title: Poly(D,L-lactide-co-glycolide acid) nanoparticles for DNA delivery: waiving preparation complexity and increasing efficiency.
abstract: When designing a nonviral gene delivery system based on polymeric nanoparticles (NPs), it is important to keep in mind obstacles associated with future clinical applications. Simplifying the procedure of NPs production and taking toxicity into account are the most important issues that need to be addressed. Toxicity concerns in clinical trials may be raised when using additives such as cationic polymers/lipids, buffering reagents, and proteins. Therefore, the aim of this study was to simplify the formulation of poly (lactide-co-glycolide) acid NPs by shortening steps such as sonication time and by avoiding the use of additives while preserving its efficiency. NPs (300 nm) were formulated using a modified w/o/w technique with DNA entrapment efficiency of 80%. Once achieving such NPs, formulation parameters such as DNA loading, release kinetics, DNA integrity and bioactivity, uptake by cells, and toxicity were addressed. The NPs were readily taken by several cell lines and were localized mostly in their endo-lysosomal compartments. The NPs did not affect cells viability. Most importantly, transfection studies in COS-7 and Cf2th cells resulted with a 250-fold protein expression levels when compared with the control. These expression levels are higher than ones achieved with more complicated NPs systems, demonstrating the efficiency of our simplified NPs for gene delivery.
authors.affiliation: Faculty of Biotechnology and Food Engineering, Technion, Israel Institute of Technology, Haifa, Israel.
authors.forename: Koby
authors.initials: K
authors.lastname: Gvili
authors.forename: Ofra
authors.initials: O
authors.lastname: Benny
authors.forename: Dganit
authors.initials: D
authors.lastname: Danino
authors.forename: Marcelle
authors.initials: M
authors.lastname: Machluf
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004247:DNA / Q000008:administration & dosage* / Q000378:metabolism; D003692:Delayed-Action Preparations; D004285:Dogs; D018014:Gene Transfer Techniques; D006801:Humans; D007700:Kinetics; D019344:Lactic Acid / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000737:chemistry*; D014162:Transfection
chemical_list: D001672:Biocompatible Materials; D003692:Delayed-Action Preparations; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D004247:DNA