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Background Endolaser probes have been designed and sold for single-use only. However, in Brazil, they are not included in the list of single-use medical products that are prohibited from being reprocessed and could potentially be reused if safety requirements are accomplished. Therefore, this study aimed to determine and compare the quality, safety and costs of reprocessed versus original single-use endolaser probes of a specific brand and model. Methods The study, conducted at a university hospital in Sao Paulo, Brazil, was divided in two phases. The first one tested the feasibility, sterility and physical integrity of ten reprocessed laser probes. In the second phase, all vitrectomy procedures using endolaser probes (reprocessed and original ones) from August 2017 to October 2019 were evaluated. The operated cases were followed for any signs of infection and number of defective probes for each group were counted. The cost of acquiring a new probe and for all reprocessing stages were evaluated and quantified in US dollars($). Results Microbiologic, residual ethilen oxide and microscopic evaluation of integrity of reprocessed laser probes were all within acceptable range. The second phase of this study included 590 endolaser probes, of which 375 were original and 215 were reprocessed. Functionality rates differed significantly between groups. Among the original probes, 373 (99.47%) were functioning and 2 (0.53%) were non-functioning. Among the reprocessed ones, 201 (93.5%) were functioning and 14 (6.5%) were non-functioning (p Conclusions Our study showed that a single cycle endolaser probe reprocessing was safe and efficient, not associated with increase in endophthalmitis rate and proved to be significantly cost-effective, even considering a greater malfunction rate when compared to the original devices.