Department of Ophthalmology, University Medical Center, Johannes Gutenberg University, Langenbeckstr.1, 55131, Mainz, Germany.
Graefes Arch Clin Exp Ophthalmol. 2011 Feb;249(2):233-43. doi: 10.1007/s00417-010-1456-0. Epub 2010 Jul 26.
The aim of this study was to analyze and compare the effects of rigid gas permeable and soft contact lenses on the protein composition in the tear film of contact lens wearers.
Wearers of soft contact lenses (CL_S, n = 13) and rigid gas permeable contact lenses (CL_H, n = 13) were recruited for this study. Thirteen non-contact lens wearers were also included as the control. Tears were collected using Schirmer strips and frozen until use. The tears were eluted and analyzed on ProteinChips SELDI-TOF (surface-enhanced laser desorption and ionization in time of flight mass spectrometry; Bio-Rad, USA) with different chromatographic surfaces (cationic and anionic exchanger and reversed phase surface). The SELDI spectra were analyzed by multivariate statistical analysis and artificial neural networks in order to find a biomarker panel which differentiates best between the groups. In order to identify protein/peptide peaks from SELDI spectra which showed a significant difference between groups, fractionated tear samples were analyzed using MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). For validation of biomarkers, we used an antibody microarray approach.
Complex patterns of tear proteins and peptides were detected in the control group and in both contact lens groups. The tear protein composition in both wearers of rigid gas permeable (CL_H) and soft contact lenses (CL_S) differed significantly from protein composition in non-contact lens wearers (p < 0.01). The identification of biomarkers revealed an increase of Protein S100 A8 in the group of wearers of soft contact lenses (CL_S) and a decrease of a main tear protein, lysozyme, in both contact lens groups. The identified biomarker cystatin was upregulated in the group of rigid gas permeable lens wearers (CL_H), whereas the protein intensity of secretoglobin was significantly reduced in this group. Using the microarray approach, detected alterations could be confirmed.
Contact lens wear alters the protein profiles in a complex manner. This study demonstrates that significant changes can be found in wearers of soft contact lenses (CL_S) and rigid gas permeable contact lenses (CL_H). Some biomarker intensities are significantly altered only in the group of rigid gas permeable lens wearers (CL_H).
本研究旨在分析和比较硬性透气性隐形眼镜和软性隐形眼镜对隐形眼镜佩戴者泪膜蛋白组成的影响。
本研究招募了软性隐形眼镜佩戴者（CL_S，n=13）和硬性透气性隐形眼镜佩戴者（CL_H，n=13）。还招募了 13 名不戴隐形眼镜的非佩戴者作为对照。使用 Schirmer 条收集眼泪并冷冻直至使用。使用表面增强激光解吸电离飞行时间质谱（Bio-Rad，美国）的阳离子和阴离子交换剂和反相表面等不同色谱表面洗脱和分析眼泪。通过多元统计分析和人工神经网络分析 SELDI 谱，以找到最佳区分组别的生物标志物组。为了从 SELDI 谱中鉴定出显示组间差异的蛋白质/肽峰，使用 MALDI-TOF MS（基质辅助激光解吸/电离飞行时间质谱）对分级泪样进行分析。为了验证生物标志物，我们使用了抗体微阵列方法。
在对照组和硬性透气性隐形眼镜佩戴者（CL_H）和软性隐形眼镜佩戴者（CL_S）两组中均检测到复杂的泪蛋白和肽模式。硬性透气性隐形眼镜（CL_H）和软性隐形眼镜（CL_S）佩戴者的泪蛋白组成均与非隐形眼镜佩戴者的蛋白组成有显著差异（p<0.01）。生物标志物的鉴定显示，软性隐形眼镜佩戴者（CL_S）组中蛋白 S100 A8 增加，而两组接触镜佩戴者中主要泪蛋白溶菌酶减少。在硬性透气性隐形眼镜佩戴者（CL_H）组中，鉴定出的生物标志物半胱氨酸蛋白酶抑制剂上调，而在该组中，分泌球蛋白的蛋白强度显著降低。使用微阵列方法，可以证实检测到的变化。
隐形眼镜佩戴以复杂的方式改变蛋白谱。本研究表明，软性隐形眼镜佩戴者（CL_S）和硬性透气性隐形眼镜佩戴者（CL_H）均有显著变化。在硬性透气性隐形眼镜佩戴者（CL_H）组中，一些生物标志物的强度仅发生显著改变。
