Department of Veterinary Medicine, University of Perugia, Perugia, PG, Italy.
Faculty of Biomedical Sciences and Engineering & BioMediTech Institute, Bioengineering and Nanomedicine Lab, Tampere University of Technology, 33520, Tampere, Finland.
Sci Rep. 2017 Sep 4;7(1):10355. doi: 10.1038/s41598-017-09872-0.
This study investigated the location of hyaluronic acid (HA)- and chondroitin sulphate (CS)-coated gold nanoparticles in rabbit bladder and evaluated gene expression of CD44, RHAMM and ICAM-1 receptors involved in HA and CS transport into the cell. Gold nanoparticles were synthesised by reduction of gold salts with HA or CS to form HA-AuNPs and CS-AuNPs. Bladder samples were incubated with CS-AuNPs and HA-AuNPs or without glycosaminoglycans. Transmission electron microscopy, optic microscopy and scanning electron microscopy were used to determine the location of the synthesised AuNPs. Real-time PCR was used to analyse expression of urothelial cell receptors CD44, RHAMM, ICAM-1, after ex vivo administration of CS-AuNPs and HA-AuNPs. We showed that HA-AuNPs and CS-AuNPs were located in the cytoplasm and tight junctions of urothelial umbrella cells; this appearance was absent in untreated bladders. There were no significant differences in gene expression levels for CD44, RHAMM and ICAM-1 receptors in treated versus control bladder tissues. In conclusion, we clearly showed the presence of exogenous GAGs in the bladder surface and the tight junctions between umbrella cells, which is important in the regeneration pathway of the urothelium. The GAGs-AuNPs offer a promising approach to understanding the biophysical properties and imaging of urothelial tissue.
本研究调查了透明质酸(HA)和硫酸软骨素(CS)涂层金纳米粒子在兔膀胱中的位置,并评估了参与 HA 和 CS 转运进入细胞的 CD44、RHAMM 和 ICAM-1 受体的基因表达。通过用 HA 或 CS 还原金盐合成金纳米粒子,形成 HA-AuNPs 和 CS-AuNPs。用 CS-AuNPs 和 HA-AuNPs 或不使用糖胺聚糖孵育膀胱样本。透射电子显微镜、光学显微镜和扫描电子显微镜用于确定合成的 AuNPs 的位置。实时 PCR 用于分析 CS-AuNPs 和 HA-AuNPs 给药后尿路上皮细胞受体 CD44、RHAMM、ICAM-1 的表达。我们表明,HA-AuNPs 和 CS-AuNPs 位于尿路上皮伞细胞的细胞质和紧密连接中;未经处理的膀胱中不存在这种外观。与对照膀胱组织相比,处理组和对照组中 CD44、RHAMM 和 ICAM-1 受体的基因表达水平没有显着差异。总之,我们清楚地表明了外源性 GAGs 存在于膀胱表面和伞细胞之间的紧密连接中,这对于尿路上皮的再生途径很重要。GAGs-AuNPs 为理解尿路上皮组织的生物物理特性和成像提供了一种很有前途的方法。