In this study, multilayer nanofiber scaffolds for corneal epithelial layer tissue engineering with scaffold compounds of fibroin silk/collage-EGF was produced. The samples were prepared and identified from the perspective of engineering and biology application. The results of this study showed the success of preparing an alternative tissue with a suitable thickness and structure of nanofibers with suitable engineering and biological properties. In this study, a scaffold was prepared for tissue engineering of the corneal epithelial layer based on silk fibroin and collagen containing aloe vera and epithelial growth factor as contributing factors and stimuli for better corneal repair. For this purpose, nanofiber three-layer scaffolds are prepared by a combination of electrospinning and electrospray methods and with engineering features like scanning electron microscope (SEM), to study the degradability for their weight loss, water contact angle, and growth factor. Release and static and dynamic mechanical properties were also investigated. Biological characteristics such as cell binding and scaffold differentiation potential were investigated. The results of this study showed the success of preparing an alternative with a suitable thickness and structure of nanofibers with suitable engineering and biological properties. Therefore, the prepared product could potentially be used as a suitable alternative for repairing the damaged corneal epithelial layer.