Abstract: To enhance the anti-cell-adhesive properties of silk fibroin electrospun scaffolds and overcome the shortcomings of conventional blending or surface/interface modification strategies, this study proposes an electrospinning strategy combining “multiple chemical modifications and rapid UV-induced co-crosslinking” for fabricating anti-cell-adhesive electrospun scaffolds. Through methacrylated modification of silk fibroin and further co-crosslinking modification with poly(ethylene glycol) diacrylate (PEGDA) during the electrospinning process, silk fibroin-based electrospun scaffolds with significantly improved anti-cell-adhesive performance were successfully constructed. The physicochemical properties, anti-cell-adhesion ability of the scaffolds with different PEGDA contents, and their effects on chondrogenic differentiation of stem cells were investigated using an electronic universal testing machine, scanning electron microscope（SEM）, CCK-8 assay, and other methods. Investigation of the scaffold’s physicochemical properties showed that with increasing PEGDA content, the fiber diameter, pore size, and porosity of the prepared scaffolds gradually decreased, the elongation at break increased progressively, while the breaking strength first increased and then decreased. Stem cell response results indicated that with increasing PEGDA content, the scaffold’s anti-cell-adhesive capability gradually enhanced, and cell adhesion morphology transitioned from a spread state to an approximately spherical state, the scaffold’s ability to promote chondrogenesis of cells showed a trend of first increasing and then decreasing. This study provides important references and guidance for effectively regulating the anti-cell-adhesive performance and morphological features of the silk fibroin based or other similar electrospun scaffolds.