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This study was undertaken to determine the long-term outcomes of patients whose anal fistulas were managed by use of bioprosthetic plugs.A retrospective analysis was performed of all patients whose anal fistula was managed by use of a bioprosthetic plug between May 2005 and September 2006, who had a minimum of 1 year of follow-up since their last treatment. Patients whose fistulas were clinically healed were offered MRI to confirm healing of the fistula.The bioprosthetic fistula plug was used to treat an anal fistula in 63 patients with clinical healing of the fistula in 51 (81%). Multivariate analysis showed that tobacco smoking, posterior fistula, and history of previous failure of the bioprosthetic plug was predictive of failure of the bioprosthetic plug. Eight patients with clinical healing after a minimum of 1 year since their last treatment underwent MRI. No evidence of residual fistula tract or fluid in the area of the previous fistula was found in 6 (75%) of these patients.Bioprosthetic plugs are effective for the long-term closure of complex fistulas-in-ano. Randomized clinical trials comparing bioprosthetic plugs with other sphincter-preserving methods for fistula management need to be conducted to further determine the role of bioprosthetics in the management of anal fistulas.