Collagen-based biomimetic nanofibrous scaffolds: preparation and characterization of collagen/silk fibroin bicomponent nanofibrous structures

Electrospinning of collagen (COL)/silk fibroin (SF) blend solutions in 1,1,1,3,3,3-hexafluoro-2-propanol was investigated for fabrication of a biocompatible and biomimetic nanostructured scaffold for tissue engineering. The morphology of the electrospun COL/SF blend nanofibers was observed by scanning electron microscopy. The average diameters of COL/SF blend fibers ranged from 320 to 360 nm, irrespective of SF content in the blends. Both COL and SF components in the as-spun COL/SF blend matrices were stabilized by glutaraldehyde and water vapor, respectively, under the saturated glutaraldehyde aqueous solution at 25 degrees C. The glutaraldehyde vapor chemically stabilized the COL component via cross-linking, whereas the water vapor physically stabilized the SF component via crystallization to the beta-sheet structure. These structural changes of after-treated COL/SF blend matrices were examined using ATR-IR and CP/MAS (13)C NMR spectroscopy. To assay the cytocompatibility and cellular behavior of the COL/SF blend nanofibrous scaffolds, cell attachment and the spreading of normal human epidermal keratinocytes (NHEK) and fibroblasts (NHEF) seeded on the scaffolds were studied. In addition, both morphological changes and cellular responses of COL/SF blend nanofibrous matrices were also compared with COL/SF hybrid nanofibrous matrices. Generally similar levels of cell attachment and spreading of NHEF were shown in the COL/SF blend nanofibrous matrix compared with those of the pure COL and pure SF matrices; the cellular responses of NHEK were, however, markedly decreased in the COL/SF blend nanofibrous matrix as compared to the pure matrices. In contrast, cell attachment and spreading of NHEK on the COL/SF hybrid nanofibrous matrix were significantly higher than that of the COL/SF blend nanofibrous matrix. Our results indicate that a COL/SF hybrid nanofibrous matrix may be a better candidate than a COL/SF blend nanofibrous matrix for biomedical applications such as wound dressing and scaffolds for tissue engineering.