title: Effects of Polyamidoamine Dendrimers on a 3-D Neurosphere System Using Human Neural Progenitor Cells.
abstract: The practical application of engineered nanomaterials or nanoparticles like polyamidoamine (PAMAM) dendrimers has been promoted in medical devices or industrial uses. The safety of PAMAM dendrimers needs to be assessed when used as a drug carrier to treat brain disease. However, the effects of PAMAM on the human nervous system remain unknown. In this study, human neural progenitor cells cultured as a 3D neurosphere model were used to study the effects of PAMAM dendrimers on the nervous system. Neurospheres were exposed to different G4-PAMAM dendrimers for 72 h at concentrations of 0.3, 1, 3, and 10 μg/ml. The biodistribution was investigated using fluorescence-labeled PAMAM dendrimers, and gene expression was evaluated using microarray analysis followed by pathway and network analysis. Results showed that PAMAM dendrimer nanoparticles can penetrate into neurospheres via superficial cells on them. PAMAM-NH2 but not PAMAM-SC can inhibit neurosphere growth. A reduced number of MAP2-positive cells in flare regions were inhibited after 10 days of differentiation, indicating an inhibitory effect of PAMAM-NH2 on cell proliferation and neuronal migration. A microarray assay showed 32 dendrimer toxicity-related genes, with network analysis showing 3 independent networks of the selected gene targets. Inducible immediate early gene early growth response gene 1 (Egr1), insulin-like growth factor-binding protein 3 (IGFBP3), tissue factor pathway inhibitor (TFPI2), and adrenomedullin (ADM) were the key genes in each network, and the expression of these genes was significantly down regulated. These findings suggest that exposure of neurospheres to PAMAM-NH2 dendrimers affects cell proliferation and migration through pathways regulated by Egr1, IGFBP3, TFPI2, and ADM.
authors.affiliation: *Center for Environmental Risk Research, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8606, Japan Department of Life and Environmental Science, Major of Bioindustrial Sciences, the University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-0006, Japan.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: *Center for Environmental Risk Research, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8606, Japan.
authors.forename: Yoshika
authors.initials: Y
authors.lastname: Kurokawa
authors.affiliation: *Center for Environmental Risk Research, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8606, Japan.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zeng
authors.affiliation: Department of Life and Environmental Science, Major of Bioindustrial Sciences, the University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-0006, Japan Center for Environmental Health Sciences, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8606, Japan.
authors.forename: Tin-Tin
authors.initials: TT
authors.lastname: Win-Shwe
authors.affiliation: *Center for Environmental Risk Research, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8606, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Nansai
authors.affiliation: Department of Life and Environmental Science, Major of Bioindustrial Sciences, the University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-0006, Japan.
authors.forename: Zhenya
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: *Center for Environmental Risk Research, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8606, Japan hsone@nies.go.jp.
authors.forename: Hideko
authors.initials: H
authors.lastname: Sone
mesh_terms: D053607:Adrenomedullin / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D019295:Computational Biology; D030541:Databases, Genetic; D050091:Dendrimers / Q000378:metabolism / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D051766:Early Growth Response Protein 1 / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects; D053263:Gene Regulatory Networks / Q000187:drug effects; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000235:genetics / Q000378:metabolism; D053758:Nanoparticles; D058953:Neural Stem Cells / Q000187:drug effects* / Q000378:metabolism; D055495:Neurogenesis / Q000187:drug effects; D009757:Nylons / Q000378:metabolism / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D018874:Spheroids, Cellular; D013997:Time Factors; D014018:Tissue Distribution; D059467:Transcriptome / Q000187:drug effects
chemical_list: C507862:ADM protein, human; D050091:Dendrimers; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D006023:Glycoproteins; C515497:IGFBP3 protein, human; D018972:Insulin-Like Growth Factor Binding Protein 3; D009757:Nylons; C104700:PAMAM Starburst; C520550:PAMAM-G4; C021240:tissue-factor-pathway inhibitor 2; D053607:Adrenomedullin
keywords: PAMAM dendrimers; human neural progenitor cells; nano toxicology; neural development