摘要
背景:泪液取样对泪液分析的重复性至关重要。但是,一种可操作、高效、稳定的泪液收集方法尚未得到广泛的应用。这项研究评估了聚醚砜膜的效用(pesms)撕裂的收集和它的两个常用方法相比:Schirmer试纸的使用(STP)和毛细管(猫)。 方法:STP和pesms(0.45和0.65μm)是利用扫描电子显微镜(SEM),用于吸收水、牛血清白蛋白和细胞裂解物,并采用兔眼泪液收集。用聚丙烯酰胺凝胶电泳(PAGE)凝胶电泳分析了细胞裂解液和泪液中的蛋白质。Western blot分析与纽蛋白和溶菌酶C抗体进行兔泪液采样与猫,STP和PESM。在不同条件下评价兔眼表面和眨眼情况。 结果:扫描电镜观察表明,pesms表现出比STP更平滑的表面和毛孔变小。pesm65表现出最高的水吸收和体外的水回收。虽然SDS-PAGE显示细胞或泪液蛋白带型没有明显的差异,在差异中峰高度最高的骰子的相似性系数和最小方差(即带强度)在pesm65样本观察。Western blot结果显示,纽蛋白在泪液的谱带强度获得使用猫,STP和pesm65在猫< STP = pesm65秩序,而溶菌酶C是猫> >为pesm65 STP。pesm65表现出比STP不眨眼和结膜损伤兔的眼泪中取样。 结论:采用PESM在蛋白成分的方差较小撕裂收集和操作简便比传统的STP。PESm具有诊断撕裂检测的临床利用的潜力。
关键词: 泪液蛋白,聚醚砜膜、眼、SDS-聚丙烯酰胺凝胶电泳,泪液收集。
Current Molecular Medicine
Title:Characterization of Polyethersulfone Membranes for Tear Fluid Collection
Volume: 17 Issue: 4
关键词: 泪液蛋白,聚醚砜膜、眼、SDS-聚丙烯酰胺凝胶电泳,泪液收集。
摘要: Background: The sampling of tear fluids is essential for the reproducibility of tear assays. But, an operable, efficient and stable method for tear collection has not been widely established. This study evaluated the utility of polyethersulfone membranes (PESms) for tear collection and compared it with two frequently employed approaches: the use of Schirmer test paper (STP) and capillary tube (CaT).
Methods: STP and PESms (0.45 and 0.65 µm) were examined using scanning electron microscopy(SEM), applied to soak up water, bovine albumin and cellular lysates, and employed to collect tear fluids from rabbits. The proteins in the cellular lysates and tear fluid were characterized through band profiling of SDS-polyacrylamide electrophoresis (PAGE) gels. Western blot analyses with antibodies against vinculin and lysozyme C were used to compare rabbit tear fluids sampling with CaT, STP and PESm. Rabbit ocular surfaces and eye blinking were evaluated under different conditions.
Results: The SEM examination showed that PESms exhibited much smoother surfaces
and smaller pores than STP. PESm65 exhibited the highest water absorption and water
recovery in vitro. Although SDS-PAGE revealed no obvious differences in the cellular or
tear protein band patterns, the highest Dice's similarity coefficient and lowest variance in
the differences in peak heights(i.e. band intensities) were observed in the PESm65
samples. The Western blot showed that the band intensities of vinculin protein in tear
fluids obtained using CaT, STP and PESm65 are in order of CaT
Conclusion: Tear collection using PESm has less variance in protein components and is easily operable than traditional STP. PESm possesses the potential for clinical utilization in diagnostic tear assays.
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Cite this article as:
Characterization of Polyethersulfone Membranes for Tear Fluid Collection, Current Molecular Medicine 2017; 17 (4) . https://dx.doi.org/10.2174/1566524017666171101164244
DOI https://dx.doi.org/10.2174/1566524017666171101164244 |
Print ISSN 1566-5240 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5666 |
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