Development and optimization of nanosomal formulations for siRNA delivery to the liver

Abstract

The objective of this study is to develop an effective siRNA delivery system for successful delivery to the liver for the treatment of HCV. Nanosize liposomes (nanosomes) have been prepared using a mixture of cholesterol and DOTAP. A functional siRNA was encapsulated into nanosomes following condensation with protamine sulfate. The delivery of siRNA was optimized in an in vitro cell culture system. The efficacy of the formulations was evaluated by measuring functional gene silencing and cytotoxicity. Encapsulation of siRNA ≥ 7.4 nM resulted in successful delivery of siRNA to nearly 100% of cells. The formulations containing lipids to siRNA ratio ≥ 10.56:1 instantly cleared approximately 85% of HCV while maintaining cell viability at about 90%. The formulations were sonicated to further reduce the particle size. The size of these formulations was decreased up to 100 nm. However, there were no significant changes observed in zeta potential, or in siRNA encapsulation and integrity following sonication. The sonicated formulations also showed higher liver hepatocytes deposition and gene silencing properties. This study therefore provides a novel approach of siRNA delivery to liver hepatocytes which can also be applied to treat HCV in chronic liver diseases.

Stealth monoolein-based nanocarriers for delivery of siRNA to cancer cells

“…4A and B). The percentages of siRNA encapsulation are in agreement with the values found in literature [12,46]. As Fig.…”

“…For non-pegylated lipoplexes the presence of a helper lipid in the formulation helps on the release of the genetic material from endosomes [26,[46][47][48], while the presence of PEG at the nanocarrier surface blocks this release by more than one mechanism. First, PEG favors a stable lamellar lipid organization, impairing the modification into another structural organization required for endosomal membrane destabilization [25,49]; second, PEG also prevents the close interaction between lipoplexes and endosomal membranes required for destabilization and nucleic acids release [50].…”

Oncogene Knockdown via Active Loading of Small RNAs into Extracellular Vesicles by Sonication

“…Sonication has previously been employed to generate nanoparticles for siRNA delivery 12, 20 and to load EVs with small molecules 18 and protein cargo 17 , yet its utility as a small RNA loading technique for EVs is unknown. Parameters of sonication and its effects on EV and nucleic acid integrity as well as siRNA aggregation were assessed.…”

Targeting Splicing in the Treatment of Human Disease

“…There are a variety of different nanoparticle types, depending on manufacturing processes and components (Figure 5) [103]. The nanoparticle formula can be designed to produce carriers for oral [104], skin [105], liver [106], pulmonary [107], brain [108], or cancer targeting [109]. Nanoparticles can be used to deliver a combination of biomolecules to enhance the therapeutic effect against disease.…”