Effect of Pendant Group on pDNA Delivery by Cationic-β-Cyclodextrin:Alkyl-PVA-PEG Pendant Polymer Complexes

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• 作者：Aditya Kulkarni ; Vivek Badwaik ; Kyle DeFrees ; Ryan A. Schuldt ; Dinara S. Gunasekera ; Cory Powers ; Alexander Vlahu ; Ross VerHeul ; David H. Thompson
• 刊名：Biomacromolecules
• 出版年：2014
• 出版时间：January 13, 2014
• 年：2014
• 卷：15
• 期：1
• 页码：12-19
• 全文大小：435K
• ISSN：1526-4602

文摘

We have previously shown that cationic-尾-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 尾-CD cavity. Cytotoxicity studies revealed that all of the cationic-尾-CD:pendant polymer host:guest complexes have 100鈥?000-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鈥?70 nm range and 味-potentials of 15鈥?5 mV. Gel shift and heparin challenge experiments showed that the complexes are most stable at N/P 鈮?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-尾-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.