The development of artificial tympanic membrane and middle ear mucosa by reproductive tissue engineering to be aimed for new treatment of adhesive otitis media.

首先，我们利用温敏培养皿从白兔外耳道获取培养的角质形成细胞，研制了人工鼓膜上皮片。另一方面，我们研制了由细胞外基质和培养的中耳粘膜组成的人工中耳粘膜片，该黏膜片取自同种兔的中耳大泡。我们最终将人工中耳粘膜膜与鼓膜上皮膜融合，制成了人工鼓膜。经电子显微镜和免疫组织化学染色证实，人工鼓膜具有与正常鼓膜相似的病理组织学特征。虽然我们试图用人工鼓膜建立实验性粘连性中耳炎模型，但仅靠某些细胞因子和生长因子的刺激是无法完成的。

At first, we developed artificial tympanic membrane epithelium sheets from cultured keratinocyotes obtained from external ear canal of a white rabbit by using the temperature-responsive culture dish. On the other hand, we developed artificial middle ear mucosal sheets consist of extra cellular matrix and cultured middle ear mucosa that was obtained from middle ear bulla of a homogeneous rabbit. We finally manufactured an artificial tympanic membrane by fusion of artificial middle ear mucosal sheets and tympanic membrane epithelium sheets. As we confirmed the pathohistologic characteristics of the manufactured artificial tympanic membrane by electron microscope and immunohistochemical staining, it resembled characteristics of normal tympanic membrane. Although we attempted to develop an experimental adhesive otitis media model by using the artificial tympanic membrane, we could not accomplish to develop it only by the stimulation of some cytokines and growth factors.