Reduced Graphene Oxide Incorporated GelMA Hydrogel Promotes Angiogenesis For Wound Healing Applications

Syed Raza ur Rehman,1,2 Robin Augustine,1,2 Alap Ali Zahid,1,2 Rashid Ahmed,1,2 Muhammad Tariq,3 Anwarul Hasan1,2 1Department of Mechanical and Industrial Engineering, College of Engineering, Qatar University, Doha 2713, Qatar; 2Biomedical Research Center, Qatar University, Doha 2713, Qatar; 3Department of Biotechnology, Faculty of Science, Mirpur University of Science and Technology, Mirpur 10250, AJK, PakistanCorrespondence: Anwarul HasanDepartment of Mechanical and Industrial Engineering, College of Engineering, Qatar University, Doha 2713, QatarTel +97470569169Email hasan.anwarul.mit@gmail.comPurpose: Non-healing or slow healing chronic wounds are among serious complications of diabetes that eventually result in amputation of limbs and increased morbidities and mortalities. Chronic diabetic wounds show reduced blood vessel formation (lack of angiogenesis), inadequate cell proliferation and poor cell migration near wounds. In this paper, we report the development of a hydrogel-based novel wound dressing material loaded with reduced graphene oxide (rGO) to promote cell proliferation, cell migration and angiogenesis for wound healing applications.Methods: Gelatin-methacryloyl (GelMA) based hydrogels loaded with different concentrations of rGO were fabricated by UV crosslinking. Morphological and physical characterizations (porosity, degradation, and swelling) of rGO incorporated GelMA hydrogel was performed. In vitro cell proliferation, cell viability and cell migration potential of the hydrogels were analyzed by MTT assay, live/dead staining, and wound healing scratch assay respectively. Finally, in vivo chicken embryo angiogenesis (CEO) testing was performed to evaluate the angiogenic potential of the prepared hydrogel.Results: The experimental results showed that the developed hydrogel possessed enough porosity and exudate-absorbing capacity. The biocompatibility of prepared hydrogel on three different cell lines (3T3 fibroblasts, EA.hy926 endothelial cells, and HaCaT keratinocytes) was confirmed by in vitro cell culture studies (live/dead assay). The GelMA hydrogel containing 0.002% w/w rGO considerably increased the proliferation and migration of cells as evident from MTT assay and wound healing scratch assay. Furthermore, rGO impregnated GelMA hydrogel significantly enhanced the angiogenesis in the chick embryo model.Conclusion: The positive effect of 0.002% w/w rGO impregnated GelMA hydrogels on angiogenesis, cell migration and cell proliferation suggests that these formulations could be used as a functional wound healing material for the healing of chronic wounds.Keywords: GelMA hydrogel, reduced graphene oxide, nanocomposite hydrogel, angiogenesis, wound healing