Purpose To evaluate the clinical efficacy and histopathological tolerance of 2-octyl cyanoacrylate versus conventional suture materials for eyelid wound closure in rabbits. Methods We performed an experimental study on 16 eyes of eight New Zealand albino rabbits. Eyelid incisions of 15 mm were done 4mm from the upper eyelid margin in both eyes. The eyes of the rabbits were divided into two groups: eyelid incisions of the right eye were closed by a 2-octyl cyanoacrylate adhesive (group A) and eyelid incisions of the left eye were closed by 7-0 nylon sutures (group B). At 1, 2, 4, and 8 weeks after surgery, the rabbits were macroscopically examined and then sacrificed. The specimens of their eyelid tissues were stained by a hematoxylin and eosin stain and Masson-trichrome stain, and were observed under microscope. Results Both eyelid surgical closure methods were found to be equally efficacious in fixing the eyelids of groups A and B, and their clinical efficacy was similar. Histopathological findings of the hematoxylin and eosin stain of group A showed less inflammatory infiltration than group B at 2 weeks. There were no significant histopathological differences between the two groups at 1, 4, and 8 weeks. The degree of fibrosis of the Masson-trichrome stain was similar between the two groups at 8 weeks. Conclusions The 2-octyl cyanoacrylate adhesive proved to be an effective eyelid closure method and was very well tolerated by the skin surface. 2-Octyl cyanoacrylate could be used as an alternative tissue adhesive for eyelid wound closure along with conventional suture materials.