Biochimica et Biophysica Acta 2014-12-01

Diatomite biosilica nanocarriers for siRNA transport inside cancer cells.

Ilaria Rea, Nicola M Martucci, Luca De Stefano, Immacolata Ruggiero, Monica Terracciano, Principia Dardano, Nunzia Migliaccio, Paolo Arcari, Rosarita Taté, Ivo Rendina, Annalisa Lamberti

Index: Biochim. Biophys. Acta 1840(12) , 3393-403, (2014)

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Abstract

Diatomite is a natural porous biomaterial of sedimentary origin, formed by fragments of diatom siliceous skeletons, called "frustules". Due to large availability in many areas of the world, chemical stability, and non-toxicity, these fossil structures have been widespread used in lot of industrial applications, such as food production, water extracting agent, production of cosmetics and pharmaceutics. However, diatomite is surprisingly still rarely used in biomedical applications. In this work, we exploit diatomite nanoparticles for small interfering ribonucleic acid (siRNA) transport inside human epidermoid cancer cells (H1355).Morphology and composition of diatomite microfrustules (average size lower than 40μm) are investigated by scanning electron microscopy equipped by energy dispersive X-ray spectroscopy, Fourier transform infrared analysis, and photoluminescence measurements. Nanometric porous particles (average size lower than 450nm) are obtained by mechanical crushing, sonication, and filtering of micrometric frustules. siRNA bioconjugation is performed on both micrometric and nanometric fragments by silanization.In-vitro experiments show very low toxicity on exposure of the cells to diatomite nanoparticle concentration up to 300μg/ml for 72h. Confocal microscopy imaging performed on cancer cells incubated with siRNA conjugated nanoparticles demonstrates a cytoplasmatic localization of vectors. Gene silencing by delivered siRNA is also demonstrated.Our studies endorse diatomite nanoparticles as non-toxic nanocarriers for siRNA transport in cancer cells.siRNA is a powerful molecular tool for cancer treatment but its delivery is inefficient due to the difficulty to penetrate the cell membrane. siRNA-diatomite nanoconjugate may be well suited for delivery of therapeutic to cancer cells.Copyright © 2014 Elsevier B.V. All rights reserved.