Background: Postsurgical adhesions can occur following virtually all types of surgery, resulting in serious clinical complications. Therefore, prevention of adhesions is an important goal of surgical practice. A rat uterine horn model was used to investigate the efficacy of NO-carboxymethylchitosan (NOCC) and spermine NONOate (SPER/NO) alone and in combination in preventing adhesion formation. Methods: Sixty Wistar albino rats underwent bilateral uterine horn injury with a unipolar cautery. Study groups were as follows: (i) control group, no adjuvant therapy; and those with adjuvant applied, (ii) normal saline group, 2 ml of normal saline was given; (iii) NOCC group, 2 ml of 2% NOCC gel was given; (iv) SPER/NO group, 2 ml of SPER/NO (0.5 mg/ml) was given, and (v) NOCC plus SPER/NO group, 2 ml of 2% NOCC gel including SPER/NO (0.5 mg/ml) was given. After 14 days, all animals were euthanatized, and a standard adhesion scoring system including extent and severity scores was applied by a blinded examiner. Results: The extent score in NOCC plus SPER/NO group was significantly lower than those of control and normal saline groups (p<0.05). The extent score in NOCC group was significantly lower than that of normal saline group (p<0.05). The extent score in NOCC plus SPER/NO group was significantly lower than that of SPER/NO group (p<0.05). The severity score was significantly lower in NOCC plus SPER/NO and NOCC groups than that of control group (p<0.05). The severity score was significantly lower in NOCC plus SPER/NO group than that of SPER/NO group (p<0.05). Conclusions: Postoperative administration of NOCC gel and SPER/NO alone and especially in combination to the site of peritoneal injury reduces the formation of adhesions in the rat uterine horn model.