This study evaluates the efficacy and safety of non-ablative diode laser therapy for genitourinary syndrome of menopause (GSM) in post-menopausal women unable to use hormonal therapies. A pilot prospective study was conducted from September 2023 to April 2024, involving 22 post-menopausal women aged 45-73years. Participants underwent three sessions of diode laser treatment with the Leonardo® dual-wavelength Diode laser. Assessments were made at baseline, 3months, and 6months post-treatment. Main outcome measures included Visual Analog Scale (VAS) scores for GSM symptoms, Vaginal Health Index Score (VHIS), and sexual function evaluated using the Female Sexual Function Index (FSFI-6), Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12), and Sexual Quality of Life-Female (SQOL-F) questionnaire. Significant improvements were observed in VHIS, increasing from 12 to 19.27 at 6months (p<0.001). GSM symptoms improved significantly: vaginal dryness scores decreased from 7.72±2.37 to 3.72±2.53, burning sensation scores dropped from 6.00±3.22 to 1.90±1.81, and dyspareunia scores reduced from 8.09±2.11 to 3.90±2.58 (all p<0.016). Sexual function improved, indicated by FSFI-6 scores increasing from 12.27±7.29 to 19.30±6.24 (p<0.016) and SQOL-F scores rising from 63.18±22.93 to 71.45±23.31. No adverse events were reported. Non-ablative diode laser therapy is effective and safe for managing GSM symptoms in post-menopausal women, offering significant symptom relief and enhancing sexual health without serious side effects. Further research with a larger cohort and extended follow-up is needed to confirm these findings. Clinical Trials ID NCT06503003.