Clinical Biochemistry and Pharmacology Department, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
J Cell Mol Med. 2021 Apr;25(7):3339-3347. doi: 10.1111/jcmm.16408. Epub 2021 Mar 1.
The present research aims to determine whether the application of non-pigmented ciliary epithelium cells derived extracellular vesicles to human trabecular meshwork cells affects the formation and secretion of collagen type I to the extracellular matrix formation. Following the extraction of non-pigmented ciliary epithelium derived extracellular vesicles by a precipitation method, their size and concentration were determined using tunable resistive pulse sensing technology. Extracellular vesicles were incubated with trabecular meshwork cells for 3 days. Morphological changes of collagen type I in the extracellular matrix of trabecular meshwork cells were visualized using confocal microscopy and scanning electron microscopy. A Sirius Red assay was used to determine the total amount of collagen. Finally, collagen type I expression levels in the extracellular matrix of trabecular meshwork cells were quantified by cell western analysis. We found that non-pigmented ciliary epithelium extracellular vesicles were very effective at preventing collagen fibres formation by the trabecular meshwork cells, and their secretion to the extracellular matrix was significantly reduced (P < .001). Morphological changes in the extracellular matrix of trabecular meshwork cells were observed. Our study indicates that non-pigmented ciliary epithelium extracellular vesicles can be used to control collagen type I fibrillogenesis in trabecular meshwork cells. These fibrils net-like structure is responsible for remodelling the extracellular matrix. Moreover, we suggest that targeting collagen type I fibril assembly may be a viable treatment for primary open-angle glaucoma abnormal matrix deposition of the extracellular matrix.
本研究旨在确定非色素睫状上皮细胞衍生的细胞外囊泡应用于人眼小梁细胞是否会影响 I 型胶原的形成和分泌以及细胞外基质的形成。通过沉淀法提取非色素睫状上皮衍生的细胞外囊泡后,使用可调电阻脉冲感应技术测定其大小和浓度。将细胞外囊泡与小梁细胞共培养 3 天。使用共聚焦显微镜和扫描电子显微镜观察小梁细胞细胞外基质中 I 型胶原的形态变化。使用天狼猩红测定法测定胶原总量。最后,通过细胞西方分析定量测定小梁细胞细胞外基质中 I 型胶原的表达水平。我们发现非色素睫状上皮细胞外囊泡非常有效地阻止小梁细胞形成胶原纤维,并且显著减少其向细胞外基质的分泌(P<0.001)。观察到小梁细胞细胞外基质的形态变化。我们的研究表明,非色素睫状上皮细胞外囊泡可用于控制小梁细胞中 I 型胶原的原纤维形成。这些纤维的网状结构负责重塑细胞外基质。此外,我们认为靶向胶原纤维组装可能是治疗原发性开角型青光眼细胞外基质异常沉积的一种可行方法。