Development of modified tympanoplasty combined with transplantation of artificial middle ear mucosa and drug delivery system

我们成功地制备了具有三维结构的中耳粘膜上皮细胞片，与正常中耳粘膜相似。此外，我们清楚地发现，将自体细胞形成的细胞片移植到兔中耳粘膜缺失区可以促进粘膜的快速再生。此外，还成功制备了温度响应型培养插入物的人鼻上皮细胞片。通过对人造人鼻上皮细胞片的组织学研究，我们证实了皮层中存在表皮细胞样细胞，基质中存在成纤维细胞样细胞。膜片由多层粘膜上皮和下结缔组织组成，与正常中耳粘膜相似。扫描电镜显示鹅卵石样的细胞，有无数的微绒毛和纤毛样的结构。我们还证实了基膜结构，包括紧密连接、桥粒和表皮层之间的细胞粘附结构。该人工粘膜与正常中耳粘膜结构相似。因此，中耳粘膜细胞片被认为是中耳炎手术粘膜再生的有效移植材料，有望在未来应用于人体临床。

We have developed successfully middle ear mucosal epithelial cell sheets with three-dimensional structure, analogous to normal middle ear mucosa. Furthermore, we found it clear that a rapid mucosal regeneration is boosted by transplanting cell sheet created with autologous cells on to the rabbit's middle ear mucosa missing area. In addition, autologous human nasal epithelial cell sheet using temperature-responsive culture insert was made successfully. By histologically studying artificial human nasal epithelial cell sheets, we confirmed the presence of epidermal cell-like cells in the cortex and fibroblast-like cells in the substratum. The sheets consisted of multi-layered mucosal epithelia and lower connective tissue and were similar to normal middle ear mucosa. Scanning electron microscopy revealed a cobblestone-like pattern of cells with innumerable microvilli and a cilium-like configuration. We also confirmed the basal membrane configuration that included tight junctions, desmosomes, and cell adhesion structures among the epidermal layers.. This artificial mucosa had a similar configuration to that of normal middle ear mucosa.For these reasons, middle ear mucosal cell sheet is considered as an effective transplantation material for mucosal regeneration in otitis media surgery, and it is expected to apply for human clinical use in the future.