Sample records for nanoparticles exhibits cell

  1. Metallic Nickel Nanoparticles May Exhibit Higher Carcinogenic Potential than Fine Particles in JB6 Cells

    PubMed Central

    Bowman, Linda; Zou, Baobo; Mao, Guochuan; Xu, Jin; Castranova, Vincent; Zhao, Jinshun; Ding, Min

    2014-01-01

    While numerous studies have described the pathogenic and carcinogenic effects of nickel compounds, little has been done on the biological effects of metallic nickel. Moreover, the carcinogenetic potential of metallic nickel nanoparticles is unknown. Activator protein-1 (AP-1) and nuclear factor-?B (NF-?B) have been shown to play pivotal roles in tumor initiation, promotion, and progression. Mutation of the p53 tumor suppressor gene is considered to be one of the steps leading to the neoplastic state. The present study examines effects of metallic nickel fine and nanoparticles on tumor promoter or suppressor gene expressions as well as on cell transformation in JB6 cells. Our results demonstrate that metallic nickel nanoparticles caused higher activation of AP-1 and NF-?B, and a greater decrease of p53 transcription activity than fine particles. Western blot indicates that metallic nickel nanoparticles induced a higher level of protein expressions for R-Ras, c-myc, C-Jun, p65, and p50 in a time-dependent manner. In addition, both metallic nickel nano- and fine particles increased anchorage-independent colony formation in JB6 P+ cells in the soft agar assay. These results imply that metallic nickel fine and nanoparticles are both carcinogenetic in vitro in JB6 cells. Moreover, metallic nickel nanoparticles may exhibit higher carcinogenic potential, which suggests that precautionary measures should be taken in the use of nickel nanoparticles or its compounds in nanomedicine. PMID:24691273

  2. Magnetofluorescent nanoparticles for bimodal detection of breast cancer cells

    Microsoft Academic Search

    Silvia Ronchi; Miriam Colombo; Paolo Verderio; Serena Mazzucchelli; Fabio Corsi; Clara de Palma; Raffaele Allevi; Emilio Clementi; Davide Prosperi

    2010-01-01

    Silica-encapsulated iron oxide composite nanoparticles were synthesized and characterized in terms of morphological and physico-chemical properties. These nanoparticles exhibited both fluorescent and magnetic properties useful for labeling of breast cancer cells. The mechanism of uptake by tumor cells, the pathway of degradation and the potential toxicity of these magnetofluorescent nanoparticles were investigated, suggesting that they could be developed as an

  3. Probing nanoparticle interactions in cell culture media.

    PubMed

    Sabuncu, Ahmet C; Grubbs, Janna; Qian, Shizhi; Abdel-Fattah, Tarek M; Stacey, Michael W; Beskok, Ali

    2012-06-15

    Nanoparticle research is often performed in vitro with little emphasis on the potential role of cell culture medium. In this study, gold nanoparticle interactions with cell culture medium and two cancer cell lines (human T-cell leukemia Jurkat and human pancreatic carcinoma PANC1) were investigated. Gold nanoparticles of 10, 25, 50, and 100 nm in diameter at fixed mass concentration were tested. Size distributions and zeta potentials of gold nanoparticles suspended in deionized (DI) water and Dulbecco's Modified Eagle's Media (DMEM) supplemented with fetal calf serum (FCS) were measured using dynamic light scattering (DLS) technique. In DI water, particle size distributions exhibited peaks around their nominal diameters. However, the gold nanoparticles suspended in DMEM supplemented with FCS formed complexes around 100 nm, regardless of their nominal sizes. The DLS and UV-vis spectroscopy results indicate gold nanoparticle agglomeration in DMEM that is not supplemented by FCS. The zeta potential results indicate that protein rich FCS increases the dispersion quality of gold nanoparticle suspensions through steric effects. Cellular uptake of 25 and 50 nm gold nanoparticles by Jurkat and PANC1 cell lines were investigated using inductively coupled plasma-mass spectroscopy. The intracellular gold level of PANC1 cells was higher than that of Jurkat cells, where 50 nm particles enter cells at faster rates than the 25 nm particles. PMID:22421416

