Biofunctionalized Nanofibrous Bilayer Scaffolds for Enhancing Cell Adhesion, Proliferation and Osteogenesis

A simple and efficient method for fabricating functionalized multilayered nanofibrous scaffolds has been developed by combining electrospinning and thermally induced phase separation (TIPS) techniques. In this investigation, functionalized bilayer scaffolds were constructed using this method for bone tissue engineering, which consisted of a nanofibrous poly(lactic acid-co-glycolic acid) (PLGA) membrane as the base and a nanofibrous chitosan (CS) or gelatin (Gel) mesh as the surface layer, with the PLGA nanofibers having a biomimetic polydopamine (PDA) coating. It was shown that the PDA coating strongly bonded TIPS-formed CS or Gel networks onto PDA-coated electrospun PLGA membranes. The nanofibrous PLGA membrane provided adequate mechanical support for the whole structure, and the nanofibrous CS or Gel networks enhanced cell growth and maturation. The bioinspired surface modification of PLGA scaffolds through PDA coating could not only provide strong adhesion between the two scaffold layers but also improve biological properties of scaffolds. It was demonstrated that functionalized bilayer scaffolds could promote cell adhesion, spreading and proliferation of mouse preosteoblastic MC3T3-E1 cells and rat bone-marrow-derived stromal cells (rBMSCs). Furthermore, immunofluorescence staining and calcium deposition studies revealed that functionalized bilayer scaffolds could enhance osteogenic differentiation of MC3T3-E1 cells and rBMSCs in comparison with simple electrospun PLGA scaffolds. The functionalized bilayer scaffolds are promising structures for bone tissue engineering.