Introduction Orthodontic discomfort is a major barrier to treatment compliance, with pain being a common reason for patients to discontinue. Pain often originates from the periodontal ligament due to pressure, ischemia, and swelling, and is influenced by individual factors like age, gender, and stress. Vitamin D and Platelet-Rich Fibrin (PRF) have emerged as non-invasive methods to accelerate orthodontic tooth movement while potentially reducing treatment pain. Materials and Methods This split-mouth, randomized control trial involved 20 adult patients undergoing canine retraction post-premolar extraction. PRF and Vitamin D were administered in different quadrants, with saline serving as the control. PRF was prepared using the double centrifugation technique and injected into the periodontal ligament. Vitamin D was prepared as a 50 pg dose and administered similarly. Canine retraction forces were applied using 150 grams of elastomeric chains, and pain was assessed using a Visual Analog Scale (VAS) immediately, after 24 hours, and at one week. Results PRF showed significantly higher pain scores than the control group after 24 hours and one week. In contrast, no significant difference in pain perception was observed between Vitamin D and control groups. Additionally, there was no significant difference in pain between PRF and Vitamin D groups at any time point.