  4. Nanoparticle–polymer photovoltaic cells

    Microsoft Academic Search

    Brian R. Saunders; Michael L. Turner

    2008-01-01

    The need to develop and deploy large-scale, cost-effective, renewable energy is becoming increasingly important. In recent years photovoltaic (PV) cells based on nanoparticles blended with semiconducting polymers have achieved good power conversion efficiencies (PCE). All the nanoparticle types used in these PV cells can be considered as colloids. These include spherical, rod-like or branched organic or inorganic nanoparticles. Nanoparticle–polymer PV

  5. Magnetofluorescent nanoparticles for bimodal detection of breast cancer cells

    NASA Astrophysics Data System (ADS)

    Ronchi, Silvia; Colombo, Miriam; Verderio, Paolo; Mazzucchelli, Serena; Corsi, Fabio; De Palma, Clara; Allevi, Raffaele; Clementi, Emilio; Prosperi, Davide

    2010-10-01

    Silica-encapsulated iron oxide composite nanoparticles were synthesized and characterized in terms of morphological and physico-chemical properties. These nanoparticles exhibited both fluorescent and magnetic properties useful for labeling of breast cancer cells. The mechanism of uptake by tumor cells, the pathway of degradation and the potential toxicity of these magnetofluorescent nanoparticles were investigated, suggesting that they could be developed as an efficient and safe bimodal contrast agent for detection of breast cancer cells.

  6. Thymoquinone poly (lactide-co-glycolide) nanoparticles exhibit enhanced anti-proliferative, anti-inflammatory, and chemosensitization potential.

    PubMed

    Ravindran, Jayaraj; Nair, Hareesh B; Sung, Bokyung; Prasad, Sahdeo; Tekmal, Rajeshwar R; Aggarwal, Bharat B

    2010-06-01

    Thymoquinone (TQ), derived from the medicinal spice Nigella sativa (also called black cumin), has been shown to exhibit anti-inflammatory and anti-cancer activities. In this report we employed polymer-based nanoparticle approach to improve upon its effectiveness and bioavailability. TQ was encapsulated with 97.5% efficiency in biodegradable nanoparticulate formulation based on poly (lactide-co-glycolide) (PLGA) and the stabilizer polyethylene glycol (PEG)-5000. Dynamic laser light scattering and transmission electron microscopy confirmed particle diameter between 150 and 200nm. Electrophoretic gel shift mobility assay showed that TQ nanoparticles (NP) were more active than TQ in inhibiting NF-kappaB activation and in suppressing the expression of cyclin D1, matrix metalloproteinase (MMP)-9, vascular endothelial growth factor (VEGF), those are markers of cell proliferation, metastasis and angiogenesis, respectively. TQ-NP were also more potent than TQ in suppressing proliferation of colon cancer, breast cancer, prostate cancer, and multiple myeloma cells. Esterase staining for plasma membrane integrity revealed that TQ-NP were more potent than TQ in sensitizing leukemic cells to TNF- and paclitaxel-induced apoptosis. Overall our results demonstrate that encapsulation of TQ into nanoparticles enhances its anti-proliferative, anti-inflammatory, and chemosensitizing effects. PMID:20105430

  7. Dielectric spectroscopy of metal nanoparticle doped liquid crystal displays exhibiting frequency modulation response

    Microsoft Academic Search

    Shunsuke Kobayashi; Tomohiro Miyama; Naoto Nishida; Yoshio Sakai; Hiroyuki Shiraki; Yukihide Shiraishi; Naoki Toshima

    2006-01-01

    Twisted nematic liquid crystal displays (TN-LCDs), doped with the nanoparticles of metal, such as Pd, Ag, or Ag-Pd, which are protected with ligand molecules, such as nematic liquid crystal, exhibit a frequency modulation (FM) electro-optical (EO) response with short response time of milliseconds (ms) or sub-ms order together with the ordinary rms voltage response. These devices are called FM\\/AM-TN-LCDs; they

  8. Shear-regulated uptake of nanoparticles by endothelial cells and development of endothelial-targeting nanoparticles.

    PubMed

    Lin, Arthur; Sabnis, Abhimanyu; Kona, Soujanya; Nattama, Sivaniaravindapriya; Patel, Hemang; Dong, Jing-Fei; Nguyen, Kytai T

