Aim: To evaluate the efficacy and safety of mesenchymal stem cell (MSC) therapy for pelvic floor reconstruction and urinary continence restoration in patients with pelvic floor dysfunction.Material and Methods: This prospective, interventional study involved 80 participants aged 30–70 years diagnosed with pelvic floor dysfunction or urinary incontinence. MSCs were harvested from autologous bone marrow or adipose tissue, characterized for viability and surface markers, and injected into the pelvic floor muscles or periurethral tissues under ultrasound guidance. Primary outcomes included improvements in pelvic floor function assessed by Pelvic Floor Distress Inventory (PFDI) scores and secondary outcomes involved urinary continence restoration measured by the International Consultation on Incontinence Questionnaire (ICIQ). MRI evaluations and adverse event monitoring were conducted over 12 months to assess tissue regeneration and safety.Results: The study demonstrated significant improvements in PFDI and ICIQ scores, with a 56% and 60% improvement, respectively, at 12 months (p < 0.001). MRI evaluations revealed a 46% increase in pelvic floor muscle volume over the same period (p < 0.001). Adverse events were minimal, with 10% reporting mild localized pain and 2.5% experiencing moderate infections, both resolving without long-term complications. The therapy exhibited a favorable safety profile.Conclusion: MSC therapy significantly improved pelvic floor function, urinary continence, and muscle integrity with sustained benefits over 12 months. The procedure was safe and well-tolerated, underscoring its potential as a regenerative treatment for pelvic floor dysfunction. Further research is warranted to validate these findings in larger cohorts and optimize therapeutic protocols.