- 作者: Cario Mischiati; Alessia Sereni; Alessia finotti; Laura Breda; Rita Cortesi; Claudio Nastruzzi; Alessandra romanelli; Michele Saviano; Nicoletta Bianchi; Carlo Pedone; Monica Borgatti; Roberto Gambari
- 中文摘要: --
- 英文摘要: The major aim of this paper was to determine whethercationic microspheres(CM),consisting of the permeablepolymer EudragitR RS 100 plus the cationic surfactantdioctadecyl-dimethyl-ammonium bromide(DDAB18),could bind to double-stranded peptide nucleic acid PNA-DNA-PNA(PDP) chimeras exhibiting decoy activityagainst NF-KB transcription factors. Microspheres wereproduced by the 'solvent evaporation method' and cen-trifugation at 500, 1,000 and 3,000 rpm to obtain differ-ent-sized microparticles. Microsphere morphology, sizeand size distribution were determined by optical andelectron microscopy observations. In order to determinetheir binding activity, double-stranded DNA-based andPDP-based decoy molecules were incubated with differ-ent amounts of microparticles in the presence of 100 ngof either 32P-labeled DNA-DNA or DNA-PDP hybrid mole-cules or cold PDP-PDP hybrids. The complexes were ana-lyzed by agarose gel electrophoresis. The resistance of32P-labeled DNA-DNA and DNA-PDP molecules in thepresence of serum or cellular extracts was evaluatedafter binding to CM by gel electrophoresis analysis.DDAB18 Eudragit RS 100 microspheres are able to bind toDNA-PDP and PDP-PDP hybrids, to deliver these mole-cules to target cells and to protect DNA-PDP moleculesfrom enzymatic degradation in simulated biologicalfluids. In addition,when assayed in ex vivo conditions,DDABi8 Eudragit RS 100 microspheres exhibited low tox-icity.The results presented in this paper demonstratethat CM can be considered suitable formulations forpharmacogenomic therapy employing double-strandedPDP chimeras.
- 中文關鍵字: --
- 英文關鍵字: Peptide nucleic acid-DNA chimeras; Microspheres; Gene therapy