Gelatin–Polyvinyl Alcohol Microspheres for Controlled and Sustained Release of BMP-2 and VEGF Enhance Osteogenic and Angiogenic Cell Differentiation

Bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) play a pivotal role in promoting osteogenesis and angiogenesis that concurrently take place during bone regeneration. The rapid degradation and diffusion of these growth factors, combined with the potential side effects associated with their exogenous insertion, limit their applications. To overcome these shortcomings, we developed a controlled release system for BMP-2 and VEGF on microspheres comprising gelatin (Gel) and polyvinyl alcohol (PVA). We fabricated Gel–PVA microspheres using a constant Gel concentration of 10% w/v and a varied PVA concentration of 0, 5, and 10% w/v (Gel–PVA0%, Gel–PVA5%, and Gel–PVA10%, respectively). The microspheres were loaded with the model protein bovine serum albumin (BSA) first. The Gel–PVA10% microspheres demonstrated significantly higher loading capacity and encapsulation efficiency, as well as lower cumulative release rate, compared to the Gel–PVA5% ones when loaded with BSA. Thus, the microspheres with the Gel–PVA10% composition were selected for loading with BMP-2 and VEGF. Kinetic studies of BMP-2 and VEGF loaded into Gel–PVA10% microspheres indicated similar results to those with BSA. The microsphere concentration with the optimal cytocompatibility was 0.5 mg/mL, and it was applied for the assessment of the osteogenic differentiation using bone marrow-derived mesenchymal stem cells (MSCs), and for the angiogenic differentiation in Wharton jelly and adipose-derived MSCs. Alkaline phosphatase activity, collagen secretion, and calcium mineralization were significantly upregulated in the presence of BMP-2-loaded microspheres, while tubular formation and PECAM-1 secretion were significantly higher in VEGF-loaded microspheres compared to the unloaded control, demonstrating their effectiveness as drug delivery carriers.