Synthesis, Functionalization, and Characterization of Fusogenic Porous Silicon Nanoparticles for Oligonucleotide Delivery

Byungji Kim; Michael  J. Sailor

doi:10.3791/59440

Síntesis, funcionalización y caracterización de nanopartículas de silicio poroso fusogénicas para...

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JoVE Journal Engineering

Synthesis, Functionalization, and Characterization of Fusogenic Porous Silicon Nanoparticles for Oligonucleotide Delivery

Síntesis, funcionalización y caracterización de nanopartículas de silicio poroso fusogénicas para entrega de oligonucleótidos

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08:53 min

April 16, 2019

DOI: 10.3791/59440-v

Byungji Kim¹, Michael J. Sailor^1,2

¹Materials Science and Engineering Program,University of California, San Diego, ²Department of Chemistry and Biochemistry,University of California, San Diego

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Overview

This article presents a method for synthesizing fusogenic porous silicon nanoparticles designed for efficient oligonucleotide delivery in vitro and in vivo. The nanoparticles are engineered to carry siRNA and are coated with fusogenic lipids, enhancing their delivery capabilities.

Key Study Components

Area of Science

Nanoparticle synthesis
Gene delivery systems
Therapeutic applications

Background

Fusogenic nanoparticles can silence genes effectively.
Targeting peptides and siRNA payloads can be customized.
Previous studies have shown efficacy against bacterial infections.
Chronic inflammation can be mitigated by gene modulation.

Purpose of Study

To develop a nanoparticle system for in vivo gene silencing.
To explore therapeutic formulations for diseases requiring gene modulation.
To demonstrate the versatility of the nanoparticle platform.

Methods Used

Assembling a Teflon etch cell with a silicon wafer.
Using hydrofluoric acid for etching the silicon.
Applying alternating current with specific parameters for nanoparticle formation.
Rinsing the etch cell to prepare for further use.

Main Results

Successful synthesis of fusogenic porous silicon nanoparticles.
High efficiency in delivering siRNA for gene silencing.
Potential for therapeutic applications in various diseases.
Demonstrated ability to terminate chronic inflammation in macrophages.

Conclusions

Fusogenic nanoparticles are promising for gene therapy.
Customizable delivery systems can target specific diseases.
Further research is warranted to explore additional applications.

Frequently Asked Questions

What are fusogenic nanoparticles?

Fusogenic nanoparticles are engineered particles that enhance the delivery of genetic material into cells.

How do these nanoparticles deliver siRNA?

They encapsulate siRNA and utilize fusogenic lipids to facilitate cellular uptake.

What diseases could benefit from this technology?

Diseases requiring gene modulation, including chronic inflammation and infections, may benefit.

What is the significance of using hydrofluoric acid in the process?

Hydrofluoric acid is used to etch silicon wafers, creating the porous structure necessary for nanoparticle formation.

Can the targeting peptide be modified?

Yes, the targeting peptide can be exchanged to tailor the nanoparticles for specific applications.

Demostramos la síntesis de nanopartículas de silicio poroso fusogénicas para la entrega eficaz de oligonucleótidos in vitro e in vivo. Nanopartículas de silicio poroso están cargadas de siRNA para formar el núcleo, que es cubierto por fusogénicas lípidos a través de la protuberancia para formar la cáscara. Funcionalización de parte de la segmentación y caracterización de partículas están incluidos.

Este protocolo describe la síntesis de un sistema de nanopartículas que es capaz de silenciar los genes in vivo mediante la entrega de siRNAs con alta eficiencia. Simplemente intercambiando el péptido de orientación y la carga útil del aresmo, las nanopartículas fusogénicas pueden utilizarse como una formulación terapéutica potencial para enfermedades que requieren modulación génica in vivo. Hemos publicado previamente resultados utilizando la nanoplataforma fusógena contra las infecciones bacterianas grampositivas al silenciar un gen que codifica para que la inflamación en los macrófagos termine la inflamación crónica.

Antes de realizar este procedimiento, ensamble una célula de grabado de teflón que contenga una oblea de silicio en una solución de ácido fluorhídrico de tres a uno. Ejecute una corriente alterna de una forma de onda cuadrada con una densidad de corriente baja de 50 miliamperios por centímetro cuadrado durante 0,6 segundos y una alta densidad de corriente de 400 miliamperios por centímetro cuadrado durante 0,36 segundos repetidos durante 500 ciclos. Una vez terminado el grabado, retire la célula del circuito y enjuague cuidadosamente la solución de ácido fluorhídrico con una jeringa.

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