Matrix-assisted laser desorption ionizationimaging mass spectrometry: A new methodology to study human osteoarthritic cartilage
Matrix-assisted laser desorption ionizationimaging mass spectrometry: A new methodology to study human osteoarthritic cartilage
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DOI：
10.1002/art.37799
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发表时间：
2013-03-01
影响因子：
--
通讯作者：
Heeren, Ron M. A.
中科院分区：
文献类型：
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作者：
Cillero-Pastor, Berta;Eijkel, Gert B.;Heeren, Ron M. A.
目的了解蛋白质在风湿性疾病不同阶段的分布及其调节作用，对于了解风湿性疾病的发生发展至关重要。我们进行了这项研究，以证明实用程序的质谱（MS）为基础的分子成像研究的空间分布的不同组成部分在人类关节软骨切片。方法我们比较了多肽和蛋白质在人对照和骨关节炎（OA）软骨中的分布。切割人对照和OA软骨切片并沉积在导电载玻片上。胰蛋白酶消化后，我们进行了基质辅助激光解吸电离成像质谱（MALDI-IMS）的实验在MALDI-IMS四极杆飞行时间质谱仪。采用多元统计方法和Mascot蛋白质数据库查询的组合进行蛋白质鉴定。进行苏木精-伊红染色和免疫组织化学以验证结果。结果我们创建了肽分布图，在150米光栅大小从控制和OA人软骨。蛋白质，如双糖蛋白聚糖，prolargin，核心蛋白聚糖和聚集蛋白聚糖核心蛋白进行了鉴定和定位。软骨寡聚基质蛋白和纤连蛋白的特异性蛋白质标记物仅在OA软骨样品中发现。其分布表现为深部强度大于浅部。在OA软骨深部发现了新的初步OA标志物。结论MALDI-IMS可识别和定位软骨中的疾病特异性肽和蛋白。所有的OA相关肽和蛋白检测显示更强的强度在深层软骨。基于MS的分子成像被证明是研究OA病理学的创新方法。
Objective Information about the distribution of proteins and the modulation that they undergo in the different phases of rheumatic pathologies is essential to understanding the development of these diseases. We undertook this study to demonstrate the utility of mass spectrometry (MS)based molecular imaging for studying the spatial distribution of different components in human articular cartilage sections. Methods We compared the distribution of peptides and proteins in human control and osteoarthritic (OA) cartilage. Human control and OA cartilage slices were cut and deposited on conductive slides. After tryptic digestion, we performed matrix-assisted laser desorption ionizationimaging MS (MALDI-IMS) experiments in a MALDIquadrupole time-of-flight mass spectrometer. Protein identification was undertaken with a combination of multivariate statistical methods and Mascot protein database queries. Hematoxylin and eosin staining and immunohistochemistry were performed to validate the results. Results We created maps of peptide distributions at 150-m raster size from control and OA human cartilage. Proteins such as biglycan, prolargin, decorin, and aggrecan core protein were identified and localized. Specific protein markers for cartilage oligomeric matrix protein and fibronectin were found exclusively in OA cartilage samples. Their distribution displayed a stronger intensity in the deep area than in the superficial area. New tentative OA markers were found in the deep area of the OA cartilage. Conclusion MALDI-IMS identifies and localizes disease-specific peptides and proteins in cartilage. All the OA-related peptides and proteins detected display a stronger intensity in the deep cartilage. MS-based molecular imaging is demonstrated to be an innovative method for studying OA pathology.