Abstract
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.
Highlights
Bladder epithelium, known as urothelium or transitional epithelium, is a highly specialised tissue that plays a key role in the defence of the bladder wall against various toxins
We investigated the location of Hyaluronic acid (HA) and chondroitin sulphate (CS) after ex vivo incubation of rabbit bladder tissue with HA-AuNPs and CS-AuNPs to determine whether the benefits of HA and CS replacement therapy might be due to coating of the urothelium with GAGs
We evaluated relative gene expression of receptors involved in HA and CS transport into the cell (CD44, RHAMM and intercellular adhesion molecule-1 (ICAM-1)) in order to understand whether incubation with GAG nanoparticles modifies receptor expression
Summary
Known as urothelium or transitional epithelium, is a highly specialised tissue that plays a key role in the defence of the bladder wall against various toxins. There is strong evidence that an absence of GAGs is linked to loss of normal urothelial function This loss of function results in “storage symptoms” such as frequency, which is an innate protective mechanism designed to minimise contact between urine and the damaged bladder wall[5]. We investigated the location of HA and CS after ex vivo incubation of rabbit bladder tissue with HA-AuNPs and CS-AuNPs to determine whether the benefits of HA and CS replacement therapy might be due to coating of the urothelium with GAGs. In addition, we evaluated relative gene expression of receptors involved in HA and CS transport into the cell (CD44, RHAMM and ICAM-1) in order to understand whether incubation with GAG nanoparticles modifies receptor expression
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