Merck
  • Home
  • Search Results
  • Diatomite biosilica nanocarriers for siRNA transport inside cancer cells.

Diatomite biosilica nanocarriers for siRNA transport inside cancer cells.

Biochimica et biophysica acta (2014-09-17)
Ilaria Rea, Nicola M Martucci, Luca De Stefano, Immacolata Ruggiero, Monica Terracciano, Principia Dardano, Nunzia Migliaccio, Paolo Arcari, Rosarita Taté, Ivo Rendina, Annalisa Lamberti
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sulfuric acid, 99.999%
Supelco
Sulfuric acid, for the determination of nitrogen, ≥97.0%
Sigma-Aldrich
Sulfo-N-succinimidyl 4-maleimidobutyrate sodium salt, ~90%
Sigma-Aldrich
Tris(2-carboxyethyl)phosphine hydrochloride, powder
Sigma-Aldrich
Tris(2-carboxyethyl)phosphine hydrochloride, BioUltra, ≥98% (NMR)
Supelco
Dehydrated Alcohol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Tris(2-carboxyethyl)phosphine hydrochloride, BioUltra, suitable for electrophoresis, SDS-PAGE tested
Sigma-Aldrich
MISSION® esiRNA, targeting mouse Gapdh
Supelco
Ethanol solution, certified reference material, 2000 μg/mL in methanol
Sigma-Aldrich
MISSION® esiRNA, targeting human GAPDH
Sigma-Aldrich
Sulfuric acid, puriss., meets analytical specification of Ph. Eur., BP, 95-97%
Sigma-Aldrich
Sulfuric acid, puriss. p.a., for determination of Hg, ACS reagent, reag. ISO, reag. Ph. Eur., 95.0-97.0%
Sigma-Aldrich
Sulfuric acid, ACS reagent, 95.0-98.0%
Sigma-Aldrich
Sulfuric acid solution, puriss. p.a., ≥25% (T)
Sigma-Aldrich
Phenol Red, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
(3-Aminopropyl)triethoxysilane, ≥98%
Sigma-Aldrich
Ethanol, purum, secunda spirit, denaturated with 2% 2-butanone and 0.5% 4-methyl-2-pentanone, S15, ~96% (based on denaturant-free substance)
Supelco
Ethanol, standard for GC
Sigma-Aldrich
(3-Aminopropyl)triethoxysilane, packaged for use in deposition systems, ≥98%
Supelco
Sulfuric acid concentrate, 0.1 M H2SO4 in water (0.2N), eluent concentrate for IC
Sigma-Aldrich
Ethanol, purum, absolute ethanol, denaturated with 4.8% isopropanol, A15 IPA1, ≥99.8% (based on denaturant-free substance)
Sigma-Aldrich
Ethanol, ACS reagent, prima fine spirit, without additive, F15 o1
Sigma-Aldrich
Ethanol, purum, fine spirit, denaturated with 2% 2-butanone, F25 MEK1, ~96% (based on denaturant-free substance)
Sigma-Aldrich
Ethanol, purum, fine spirit, denaturated with 4.8% methanol, F25 METHYL1, ~96% (based on denaturant-free substance)
Sigma-Aldrich
Ethanol, purum, absolute ethanol, denaturated with 1% cyclohexane, A15 CYCLO1, ≥99.8% (based on denaturant-free substance)
Sigma-Aldrich
Ethanol, purum, absolute ethanol, denaturated with 2% 2-butanone, A15 MEK1, ≥99.8% (based on denaturant-free substance)
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, HPLC/spectrophotometric grade
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, ACS reagent, ≥99.5%
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, meets USP testing specifications
Sigma-Aldrich
Ethyl alcohol, Pure, 190 proof, for molecular biology