title: Effect of pendant group on pDNA delivery by cationic-β-cyclodextrin:alkyl-PVA-PEG pendant polymer complexes.
abstract: We have previously shown that cationic-β-cyclodextrin:R-poly(vinyl alcohol)-poly(ethylene glycol) (CD+:R-PVA-PEG) pendant polymer host:guest complexes are safe and efficient vehicles for nucleic acid delivery, where R = benzylidene-linked adamantyl or cholesteryl esters. Herein, we report the synthesis and biological performance of a family of PVA-PEG pendant polymers whose pendant groups have a wide range of different affinities for the β-CD cavity. Cytotoxicity studies revealed that all of the cationic-β-CD:pendant polymer host:guest complexes have 100-1000-fold lower toxicity than branched polyethylenimine (bPEI), with pDNA transfection efficiencies that are comparable to bPEI and Lipofectamine 2000. Complexes formed with pDNA at N/P ratios greater than 5 produced particles with diameters in the 100-170 nm range and ζ-potentials of 15-35 mV. Gel shift and heparin challenge experiments showed that the complexes are most stable at N/P ≥ 10, with adamantyl- and noradamantyl-modified complexes displaying the best resistance toward heparin-induced decomplexation. Disassembly rates of fluoresceinated-pDNA:CD(+):R-PVA-PEG-rhodamine complexes within HeLa cells showed a modest dependence on host:guest binding constant, with adamantyl-, noradamantyl-, and dodecyl-based complexes showing the highest loss in FRET efficiency 9 h after cellular exposure. These findings suggest that the host:guest binding constant has a significant impact on the colloidal stability in the presence of serum and cellular uptake efficiency, whereas endosomal disassembly and transfection performance of cationic-β-CD:R-poly(vinyl alcohol)-poly(ethylene glycol) pendant polymer complexes appears to be controlled by the hydrolysis rates of the acetal grafts onto the PVA main chain.
authors.affiliation: Department of Chemistry, Purdue University , 560 Oval Drive, West Lafayette, Indiana, 47907, United States.
authors.forename: Aditya
authors.initials: A
authors.lastname: Kulkarni
authors.forename: Vivek
authors.initials: V
authors.lastname: Badwaik
authors.forename: Kyle
authors.initials: K
authors.lastname: DeFrees
authors.forename: Ryan A
authors.initials: RA
authors.lastname: Schuldt
authors.forename: Dinara S
authors.initials: DS
authors.lastname: Gunasekera
authors.forename: Cory
authors.initials: C
authors.lastname: Powers
authors.forename: Alexander
authors.initials: A
authors.lastname: Vlahu
authors.forename: Ross
authors.initials: R
authors.lastname: VerHeul
authors.forename: David H
authors.initials: DH
authors.lastname: Thompson
mesh_terms: D003505:Cyclodextrins / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D009930:Organic Chemicals / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D011145:Polyvinyls / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology
chemical_list: D003505:Cyclodextrins; C519734:Kollicoat IR; D009930:Organic Chemicals; C099476:PicoGreen; D011108:Polymers; D011145:Polyvinyls; D004247:DNA