Estimation of Urinary Nanocrystals in Humans using Calcium Fluorophore Labeling and Nanoparticle Tracking Analysis.
Estimation of Urinary Nanocrystals in Humans using Calcium Fluorophore Labeling and Nanoparticle Tracking Analysis.
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DOI：
10.3791/62192
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发表时间：
2021-02-09
期刊：
影响因子：
--
通讯作者：
Mitchell T
中科院分区：
文献类型：
--
作者：
Kumar P;Bell A;Mitchell T
在成人和儿童中，肾结石在全球越来越普遍。肾结石的最常见类型是草酸钙（CAOX）晶体。当尿液用矿物质（例如钙，草酸盐，磷酸盐）过饱和并在肾结石形成之前，就会发生结晶。评估石材形成器中晶状体的标准方法包括显微镜，过滤和离心。但是，这些方法主要检测微晶，而不是纳米晶体。纳米晶体已被认为对肾脏上皮细胞的危害比体外微晶体更有害。在这里，我们描述了纳米颗粒跟踪分析（NTA）检测人尿纳米晶体的能力。在饮用草酸盐负荷之前，健康的成年人被喂食草酸盐饮食，以刺激尿纳米晶体。在草酸盐负荷前后，收集尿液24小时。处理样品并用乙醇洗涤以净化样品。用钙结合荧光团（Fluo-4 AM）对尿纳米晶体进行染色。染色后，使用NTA确定纳米晶体的大小和计数。这项研究的发现表明，NTA可以有效地检测健康成年人的纳米晶体鲁里亚。这些发现表明，NTA可能是肾脏疾病患者中纳米晶状体的有价值的早期检测方法。
这项研究的目的是确定纳米颗粒跟踪分析（NTA）是否可以检测并量化含有健康成年人的纳米晶体的尿钙。当前研究的发现表明，NTA可能是估计肾结石期间尿纳米晶体的潜在工具。
Kidney stones are becoming more prevalent worldwide in adults and children. The most common type of kidney stone is comprised of calcium oxalate (CaOx) crystals. Crystalluria occurs when urine becomes supersaturated with minerals (e.g. calcium, oxalate, phosphate) and precedes kidney stone formation. Standard methods to assess crystalluria in stone formers include microscopy, filtration, and centrifugation. However, these methods primarily detect microcrystals and not nanocrystals. Nanocrystals have been suggested to be more harmful to kidney epithelial cells than microcrystals in vitro. Here, we describe the ability of Nanoparticle Tracking analysis (NTA) to detect human urinary nanocrystals. Healthy adults were fed a controlled oxalate diet prior to drinking an oxalate load to stimulate urinary nanocrystals. Urine was collected for 24 hours before and after the oxalate load. Samples were processed and washed with ethanol to purify samples. Urinary nanocrystals were stained with the calcium binding fluorophore, Fluo-4 AM. After staining, the size and count of nanocrystals were determined using NTA. The findings from this study show NTA can efficiently detect nanocrystalluria in healthy adults. These findings suggest NTA could be a valuable early detection method of nanocrystalluria in patients with kidney stone disease. The objective of this study was to determine whether nanoparticle tracking analysis (NTA) could detect and quantify urinary calcium containing nanocrystals from healthy adults. The findings from the current study suggest NTA could be a potential tool to estimate urinary nanocrystals during kidney stone disease.
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影响因子：
19.6
作者：
Huang, MY;Chaturvedi, LS;Koul, HK
通讯作者：
Koul, HK
影响因子：
2.5
作者：
BALCKE, P;ZAZGORNIK, J;SCHMIDT, P
通讯作者：
SCHMIDT, P
DOI：
10.1016/j.ymeth.2015.03.028
发表时间：
2015-10-01
期刊：
Methods (San Diego, Calif.)
影响因子：
--
作者：
Dragovic RA;Collett GP;Hole P;Ferguson DJ;Redman CW;Sargent IL;Tannetta DS
通讯作者：
Tannetta DS
影响因子：
4.5
作者：
Grover PK;Thurgood LA;Wang T;Ryall RL
通讯作者：
Ryall RL
影响因子：
11.4
作者：
Patel M;Yarlagadda V;Adedoyin O;Saini V;Assimos DG;Holmes RP;Mitchell T
通讯作者：
Mitchell T