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Development of polymeric-cationic peptide composite nanoparticles, a nanoparticle-in-nanoparticle system for controlled gene delivery.

Jain AK, Massey A, Yusuf H, McDonald DM, McCarthy HO, Kett VL - Int J Nanomedicine (2015)

Bottom Line: The best formulation was selected and was able to transfect cells while maintaining viability.The effect of transferrin-appended composite nanoparticles was also studied.Thus, we have demonstrated the manufacture of composite nanoparticles for the controlled delivery of DNA.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, Northern Ireland, UK ; Weatherall Institute of Molecular Medicine, MRC Molecular Haematology Unit, University of Oxford, John Radcliffe Hospital, Oxford, UK.

ABSTRACT
We report the formulation of novel composite nanoparticles that combine the high transfection efficiency of cationic peptide-DNA nanoparticles with the biocompatibility and prolonged delivery of polylactic acid-polyethylene glycol (PLA-PEG). The cationic cell-penetrating peptide RALA was used to condense DNA into nanoparticles that were encapsulated within a range of PLA-PEG copolymers. The composite nanoparticles produced exhibited excellent physicochemical properties including size <200 nm and encapsulation efficiency >80%. Images of the composite nanoparticles obtained with a new transmission electron microscopy staining method revealed the peptide-DNA nanoparticles within the PLA-PEG matrix. Varying the copolymers modulated the DNA release rate >6 weeks in vitro. The best formulation was selected and was able to transfect cells while maintaining viability. The effect of transferrin-appended composite nanoparticles was also studied. Thus, we have demonstrated the manufacture of composite nanoparticles for the controlled delivery of DNA.

No MeSH data available.


Formulation optimization of RNPs.Notes: (A and B) Gel retardation assay. Numbers denote the N:P ratio. (C) Particle size analysis and zeta potential of RNPs made at different N:P ratios. Error bars show ±SD, n=3.Abbreviations: L, 1 kb plus DNA ladder; N, native pDNA; OC, open circular/relaxed plasmid DNA; SC, supercoiled plasmid DNA; RNPs, RALA nanoparticles; pDNA, plasmid DNA; SD, standard deviation; PDI, polydispersity index.
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f2-ijn-10-7183: Formulation optimization of RNPs.Notes: (A and B) Gel retardation assay. Numbers denote the N:P ratio. (C) Particle size analysis and zeta potential of RNPs made at different N:P ratios. Error bars show ±SD, n=3.Abbreviations: L, 1 kb plus DNA ladder; N, native pDNA; OC, open circular/relaxed plasmid DNA; SC, supercoiled plasmid DNA; RNPs, RALA nanoparticles; pDNA, plasmid DNA; SD, standard deviation; PDI, polydispersity index.

Mentions: Figure 2A shows a representative image of the gel retardation assay performed to determine the N:P ratio required to condense DNA into the RNPs. Lanes corresponding to nanoparticles prepared with N:P ratios of 0.5–2.5 exhibit a DNA band at the same distance as pDNA (labeled D), indicating that in these formulations, the amount of RALA peptide was not sufficient to condense the DNA completely. At N:P ratio ≥3, no migration of DNA into the gel was seen indicating that beyond this ratio the DNA was condensed with the peptide. Lanes corresponding to N:P ratios of 3–4 showed some fluorescence in the well indicating formation of DNA-peptide aggregates, while at higher N:P ratios no DNA fluorescence was seen, confirming that in these lanes, the DNA is buried within the RNPs.


Development of polymeric-cationic peptide composite nanoparticles, a nanoparticle-in-nanoparticle system for controlled gene delivery.

Jain AK, Massey A, Yusuf H, McDonald DM, McCarthy HO, Kett VL - Int J Nanomedicine (2015)

Formulation optimization of RNPs.Notes: (A and B) Gel retardation assay. Numbers denote the N:P ratio. (C) Particle size analysis and zeta potential of RNPs made at different N:P ratios. Error bars show ±SD, n=3.Abbreviations: L, 1 kb plus DNA ladder; N, native pDNA; OC, open circular/relaxed plasmid DNA; SC, supercoiled plasmid DNA; RNPs, RALA nanoparticles; pDNA, plasmid DNA; SD, standard deviation; PDI, polydispersity index.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664533&req=5

f2-ijn-10-7183: Formulation optimization of RNPs.Notes: (A and B) Gel retardation assay. Numbers denote the N:P ratio. (C) Particle size analysis and zeta potential of RNPs made at different N:P ratios. Error bars show ±SD, n=3.Abbreviations: L, 1 kb plus DNA ladder; N, native pDNA; OC, open circular/relaxed plasmid DNA; SC, supercoiled plasmid DNA; RNPs, RALA nanoparticles; pDNA, plasmid DNA; SD, standard deviation; PDI, polydispersity index.
Mentions: Figure 2A shows a representative image of the gel retardation assay performed to determine the N:P ratio required to condense DNA into the RNPs. Lanes corresponding to nanoparticles prepared with N:P ratios of 0.5–2.5 exhibit a DNA band at the same distance as pDNA (labeled D), indicating that in these formulations, the amount of RALA peptide was not sufficient to condense the DNA completely. At N:P ratio ≥3, no migration of DNA into the gel was seen indicating that beyond this ratio the DNA was condensed with the peptide. Lanes corresponding to N:P ratios of 3–4 showed some fluorescence in the well indicating formation of DNA-peptide aggregates, while at higher N:P ratios no DNA fluorescence was seen, confirming that in these lanes, the DNA is buried within the RNPs.

Bottom Line: The best formulation was selected and was able to transfect cells while maintaining viability.The effect of transferrin-appended composite nanoparticles was also studied.Thus, we have demonstrated the manufacture of composite nanoparticles for the controlled delivery of DNA.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, Northern Ireland, UK ; Weatherall Institute of Molecular Medicine, MRC Molecular Haematology Unit, University of Oxford, John Radcliffe Hospital, Oxford, UK.

ABSTRACT
We report the formulation of novel composite nanoparticles that combine the high transfection efficiency of cationic peptide-DNA nanoparticles with the biocompatibility and prolonged delivery of polylactic acid-polyethylene glycol (PLA-PEG). The cationic cell-penetrating peptide RALA was used to condense DNA into nanoparticles that were encapsulated within a range of PLA-PEG copolymers. The composite nanoparticles produced exhibited excellent physicochemical properties including size <200 nm and encapsulation efficiency >80%. Images of the composite nanoparticles obtained with a new transmission electron microscopy staining method revealed the peptide-DNA nanoparticles within the PLA-PEG matrix. Varying the copolymers modulated the DNA release rate >6 weeks in vitro. The best formulation was selected and was able to transfect cells while maintaining viability. The effect of transferrin-appended composite nanoparticles was also studied. Thus, we have demonstrated the manufacture of composite nanoparticles for the controlled delivery of DNA.

No MeSH data available.