Aim: This study aimed to investigate the relationship between dry eye syndrome (DES) and contact lens use by comparing clinical and subjective measures of tear film function and ocular surface health in contact lens users and non-users. Materials and Methods: A prospective, observational study was conducted on 110 participants (55 contact lens users and 55 non-contact lens users) at a tertiary care hospital. All participants underwent Schirmer’s test, Tear Break-Up Time (TBUT) test, Ocular Surface Disease Index (OSDI) questionnaire, and fluorescein staining. Statistical analysis was performed using SPSS version 25.0, with t-tests, Chi-square tests, and Mann-Whitney U tests applied as appropriate. A p-value < 0.05 was considered statistically significant. Results: Contact lens users had significantly lower tear production (Schirmer’s test: 8.92 ± 3.25 mm vs. 13.48 ± 3.87 mm, p = 0.0012) and reduced tear film stability (TBUT: 7.89 ± 2.41 sec vs. 12.34 ± 3.01 sec, p = 0.0008) compared to non-users. The OSDI score was significantly higher in contact lens users (32.18 ± 6.92 vs. 12.45 ± 4.67, p = 0.0004), with 81.82% experiencing moderate-to-severe dry eye symptoms. Fluorescein staining showed higher corneal damage trends in contact lens users (54.54%) compared to non-users (41.82%), though this difference was not statistically significant (p = 0.5763). Conclusion: Contact lens wear is significantly associated with dry eye syndrome, as evidenced by reduced tear production, lower tear film stability, increased subjective symptoms, and ocular surface damage. These findings highlight the need for regular monitoring, improved lens hygiene, and preventive strategies to reduce dry eye complications in contact lens users.