Background Caused by age or trauma, collapsed connective tissue can cause nerve entrapment and damage within the tarsal tunnel. With an unknown incidence rate, tarsal tunnel syndrome is relatively underdiagnosed, and current literature is inconclusive on best practices for patient care. While most standard treatments involve symptom management, this retrospective case series highlights a novel approach, targeting the damaged tissues surrounding the nerves and replacing the structural cushioning with a Wharton’s jelly tissue allograft. Methods This cohort was selected from the retrospective repository at Regenativelabs. The eight selected patients had tarsal tunnel-related defects from four clinical sites. Patient outcomes were tracked on a 90-day calendar utilizing the Numeric Pain Rating Scale (NPRS) and the Western Ontario and McMaster University Arthritis Index (WOMAC). All patients had failed standard care practices for at least six weeks. Each patient received one 2mL application of 150mg minimally manipulated Wharton’s jelly tissue allograft to strategic sites around the affected tarsal tunnel. No patients experienced adverse reactions. Red light and laser therapies were recommended post-application. Results The percent change of improvement in patient pain scales was calculated with the cohort averages at initial application, 30-day follow-up, and 90-day follow-up appointments. All patients reported decreased pain on both scales. Percent change calculated from the initial application to the 90-day follow-up showed an improvement of 59.43% in NPRS and a 37.58% improvement in WOMAC. Conclusions Given the reported pain improvements on various pain rating scales, this study provides evidence that WJ allograft applications are safe, minimally invasive, and efficacious for patients who have failed standard care treatments for connective tissue defects associated with Tarsal Tunnel syndrome. The results of this study warrant further research to confirm the efficacy of Wharton's jelly added to conservative care protocols to clarify WJ allograft application's optimal dose, protocol, and durability.