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.

Keywords::

• chitosan
• gene delivery
• inhalation
• microencapsulation
• plasmid
• solid lipid nanoparticles
• spray-drying

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.tandfonline.com/doi/full/10.2217/nnm-2018-0270

Financial & competing interests disclosure

This work was supported by Fundação para a Ciência e Tecnologia, Portugal (projects UID/DTP/04138/2013 and PTDC/DTP-FTO/0094/2012, and grants SFRH/BD/89520/2012 and SFRH/BSAB/1210/2011). This work was also supported by Spanish Government (PN de I+D 2008-2011, ISCIII-Subdirección General de Evaluación y Fomento de la Investigación, Acción Estratégica de Salud, Projects FIS, PS09/00816, and MAT2013-40971-R), Xunta de Galicia (Project Competitive Reference Groups-FEDER Funds Ref. 2014/043 and Project Competitive Emergent Groups-FEDER Funds EM2013-046). Jorge Vitor’s lab has been financed by New England Biolabs, Inc. (USA). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Author contributions

DP Gaspar developed all formulation described in the study and wrote the manuscript; MC Leiva helped to develop the DNA-loaded nanoparticles; J Vital and J Vítor characterized the plasmid association with nanoparticles and its integrity; LMD Gonçalves performed in vitro studies using pulmonary cell lines; P Taboada was responsible for physical characterization, particularly ITC; C Remuñán-López was responsible for spray-drying and dry powder development; AJ Almeida was responsible for nanoformulation development and writing up.

Acknowledgments

The authors thank María José, Raquel Antón and Rafael Romero (Universidad de Santiago de Compostela, Spain) for the transmission electron microscopy and scanning electron microscopy images, and for technical help, as well as Raul Ortiz (IBIMER, CIBM, Granada, Spain), for the kind gift of the plasmid.

Additional information

Funding

This work was supported by Fundação para a Ciência e Tecnologia, Portugal (projects UID/DTP/04138/2013 and PTDC/DTP-FTO/0094/2012, and grants SFRH/BD/89520/2012 and SFRH/BSAB/1210/2011). This work was also supported by Spanish Government (PN de I+D 2008-2011, ISCIII-Subdirección General de Evaluación y Fomento de la Investigación, Acción Estratégica de Salud, Projects FIS, PS09/00816, and MAT2013-40971-R), Xunta de Galicia (Project Competitive Reference Groups-FEDER Funds Ref. 2014/043 and Project Competitive Emergent Groups-FEDER Funds EM2013-046). Jorge Vitor’s lab has been financed by New England Biolabs, Inc. (USA). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.