Although previous studies have shown that vaginal natural orifice transluminal endoscopic surgery (vNOTES) has the advantages of causing less pain, faster recovery, and better concealment of surgical incisions, which aligns with the concept of the day-care procedure, this approach poses a greater risk of damaging adjacent organs (i. e., rectum and bladder) due to its anatomical specificity. Moreover, the day-care procedure may lead to relatively less preoperative evaluation and postoperative care. Hence, it is necessary to explore the safety and effectiveness of vNOTES for ovarian cystectomy in the day-care procedure, to provide a theoretical basis for the wider development of vNOTES surgery.This retrospective study included 131 patients at our hospital who underwent ovarian cystectomy from September 2021 to October 2022. Based on the surgical approach, patients were classified into transumbilical laparoendoscopic single-site surgery (LESS) and vNOTES groups. The patients' demographic characteristics and follow-up data were collected during the perioperative period and 1-month postoperatively.Vaginal natural orifice transluminal endoscopic surgery has less postoperative exhaust time, a lower postoperative 6-hour pain score, and a lower incidence of analgesic drug use, with higher surgical conversion incidence. Multiple linear regression analysis showed that the surgical conversion, chocolate cyst, bilateral cyst, and pelvic adhesion increased the operation duration by ~43 (95% CI: 10.309, 68.152, p < 0.001), 15 (95% CI: 6.342, 45.961, p = 0.036), 10 (95% CI: 3.07, 40.166, p = 0.019), and 8 (95% CI: 4.555, 26.779, p = 0.035) min, respectively. Interestingly, vNOTES decreased the operation duration by ~8.5 min (95% CI: -18.313, -2.699, p = 0.033).Vaginal natural orifice transluminal endoscopic surgery was equally safe and effective for ovarian cystectomy compared to LESS. vNOTES aligned with the concept of the day-care procedure due to its reduced postoperative pain, shorter exhaust time, and absence of scarring. However, surgeons should conduct a comprehensive preoperative evaluation and exclude patients suspected to have severe pelvic adhesions.Copyright © 2023 Xie, Li, Huang, Wang, Liu, Wang, Liao, Yu, Yan, Zhang, Huang, Liu, Li, Lin, Jia and Gan.