Lai Chun-Ting, Yao Wei-Chieng, Lin Szu-Yuan, Liu Hsin-Yu, Chang Huai-Wen, Hu Fung-Rong, Chen Wei-Li
Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan.
Department of Anesthesia, Min-Sheng General Hospital, Tao-Yuan City, Taiwan.
PLoS One. 2015 Sep 3;10(9):e0137186. doi: 10.1371/journal.pone.0137186. eCollection 2015.
To evaluate the ocular surface change and the inflammatory response in a rabbit model of short-term exposure keratopathy.
Short term exposure keratopathy by continuous eyelid opening was induced in New Zealand white rabbits for up to 4 hours. Ultrasound pachymetry was used to detect central total corneal thickness. In vivo confocal microscopy and impression cytology were performed to evaluate the morphology of ocular surface epithelium and the infiltration of inflammatory cells. Immunohistochemistry for macrophage,neutrophil, CD4(+) T cells, and CD8(+) T cells were performed to classify the inflammatory cells. Scanning electron microscopy(SEM) was performed to detect ocular surface change.The concentrations of IL-8, IL-17, Line and TNF-αwere analyzed by multiplex immunobead assay. TUNEL staining was performed to detect cellular apoptosis.
Significant decrease ofcentral total cornealthickness were found within the first 5 minutes and remained stable thereafter, while there were no changes of corneal epithelial thickness.No significant change of corneal, limbal and conjunctival epithelial morphology was found by in vivo confocal microscopy except the time dependent increase of superficial cellular defects in the central cornea. Impression cytology also demonstrated time dependent increase of sloughing superficial cells of the central cornea. Aggregations ofinflammatory cells were found at 1 hour in the limbal epithelium, 2 hours in the perilimbal conjunctival epithelium, and 3 hours in the peripheral corneal epithelium.In eyes receiving exposure for 4 hours, the infiltration of the inflammatory cells can still be detected at 8 hours after closing eyes.Immunohistochemical study demonstrated the cells to be macrophages, neutrophils, CD4-T cells and CD-8 T cells.SEM demonstrated time-depending increase of intercellular border and sloughing of superficial epithelial cells in corneal surface. Time dependent increase of IL-8, IL-17 and TNF-α in tear was found.TUNEL staining revealed some apoptotic cells in the corneal epithelium and superficial stroma at 3 hours after exposure.
Short term exposure keratopathy can cause significant changes to the ocular surface and inflammatory response. Decrease of central total corneal thickness, aggregation of inflammatory cells, and cornea epithelial cell and superficial keratocyte apoptosis were found no less than 4 hours following the insult.
评估短期暴露性角膜病变兔模型的眼表变化和炎症反应。
通过持续睁眼诱导新西兰白兔发生短期暴露性角膜病变，持续时间长达4小时。使用超声角膜测厚仪检测中央角膜总厚度。进行活体共聚焦显微镜检查和印迹细胞学检查，以评估眼表上皮形态和炎症细胞浸润情况。对巨噬细胞、中性粒细胞、CD4(+) T细胞和CD8(+) T细胞进行免疫组织化学染色，以对炎症细胞进行分类。进行扫描电子显微镜（SEM）检查以检测眼表变化。通过多重免疫珠测定法分析IL-8、IL-17、白细胞介素和TNF-α的浓度。进行TUNEL染色以检测细胞凋亡。
在最初5分钟内发现中央角膜总厚度显著降低，此后保持稳定，而角膜上皮厚度无变化。活体共聚焦显微镜检查未发现角膜、角膜缘和结膜上皮形态有明显变化，但中央角膜浅表细胞缺损随时间增加。印迹细胞学检查也显示中央角膜浅表脱落细胞随时间增加。在角膜缘上皮1小时、角膜缘周围结膜上皮2小时和周边角膜上皮3小时发现炎症细胞聚集。在暴露4小时的眼睛中，闭眼8小时后仍可检测到炎症细胞浸润。免疫组织化学研究表明这些细胞为巨噬细胞、中性粒细胞、CD4-T细胞和CD-8 T细胞。SEM显示角膜表面细胞间边界随时间增加以及浅表上皮细胞脱落。发现泪液中IL-8、IL-17和TNF-α随时间增加。TUNEL染色显示暴露后3小时角膜上皮和浅表基质中有一些凋亡细胞。
短期暴露性角膜病变可导致眼表和炎症反应发生显著变化。在损伤后不少于4小时发现中央角膜总厚度降低、炎症细胞聚集以及角膜上皮细胞和浅表角膜细胞凋亡。