    2010-06-01

    The purpose of this research project was to develop nanoparticles with improved targeting, adhesion, and cellular uptake to activated or inflamed endothelial cells (ECs) under physiological flow conditions. Our hypothesis is that by mimicking platelet binding to activated ECs through the interaction between platelet glycoprotein Ibalpha (GP Ibalpha) and P-selectin on activated endothelial cells, GP Ibalpha-conjugated nanoparticles could exhibit increased targeting and higher cellular uptake in injured or activated endothelial cells under physiological flow conditions. To test this hypothesis, fluorescent-carboxylated polystyrene nanoparticles were selected for the study as a model particle because of its narrow size distribution as a "proof-of-concept." Using confocal microscopy, fluorescent measurements, and protein assays, cellular uptake properties were characterized for these polystyrene nanoparticles. The study also found that conjugation of 100-nm polystyrene nanoparticles with glycocalicin (the extracellular segment of GP Ibalpha) significantly increased the particle adhesion on P-selectin-coated surfaces and cellular uptake of nanoparticles by activated endothelial cells under physiological flow conditions. The results demonstrate that these novel endothelial-targeting nanoparticles could be the first step toward developing a targeted and sustained drug delivery system that can improve shear-regulated particle adhesion and cellular uptake. PMID:19653303

  9. Novel, silver-ion-releasing nanofibrous scaffolds exhibit excellent antibacterial efficacy without the use of silver nanoparticles.

    PubMed

    Mohiti-Asli, Mahsa; Pourdeyhimi, Behnam; Loboa, Elizabeth G

    2014-05-01

    Nanofibers, with their morphological similarities to the extracellular matrix of skin, hold great potential for skin tissue engineering. Over the last decade, silver nanoparticles have been extensively investigated in wound-healing applications for their ability to provide antimicrobial benefits to nanofibrous scaffolds. However, the use of silver nanoparticles has raised concerns as these particles can penetrate into the stratum corneum of skin, or even diffuse into the cellular plasma membrane. We present and evaluate a new silver ion release polymeric coating that we have found can be applied to biocompatible, biodegradable poly(l-lactic acid) nanofibrous scaffolds. Using this compound, custom antimicrobial silver-ion-releasing nanofibers were created. The presence of a uniform, continuous silver coating on the nanofibrous scaffolds was verified by XPS analysis. The antimicrobial efficacy of the antimicrobial scaffolds against Staphylococcus aureus and Escherichia coli bacteria was determined via industry-standard AATCC protocols. Cytotoxicity analyses of the antimicrobial scaffolds toward human epidermal keratinocytes and human dermal fibroblasts were performed via quantitative analyses of cell viability and proliferation. Our results indicated that the custom antimicrobial scaffolds exhibited excellent antimicrobial properties while also maintaining human skin cell viability and proliferation for silver ion concentrations below 62.5?gml(-1) within the coating solution. This is the first study to show that silver ions can be effectively delivered with nanofibrous scaffolds without the use of silver nanoparticles. PMID:24365706

  10. Fabricating solar cells with silicon nanoparticles

    DOEpatents

    Loscutoff, Paul; Molesa, Steve; Kim, Taeseok

    2014-09-02

    A laser contact process is employed to form contact holes to emitters of a solar cell. Doped silicon nanoparticles are formed over a substrate of the solar cell. The surface of individual or clusters of silicon nanoparticles is coated with a nanoparticle passivation film. Contact holes to emitters of the solar cell are formed by impinging a laser beam on the passivated silicon nanoparticles. For example, the laser contact process may be a laser ablation process. In that case, the emitters may be formed by diffusing dopants from the silicon nanoparticles prior to forming the contact holes to the emitters. As another example, the laser contact process may be a laser melting process whereby portions of the silicon nanoparticles are melted to form the emitters and contact holes to the emitters.

  11. Altered cytoskeletal structures in transformed cells exhibiting obviously metastatic capabilities

    Microsoft Academic Search

    Zhongxiang Lin; Yaling Han; Bingquan Wu; Weigang Fang

    1990-01-01

    Cytoskeletal changes in transformed cells (LM-51) exhibiting obviously metastatic capabilities were investigated by utilization of double-fluorescent labelling through combinations of: (1) tubulin indirect immunofluoreseonce plus Rhodamine-phalloidin staining of F-actins; (2) indirect immunofluorescent staining with ?-actinin polyclonal- and vinculin monoclonal antibodies. The LM-51 cells which showed metastatic index of >50% were derived from lung metastasis in nude mice after subcutaneous inoculation

