The developmentof artificial tympanic membrane by tissue engineering for reproduction and pathologic elucidation of middle ear cholesteatoma.

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We developed artificial middle ear mucosal sheets from cultured middle ear mucosa that was taken from middle ear bulla of a white rabbit by using the temperature-responsive culture dish. And also, we developed tympanic membrane epithelium sheets consist of extra cellular matrix and keratinocyte that was taken from external ear canal of the same rabbit. After that, we finally manufactured an artificial tympanic membrane by fusion of artificial middle ear mucosal sheets and tympanic membrane epithelium sheets. We confirmed this artificial tympanic membrane was resembled normal tympanic membrane from the viewpoint of pathohistologic characteristics by immunohistochemical staining and electron microscope findings. Although, we attempted to develop an experimental middle ear cholesteatoma model by using the artificial tympanic membrane, it was impossible to develop it only by the stimulation of some chemical mediators and cytokines.On the other hand, we confirmed the suppression of bone and granulation tissue hyperplasia in the middle ear bulla after implantation of artificial middle ear sheets.