A nanofibrous beta-chitin web was fabricated via electrospinning for use as a novel wound dressing material. beta-chitin was extracted from cuttlefish bone using deproteinization and demineralization. First, cuttlefish bone was alkali-treated to remove the proteins and was then treated with the acid for demineralization. The extracted beta-chitin was dissolved in formic acid as solvent to evaluate its electrospinnability, and the electrospinnability increased remarkably when beta-chitin was blended with poly(ethylene oxide) (PEO) than without. The blended beta-chitin/PEO nanofibers had a fiber diameter of about 400 nm, and the diameter decreased after soaking in water to remove the PEO. The structural and physical properties of the beta-chitin material and its nanofibers were characterized using Attenuated total reflectance infrared spectroscopy (ATR-IR), Proton nuclear magnetic resonance (H-1 NMR), Scanning electron microscopy/Energy dispersive spectroscopy (SEM/EDS), X-ray diffraction (XRD), texturometry, viscometry and contact angle measurements, and an animal test was conducted to investigate the wound healing effect. The beta-chitin nanofibers were found to have great potential as nanomaterials for wound healing.