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2 results on '"Vanessa Ocana"'

1. Efficient, non-toxic hybrid PPV-PAMAM dendrimer as a gene carrier for neuronal cells

Author

Vanessa Ocana, Silvia Monteagudo, Julián Rodríguez-López, Francisco C. Pérez-Martínez, Prado Sánchez-Verdú, Iván Rivilla, Valentín Ceña, Javier Guerra, Joaquín C. García-Martínez, Sonia Merino, and Ana C. Rodrigo

Subjects
Small interfering RNA, Dendrimers, Spectrometry, Mass, Electrospray Ionization, Magnetic Resonance Spectroscopy, Polymers and Plastics, Spectrophotometry, Infrared, Blotting, Western, Genetic Vectors, Bioengineering, Gene delivery, Conjugated system, Biomaterials, Rats, Sprague-Dawley, PAMAM dendrimer, Dendrimer, Materials Chemistry, Animals, RNA, Small Interfering, Gene, Cells, Cultured, Neurons, Chemistry, Reverse Transcriptase Polymerase Chain Reaction, Molecular biology, Rats, Toxicity, Biophysics, NMDA receptor
Abstract

A novel hybrid dendrimer (TRANSGEDEN) that combines a conjugated rigid polyphenylenevinylene (PPV) core with flexible polyamidoamine (PAMAM) branches at the surface was synthesized and characterized. The potential of this material as a nonviral gene delivery system was also examined, and it was observed that dendriplexes formed by TRANSGEDEN and small interfering ribonucleic acids (siRNAs) can be incorporated into >90% of neuronal cells without any toxicity up to a dendrimer concentration of 3 μM. TRANSGEDEN was used to deliver a specific siRNA to rat cerebellar granular neurons (CGNs) to knock down the cofilin-1 protein. Cofilin-1 removal partially protects CGNs from N-methyl D-aspartate (NMDA)-mediated neuronal death.

Published
2011

2. Role of Generation, Architecture, pH and Ionic Strength on Successful siRNA Delivery and Transfection by Hybrid PPV-PAMAM Dendrimers

Author

Giovanni M. Pavan, Blanca Carrión, Francisco C. Pérez-Martínez, Silvia Monteagudo, Julián Rodríguez-López, Javier Guerra, Andrea Danani, Vanessa Ocana, and Valentín Ceña

Subjects
Models, Molecular, Small interfering RNA, Dendrimers, SiRNA binding, Nanotechnology, Molecular Dynamics Simulation, Transfection, Biochemistry, Rats, Sprague-Dawley, Dendrimer, Drug Discovery, Animals, Humans, RNA, Small Interfering, Cells, Cultured, Pharmacology, Cerebral Cortex, Neurons, Pamam dendrimers, Chemistry, Organic Chemistry, Osmolar Concentration, Glyceraldehyde-3-Phosphate Dehydrogenases, Biological activity, Hydrogen-Ion Concentration, Rats, Ionic strength, Gene Knockdown Techniques, Biophysics, Molecular Medicine, Thermodynamics, Nanocarriers
Abstract

Small interfering RNA (siRNA) constitutes an excellent way of knocking down genes. However, it requires the use of delivery systems to reach the target cells, especially to neuronal cells. Dendrimers are one of the most widely used synthetic nanocarriers for siRNA delivery. However, due to the complexity of the dendrimer-siRNA interactions, when a new dendritic carrier is designed it is difficult to predict its efficiency to bind and to deliver siRNA. At the same time it is not easy to understand the origin of eventual limited functionalities. We have modeled the interactions between two dendrimers (TDG-G1 and TDG-G2) and siRNA using molecular dynamics (MD) simulation. The results were compared to experimental physico-chemical parameters such as siRNA complexation, complex stability, size, and zeta potentials and biological effects such as down-regulation of a specific RNA expression in cortical neurons in culture. Data indicate that the combination of rigid core and flexible branches guarantees strong siRNA binding, which is important to have a good transfection profile. However, the successful nanocarrier for siRNA delivery (TDG-G1) is identified not only by a high affinity for siRNA, but by a favorable equilibrium between a strong binding and the ability to release siRNA to exert its biological action. The conditions under which the dendriplex is formed are also relevant for transfection efficiency and biological activity.

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