屈光手術 Wet Lab Lecture | Refractive Wet Lab Instruction Course

張朝凱 Chao-Kai ChangTaiwan Moderator

馬惠康 David Hui-Kang MaTaiwan Moderator Cultivated Oral/Nasal Mucosal Epithelial Sheets for Ocular Surface Reconstruction in TaiwanBackground To investigate the in vivo epithelial phenotypes of cultivated nasal mucosal epithelial cell (CUNMEC) sheets generated by a microspheroidal suspension culture technique. Methods Human nasal mucosal tissues were obtained during DCR. The tissues were digested with 2 mg/mL collagenase A, then the cell aggregates were seeded onto a de-epithelialized AM and cultivated for 2 weeks. The CUNMEC sheets were then transplanted into New Zealand albino rabbits. The animals were subjected to immunosuppression for two weeks and then sacrificed. The samples were subjected to light and electron microscopy (EM) and immunoconfocal microscopy examinations. Results The presence of CUNMECs in rabbit eyes was confirmed by anti-human cytoplasmic antibody staining. Keratins 3, 4, and 13 were positively expressed in the suprabasal region, whereas p63 expression was observed only in the basal layer. Transmission EM revealed stratified epithelium with the formation of desmosomes and hemidesmosomes. Scanning EM revealed a cobble stone-like epithelial surface without cilia formation. α-Tubulin, a major component of cilia, was expressed only in the apical region of the nasal mucosa and showed diffuse cytoplasmic staining in the CUNMEC sheets. ZO-1, a component of tight junctions, was expressed in the apical region of the nasal mucosa and was expressed at intercellular borders in CUNMECs. Conclusion Following transplantation, the CUNMEC sheet presented a stratified epithelial layer containing GCs but without cilia and expressed progenitor cell markers, which are morphologically similar to those of ocular surface epithelia and may have the potential to function as a surrogate epithelium for ocular surface reconstruction, especially in dry eye conditions.