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Adv Pharm Bull. 2024;14(3): 613-622.
doi: 10.34172/apb.2024.050
PMID: 39494265
PMCID: PMC11530889
  Abstract View: 321
  PDF Download: 154

Research Article

NLC Delivery of EGFP Plasmid to TM4 Cell Nuclei for Targeted Gene Therapy

Nurul Jummah 1,2 ORCID logo, Satrialdi Satrialdi 1 ORCID logo, Aluicia Anita Artarini 3 ORCID logo, Anindyajati Anindyajati 3 ORCID logo, Diky Mudhakir 1* ORCID logo

1 Department of Pharmaceutics, School of Pharmacy, Institut Teknologi Bandung (ITB), Bandung 40132, Indonesia.
2 Department of Pharmacy, Faculty of Mathematics and Natural Science, Universitas Islam Makassar, Makassar 90245, Indonesia.
3 Biotechnology Laboratory, Department of Pharmaceutics, School of Pharmacy, Institut Teknologi Bandung (ITB), Bandung 40132, Indonesia.
*Corresponding Author: Diky Mudhakir, Email: mudhakir@itb.ac.id

Abstract

Purpose: This study evaluated whether a nanostructured lipid carrier (NLC) delivery system could safely and accurately deliver nucleic acids to the cell nucleus using the enhanced green fluorescent protein (EGFP)-C1 plasmid model.

Methods: The NLC was formulated using the emulsification method and equipped for cationic lipid-mediated transfection with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), which interacts electrostatically with nucleic acid. The NLC attributes, including size, polydispersity index, and zeta potential, were assessed by dynamic light scattering (DLS). The morphological structure was analyzed using transmission electron microscopy. Entrapment efficiency was evaluated by a direct method. Cellular uptake mechanisms of pEGFP-C1-NLC and the ability of pEGFP-C1 to penetrate the nucleus of TM4 cells to express EGFP were observed using confocal microscopy.

Results: pEGFP-C1-NLC exhibited particle sizes in the range 56-88 nm with a particle charge range of -6.0 to+1.3 mV. The polydispersity index<0.5 showed good size uniformity, and entrapment efficiency of pEGFP-C1in the NLC was 92.06±2.295%. The NLC formulation was internalized predominantly via caveolae-mediated endocytosis, as indicated by EGFP expression following successful delivery of pEGFP by the NLC into the cells.

Conclusion: NLC formulation could deliver genetic material to the nucleus and could be considered a gene therapy candidate for spermatogenesis.

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Submitted: 09 Oct 2023
Revision: 21 May 2024
Accepted: 19 Jun 2024
ePublished: 22 Jun 2024
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