Gene delivery potential of biofunctional carbonate apatite nanoparticles in lungs.

Rozita Rosli,Ezharul Houque Chowdhury,Suleiman Yusuf Alhaji,Syahril Abdullah,Fatma Hassan

doi:10.1155/2014/646787

Abstract

Existing nonviral gene delivery systems to lungs are inefficient and associated with dose limiting toxicity in mammalian cells. Therefore, carbonate apatite (CO3Ap) nanoparticles were examined as an alternative strategy for effective gene delivery to the lungs. This study aimed to (1) assess the gene delivery efficiency of CO3Ap in vitro and in mouse lungs, (2) evaluate the cytotoxicity effect of CO3Ap/pDNA in vitro, and (3) characterize the CO3Ap/pDNA complex formulations. A significantly high level of reporter gene expression was detected from the lung cell line transfected with CO3Ap/pDNA complex prepared in both serum and serum-free medium. Cytotoxicity analysis revealed that the percentage of the viable cells treated with CO3Ap to be almost similar to the untreated cells. Characterization analyses showed that the CO3Ap/pDNA complexes are in a nanometer range with aggregated spherical structures and tended to be more negatively charged. In the lung of mice, highest level of transgene expression was observed when CO3Ap (8 μL) was complexed with 40 μg of pDNA at day 1 after administration. Although massive reduction of gene expression was seen beyond day 1 post administration, the level of expression remained significant throughout the study period.

Highlights

Current understanding of genetic approaches in the treatment of lung genetic diseases reveals that enhancing the transportation of gene into the airway cells is a critical step for improving lung gene therapy [1]
H1299 was successfully transfected with the various formulations of CO3Ap/plasmid DNA (pDNA)
The results showed evidence of overall decline in gene expression to about 30% relative light units (RLUs)/mg protein (Figure 1(b)) compared to transfections using a CO3Ap complex formulation in medium with serum (Figure 1(a))

Summary

Introduction

Current understanding of genetic approaches in the treatment of lung genetic diseases reveals that enhancing the transportation of gene into the airway cells is a critical step for improving lung gene therapy [1]. The physical barriers to the airway gene transfer and enzymatic activities in the lung that may render the gene transfer ineffective have to be overcome. Cationic lipids are generally found to be ineffective in gene delivery to the mammalian cells [2], especially in the presence of serum [3]. Only a few have impressive activity in vivo [4]. Pulmonary hypotension, induction of inflammatory cytokines, and tissue infiltration of neutrophils have been reported following aerosolisation of lipid formulations into the lungs [6]

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Methods

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Discussion

Conclusion