OBJECTIVE The goal of this experimental study was to investigate the specific effect of myringosclerosis on tympanograms in the tympanic membranes of myringotomized rats by using otomicroscopy, tympanometry, and histopathology. BACKGROUND Myringosclerosis is a common sequela of ventilation tube treatment of otitis media with effusion. The condition involves the hyalinization and calcification of the collagen layer in certain areas of the tympanic membrane. Previous animal experiments suggest an intimate relationship between the formation of myringosclerosis and an increased oxygen concentration in the environment of the wound after myringotomy. The result of a myringotomy therefore is an increased production of free oxygen radicals, initiating irreversible tissue damage involving fibrosis, hyalin degeneration, and finally apoptosis as observed in myringosclerosis. We propose an experimental model specific for creating sclerotic plaques solely on the tympanic membrane and for performing tympanometric measurements on this pure myringosclerosis model without creating any abnormality in the middle ear to test in what proportion myringosclerosis contributes to decrease of amplitude in tympanograms. METHODS To assess the normal tympanometric values of Wistar albino rats, the pressure and peak admittance of the left middle ears were measured using a semiquantitative computerized clinical admittance meter using a sound frequency of 226 Hz. Twelve animals were randomly selected for the myringotomy group and perforations in the left ears were created. All tympanic membrane perforations in this group had healed and closed prior to the otomicroscopic examination and no pathologic reaction was observed in the external ear canals of rats. Otomicroscopic and tympanometric measurements were carried out on Day 15 and the degree of myringosclerosis was noted before the animals were killed. Twelve specimens in the myringotomy group were histopathologically examined for the presence of myringosclerotic plaques. RESULTS Under light microscopy, extensive sclerotic lesions were found in the tympanic membranes of the myringotomy group, and these sclerotic deposits were located in the lamina propria. The myringosclerosis occurred predominantly adjacent to the handle of the malleus, but also near the annular region. In all ears with myringosclerosis, the magnitude of the maximum admittance reduced to approximately 50% of the Day-0 values, and this reduction was statistically significant (Z=-3.061, p=0.002). CONCLUSIONS The present findings in this study are consistent with the fact that the movement of the tympanic membrane is hampered by lesions of sclerotic material, resulting in a decrease of amplitude in tympanograms (such as Type As) without any effusion or inflammation in the middle ear.