Purpose: To evaluate the efficacy of different autologous serum eye drop (ASED) concentrations in treating ocular surface diseases. Methods: All articles assessing the efficacy of 20%, 50%, or 100% ASED were retrieved from PubMed, Cochrane, ScienceDirect, and EBSCO electronic databases and checked for applicability. Efficacy was evaluated based on symptom scoring, corneal staining, and tear film stability, which includes tear break-up time (TBUT) and the Schirmer’s test. Results: Seven randomized clinical trials, three cohort studies, and one prospective study were included in this review. ASED significantly improved subjective ocular symptoms (overall standardized mean differences [SMD]: −2.99; 95% confidence interval [CI]: −4.38 to −1.60), without significant difference between 20%, 50%, and 100% concentrations (P = 0.617). Overall, TBUT also showed a considerable increase (mean differences [MD]: 1.68 s; 95% CI: 0.43–2.94), remarkably with 20% ASED (MD: 2.80; 95% CI: 1.61–4.00). In contrast, the Schirmer’s test showed no significant trend toward improvement (MD: 0.53; 95% CI: −0.78–1.84). A remarkable effect was observed in keratoepitheliopathy (overall SMD: −1.08; 95% CI: −1.85 to −0.32), especially in the 50% ASED subgroup (SMD: −1.54; 95% CI: −2.73 to −0.35). All outcomes showed high heterogeneity (I 2 = 79.6%–97.3%). Sensitivity analyses limited to randomized controlled trials showed consistent findings, supporting the robustness of the results. Conclusion: ASED was effective in improving subjective ocular symptoms, tear break-up time, and keratoepitheliopathy across different concentrations but did not significantly affect Schirmer’s test. © 2025 Journal of Current Ophthalmology.