Chitosan-Graft-Polyethylenimine/DNA Nanoparticles as Novel Non-Viral Gene Delivery Vectors Targeting Osteoarthritis

Abstract

The development of safe and efficient gene carriers is the key to the clinical success of gene therapy. The present study was designed to develop and evaluate the chitosan-graft-polyethylenimine (CP)/DNA nanoparticles as novel non-viral gene vectors for gene therapy of osteoarthritis. The CP/DNA nanoparticles were produced through a complex coacervation of the cationic polymers with pEGFP after grafting chitosan (CS) with a low molecular weight (Mw) PEI (Mw = 1.8 kDa). Particle size and zeta potential were related to the weight ratio of CP:DNA, where decreases in nanoparticle size and increases in surface charge were observed as CP content increased. The buffering capacity of CP was significantly greater than that of CS. The transfection efficiency of CP/DNA nanoparticles was similar with that of the Lipofectamine™ 2000, and significantly higher than that of CS/DNA and PEI (25 kDa)/DNA nanoparticles. The transfection efficiency of the CP/DNA nanoparticles was dependent on the weight ratio of CP:DNA (w/w). The average cell viability after the treatment with CP/DNA nanoparticles was over 90% in both chondrocytes and synoviocytes, which was much higher than that of PEI (25 kDa)/DNA nanoparticles. The CP copolymers efficiently carried the pDNA inside chondrocytes and synoviocytes, and the pDNA was detected entering into nucleus. These results suggest that CP/DNA nanoparticles with improved transfection efficiency and low cytotoxicity might be a safe and efficient non-viral vector for gene delivery to both chondrocytes and synoviocytes.

Design and Characterization of Endostatin-Loaded Nanoparticles for In Vitro Antiangiogenesis in Squamous Cell Carcinoma

“…The presence of peaks at = 2.5-3.2 ppm was assigned to methylene protons of PEI (-NHCH 2 CH 2 -) and confirmed that PEI was successfully grafted onto the CHT chain. Similar results were reported by Sarkar and coworkers [19] for the preparation of fluorescent chitosan-graft-polyethyleneimine and Lui and coworkers [20] for chitosan-Graft-polyethylenimine/DNA nanoparticles. After the reaction between CHT-g-PEI and COOH-PEG-NH 2 , there were new peaks in the 1 H spectrum at = 3.6 ppm which belonged to the methylene protons of PEG (-OCH 2 CH 2 -) (relative to Figure 2(a)).…”

“…Similarly, amidation reaction between the carboxyl group on the bifunctional PEG and the free amine on the grafted CHT-g-PEI through NHS/EDC cross linkers was employed to synthesize the amino terminal CHT-PEI-PEG conjugate for endostatin encapsulation. The structural and functional modification of both CHT-g-PEI and CHT-g-PEI-PEG-NH 2 conjugates were confirmed by both 1 H NMR and FT-IR spectra as presented in Figures 2 and 3, respectively [19][20][21][22][23][24].…”

Molecular targets in arthritis and recent trends in nanotherapy

“…The CS-graft-PEI copolymers efficiently carried the pDNA inside chondrocytes and synoviocytes without lowering the cell viability. 87 Diacereinloaded SLNs modified with chondroitin sulfate (ChS) have also been used in chemically induced rat model for OA was used for the study. It was observed that the presence of drug in joints was higher in rats fed on NP diet when compared with the free drug.…”

Cross-linked polyethylenimine–tripolyphosphate nanoparticles for gene delivery

“…35 In this study, we showed that neutralizing the charge on PEI with TPP resulted in significantly decreased cytotoxicity in HeLa, HEK293T, HepG2, and MCF-7 cells. Further, nanoparticles containing less TPP did more harm to cells than those with more.…”