Fabrication Of Gold Nanoparticles In Absence Of Surfactant As In Vitro Carrier Of Plasmid DNA

Int J Nanomedicine. 2019 Oct 22:14:8399-8408. doi: 10.2147/IJN.S226498. eCollection 2019.

Abstract

Purpose: This work aimed to synthesize surfactant-free AuNPs for targeted delivery of plasmid DNA encoded p53 gene and to avoid conventional production method of Gold nanoparticles (AuNPs) which may adversely affect the final shape, diversity, and size due to accumulation of the formulated surfactant - gold complex to the surface.

Methods: The AuNPs were fabricated using seeded-growth method with L-Cystine methyl ester hydrochloride as capping agent, then loaded with plasmid DNA encoded p53 gene. The resultant AuNPs and AuNPs-p53 complex were evaluated for physical characteristics and morphology. Confirmation of complex formation was performed using gel electrophoresis. Furthermore, the efficient delivery and cytotoxicity behavior of the encoded gene were examined on both healthy lung cells (WI38) and cancerous lung cells (A549).

Results: L-cysteine methyl ester hydrochloride AuNPs showed acceptable physical characteristics (30 nm, +36.9 mv, and spherical morphology). P53 attachment to AuNPs was confirmed by gel electrophoresis. The RT-PCR proved the overexpression of p53 by the fabricated AuNPs-p53 complex. The high percentage of cell viability in normal lung cell line (WI 38) proved the safety of L-cysteine methyl ester functionalized AuNPs. Additionally, the apoptotic effect due to expression of p53 gene loaded on AuNPs was only prominent in lung cancer cell line (A549), revealing selectivity and targeting efficiency of anticancer AuNPs-p53 complex.

Conclusion: AuNPs can be considered as a potential delivery system for effective transfection of plasmid DNA which can be used for successful treatment of cancer.

Keywords: gene delivery; gold nanoparticles; human lung cancer; plasmid DNA; transfection.

MeSH terms

  • A549 Cells
  • Cell Death
  • Cell Survival
  • DNA / genetics*
  • Dynamic Light Scattering
  • Gold / chemistry*
  • Humans
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / ultrastructure
  • Microtechnology / methods*
  • Plasmids / genetics*
  • Surface-Active Agents / chemistry*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Surface-Active Agents
  • Tumor Suppressor Protein p53
  • Gold
  • DNA