Ilaria Rea1, Nicola M Martucci2, Luca De Stefano3, Immacolata Ruggiero2, Monica Terracciano4, Principia Dardano3, Nunzia Migliaccio2, Paolo Arcari2, Rosarita Taté5, Ivo Rendina3, Annalisa Lamberti2. 1. Institute for Microelectronics and Microsystems, National Council of Research, Naples, Italy. Electronic address: ilaria.rea@na.imm.cnr.it. 2. Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy. 3. Institute for Microelectronics and Microsystems, National Council of Research, Naples, Italy. 4. Institute for Microelectronics and Microsystems, National Council of Research, Naples, Italy; Department of Pharmacy, University of Naples Federico II, Naples, Italy. 5. Institute of Genetics and Biophysics, National Council of Research, Naples, Italy.
Abstract
BACKGROUND: 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). METHODS: 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. RESULTS: 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. CONCLUSION: Our studies endorse diatomite nanoparticles as non-toxic nanocarriers for siRNA transport in cancer cells. GENERAL SIGNIFICANCE: 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.
BACKGROUND: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 humanepidermoid cancer cells (H1355). METHODS: 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. RESULTS: 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. CONCLUSION: Our studies endorse diatomite nanoparticles as non-toxic nanocarriers for siRNA transport in cancer cells. GENERAL SIGNIFICANCE: 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.
Authors: Sherouk M Ibrahim; May N Bin Jumah; Sarah I Othman; Reem Saleh Alruhaimi; Nora Al-Khalawi; Yasser F Salama; Ahmed A Allam; Mostafa R Abukhadra Journal: ACS Omega Date: 2021-05-13
Authors: Ilaria Rea; Monica Terracciano; Soundarrajan Chandrasekaran; Nicolas H Voelcker; Principia Dardano; Nicola M Martucci; Annalisa Lamberti; Luca De Stefano Journal: Nanoscale Res Lett Date: 2016-09-15 Impact factor: 4.703
Authors: Meerambika Mishra; Ananta P Arukha; Tufail Bashir; Dhananjay Yadav; G B K S Prasad Journal: Front Microbiol Date: 2017-07-05 Impact factor: 5.640