Creating growth factor gradients in three dimensional porous matrix by centrifugation and surface immobilization

Polycaprolactone (PCL)/Pluronic F127 cylindrical scaffolds with gradually increasing growth factor concentrations were fabricated by the centrifugation of fibril-like PCLs and the subsequent fibril surface immobilization of growth factors. The cylindrical scaffolds exhibited gradually increasing surface areas along the longitudinal direction [from 3.17 ± 0.05 m(2)/g (top position) to 5.42 ± 0.01 m(2)/g (bottom position)]. The growth factors (BMP-7, TGF-β(2) and VEGF(165)) as model bioactive molecules were immobilized onto the fibril surfaces of the scaffolds via heparin binding to produce scaffolds with gradually increasing concentrations of growth factors from the top position (BMP-7, 60.89 ± 2.51; TGF-β(2), 42.85 ± 2.00; VEGF(165), 42.52 ± 3.22 ng/scaffold section) to the bottom position (BMP-7, 181.07 ± 3.21; TGF-β(2), 142.08 ± 2.91; VEGF(165), 112.00 ± 4.00 ng/scaffold section). The released amount of growth factor (VEGF(165)) from the cylindrical scaffold gradually decreased along the longitudinal direction in a sustained manner for up to 35 days, which can allow for a minutely controlled spatial distribution of growth factors in a 3D environment. The 3D porous scaffold with a concentration gradient of growth factors may become a useful tool for basic studies, including in vitro investigations of 3D chemotaxis/haptotaxis for the control of specific biological process. It may also be used as a tissue engineering scaffolding system for a variety of tissues/organs requiring the spatial regulation of growth factors for effective regeneration.