PROJECT SUMMARY In this project, we propose to design a hybrid scaffold from the integration of gelatin glycidyl methacrylate (GELGYM) with exfoliated pristine graphene (EG), accompanied by double crosslinking technique, that can function as a suturable artificial cornea. We will determine (1) whether hybrid GELGYM membranes acts as a biocompatible artificial cornea that supports cell growth and differentiation;; and (2) the capability of the hybrid scaffold to function as a corneal substitute. The differentiation and biocompatibility studies proposed here will offer quantitative information on the degree of biointegration of the hybrid scaffold in contact with the host’s corneal cells and extracellular matrix under in vitro and in vivo conditions. We expect that exfoliation of graphene, its integration in the GELGYM hydrogel architecture and subsequent double crosslinking can create an nano-microspherical cavities in the hydrogel microstructure covered with the graphene create anchoring points that can withstand against tearing, after applying the physical stress to the suture passing the construct. This can diminish the tendency to form notches during suturing, while enhancing mechanical properties of the construct and facilitate its implantation in patients. Moreover, we expect integration will further promote cell proliferation and differentiation of corneal cells, allowing for the appropriate diffusion of glucose and oxygen. Additionally, I anticipate that integration of exfoliated graphene will positively regulate epithelial-stromal and endothelial stromal cell interactions. The success of this approach can reduce or eliminate the need for donor corneas and positively impact the quality of life for many patients. The research aims are supported by a training plan focused on the acquisition of relevant multidisciplinary expertise in the fields of cellular and molecular biology, pathophysiology, tissue-engineering and material science. To this end, a “Mentoring Team” which includes the leading mentor, Dr. Chodosh, along with two co- mentors: Dr. Gonzalez and Dr. Kong has been assembled. Moreover, Dr. Paschalis as an independent collaborator will support on technical training on animal handling, surgery, and postoperative care along with data analysis and interpretation of animal findings following implantation of the scaffold into rabbit eyes. This group of mentors will guide me during the K99 mentored phase to achieve my long-term career goal of becoming a scientist-leader in my field with a strong independent laboratory. I envision that this award will provide me with an excellent platform from which to have a successful transition to independent research fac...