Purpose: We used a noninvasive ocular surface analyzer to explore changes in the ocular surface parameters of patients who underwent 25-gauge pars plana vitrectomy (PPV).Methods: The medical records of 82 patients (163 eyes) who underwent PPV were retrospectively reviewed. The non-invasive tear film break-up time (NIBUT), the lipid layer thickness, meibomian gland loss status, and the tear meniscus height were measured before, and 1 and 3 months after treatment. Consecutive changes in the ocular surface indices of operative and non-operative fellow eyes were statistically compared. Also, factors that reduced the NIBUT at 1 month after surgery were analyzed.Results: One month after surgery, the NIBUT was significantly decreased in the operative group compared to before surgery (<i>p</i> = 0.006). Compared to non-operative eyes, the NIBUT of operative eyes decreased significantly 1 month after surgery (<i>p</i> = 0.003). Older age and lower preoperative NIBUT (<i>p</i> = 0.029, <i>p</i> = 0.002) were significantly associated with lower NIBUT 1 month after surgery.Conclusions: Twenty-five gauge PPV significantly reduces the NIBUT 1 month after surgery.