建立小鼠模型的泪液拉丝试验方案。
Establishment of a Tear Ferning Test Protocol in the Mouse Model.
机构信息
Institute of Biochemistry, Microbiology, and Immunology, Chung Shan Medical University, Taichung, Taiwan.
Department of Nutrition, Chung Shan Medical University, Taichung, Taiwan.
出版信息
Transl Vis Sci Technol. 2020 Dec 1;9(13):1. doi: 10.1167/tvst.9.13.1. eCollection 2020 Dec.
PURPOSE
Analysis of ferning formation after tear drop desiccation on a glass slide has been applied as a simple method to examine tear normality and is referred to as the tear ferning (TF) test. Despite use of the TF test in clinical settings and in some animals, thus far no TF test protocol has been developed for the mouse model. This study aimed to establish a mouse TF test protocol that can be used for dry eye research using the mouse as the study model.
METHODS
Tear samples were collected from 24 healthy mice after repeated flushes with 2, 5, 10, or 20 µL wash solutions, either 0.9% NaCl saline or sterile water, on the ocular surface. After sample collection, TF tests were performed at variable drop volumes (2-20 µL), at a relative humidity of either 46% ± 2% or 53% ± 2%, and with temperature fixed at 24°C ± 2°C for comparison. Moreover, the influence of osmolarity (between 280 and 360 mOsm/L) and pH values (6.5-8.0) and the effect of centrifugation (4000 rpm, 10 minutes) on ferning formation were examined. Reproducibility and ferning storage stability were also determined.
RESULTS
An optimized protocol was established with relative humidity at 46% ± 2% and drop aliquot at 2 µL, using 0.9% NaCl saline as the wash solution. Using sterilized water as the wash solution did not result in any crystalloid formation. Centrifugation did not aid ferning formation in any of the samples. Higher osmolarity increased ferning formation from grades between 0 to 1 to grades between 2 to 3, but pH values that varied between 6.5 and 8.0 did not affect ferning formation. The established mouse TF test protocol also displayed reproducibility and storage stability.
CONCLUSIONS
A TF test protocol for the mouse model was established that could be used for comparative analyses under various ocular surface disease conditions.
TRANSLATIONAL RELEVANCE
This mouse TF test protocol will facilitate the application of basic research into the mouse model to clinical care.
目的
在玻璃载玻片上进行泪滴干燥分析，即泪滴蕨类样（TF）形成分析，已被用作一种简单的方法来检查泪液的正常性，这种方法被称为泪滴蕨类样（TF）测试。尽管在临床环境和一些动物中使用了 TF 测试，但迄今为止，还没有为小鼠模型开发 TF 测试协议。本研究旨在建立一种用于研究干眼症的小鼠 TF 测试协议，该协议使用小鼠作为研究模型。
方法
用 2 、 5 、 10 或 20 μL 冲洗液（0.9%NaCl 生理盐水或无菌水）重复冲洗 24 只健康小鼠的眼表，收集泪液样本。收集样本后，在相对湿度为 46%±2%或 53%±2%、温度固定在 24°C±2°C的条件下，用 2-20 μL 的不同液滴体积进行 TF 测试。此外，还研究了渗透压（280-360mOsm/L 之间）和 pH 值（6.5-8.0）以及离心（4000rpm ，10 分钟）对蕨类样形成的影响。还确定了重复性和蕨类样储存稳定性。
结果
建立了一个优化的协议，相对湿度为 46%±2%，液滴等分试样为 2 μL ，使用 0.9%NaCl 生理盐水作为冲洗液。使用无菌水作为冲洗液不会导致任何结晶形成。在任何样本中，离心都不会促进蕨类样形成。较高的渗透压使蕨类样形成从 0-1 级增加到 2-3 级，但 pH 值在 6.5-8.0 之间变化不会影响蕨类样形成。建立的小鼠 TF 测试协议还显示出重复性和储存稳定性。
结论
建立了一种用于小鼠模型的 TF 测试协议，可用于各种眼表面疾病条件下的比较分析。
翻译
林大闲人
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