TY - JOUR
T1 - Poly(D,L-lactide-co-glycolide acid) nanoparticles for DNA delivery
T2 - Waiving preparation complexity and increasing efficiency
AU - Gvili, Koby
AU - Benny, Ofra
AU - Danino, Dganit
AU - Machluf, Marcelle
PY - 2007/4/15
Y1 - 2007/4/15
N2 - 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 endolysosomal compartments. The NPs did not affect cells viability. Most importantly, transfectton 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.
AB - 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 endolysosomal compartments. The NPs did not affect cells viability. Most importantly, transfectton 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.
KW - Gene delivery
KW - Nanoparticles
KW - PLGA
KW - Transfection efficiency
UR - http://www.scopus.com/inward/record.url?scp=34247516346&partnerID=8YFLogxK
U2 - 10.1002/bip.20697
DO - 10.1002/bip.20697
M3 - 文章
C2 - 17266128
AN - SCOPUS:34247516346
SN - 0006-3525
VL - 85
SP - 379
EP - 391
JO - Biopolymers
JF - Biopolymers
IS - 5-6
ER -