Background: Medial tibial stress syndrome (MTSS) is a common overuse injury characterized by pain along the distal two-thirds of the medial tibial border. It predominantly affects athletes engaged in weight-bearing activities, including football players. Despite its prevalence, there is limited research focusing on MTSS specifically within the context of football, particularly in relation to unilateral versus bilateral occurrences and the implications of limb dominance. Objective: The study aimed to measure the frequency of unilateral versus bilateral MTSS in footballers and to provide insights into the relationship between MTSS and limb dominance. Methods: This comparative cross-sectional study was conducted over six months from July 2021 to December 2021 across various football clubs in Lahore, Pakistan. Ethical approval was obtained from the Lahore College of Physical Therapy, and permissions were secured from club administrations. The sample comprised 140 recreational footballers aged 20-29 years, selected using non-probability convenience sampling. Participants were diagnosed using the MTSS criteria established by Marinus Winters in 2017, which included history taking and physical examination. Data collection involved a detailed questionnaire covering demographics, medical history, and sporting history. The study excluded individuals with previous surgical history, systemic diseases, or tumors. Data analysis was performed using SPSS version 25, employing descriptive and inferential statistics to examine relationships between variables. Results: The mean age of participants was 21.3 years (SD = 2.97), with a mean weight of 62.2 kg (SD = 8.1) and mean height of 5.5 feet (SD = 0.39). The sample included 97 males (69.3%) and 43 females (30.7%). MTSS was reported unilaterally in 82.2% of participants, with 67.9% (n = 95) experiencing symptoms in the right leg and 14.3% (n = 20) in the left leg. Bilateral MTSS was observed in 17.9% (n = 25) of participants. Conclusion: The majority of athletes reported unilateral MTSS, with the right leg being more commonly affected than the left. These findings suggest the need for comprehensive bilateral assessment in the diagnosis and treatment of MTSS in footballers to optimize performance and prevent recurrence.