The current treatment of choice for chronic tympanic membrane perforations is surgery. Recent studies using various polypeptide growth factors to accelerate closure of tympanic membrane perforations in model systems have produced mixed results. This study evaluates the effect of acidic fibroblast growth factor (AFGF) and live yeast cell derivative (LYCD) on the rate of healing of acute tympanic membrane perforations in a rat model. Thirty-seven rats had both ears separately randomized in a blinded fashion to receive AFGF in one of three concentrations, LYCD, or a control solution. The rats initially underwent subtotal removal of the tympanic membranes bilaterally. Solutions were applied to the randomized ears daily for 3 days, starting at the time of the surgical perforation. The ears were photographed every 3 to 8 days for 35 days. The photographs were digitally scanned and a computer analysis was used to calculate the percentage of residual perforation. No significant difference in the rate of healing was observed for ears treated with AFGF or LYCD versus the controls. Given the potential advantages of medical treatment of tympanic membrane perforations and the established efficacy of growth factors in other model systems, however, further research is warranted.