Fabrication and characterization of apigenin-loaded chitosan/gelatin membranes for bone tissue engineering applications

Fabricating degradable polymer-based membranes has attracted much attention for guided bone regeneration. Chitosan/gelatin (Cs/Gel) composites are among the most known scaffolds with structural similarity to bone matrix and a high potential to support cell attachment and proliferation. Recently, plant-derived phenolic compound apigenin has been identified to direct the osteogenic differentiation of mesenchymal stem cells and retain osteoblast metabolic functions. We incorporated apigenin into Cs/Gel membranes to improve apigenin bioavailability and get proper concentrations for efficient biological activities. Apigenin-loaded Cs/Gel membranes were prepared using a solution casting method with various apigenin contents (0, 10, 25, 50, and 100 µM). Chemical composition, morphological characteristics, swelling behavior, degradation rate, and apigenin release from membranes were evaluated. Saos-2 osteoblasts were cultured on membranes to investigate cell-membrane interaction, proliferation, viability, and mineralization under the osteogenic culture condition. The results showed that membranes had homogeneous and moderate rough surfaces, facilitating osteoblast attachment and expansion. Swelling ratios exceeded 200%, reaching a stable rate in 24 h. Apigenin-loaded membranes degraded slower in vitro. Membranes containing lower apigenin concentrations exhibited a higher cargo release profile over 21 days. Apigenin improved osteoblast proliferation and viability, but the mineralization depended on apigenin dose, with optimized values at low concentrations. These data suggested that Cs/Gel membranes loaded with low apigenin contents improved osteoblast survival, proliferation, and mineralization.