxidized (2%) to increase its degradability according to published methods in the literature.30 (link),31 The prepared alginate was concentrated, freeze-dried under reduced pressure, and mixed with a cocktail containing 10 µM forskolin (FSK) (Sigma), 10 µM MeBIO (Santa Cruz Biotechnology), and 10 ng/ml recombinant human bFGF (R&D Systems).
To study the effects of elasticity of the alginate hydrogel on myogenic differentiation capacity of encapsulated GMSCs the elastic modulus (E) of alginate hydrogels in presence of different concentrations of CaCl2 (10, 25, 50, 75, and 100 mM) was measured using an Instron mechanical testing machine (Norwood, PA) at a compression rate of 0.5 mm.min−1 according to the methods already in literature.18 (link)To characterize the release profile of all the components of the cocktail, the hydrogel microspheres were loaded at the abovementioned concentrations and at each selected time interval (1, 3, 5, 7, 10, 12, and 14 days), the amounts of medium were collected and analyzed for released of FSK and MeBIO, using a UV spectrophotometer at 320 nm (Beckman, Brea, CA). Moreover, the release profile of bFGF was characterized using human b-FGF Immunoassay kit (BioSource International Inc. Camerillo, CA).
To study the effects of elasticity of the alginate hydrogel on myogenic differentiation capacity of encapsulated GMSCs the elastic modulus (E) of alginate hydrogels in presence of different concentrations of CaCl2 (10, 25, 50, 75, and 100 mM) was measured using an Instron mechanical testing machine (Norwood, PA) at a compression rate of 0.5 mm.min−1 according to the methods already in literature.18 (link)To characterize the release profile of all the components of the cocktail, the hydrogel microspheres were loaded at the abovementioned concentrations and at each selected time interval (1, 3, 5, 7, 10, 12, and 14 days), the amounts of medium were collected and analyzed for released of FSK and MeBIO, using a UV spectrophotometer at 320 nm (Beckman, Brea, CA). Moreover, the release profile of bFGF was characterized using human b-FGF Immunoassay kit (BioSource International Inc. Camerillo, CA).
