Pollins Alonda C, Kim Justine S, Boyer Richard B, Thayer Wesley P
Department of Plastic Surgery, Vanderbilt University Medical Center, Nashville, TN, USA.
Vanderbilt University School of Medicine, Nashville, TN, USA.
J Mater Sci Mater Med. 2017 Jan;28(1):20. doi: 10.1007/s10856-016-5834-y. Epub 2016 Dec 23.
Peripheral nerve repair using nerve grafts has been investigated for several decades using traditional techniques such as histology, immunohistochemistry, and electron microscopy. Recent advances in mass spectrometry techniques have made it possible to study the proteomes of complex tissues, including extracellular matrix rich tissues similar to peripheral nerves. The present study comparatively assessed three previously described processing methods for generating acellular nerve grafts by mass spectrometry. Acellular nerve grafts were additionally examined by F-actin staining and nuclear staining for debris clearance. Application of newer techniques allowed us to detect and highlight differences among the 3 treatments. Isolated proteins were separated by mass on polyacrylamide gels serving 2 purposes. This further illustrated that these treatments differ from one another and it allowed for selective protein extractions within specific bands/molecular weights. This approach resulted in small pools of proteins that could then be analyzed by mass spectrometry for content. In total, 543 proteins were identified, many of which corroborate previous findings for these processing methods. The remaining proteins are novel discoveries that expand the field. With this pilot study, we have proven that mass spectrometry techniques complement and add value to peripheral nerve repair studies.
几十年来，人们一直使用组织学、免疫组织化学和电子显微镜等传统技术研究利用神经移植物进行周围神经修复。质谱技术的最新进展使得研究复杂组织的蛋白质组成为可能，包括类似于周围神经的富含细胞外基质的组织。本研究通过质谱对三种先前描述的制备脱细胞神经移植物的处理方法进行了比较评估。另外，通过F-肌动蛋白染色和核染色检查脱细胞神经移植物的碎片清除情况。应用更新的技术使我们能够检测并突出三种处理方法之间的差异。分离出的蛋白质在聚丙烯酰胺凝胶上按分子量分离，有两个目的。这进一步表明这些处理方法彼此不同，并且能够在特定条带/分子量范围内进行选择性蛋白质提取。这种方法得到了少量蛋白质池，然后可通过质谱分析其成分。总共鉴定出543种蛋白质，其中许多证实了这些处理方法先前的研究结果。其余蛋白质是拓展该领域的新发现。通过这项初步研究，我们证明了质谱技术对周围神经修复研究具有补充作用并能增加其价值。
