The purpose of this study is to evaluate outflow pathways from subconjunctival blebs and to identify their identity. Post-mortem porcine (n = 20), human (n = 1), and bovine (n = 1) eyes were acquired, and tracers (fluorescein, indocyanine green, or fixable/fluorescent dextrans) were injected into the subconjunctival space to create raised blebs where outflow pathways were visualized qualitatively and quantitatively. Rodents with fluorescent reporter transgenes were imaged for structural comparison. Concurrent optical coherence tomography (OCT) was obtained to study the structural nature of these pathways. Using fixable/fluorescent dextrans, tracers were trapped to the bleb outflow pathway lumen walls for histological visualization and molecular identification using immunofluorescence against lymphatic and blood vessel markers. Bleb outflow pathways could be observed using all tracers in all species. Quantitative analysis showed that the nasal quadrant had more bleb-related outflow pathways compared to the temporal quadrant (nasal: 1.9±0.3 pathways vs. temporal: 0.7±0.2 pathways; p = 0.003). However, not all blebs resulted in an outflow pathway (0-pathways = 18.2%; 1-pathway = 36.4%; 2-pathways = 38.6%; and 3-pathways = 6.8%). Outflow signal was validated as true luminal pathways using optical coherence tomography and histology. Bicuspid valves were identified in the direction of flow in porcine eyes. Immunofluorescence of labeled pathways demonstrated a lymphatic (Prox-1 and podoplanin) but not a blood vessel (CD31) identity. Therefore, subconjunctival bleb outflow occurs in discrete luminal pathways. They are lymphatic as assessed by structural identification of valves and molecular identification of lymphatic markers. Better understanding of lymphatic outflow may lead to improved eye care for glaucoma surgery and ocular drug delivery.

中文翻译：

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结膜下滤泡淋巴流出的功能、结构和分子鉴定。


本研究的目的是评估结膜下滤泡的流出途径并确定其身份。获得死后猪 (n = 20)、人 (n = 1) 和牛 (n = 1) 的眼睛，并将示踪剂（荧光素、吲哚青绿或可固定/荧光右旋糖酐）注射到结膜下空间以产生凸起的气泡，其中流出路径可以定性和定量地可视化。对带有荧光报告基因转基因的啮齿动物进行成像以进行结构比较。获得并行光学相干断层扫描（OCT）来研究这些通路的结构性质。使用可固定/荧光右旋糖酐，示踪剂被捕获到滤泡流出通路管腔壁上，以便使用针对淋巴和血管标记物的免疫荧光进行组织学可视化和分子鉴定。可以使用所有物种的所有示踪剂来观察泡流出路径。定量分析显示，与颞象限相比，鼻象限具有更多与滤泡相关的流出路径（鼻：1.9±0.3 路径 vs. 颞：0.7±0.2 路径；p = 0.003）。然而，并非所有气泡都会导致流出途径（0 途径 = 18.2%；1 途径 = 36.4%；2 途径 = 38.6%；3 途径 = 6.8%）。使用光学相干断层扫描和组织学将流出信号验证为真实的管腔路径。在猪眼中的血流方向上发现了二尖瓣。标记通路的免疫荧光显示了淋巴管（Prox-1 和 podoplanin），但没有显示血管（CD31）特性。因此，结膜下水泡流出发生在离散的管腔路径中。通过瓣膜的结构鉴定和淋巴标记物的分子鉴定来评估它们是淋巴管。 更好地了解淋巴流出可能会改善青光眼手术和眼部药物输送的眼部护理。