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Diatomite biosilica nanocarriers for siRNA transport inside cancer cells
Affiliation:1. Institute for Microelectronics and Microsystems, National Council of Research, Naples, Italy;2. Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy;3. Department of Pharmacy, University of Naples Federico II, Naples, Italy;4. Institute of Genetics and Biophysics, National Council of Research, Naples, Italy
Abstract:BackgroundDiatomite 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).MethodsMorphology and composition of diatomite microfrustules (average size lower than 40 μm) 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 450 nm) are obtained by mechanical crushing, sonication, and filtering of micrometric frustules. siRNA bioconjugation is performed on both micrometric and nanometric fragments by silanization.ResultsIn-vitro experiments show very low toxicity on exposure of the cells to diatomite nanoparticle concentration up to 300 μg/ml for 72 h. 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.ConclusionOur studies endorse diatomite nanoparticles as non-toxic nanocarriers for siRNA transport in cancer cells.General significancesiRNA 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.
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