Recently, corneal transplantation has been proposed as an effective treatment for irreversible corneal damage which facilitates access to healthy corneas; however, lack of allografts made the treatment subject to many limitations in medicine. In this regard, this study aimed to produce multilayer nanofiber scaffolds for corneal epithelial layer tissue engineering with scaffold compounds of fibroin silk/collage-EGF. The samples were first prepared and identified from the perspective of engineering and biology applications. The results of this study showed that an alternative tissue with a suitable thickness and structure of nanofibers with suitable engineering and biological properties was successfully prepared. In addition, a scaffold was prepared in this research for tissue engineering of the corneal epithelial layer based on silk fibroin and collagen containing aloe vera and epithelial growth factor as the contributing factors and stimuli for better corneal repair. For this purpose, nanofiber three-layer scaffolds were prepared by a combination of electrospinning and electrospray methods characterized by engineering features, such as Scanning Electron Microscope (SEM), to study the degradability for their weight loss, water contact angle, and growth factor. Release as well as static and dynamic mechanical properties were also investigated. Biological characteristics such as cell binding and scaffold differentiation potential were further explored. The results of this study showed the successful preparation of an alternative with a suitable thickness and structure of nanofibers with suitable engineering and biological properties. The obtained results confirmed the production of a proper scaffold with suitable thickness and nanofiber structure. Therefore, the prepared product could potentially be used as a suitable alternative for repairing the damaged corneal epithelial layer.