Kinetic analysis of cell proliferation using bromodeoxyuridine labeling and in situ detection of dying cells in the tympanic membrane and middle ear cholesteatoma.

Using specimens from the posterior-superior quadrant of the human tympanic membrane, meatal skin, retroauricular skin and middle ear cholesteatoma, epidermal cell proliferation was studied by cultures in FC43 emulsion containing bromodeoxyuridine (BrdU), and cell death was detected by in situ labeling of nuclear DNA fragmentation (TUNEL staining). The culture of specimens with BrdU revealed labeling in the basal cell layer and/or the layer immediately above it. The counts of BrdU-labeled cells both at and beside the malleus handle and at the annulus were significantly higher than those in the tympanic membrane, meatal skin, retroauricular skin and cholesteatoma, indicating the existence of epidermal proliferation centers in the annulus and malleus handle. TUNEL-positive cells were observed in the uppermost layer of the epidermis, and counts of dying cells did not show any significant differences among specimens from the different areas. From these observations, we conclude that addition of newly proliferated cells at the proliferation center and uniform cell death cause epidermal cell migration over the tympanic membrane and ear canal. In addition, no proliferation center was seen in the epidermis of cholesteatoma, suggesting a disturbance of epidermal cell migration. Furthermore, BrdU-labeling at the margin of persistently perforated tympanic membranes from patients with chronic otitis media revealed that, at the perforation margin, the counts of BrdU-labeled cells were not higher than those of the normal tympanic membrane. In addition, a few BrdU-labeled cells were observed in the lamina propria and mucosal cell layer, indicating that persistent perforation of the tympanic membrane results from the failure of proliferating cells to increase at the margin of the perforation.