  12. Apoptotic HPV Positive Cancer Cells Exhibit Transforming Properties

    PubMed Central

    Gaiffe, Emilie; Prétet, Jean-Luc; Launay, Sophie; Jacquin, Elise; Saunier, Maëlle; Hetzel, Geneviève; Oudet, Pierre; Mougin, Christiane

    2012-01-01

    Previous studies have shown that DNA can be transferred from dying engineered cells to neighboring cells through the phagocytosis of apoptotic bodies, which leads to cellular transformation. Here, we provide evidence of an uptake of apoptotic-derived cervical cancer cells by human mesenchymal cells. Interestingly, HeLa (HPV 18+) or Ca Ski (HPV16+) cells, harboring integrated high-risk HPV DNA but not C-33 A cells (HPV-), were able to transform the recipient cells. Human primary fibroblasts engulfed the apoptotic bodies effectively within 30 minutes after co-cultivation. This mechanism is active and involves the actin cytoskeleton. In situ hybridization of transformed fibroblasts revealed the presence of HPV DNA in the nucleus of a subset of phagocytosing cells. These cells expressed the HPV16/18 E6 gene, which contributes to the disruption of the p53/p21 pathway, and the cells exhibited a tumorigenic phenotype, including an increased proliferation rate, polyploidy and anchorage independence growth. Such horizontal transfer of viral oncogenes to surrounding cells that lack receptors for HPV could facilitate the persistence of the virus, the main risk factor for cervical cancer development. This process might contribute to HPV-associated disease progression in vivo. PMID:22574222

  13. Green synthesis of silver nanoparticles using Delphinium denudatum root extract exhibits antibacterial and mosquito larvicidal activities

    NASA Astrophysics Data System (ADS)

    Suresh, Gopal; Gunasekar, Poosali Hariharan; Kokila, Dhanasegaran; Prabhu, Durai; Dinesh, Devadoss; Ravichandran, Nagaiya; Ramesh, Balasubramanian; Koodalingam, Arunagirinathan; Vijaiyan Siva, Ganesan

    2014-06-01

    Green synthesis of silver nanoparticles (AgNPs) using aqueous root extract of Delphinium denudatum (Dd) by reduction of Ag+ ions from silver nitrate solution has been investigated. The synthesized DdAgNPs were characterized by using UV-Vis spectroscopy, X-ray diffraction (XRD), Field emission scanning electron microscope (FESEM) and Fourier transform infrared spectroscopy (FTIR). The prepared DdAgNPs showed maximum absorbance at 416 nm and particles were polydispersed in nature, spherical in shape and the size of the particle obtained was ?85 nm. The DdAgNPs exhibited antibacterial activity against Staphylococcus aureus ATCC 6538, Bacillus cereus NCIM 2106, Escherichia coli ATCC 8739 and Pseudomonas aeruginosa ATCC 9027. The DdAgNPs showed potent larvicidal activity against second instar larvae of dengue vector Aedes aegypti with a LC50 value of 9.6 ppm.

  14. Green synthesis of silver nanoparticles using Delphinium denudatum root extract exhibits antibacterial and mosquito larvicidal activities.

    PubMed

    Suresh, Gopal; Gunasekar, Poosali Hariharan; Kokila, Dhanasegaran; Prabhu, Durai; Dinesh, Devadoss; Ravichandran, Nagaiya; Ramesh, Balasubramanian; Koodalingam, Arunagirinathan; Vijaiyan Siva, Ganesan

    2014-06-01

    Green synthesis of silver nanoparticles (AgNPs) using aqueous root extract of Delphinium denudatum (Dd) by reduction of Ag(+) ions from silver nitrate solution has been investigated. The synthesized DdAgNPs were characterized by using UV-Vis spectroscopy, X-ray diffraction (XRD), Field emission scanning electron microscope (FESEM) and Fourier transform infrared spectroscopy (FTIR). The prepared DdAgNPs showed maximum absorbance at 416nm and particles were polydispersed in nature, spherical in shape and the size of the particle obtained was?85nm. The DdAgNPs exhibited antibacterial activity against Staphylococcus aureus ATCC 6538, Bacillus cereus NCIM 2106, Escherichia coli ATCC 8739 and Pseudomonas aeruginosa ATCC 9027. The DdAgNPs showed potent larvicidal activity against second instar larvae of dengue vector Aedes aegypti with a LC50 value of 9.6ppm. PMID:24632157

