Background: To compare the vaginal route and laparoscopic suture for vaginal cuff closure (VCC) in patients undergoing a total laparoscopic hysterectomy (TLH). Methods: A total of 471 women who required hysterectomy were allocated to two groups. 261 women had TLH via VCC by the vaginal route and 210 women had TLH via VCC by laparoscopic suture. All TLHs were performed by the same laparoscopic surgeon. Results: The cuff-related complications included vaginal disruption (3.4%), dehiscence (1.27%), vaginal vault bleeding (1.91%), vaginal spotting (19.32%), granulation (1.27%), cuff infection (1.49%), and yellowish vaginal discharge (6.16%). No difference in vaginal cuff complications was found between the laparoscopic and vaginal approach. The median operation time was significantly shorter for the laparoscopic suture (76.74 min, range 40–220; 95% CI 74.84–83.45) than the vaginal route for VCC (85.77 min, range 45–290; 95% CI 86.87–95.36) after hysterectomy (p < 0.001). Conclusion: For VCC with TLH, laparoscopic suture was a safe and less time-consuming procedure. The cuff-related complications were similar in the two groups.