The usage of materials with the potential to accelerate wound healing is a great benefit for patients and health care systems. This study evaluated the impact of using graphene oxide (GO)–cellulose nanocomposite on skin wound healing via in vitro and in vivo investigations. The nanomaterial was synthesized and characterized. Cytocompatibility performance of the GO-cellulose was investigated through in vitro testing based on MTT and live/dead assays by EA.hy926 human endothelial cells (ECs). Additionally, the effect of GO-cellulose on induced wound scratch model using EA.hy926 ECs was investigated. Finally, the therapeutic effect of GO-cellulose was evaluated in vivo after the creation of two full-thickness wounds in the dorsum of rats (8 mm diameter). These wounds were randomly placed into two groups, the control group (10 wounds) and the GO-cellulose group (10 wounds), and monitored for gross and histopathological changes at 7 and 21 days after wound induction. MTT and Live/Dead assays showed excellent GO-cellulose cytocompatibility, whereas no difference in ECs viability was observed after culturing using conditioned media. GO-cellulose nanocomposite enhanced cell migration in the in vitro wound scratch assay. As compared to the control group, the GO-cellulose nanocomposite group's wound healing process was promoted in the in vivo rat skin wounds. Interestingly, wound re-epithelization and neovascularization were significantly accelerated in the GO-cellulose-treated rats. Furthermore, thick granulation tissue formation and intense collagen deposition were found in the GO-cellulose group. These findings showed that GO-cellulose has a promoting effect on skin wound healing, suggesting its promising and potential application in tissue regeneration.