Advanced in vitro exposure systems.

Hybrid Lipid/Polymer Nanoparticles for Pulmonary Delivery of siRNA: Development and Fate Upon In Vitro Deposition on the Human Epithelial Airway Barrier

26. Jul. 2018

DOI: 10.1089/jamp.2017.1364

Ivana d’Angelo, PhD,1 Gabriella Costabile, PhD,2,3 Estelle Durantie, PhD,3 Paola Brocca, PhD,4 Valeria Rondelli, PhD,4 Annapina Russo, PhD,5 Giulia Russo, PhD,5 Agnese Miro, PharmD,2 Fabiana Quaglia, PhD,2 Alke Petri-Fink, PhD,3 Barbara Rothen-Rutishauser, PhD,3 and Francesca Ungaro, PhD2
1 Di.S.T.A.Bi.F., University of Campania ‘‘Luigi Vanvitelli,’’ Caserta, Italy.
2 Laboratory of Drug Delivery, Department of Pharmacy, University of Napoli Federico II, Napoli, Italy.
3 Adolphe Merkle Institute, University of Fribourg, Fribourg, Switzerland.
4 Applied Physics, Department of Medical Biotechnology and Translational Medicine, University of Milano, Milano, Italy.
5 Laboratory of Biochemistry, Department of Pharmacy, University of Napoli Federico II, Napoli, Italy.

The Vitrocell Cloud was used for the exposure to aerosolized nanoparticles on a in vitro three-dimensional cell cluture, mimicking the human epithelial airway barrier comprising human bronchial epithelial cells,human blood monocyte-derived macrophages and dendritic cells to show the optimized siRNA transport and delivery at lung.


Background: Nowadays, the downregulation of genes involved in the pathogenesis of severe lung diseases through local siRNA delivery appears an interesting therapeutic approach. In this study, we propose novel hybrid lipid-polymer nanoparticles (hNPs) consisting of poly(lactic-co-glycolic) acid (PLGA) and dipalmitoyl phosphatidylcholine (DPPC) as siRNA inhalation system.
Methods: A panel of DPPC/PLGA hNPs was prepared by emulsion/solvent diffusion and fully characterized. A combination of model siRNAs against the sodium transepithelial channel (ENaC) was entrapped in optimized hNPs comprising or not poly(ethylenimine) (PEI) as third component. siRNA-loaded hNPs were characterized for encapsulation efficiency, release kinetics, aerodynamic properties, and stability in artificial mucus (AM). The fate and cytotoxicity of hNPs upon aerosolization on a triple cell co-culture model (TCCC) mimicking human epithelial airway barrier were assessed. Finally, the effect of siRNA-loaded hNPs on ENaC protein expression at 72 hours was evaluated in A549 cells.
Results: Optimized muco-inert hNPs encapsulating model siRNA with high efficiency were produced. The developed hNPs displayed a hydrodynamic diameter of *150 nm, a low polydispersity index, a negative f potential close to -25 mV, and a peculiar triphasic siRNA release lasting for 5 days, which slowed down in the presence of PEI. siRNA formulations showed optimal in vitro aerosol performance after delivery with a vibrating mesh nebulizer. Furthermore, small-angle X-ray scattering analyses highlighted an excellent stability upon incubation with AM, confirming the potential of hNPs for direct aerosolization on mucus-lined airways. Studies in TCCC confirmed that fluorescent hNPs are internalized inside airway epithelial cells and do not exert any cytotoxic or acute proinflammatory effect. Finally, a prolonged inhibition of ENaC protein expression was observed in A549 cells upon treatment with siRNA-loaded hNPs. 
Conclusions: Results demonstrate the great potential of hNPs as carriers for pulmonary delivery of siRNA, prompting toward investigation of their therapeutic effectiveness in severe lung diseases.

Read research article

Comments (0)Number of views (678)

Back to Top