A novel anti-adhesion nonwoven mat mainly composed of silk fibroin protein (SFP) was fabricated via the single-spinneret electrospinning technique. A series of SFP-based electrospun nonwoven mats containing additives of different synthetic polymer ratios, such as pure SFP, SFP/poly(vinyl alcohol) (PVA), SFP/polyethylene glycol (PEG), and SFP/polyethylene oxide (PEO) were produced and compared. All membranes were porous and had diameters of 324.02+/-113.7, 308.86+/-74.02, 366.22+/-115.81, and 341.82+/-119.42 nm, respectively. The average pore size for each membrane was 1.132+/-0.99, 0.811+/-0.424, 0.975+/-0.741, and 0.784+/-0.497 mu m(2). No nonwoven mats showed significant cytotoxicity toward fibroblast cells based on the results of MTT assays. Surprisingly, for all groups of SFP-based nonwoven mats, nitrate formation was reduced by up to 94.55+/-14.50%, 92.16+/-19.38%, 91.28+/-28.375%, and 92.00+/-12.64% in lipopolysaccharide-induced RAW 264.7 macrophages model. Tissue anti-adhesion potential was evaluated in an in vitro fibroblast cell adhesion model and in vivo wounded mice model. In vitro, the mean cell anti-adhesion percentage of fibroblast cells changed over time in the following order: PVA/SFP>SFP>PEG/SFP similar to PEO/SFP. In vivo, SFP and PVA/SFP-treated groups both showed superior collagen regeneration and wound closure. (C) 2017 Wiley Periodicals, Inc.