For hydrogel materials used in surgery to prevent post-operative adhesion formation, the ability to reduce adhesion formation effectively through ease of application is the most outstanding attribute for improving their clinical utility. In this study, hydrogel formulation with carboxymethyl cellulose (CMC) and water soluble sodium oxidized regenerated cellulose (ORC) powder was developed. The formulation was achieved with different molecular weights and degrees of substitution of the CMC to investigate the effects of these two variables on adhesion prevention. In vivo studies showed that hydrogel formulations with medium molecular weight and a higher degree of substitution gave the best anti-adhesion performance. Histological analyses indicated the materials did not damage the tissue at the surgery area. Promising results were obtained for the development of ORC containing hydrogel formulations for post-operative adhesion prevention applications.