Polyphosphate coated nanoparticles: Enzyme-activated charge-reversal gene delivery systems

Zeynep Burcu Akkuş-Dağdeviren; Sema Arısoy; Julian David Friedl; Andrea Fürst; Ahmad Saleh; Andreas Bernkop-Schnürch

doi:10.1016/j.ijpharm.2023.123474

Polyphosphate coated nanoparticles: Enzyme-activated charge-reversal gene delivery systems

Int J Pharm. 2023 Nov 5:646:123474. doi: 10.1016/j.ijpharm.2023.123474. Epub 2023 Oct 2.

Authors

Zeynep Burcu Akkuş-Dağdeviren¹, Sema Arısoy², Julian David Friedl¹, Andrea Fürst¹, Ahmad Saleh³, Andreas Bernkop-Schnürch⁴

Affiliations

¹ Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
² Department of Pharmaceutical Technology, Selcuk University, Faculty of Pharmacy, Konya, Turkey.
³ Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria; Department of Pharmacy, Universitas Mandala Waluya, A.H. Nasution, Kendari 93231, Southeast Sulawesi, Republic of Indonesia.
⁴ Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria. Electronic address: andreas.bernkop@uibk.ac.at.

PMID: 37793466
DOI: 10.1016/j.ijpharm.2023.123474

Abstract

Aim: The current study aimed to develop enzyme-activated charge-reversal lipid nanoparticles (LNPs) as novel gene delivery systems.

Methods: Palmitic acid was covalently bound to protamine being utilised as transfection promoter to anchor it on the surfaces of LNPs. Green fluorescent protein (GFP) encoding plasmid DNA (pDNA) was ion paired with various cationic counter ions to achieve high encapsulation in LNPs. Protamine-decorated LNPs were prepared by solvent injection method followed by coating with sodium tripolyphosphate (TPP) to generate a bio-inert anionic outer surface. Resulting LNPs were characterised regarding size, polydispersity, zeta potential and encapsulation efficiency. Enzyme-triggered charge-reversal of LNPs was investigated using isolated alkaline phosphatase (ALP) monitoring changes in zeta potential as well as monophosphate release. Furthermore, monophosphate release, cell viability and transfection efficiency were evaluated on a human alveolar epithelial (A549) cell line.

Results: Protamine-decorated and TPP-coated (Prot-pDNA/DcChol-TPP) LNPs displayed a mean size of 298.8 ± 17.4 nm and a zeta potential of -13.70 ± 0.61 mV. High pDNA encapsulation was achieved with hydrophobic ion pairs of pDNA with 3ß-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride (DcChol). Zeta potential of Prot-pDNA/DcChol-TPP LNPs reversed to positive values with a total Δ26.8 mV shift upon incubation with ALP. Conformably, a notable amount of monophosphate was released upon incubation of Prot-pDNA/DcChol-TPP LNPs with isolated as well as cell-associated ALP. A549 cells well tolerated LNPs displaying more than 95 % viability. Compared with naked pDNA, unmodified LNPs and control LNPs, Prot-pDNA/DcChol-TPP LNPs showed a significantly increased transfection efficiency.

Conclusion: Prot-pDNA/DcChol-TPP LNPs can be regarded as promising gene delivery systems.

Keywords: Charge-reversal; Gene delivery; Lipid nanoparticles; Polyphosphate coating; Protamine; Transfection.

MeSH terms

DNA
Gene Transfer Techniques*
Humans
Nanoparticles* / chemistry
Plasmids
Protamines
Transfection

Substances

DNA
Protamines