Green, fast and cheap paper-based method for estimating equivalence ratio of cationic carriers to DNA in gene delivery formulations

Phadungcharoen, Noppharat; Plianwong, Samarwadee; Srivichai, Chakrit; Chanthananon, Natcha; Kaosal, Watcharawat; Pannil, Ornkamon; Opanasopit, Praneet; Ngawhirunpat, Tanasait; Rojanarata, Theerasak

doi:10.1016/j.ejps.2018.01.030

Cited by 4 publications

(1 citation statement)

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“…Complex characterization. The complete complex generation was determined by paper-based method as previously described [10]. Briefly, the complexes were spotted on filter paper, then let them dry and lastly staining light green to yellow anionic 2',7'-dichlorofluorescein dye solution over the complexes.…”

Section: Methodsmentioning

confidence: 99%

The Effect of Spermidine and Spermine on Chitosan-Mediated Gene Delivery

Apirakaramwong

Sunthornphan

Pasuthawong

et al. 2020

KEM

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Chitosan (CS) is a biodegradable and highly positive charged polymer that can be utilized as a non-viral vector for gene delivery. However, its transfection efficiency is low. Spermidine (SPD) and spermine (SPM) are ordinary polyamines found in almost all living organisms. Their structures contain tri- and tetra-amine, respectively. The present study aims to elucidate the effect of SPD or SPM on transfection efficiency and cytotoxicity of CS/DNA complexes in human cervical carcinoma (HeLa) cells. The ternary complexes (CS, DNA and SPD or SPM) at different weight ratios and mixing orders were formulated and tested on transfection efficiency. Their particle sizes and charges were also estimated. CS that has a molecular weight of 45,000 Daltons and degree of deacetylation of 85 was able to form complete complex with plasmid DNA expressing enhanced green fluorescence protein (pEGFP-C2) at weight ratio of 4. The SPM/CS/DNA (1:4:1) and SPD/ CS/DNA (512:4:1) complexes illustrated the greatest transfection efficiency. Moreover, the transfection efficiency was affected by the mixing order between CS, DNA and SPD or SPM. For cell toxicity experiments, more than 85% the average cell survival of the complexes were detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) cell growth assay. These findings suggest that addition of either SPD or SPM to CS before forming ternary complexes with DNA may significantly improve gene delivery potential in vitro.

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Section: Methodsmentioning

confidence: 99%