Inhibition of Glomerular Mesangial Cell Proliferation by siPDGF-B- and siPDGFR-β-Containing Chitosan Nanoplexes

Abstract

Mesangioproliferative glomerulonephritis is a disease that has a high incidence in humans. In this disease, the proliferation of glomerular mesangial cells and the production of extracellular matrix are important. In recent years, the RNAi technology has been widely used in the treatment of various diseases due to its capability to inhibit the gene expression with high specificity and targeting. The objective of this study was to decrease mesangial cell proliferation by knocking down PDGF-B and its receptor, PDGFR-β. To be able to use small interfering RNAs (siRNAs) in the treatment of this disease successfully, it is necessary to develop appropriate delivery systems. Chitosan, which is a biopolymer, is used as a siRNA delivery system in kidney drug targeting. In order to deliver siRNA molecules targeted at PDGF-B and PDGFR-β, chitosan/siRNA nanoplexes were prepared. The in vitro characterization, transfection studies, and knockdown efficiencies were studied in immortalized and primary rat mesangial cells. In addition, the effects of chitosan nanoplexes on mesangial cell proliferation and migration were investigated. After in vitro transfection, the PDGF-B and PDGFR-β gene silencing efficiencies of PDGF-B and PDGFR-β targeting siRNA-containing chitosan nanoplexes were 74 and 71% in immortalized rat mesangial cells and 66 and 62% in primary rat mesangial cells, respectively. siPDGF-B- and siPDGFR-β-containing nanoplexes indicated a significant decrease in mesangial cell migration and proliferation. These results suggested that mesangial cell proliferation may be inhibited by silencing of the PDGF-B signaling pathway. Gene silencing approaches with chitosan-based gene delivery systems have promise for the efficient treatment of renal disease.

Optimization and screening of solid lipid nanoparticle production for gene delivery by factorial design and response surface methodology

“…Particle size was again found nanosized and below 80 nm. This size range is adequate for traverse into the cells [36]. Moreover, the charge of the system remained cationic.…”

TNF-α Plus IL-1β Induces Opposite Regulation of Cx43 Hemichannels and Gap Junctions in Mesangial Cells through a RhoA/ROCK-Dependent Pathway

“…Primary MCs were isolated from mouse glomeruli treated with collagenase [ 87 , 88 , 89 , 90 ]. In brief, the kidney fragments were minced with a razor blade, and a 100 μm nylon sieve was used to collect the glomeruli from the cortex homogenates of mice kidneys in aseptic conditions.…”

“…Then, it was triturated through a 21-gauge needle. Glomerular remnants were washed, and mesangial and endothelial cells were plated onto a six-well plate in complete medium and incubated at 37 °C in a humidified 5% CO 2 incubator [ 87 , 88 , 89 , 90 ].…”

Chitin and Chitosan Derivatives as Biomaterial Resources for Biological and Biomedical Applications

“…Chitosan has also been applied as siRNA delivery system targeting kidney. For instance, chitosan nanoplex effectively covered siRNA for knocking down PDGF-B and PDGFR-beta [100]. Chitosan was also used as biomaterials in combination with collagen for culturing human renal proximal tubular cells [101].…”