Transfection of Pulmonary Cells by Stable pDNA -Polycationic Hybrid Nanostructured Particles

Abstract

Aim: Cationically modified solid lipid nanoparticles (SLN) were investigated as plasmid DNA (pDNA) carriers and transfection agents for the pulmonary route. Materials & methods: pDNA-loaded SLN were produced using glyceryl dibehenate or tristearate as matrix lipids and chitosan as surface charge modifier, and encapsulated by spray-drying in mannitol and trehalose microspheres. Results: Nanoparticles of 200 nm, and zeta potential around +15 mV were produced. Electrophorectic analysis confirmed plasmid stability and integrity. The pDNA-loaded SLN were able to transfect the Calu-3 and A549 pulmonary cell lines, while showing low cytotoxicity. Microencapsulation of SLN yielded dry powders suitable for inhalation that protected pDNA from degradation. Conclusion: Microencapsulated SLN are a promising safe and effective carrier system for pulmonary gene delivery following pulmonary administration.

Recent Progress in Nucleic Acid Pulmonary Delivery toward Overcoming Physiological Barriers and Improving Transfection Efficiency

“…[ 120 ] Followed by a spray dried with L‐leucine and mannitol at an inlet temperature of 70 °C, the inhalable miRNA‐146a dry powder could effectively maintain its biological activity compared to the freshly prepared miRNA‐146a‐loaded DOTAP/PGA‐PDL nanoparticles. Other similar nano‐in‐micro designs were observed for siRNA and pDNA pulmonary delivery, preparing nanoparticles using lipid materials, polymeric materials, and peptides, followed by spray drying with mannitol, lactose, or trehalose to prepare microparticles, [ 85,128–130 ] exhibited the similar advantages in improving formulation pulmonary deposition with elevated FPF and emitted fraction (EF).…”

Liposomes or Extracellular Vesicles: A Comprehensive Comparison of Both Lipid Bilayer Vesicles for Pulmonary Drug Delivery

“…It has been demonstrated that spray drying of drug-loaded LPs is suitable for generating particles with a small MMAD, i.e., high FPF, as in situ rehydration of LPs occurs following powder deposition in the respiratory tract aqueous environment [ 224 ]. Enhancing transfection of gene therapy by introducing a spray-dried lactose solution comprising lipid-polycation-pDNA in comparison with the preparation prior to spray drying [ 225 ]. Practically speaking, pro-LPs are more suitable for pulmonary delivery via DPIs, as spray-dried phospholipid formulations can produce LPs immediately once in contact with the aquatic respiratory environment [ 212 ].…”

Treating Pulmonary Fibrosis with Non-Viral Gene Therapy: From Bench to Bedside

“…The excellent abilities of muco-adhesion and muco-penetration make chitosan a potential lung delivery vector [92]. Chitosan can effectively bind with gene drugs, partially protecting nucleic acid from nuclease degradation and facilitating the transport of gene drugs from the cytoplasm to the nucleus by escaping endosome and lysosomal systems [93]. Chitosan can be used as a carrier alone or as a surface modification to improve the transport efficiency of genetic drugs.…”