Background The staple line leakage is a dangerous complication of sleeve gastrectomy. Various strategies have been tried to reduce the leakage risk. The amniotic membrane (AmM) is the inner layer of the placental membranes and has anti-inflammatory, anti-fibrosis, and anti-scarring effects, and it also has lower immune characteristics which are another essential characteristic of AmM concerning its utility for grafting. In this study, we aimed to investigate the impact of AmM on the staple line healing process of sleeve gastrectomy model in rats. Materials and Methods We used twenty-eight Long-Evans rats in this study. Sleeve gastrectomy was performed with tristapler. Fourteen rats served as controls, AmM was applied staple line of the other fourteen. Fourteen animals were sacrificed (seven from the AmM applied group and the other seven from the control group) on the third postoperative day. And, the other fourteen animals were sacrificed (seven from the AmM applied group and seven from the control group) on the seventh postoperative day. The tissue around the staple line was evaluated microscopically and macroscopically, bursting pressures and hydroxyproline levels were also measured. Results The bursting pressure and hydroxyproline measurements of the AmM applied group was significantly higher on the seventh postoperative day (p = 0.015, p = 0.012) Fibroblast activity and neoangiogenesis of the AmM applied group was also significantly higher on the seventh postoperative day (p = 0.004, p = 0.002). Conclusion This study showed that covering of staple line of sleeve gastrectomy model in rats significantly provided higher bursting pressures and increased hydroxyproline levels, fibroblast activity, and neoangiogenesis which may potentially lead a better staple line healing. We think further investigations are needed on this issue.