Purpose: To report a meta-analysis of the literature comparing internal limiting membrane (ILM) peeling with and without indocyanine green (ICG) staining in macular hole (MH) treatment. Methods: A Pubmed search was conducted from January 1999 to June 2004. Manuscripts describing the anatomical and functional outcomes of vitrectomy plus ILM peeling with and without ICG application in MH surgery were reviewed. A statistical meta-analysis was performed including studies which defined anatomical outcomes as closure of the MH and disappearance of the fluid cuff and functional outcomes as improvement of 2 or more Snellen lines. A secondary outcome was to investigate the incidence of retinal pigment epithelium (RPE) alterations with and without ICG staining in MH surgery. Results: Results including all types of MHs in 837 eyes indicated the same anatomical success but worse functional outcomes in the group with ICG application (p = 0.0008; odds ratio = 0.587, 95% confidence interval = 0.427–0.808). A higher incidence of RPE alterations in the ICG injection group was observed. The incidence of RPE alterations was found to be 1.98%, whereas RPE changes were noted in 13.83% of 201 patients with ICG application (odds ratio = 7.998). Conclusion: This meta-analysis of previous reports comparing ILM peeling with and without intravitreal ICG application in the treatment of MHs demonstrated statistically worse functional outcomes when ICG was applied (p = 0.0008). A higher number of RPE alterations were observed in the ICG-stained group.