To investigate the efficacy of Seprafilm (Genzyme, Framingham, MA, USA) in preventing postoperative adhesion between the conjunctiva and sclera after glaucoma filtering surgery. A subconjunctival pocket was created and Seprafilm was inserted into the pocket in nine rabbits (Seprafilm group), whereas in a second group, a subconjunctival pocket was created in nine rabbits but no Seprafilm was inserted (non-Seprafilm group). The postoperative adhesion force was measured 4 weeks after surgery. For the trabeculectomy study, trabeculectomy was performed and Seprafilm placed on the scleral flap in five rabbits (Seprafilm trabeculectomy group), whereas in a second group, trabeculectomy was performed in five rabbits but no Seprafilm was placed (non-Seprafilm trabeculectomy group). Filtering bleb formation and intraocular pressure (IOP) was evaluated on days 1, 3, 5, 7, 14, 21, and 28 following surgery. The eyes were enucleated for histologic evaluation 4 weeks after surgery. The mean adhesive force between the conjunctiva and sclera in the Seprafilm group (125.6 +/- 94.5 mmHg) was lower than that of the non-Seprafilm group (263.3 +/- 79.3 mmHg) (P = 0.0041, unpaired t test). A more prominent bleb was observed in the Seprafilm trabeculectomy group than in the non-Seprafilm trabeculectomy group. Histologically, the subconjunctival space was larger in the Seprafilm trabeculectomy group than in the non-Seprafilm trabeculectomy group. Mean IOP was significantly lower in the Seprafilm trabeculectomy group (9.9 +/- 0.6 mmHg) than in the non-Seprafilm trabeculectomy group (11.9 +/- 0.7 mmHg) 4 weeks after surgery (P = 0.0044, unpaired t test). Seprafilm can reduce postoperative conjunctiva-sclera adhesion and may be a desirable antifibrotic agent for trabeculectomy in the early stages of wound repair.