title: Silencing of the scavenger receptor (Class B - Type 1) gene using siRNA-loaded chitosan nanaoparticles in a HepG2 cell model.
abstract: Gene silencing mediated by small interfering RNA (siRNA) has gained increasing interest through the past few decades. However, the partial negative charge and the susceptibility to degradation by nucleases have hampered its use in a naked form. In this study, we investigated the use of chitosan nanoparticles as non-viral delivery carriers of siRNA. As a model target, we selected the scavenger receptor (SR-B1), due to its proposed involvement in hepatitis C virus (HCV) internalization. Low molecular weight (LMW) chitosan nanoparticles were prepared by simple ionic gelation using sodium tripolyphosphate (TPP) as a cross-linking agent; a fixed chitosan and TPP concentration of 0.1% was used, and a chitosan to TPP weight ratios of 3:1, 5:1, and 9:1 were investigated. Nanoparticle uptake efficiency was measured using FITC-labeled chitosan nanoparticles and silencing of scavenger receptor class B type 1 (SR-B1) in HepG2 cell line was tested using Western blot analysis. Nanoparticles produced were spherical in shape with an optimum particle size and distribution. The uptake of FITC-labeled nanoparticles by HepG2 cells was found to be both concentration and time dependent. Furthermore, Western Blot analysis showed that SR-B1 siRNA was able to silence the scavenger receptor for up to 96 h of incubation with HepG2 cells.
authors.affiliation: Department of Pharmaceutical Technology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt.
authors.forename: Mariane M
authors.initials: MM
authors.lastname: Farid
authors.affiliation: Department of Pharmaceutical Technology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt; Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt. Electronic address: r_hathout@yahoo.com.
authors.forename: Rania M
authors.initials: RM
authors.lastname: Hathout
authors.affiliation: Department of Pharmaceutical Biology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Fawzy
authors.affiliation: Department of Pharmaceutical Biology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt.
authors.forename: Khaled
authors.initials: K
authors.lastname: Abou-Aisha
mesh_terms: D048271:Chitosan / Q000737:chemistry*; D056945:Hep G2 Cells; D016174:Hepacivirus; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D051116:Receptors, Scavenger / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D051116:Receptors, Scavenger; D048271:Chitosan
keywords: Chitosan nanoparticles; HCV; SR-B1; Scavenger receptor; siRNA