Boere J, van de Lest C H A, de Grauw J C, Plomp S G M, Libregts S F W M, Arkesteijn G J A, Malda J, Wauben M H M, van Weeren P R
Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3584CM Utrecht, The Netherlands.
Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3584CM Utrecht, The Netherlands; Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 2, 3584CM Utrecht, The Netherlands.
Vet J. 2019 Feb;244:91-93. doi: 10.1016/j.tvjl.2018.12.010. Epub 2018 Dec 12.
Extracellular vesicle (EV) concentration, characteristics and function in equine synovial fluid (SF) during normal growth and development has not previously been studied. Isolation of EVs was performed in SF from three healthy foals and two adult horses by differential ultracentrifugation (10,000g and 200,000g); EVs were purified by sucrose density gradient floatation and analysed by high-resolution flow cytometry (FCM), buoyant density and western blotting. Additionally, repeated biomarker analysis of sulphated glycosaminoglycans (GAG), matrix metalloproteinase (MMP), C-terminal crosslinked telopeptide type II collagen (CTX-II), collagenase cleaved neopeptide type II collagen (C2C) was performed in SF from 10 foals and six adult horses. In contrast with the quantitative EV profile, the biomarker profile in SF from juvenile joints was substantially different from that in SF from adult animals. However, there were qualitative differences in the high-resolution FCM scatter plots. Future in-depth functional analyses may reveal differences between juvenile and mature EVs in SF.
细胞外囊泡（EV）在马正常生长发育过程中于滑液（SF）中的浓度、特征及功能此前尚未得到研究。通过差速超速离心（10,000g和200,000g）从3匹健康幼驹和2匹成年马的滑液中分离出细胞外囊泡；通过蔗糖密度梯度浮选法纯化细胞外囊泡，并采用高分辨率流式细胞术（FCM）、浮力密度及蛋白质免疫印迹法进行分析。此外，还对10匹幼驹和6匹成年马的滑液进行了硫酸化糖胺聚糖（GAG）、基质金属蛋白酶（MMP）、II型胶原C端交联端肽（CTX-II）、胶原酶裂解的II型胶原新肽（C2C）的重复生物标志物分析。与细胞外囊泡定量概况相反，幼年关节滑液中的生物标志物概况与成年动物滑液中的生物标志物概况有很大不同。然而，在高分辨率流式细胞术散点图上存在定性差异。未来深入的功能分析可能会揭示幼年和成熟滑液细胞外囊泡之间的差异。
