Aim To verify the safety and efficacy of Ologen (OLO) implant as adjuvant compared with low-dosage mitomycin-C (MMC) in trabeculectomy. Methods This was a prospective randomized clinical trial with a 24-month follow-up. Forty glaucoma patients (40 eyes) were assigned to trabeculectomy with MMC or OLO. Primary outcome includes target IOP at r21, r17, and r15 mm Hg; complete (target IOP without medications), and qualified success (target IOP regardless of medications). Secondary outcomes include bleb evaluation, according to Moorfields Bleb Grading System (MBGS); spectral domain optical coherence tomography (SD-OCT) examination; number of glaucoma medications; and frequency of postoperative adjunctive procedures and complications. Results The mean preoperative IOP was 26.5 ( ± 5.2) in MMC and 27.3 ( ± 6.0) in OLO eyes, without statistical significance. One-day postoperatively, the IOP dropped to 5.2 ( ± 3.5) and 9.2 ( ± 5.5) mm Hg, respectively (P ¼ 0.009). The IOP reduction was significant at end point in all groups (P ¼ 0.01), with a mean IOP of 16.0 (±2.9) and 16.5 (±2.1) mm Hg in MMC and OLO, respectively. The rates and Kaplan-Meier curves did not differ for both complete and qualified success at any target IOP. The bleb height in OLO group was higher than MMC one (Po0.05). SD-OCT analysis of successful/ unsuccessful bleb in patients with or without complete success at IOP r17 mm Hg indicated a sensitivity of 83% and 73% and a specificity of 75% and 67%, respectively, for MMC and OLO groups. No adverse reaction to OLO was noted. Conclusions Our results suggest that OLO implant could be a new, safe, and effective alternative to MMC, with similar long-term success rate.