Abstract Background The purpose of this paper is to study the utility of adhesives (artificial-cyanoacrylate and biological-fibrin glue) for improving transconjunctival sutureless vitrectomy (TSV) sclerotomy closure competency. Methods Experimental and observer-masked study in which after performing TSV in cadaveric pig eyes, different adhesives were tested on sclerotomy entrances in order to determine if they improved closure competency in face of progressive intraocular pressure increase. In 76 eyes cyanoacrylate-treated sclerotomies were compared with sclerotomies in which no additional manoeuvre to aid closing was performed; in 76 eyes fibrin glue with no manoeuvre; and in the last 76 eyes, cyanoacrylate-treated sclerotomies were compared with fibrin glue-treated sclerotomies. Results A total of 228 eyes had a 23-gauge TSV performed. Both cyanoacrylate and fibrin glue treated sclerotomies achieved higher mean opening pressures when compared with nontreated sclerotomies in the same eye (p < 0.002). When cyanoacrylate was compared with biological adhesive in the same eye, no statistically significant differences were obtained (p = 0.216). Discussions This experimental study provides support for the possible role of adhesives in improving TSV sclerotomy closure competency in clinical practice.