Nanofibrous gelatine scaffolds integrated with nerve growth factor-loaded alginate microspheres for brain tissue engineering. 
Neural regeneration research is designed in part to develop strategies for therapy after nerve damage due to injury or disease.  In this study, a new gelatine-based biomimetic scaffold was fabricated for brain tissue engineering applications.  A technique combining thermally induced phase separation and porogen leaching was used to create interconnected macropores and nanofibrous structure.  To promote tissue regeneration processes, the scaffolds were integrated with nerve growth factor (NGF)-loaded alginate microspheres.  The results showed that nanofibrous matrix could only be obtained when gelatine concentration was at least 7.5% (w/v).  The scaffold with a modulus value (1.2 kPa) similar to that of brain tissue (0.5-1 kPa) was obtained by optimizing the heat treatment time, macropore size and gelatine concentration.  The encapsulation efficiencies of NGF into 0.1% and 1% alginate microspheres were 85% and 100%, respectively.  The release rate of NGF from the microspheres was controlled by the alginate concentration and the poly(L-lysine) coating.  The immobilization of the microspheres in the scaffold reduced burst release and significantly extended the release period.  The nanofibrous architecture and controlled release of NGF from the microspheres induced neurite extension of PC12 cells, demonstrating that the released NGF was in an active form.  The results suggest that the scaffolds prepared in this study may have potential applications in brain tissue engineering due to topologic and mechanical properties similar to brain tissue and pore structure suitable for cell growth and differentiation.  Copyright © 2016 John Wiley & Sons, Ltd.