A mild process to design silk scaffolds with reduced β-sheet structure and various topographies at the nanometer scale.

Three-dimensional (3-D) porous silk scaffolds with good biocompatibility and minimal immunogenicity show promise in a range of tissue regeneration applications. However, the challenge remains to effectively fabricate their microstructures and mechanical properties to satisfy the specific requirements of different tissues. In this study, silk scaffolds were fabricated to form an extracellular matrix (ECM) mimetic nanofibrous architecture using a mild process. A slowly increasing concentration process was applied to regulate silk self-assembly into nanofibers in aqueous solution. Then glycerol was blended with the nanofiber solution and induced silk crystallization in the lyophilization process, endowing freeze-dried scaffolds with water stability. The glycerol was leached from the scaffolds, leaving a similar porous structure at the micrometer scale but different topographies at the nanoscale. Compared to previous salt-leached and methanol-annealed scaffolds, the present scaffolds showed lower β-sheet content, softer mechanical property and improved cell growth and differentiation behaviors, suggesting their promising future as platforms for controlling stem cell fate and soft tissue regeneration.