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

Huading Lu,Huiqing Zhao,Yuhu Dai,Lulu Lv,Xiaoming He

doi:10.1371/journal.pone.0084703

Huading Lu, Huiqing Zhao + Show 3 more

Open Access

https://doi.org/10.1371/journal.pone.0084703

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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.

Highlights

Osteoarthritis (OA) is one of the greatest challenges for clinical therapy due to avascularity and the lack innervation of cartilage
We investigated the characteristics of CP/plasmid enhanced green fluorescent protein nanoparticles and their cytotoxicity, the transfection efficiency in chondrocytes and synoviocytes, and their ability to carry nucleic acids into the nucleus of chondrocytes and synoviocytes
CP/DNA nanoparticles were created as a novel, non-viral gene carrier targeted to OA and other joint diseases

Summary

Introduction

Osteoarthritis (OA) is one of the greatest challenges for clinical therapy due to avascularity and the lack innervation of cartilage. Gene therapy represents a promising technology for OA treatment by targeting specific disease-relevant mechanisms, and by treating the causes of OA rather than the symptoms [2]. The development of an efficient and safe gene transfer system is one of the most important factors for a successful gene therapy with practical use in the clinic. Most gene therapy protocols in clinical trials employ viral vectors due to their high transfection efficiency, their fatal drawbacks, such as immunogenicity, potential infectivity, complicated production, and oncogenic effects, may prevent their further use in the clinic [3,4,5,6]. Non-viral vectors have attracted much attention because of their potential advantages, such as ease of synthesis and modification, efficient cell/tissue targeting, low immune response, and unrestricted plasmid size, among others [4,5,6,7,8,9]

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