Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study

Oliveira, Ana V.; Ap, Silva; Bitoque, Diogo B.; Ga, Silva; Costa, Ana M. Rosa da

doi:10.4103/0975-7406.111823

Cited by 21 publications

(9 citation statements)

References 36 publications

(44 reference statements)

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“…Although transfection in ARPE-19 cells was higher for the DC-Chol/DOPE and EPC/DOPE lipoplexes, transfection efficiencies of the C20-18/DOPE and C20-20/DOPE lipoplexes are noteworthy when compared to other investigations reporting considerably lower efficiencies [ 28 ].…”

Section: Discussionmentioning

confidence: 59%

Cationic Polyene Phospholipids as DNA Carriers for Ocular Gene Therapy

Machado

Bitoque

Oliveira

et al. 2014

BioMed Research International

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Recent success in the treatment of congenital blindness demonstrates the potential of ocular gene therapy as a therapeutic approach. The eye is a good target due to its small size, minimal diffusion of therapeutic agent to the systemic circulation, and low immune and inflammatory responses. Currently, most approaches are based on viral vectors, but efforts continue towards the synthesis and evaluation of new nonviral carriers to improve nucleic acid delivery. Our objective is to evaluate the efficiency of novel cationic retinoic and carotenoic glycol phospholipids, designated C20-18, C20-20, and C30-20, to deliver DNA to human retinal pigmented epithelium (RPE) cells. Liposomes were produced by solvent evaporation of ethanolic mixtures of the polyene compounds and coformulated with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or cholesterol (Chol). Addition of DNA to the liposomes formed lipoplexes, which were characterized for binding, size, biocompatibility, and transgene efficiency. Lipoplex formulations of suitable size and biocompatibility were assayed for DNA delivery, both qualitatively and quantitatively, using RPE cells and a GFP-encoding plasmid. The retinoic lipoplex formulation with DOPE revealed a transfection efficiency comparable to the known lipid references 3β-[N-(N′,N′-dimethylaminoethane)-carbamoyl]-cholesterol (DC-Chol) and 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (EPC) and GeneJuice. The results demonstrate that cationic polyene phospholipids have potential as DNA carriers for ocular gene therapy.

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Section: Discussionmentioning

confidence: 59%

Cationic Polyene Phospholipids as DNA Carriers for Ocular Gene Therapy

Machado

Bitoque

Oliveira

et al. 2014

BioMed Research International

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“…In addition, they facilitate the complexes to avoid endosomal degradation via proton sponge effect and move into the cytoplasm [23]. CS nanoparticles have been considered a perfect candidate that can be applied in biomedical events such as tissue engineering, drug delivery and gene delivery [24]. At present, CS-based carriers have attracted more and more attention as a reliable system for delivering genes [14], so we concluded that CS nanoparticles could be used as an ideal biomaterial carrier for MB delivery, while simultaneously detecting the miRNA in living cells.…”

Section: Resultsmentioning

confidence: 99%

Chitosan Combined with Molecular Beacon for Mir-155 Detection and Imaging in Lung Cancer

Zhu

An²,

Yao

et al. 2014

Molecules

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Lung cancer is the major cause of cancer-related deaths worldwide, thus developing effective methods for its early diagnosis is urgently needed. In recent years, microRNAs (miRNAs, miR) have been reported to play important roles in carcinogenesis and have become potential biomarkers for cancer diagnosis and treatment. Molecular beacon (MB) technology is a universal technology to detect DNA/RNA expression in living cells. As a natural polymers, chitosan (CS) nanoparticles could be used as a carrier for safe delivery of nucleic acid. In this study, we developed a probe using nanoparticles of miR-155 MB self assembled with CS (CS-miR-155 MB) to image the expression of miR-155 in cancer cells. Hybridization assay showed that the locked nucleic acid (LAN) modified miR-155 MB could target miR-155 effectively and sensitively. The miR-155 MB self-assembly with CS nanoparticles formed stable complexes at the proper weight ratio. The CS nanoparticles showed higher fluorescence intensity and transfection efficiency than the lipid-based formulation transfection agent by confocal microscopy and flow cytometry analysis. The CS-MB complexes were found to be easily synthesized and exhibited strong OPEN ACCESSMolecules 2014, 19 14711 enzymatic stability, efficient cellular uptake, high target selectivity and biocompatibility. The CS-MB complexes can also be applied in other cancers just by simply changing for a targeted miRNA highly expressed in those cancer cells. Therefore, it is a promising vehicle used for detecting miRNA expression in living cells.

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“…The samples were read at 570 nm and 630 nm (background) to obtain cell viability % (optical density, OD570-OD630). The percentage of cell viability was calculated by the following formula: [OD(570 nm) -OD(630 nm) treated/ OD(570 nm) -OD(630 nm) untreated] × 100% (Oliveira et al, 2013). All experiments were performed in triplicate, and the relative cell viability (%) was expressed as a percentage relative to the untreated control cells (ctr), i.e.…”

Section: Methodsmentioning

confidence: 99%

Protamine-like proteins have bactericidal activity. The first evidence in Mytilus galloprovincialis

et al. 2018

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The major acid-soluble protein components of the mussel Mytilus galloprovincialis sperm chromatin consist of the protamine-like proteins PL-II, PL-III and PL-IV, an intermediate group of sperm nuclear basic proteins between histones and protamines. The aim of this study was to investigate the bactericidal activity of these proteins since, to date, there are reports on bactericidal activity of protamines and histones, but not on protamine-like proteins. We tested the bactericidal activity of these proteins against Gram-positive bacteria: Enterococcus faecalis and two different strains of Staphylococcus aureus, as well as Gram-negative bacteria: Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella typhmurium, Enterobacter aerogenes, Enterobacter cloacae, and Escherichia coli. Clinical isolates of the same bacterial species were also used to compare their sensitivity to these proteins. The results show that Mytilus galloprovincialis protamine-like proteins exhibited bactericidal activity against all bacterial strains tested with different minimum bactericidal concentration values, ranging from 15.7 to 250 µg/mL. Furthermore, these proteins were active against some bacterial strains tested that are resistant to conventional antibiotics. These proteins showed very low toxicity as judged by red blood cell lysis and viability MTT assays and seem to act both at the membrane level and within the bacterial cell. We also tested the bactericidal activity of the product obtained from an in vitro model of gastrointestinal digestion of protamine-like proteins on a Gram-positive and a Gram-negative strain, and obtained the same results with respect to undigested protamine-like proteins on the Gram-positive bacterium. These results provide the first evidence of bactericidal activity of protamine-like-proteins.

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Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study

Cited by 21 publications

References 36 publications

Cationic Polyene Phospholipids as DNA Carriers for Ocular Gene Therapy

Cationic Polyene Phospholipids as DNA Carriers for Ocular Gene Therapy

Chitosan Combined with Molecular Beacon for Mir-155 Detection and Imaging in Lung Cancer

Protamine-like proteins have bactericidal activity. The first evidence in Mytilus galloprovincialis

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