Tada Takeshi, Ohnishi Hiroe, Yamamoto Norio, Kuwata Fumihiko, Hayashi Yasuyuki, Okuyama Hideaki, Morino Tsunetaro, Kasai Yoshiyuki, Kojima Hiromi, Omori Koichi
Department of Otolaryngology-Head and Neck Surgery, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto City, Kyoto 606-8507, Japan.
Department of Otorhinolaryngology, Jikei University School of Medicine, 3-25-8 Nishi-Shimbashi, Minato-ku, Tokyo 105-8461, Japan.
Regen Ther. 2022 Jan 14;19:77-87. doi: 10.1016/j.reth.2022.01.001. eCollection 2022 Mar.
Early postoperative regeneration of the middle ear mucosa is essential for the prevention of postoperative refractory otitis media and recurrent cholesteatoma. As a means for intractable otitis media management, we focused on human induced pluripotent stem cell (hiPSC)-derived airway epithelial cells (AECs), which have been used in upper airway mucosal regeneration and transplantation therapy. In this study, we transplanted hiPSC-derived AECs into the middle ear of immunodeficient rats.
Following the preparation of AEC sheets from hiPSCs, the bilateral middle ear mucosa of X-linked severe combined immunodeficient rats was scraped, and the AEC sheets were transplanted in the ears unilaterally.
Human nuclear antigen (HNA)-positive ciliated cells were observed on the transplanted side of the middle ear cavity surface in three of six rats in the 1-week postoperative group and in three of eight rats in the 2-week postoperative group. No HNA-positive cells were found on the control side. The percentage of HNA-positive ciliated cells in the transplanted areas increased in the 2-week postoperative group compared with the 1-week group, suggesting survival of hiPSC-derived AECs. Additionally, HNA-positive ciliated cells were mainly located at sites where the original ciliated cells were localized. Immunohistochemical analysis showed that the transplanted AECs contained cytokeratin 5- and mucin 5AC-positive cells, indicating that both basal cells and goblet cells had regenerated within the middle ear cavity.
The results of this study are an important first step in the establishment of a novel transplantation therapy for chronic otitis media.
中耳黏膜的早期术后再生对于预防术后难治性中耳炎和复发性胆脂瘤至关重要。作为治疗难治性中耳炎的一种方法，我们将重点放在了人诱导多能干细胞（hiPSC）来源的气道上皮细胞（AEC）上，这些细胞已被用于上呼吸道黏膜再生和移植治疗。在本研究中，我们将hiPSC来源的AEC移植到免疫缺陷大鼠的中耳。
从hiPSC制备AEC片后，刮除X连锁严重联合免疫缺陷大鼠的双侧中耳黏膜，并将AEC片单侧移植到耳中。
术后1周组6只大鼠中有3只、术后2周组8只大鼠中有3只在中耳腔表面移植侧观察到人类核抗原（HNA）阳性的纤毛细胞。对照侧未发现HNA阳性细胞。与术后1周组相比，术后2周组移植区域HNA阳性纤毛细胞的百分比增加，提示hiPSC来源的AEC存活。此外，HNA阳性纤毛细胞主要位于原始纤毛细胞所在的部位。免疫组织化学分析显示，移植的AEC含有细胞角蛋白5和黏蛋白5AC阳性细胞，表明中耳腔内基底细胞和杯状细胞均已再生。
本研究结果是建立慢性中耳炎新型移植治疗方法的重要第一步。
