The human conjunctival epithelial cells (HCEC) line the inner sides of the eyelids and the anterior part of the sclera. They include goblet cells that secret mucus into the tear film that protects the ocular surface. The conjunctival epithelium is subjected to mechano-physical stimuli due to eyelid movement during blinking, during wiping and rubbing the eyes, and when exposed to wind and air currents. We cultured primary HCEC under air–liquid interface (ALI) conditions in custom-designed wells that can be disassembled for installation of the in vitro model in a flow chamber. We exposed the HCEC after ALI culture of 8–10 days to steady and oscillatory airflows. The in vitro model of HCEC was exposed to steady wall shear stresses (sWSS) of 0.5 and 1.0 dyne/cm² for lengths of 30 and 60 min and to oscillatory wall shear stresses (oWSS) of 0.5 and 0.77 dyne/cm² amplitudes for a length of 10 min. Cytoskeletal alterations and MUC5AC mucin secretion in response to WSS were investigated using immunohistochemically fluorescent staining and enzyme-linked lectin assay (ELLA), respectively. The results revealed that both exposure times and sWSS values increased the polymerization of F-actin filaments while mucin secretion decreased. However, after a recovery of 24 h in the incubator we observed a decrease of F-actin fibers and mucin secretion only for exposure of 30 min. The length of exposure was more influential on cytoskeletal alterations than the level of sWSS. The very small effect of sWSS on mucin secretion is most likely related to the much smaller amount of goblet cell than in other mucus-secreting tissue. The results for both oWSS amplitudes revealed similar trends regarding F-actin and mucin secretion. Immediately post-exposure we observed an increase in polymerization of F-actin filaments while mucin secretion decreased. However, after 24-h recovery we observed that both F-actin and mucin secretion returned to the same values as for unexposed cultures. The results of this study suggest that WSS should be considered while exploring the physiological characteristics of HCEC.

人结膜上皮细胞 (HCEC) 排列在眼睑内侧和巩膜前部。它们包括将粘液分泌到保护眼表的泪膜中的杯状细胞。结膜上皮在眨眼、擦拭和揉眼睛以及暴露在风和气流中时，由于眼睑运动而受到机械物理刺激。我们在气液界面 (ALI) 条件下在定制设计的孔中培养初级 HCEC，这些孔可拆卸以将体外模型安装在流动室中。我们在 ALI 培养 8-10 天后将 HCEC 暴露在稳定和振荡的气流中。HCEC 的体外模型暴露于 0.5 和 1.0 dyne/cm ^{2的稳定壁剪切应力 (sWSS)}对于 30 和 60 分钟的长度以及 0.5 和 0.77 达因/厘米^{2的振荡壁剪切应力 (oWSS)}幅度为 10 分钟的长度。分别使用免疫组织化学荧光染色和酶联凝集素测定 (ELLA) 研究了响应于 WSS 的细胞骨架改变和 MUC5AC 粘蛋白分泌。结果表明，暴露时间和 sWSS 值都增加了 F-肌动蛋白丝的聚合，而粘蛋白分泌减少。然而，在培养箱中恢复 24 小时后，我们观察到仅暴露 30 分钟后 F-肌动蛋白纤维和粘蛋白分泌减少。与 sWSS 水平相比，暴露时间对细胞骨架改变的影响更大。sWSS 对粘蛋白分泌的非常小的影响很可能与杯状细胞的数量比其他粘液分泌组织中的少得多有关。两个 oWSS 振幅的结果揭示了 F-肌动蛋白和粘蛋白分泌的相似趋势。暴露后，我们立即观察到 F-肌动蛋白丝聚合增加，而粘蛋白分泌减少。然而，在 24 小时恢复后，我们观察到 F-肌动蛋白和粘蛋白分泌恢复到与未暴露培养物相同的值。本研究结果表明，在探索 HCEC 的生理特征时应考虑 WSS。