Baguet J, Sommer F, Claudon-Eyl V, Duc T M
Laboratoire Meuse Optique Contact (MOC), Centre Hospitalier, Bar le Duc, France.
Biomaterials. 1995 Jan;16(1):3-9. doi: 10.1016/0142-9612(95)91089-h.
The purpose of this study was to investigate lacrymal component accumulation on a soft contact lens (SCL) surface after various periods of continuous wear, using the recently developed atomic force microscopy (AFM). AFM allowed high resolution images of unworn and worn SCL, and presented two main advantages. 1. The SCL are analysed under nearly physiological conditions without being dried or destroyed. So the same SCL was analysed at various times during a long wearing period. To identify the deposited tear proteins, a qualitative analysis of solubilized deposit by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) on 4-15% gradient minigels was performed as well. We present typical images which emphasize the importance of the coating by lacrymal components. AFM analysis of worn SCL showed the deposition on the surface of a uniform lacrymal component coating (named deposit type I) with a progressive accumulation of numerous discrete granules (named deposit type II). SDS-PAGE of extracted deposits revealed the main tear proteins as: IgA, lactoferrin, tear lipocalin and lysozyme and the unknown protein of molecular weight 30,000. There is no clear difference in the protein patterns of the two types of deposits. Furthermore, a particular mode of use of AFM is described to illustrate the potential of this technique as a local tool for measuring protein coating thickness. Thus, for analysis of protein deposits on SCL surfaces, SDS-PAGE on minigels and AFM were easy and rapid to perform. When associated, these two techniques could find use in a wide range of worn SCL evaluation and most generally in biocompatibility evaluation studies.
本研究的目的是使用最近开发的原子力显微镜(AFM),研究在不同持续佩戴时间后,软性接触镜(SCL)表面的泪液成分积聚情况。AFM能够获取未佩戴和佩戴后的SCL的高分辨率图像,并具有两个主要优点。1. 可在几乎生理条件下对SCL进行分析,而无需干燥或破坏它们。因此,在长时间佩戴期间的不同时间对同一SCL进行了分析。为了鉴定沉积的泪液蛋白质,还通过在4-15%梯度迷你胶上进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对溶解的沉积物进行了定性分析。我们展示了典型图像,强调了泪液成分涂层的重要性。对佩戴后的SCL进行AFM分析显示,在表面有一层均匀的泪液成分涂层(称为沉积物I型)沉积,同时有大量离散颗粒逐渐积聚(称为沉积物II型)。对提取沉积物进行的SDS-PAGE分析揭示了主要的泪液蛋白质为:IgA、乳铁蛋白、泪液脂质运载蛋白和溶菌酶以及分子量为30,000的未知蛋白质。两种类型沉积物的蛋白质图谱没有明显差异。此外,还描述了一种特殊的AFM使用模式,以说明该技术作为测量蛋白质涂层厚度的局部工具的潜力。因此,对于分析SCL表面的蛋白质沉积物,迷你胶上的SDS-PAGE和AFM操作简便、快速。当两者结合使用时,这两种技术可用于广泛的佩戴后SCL评估,更普遍地用于生物相容性评估研究。