The major aim of this paper was to determine whether cationic microspheres (CM), consisting of the permeable polymer Eudragit ® RS 100 plus the cationic surfactant dioctadecyl-dimethyl-ammonium bromide (DDAB18), could bind to double-stranded peptide nucleic acid PNADNA-PNA (PDP) chimeras exhibiting decoy activity against NF-KB transcription factors. Microspheres were produced by the 'solvent evaporation method' and centrifugation at 500, 1,000 and 3,000 rpm to obtain different-sized microparticles. Microsphere morphology, size and size distribution were determined by optical and electron microscopy observations. In order to determine their binding activity, double-stranded DNA-based and PDP-based decoy molecules were incubated with different amounts of microparticles in the presence of 100 ng of either 32p-labeled DNA-DNA or DNA-PDP hybrid molecules or cold PDP-PDP hybrids. The complexes were analyzed by agarose gel electrophoresis. The resistance of 32p-labeled DNA-DNA and DNA-PDP molecules in the presence of serum or cellular extracts was evaluated after binding to CM by gel electrophoresis analysis. DDAB18 Eudragit RS 100 microspheres are able to bind to DNA-PDP and PDP-PDP hybrids, to deliver these molecules to target cells and to protect DNA-PDP molecules from enzymatic degradation in simulated biological fluids. In addition, when assayed in ex vivo conditions, DDABm Eudragit RS 100 microspheres exhibited low toxicity. The results presented in this paper demonstrate that CM can be considered suitable formulations for pharmacogenomic therapy employing double-stranded PDP chimeras.
Peptide nucleic acid-DNA decoy chimeras targeting NF-kappaB transcription factors: Induction of apoptosis in human primary osteoclasts / Penolazzi, L; Borgatti, M; Lambertini, E; Mischiati, C; Finotti, A; Romanelli, Alessandra; Saviano, Michele; Pedone, Carlo; Piva, R; Gambari, R.. - In: INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE. - ISSN 1107-3756. - STAMPA. - 11:(2004), pp. 145-152. [10.1159/000079683]
Peptide nucleic acid-DNA decoy chimeras targeting NF-kappaB transcription factors: Induction of apoptosis in human primary osteoclasts
ROMANELLI, ALESSANDRA;SAVIANO, MICHELE;PEDONE, CARLO;
2004
Abstract
The major aim of this paper was to determine whether cationic microspheres (CM), consisting of the permeable polymer Eudragit ® RS 100 plus the cationic surfactant dioctadecyl-dimethyl-ammonium bromide (DDAB18), could bind to double-stranded peptide nucleic acid PNADNA-PNA (PDP) chimeras exhibiting decoy activity against NF-KB transcription factors. Microspheres were produced by the 'solvent evaporation method' and centrifugation at 500, 1,000 and 3,000 rpm to obtain different-sized microparticles. Microsphere morphology, size and size distribution were determined by optical and electron microscopy observations. In order to determine their binding activity, double-stranded DNA-based and PDP-based decoy molecules were incubated with different amounts of microparticles in the presence of 100 ng of either 32p-labeled DNA-DNA or DNA-PDP hybrid molecules or cold PDP-PDP hybrids. The complexes were analyzed by agarose gel electrophoresis. The resistance of 32p-labeled DNA-DNA and DNA-PDP molecules in the presence of serum or cellular extracts was evaluated after binding to CM by gel electrophoresis analysis. DDAB18 Eudragit RS 100 microspheres are able to bind to DNA-PDP and PDP-PDP hybrids, to deliver these molecules to target cells and to protect DNA-PDP molecules from enzymatic degradation in simulated biological fluids. In addition, when assayed in ex vivo conditions, DDABm Eudragit RS 100 microspheres exhibited low toxicity. The results presented in this paper demonstrate that CM can be considered suitable formulations for pharmacogenomic therapy employing double-stranded PDP chimeras.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.