Continuous microfluidic assembly of biodegradable poly(beta-amino ester)/DNA nanoparticles for enhanced gene delivery.
Translation of biomaterial-based nanoparticle formulations to the clinic faces significant challenges including efficacy, safety, consistency and scale-up of manufacturing, and stability during long-term storage. Continuous microfluidic fabrication of polymeric nanoparticles has the potential to alleviate the challenges associated with manufacture, while offering a scalable solution for clinical level production. Poly(beta-amino esters) (PBAE)s are a class of biodegradable cationic polymers that self-assemble with anionic plasmid DNA to form polyplex nanoparticles that have been shown to be effective for transfecting cancer cells specifically in vitro and in vivo. Here, we demonstrate the use of a microfluidic device for the continuous and scalable production of PBAE/DNA nanoparticles followed by lyophilization and long term storage that results in improved in vitro efficacy in multiple cancer cell lines compared to nanoparticles produced by bulk mixing as well as in comparison to widely used commercially available transfection reagents polyethylenimine and Lipofectamine® 2000. We further characterized the nanoparticles using nanoparticle tracking analysis (NTA) to show that microfluidic mixing resulted in fewer DNA-free polymeric nanoparticles compared to those produced by bulk mixing. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1813-1825, 2017.
Biomedical Engineering, Johns Hopkins University, Baltimore, 21231.|Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, 21231.
David R
DR
Wilson
Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.|Materials Science and Engineering, Johns Hopkins University, Baltimore, 21231.
Arman
A
Mosenia
Biomedical Engineering, Johns Hopkins University, Baltimore, 21231.|Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.
Mark P
MP
Suprenant
Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.
Rahul
R
Upadhya
Biomedical Engineering, Johns Hopkins University, Baltimore, 21231.|Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.
Denis
D
Routkevitch
Biomedical Engineering, Johns Hopkins University, Baltimore, 21231.|Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, 21231.
Randall A
RA
Meyer
Johns Hopkins University School of Medicine, Neurosurgery, Baltimore, 21231.|Oncology, Johns Hopkins University School of Medicine, Baltimore, 21231.
Alfredo
A
Quinones-Hinojosa
Biomedical Engineering, Johns Hopkins University, Baltimore, 21231.|Translational Tissue Engineering Center, Johns Hopkins of Medicine, Baltimore, 21231.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, 21231.|Johns Hopkins University School of Medicine, Neurosurgery, Baltimore, 21231.|Oncology, Johns Hopkins University School of Medicine, Baltimore, 21231.|Materials Science and Engineering, Johns Hopkins University, Baltimore, 21231.|Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, 21231.
Jordan J
JJ
Green
D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004867:Equipment Design; D005612:Freeze Drying; D018014:Gene Transfer Techniques; D006801:Humans; D056656:Lab-On-A-Chip Devices; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011108:Polymers / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D011108:Polymers; C507253:poly(beta-amino ester); D004247:DNA
gene delivery; hydrodynamic focusing; microfluidic; nanoparticle; nanoparticle tracking analysis; polymer