  15. Cytotoxicity of monodispersed chitosan nanoparticles against the Caco-2 cells

    SciTech Connect

    Loh, Jing Wen [Laboratory for Drug Delivery, Pharmacy, Characterisation and Analysis, University of Western Australia (Australia)] [Laboratory for Drug Delivery, Pharmacy, Characterisation and Analysis, University of Western Australia (Australia); Saunders, Martin [Centre for Microscopy, Characterisation and Analysis, University of Western Australia (Australia)] [Centre for Microscopy, Characterisation and Analysis, University of Western Australia (Australia); Lim, Lee-Yong, E-mail: lee.lim@uwa.edu.au [Laboratory for Drug Delivery, Pharmacy, Characterisation and Analysis, University of Western Australia (Australia) [Laboratory for Drug Delivery, Pharmacy, Characterisation and Analysis, University of Western Australia (Australia); School of Biomedical, Biomolecular and Chemical Sciences, 35 Stirling Hwy, Crawley 6009 (Australia)

    2012-08-01

    Published toxicology data on chitosan nanoparticles (NP) often lack direct correlation to the in situ size and surface characteristics of the nanoparticles, and the repeated NP assaults as experienced in chronic use. The aim of this paper was to breach these gaps. Chitosan nanoparticles synthesized by spinning disc processing were characterised for size and zeta potential in HBSS and EMEM at pHs 6.0 and 7.4. Cytotoxicity against the Caco-2 cells was evaluated by measuring the changes in intracellular mitochondrial dehydrogenase activity, TEER and sodium fluorescein transport data and cell morphology. Cellular uptake of NP was observed under the confocal microscope. Contrary to established norms, the collective data suggest that the in vitro cytotoxicity of NP against the Caco-2 cells was less influenced by positive surface charges than by the particle size. Particle size was in turn determined by the pH of the medium in which the NP was dispersed, with the mean size ranging from 25 to 333 nm. At exposure concentration of 0.1%, NP of 25 ± 7 nm (zeta potential 5.3 ± 2.8 mV) was internalised by the Caco-2 cells, and the particles were observed to inflict extensive damage to the intracellular organelles. Concurrently, the transport of materials along the paracellular pathway was significantly facilitated. The Caco-2 cells were, however, capable of recovering from such assaults 5 days following NP removal, although a repeat NP exposure was observed to produce similar effects to the 1st exposure, with the cells exhibiting comparable resiliency to the 2nd assault. -- Highlights: ? Chitosan nanoparticles reduced mitochondrial dehydrogenase activity. ? Cellular uptake of chitosan nanoparticles was observed. ? Chitosan nanoparticles inflicted extensive damage to the cell morphology. ? The transport of materials along the paracellular pathway was facilitated.

  16. Planktonic and biofilm-grown nitrogen-cycling bacteria exhibit different susceptibilities to copper nanoparticles.

    PubMed

    Reyes, Vincent C; Opot, Stephen O; Mahendra, Shaily

    2015-04-01

    Proper characterization of nanoparticle (NP) interactions with environmentally relevant bacteria under representative conditions is necessary to enable their sustainable manufacture, use, and disposal. Previous nanotoxicology research based on planktonic growth has not adequately explored biofilms, which serve as the predominant mode of bacterial growth in natural and engineered environments. Copper nanoparticle (Cu-NP) impacts on biofilms were compared with respective planktonic cultures of the ammonium-oxidizing Nitrosomonas europaea, nitrogen-fixing Azotobacter vinelandii, and denitrifying Paracoccus denitrificans using a suite of independent toxicity diagnostics. Median inhibitory concentration (IC50) values derived from adenosine triphosphate (ATP) for Cu-NPs were lower in N. europaea biofilms (19.6?±?15.3?mg/L) than in planktonic cells (49.0?±?8.0?mg/L). However, in absorbance-based growth assays, compared with unexposed controls, N. europaea growth rates in biofilms were twice as resilient to inhibition than those in planktonic cultures. Similarly, relative to unexposed controls, growth rates and yields of P. denitrificans in biofilms exposed to Cu-NPs were 40-fold to 50-fold less inhibited than those in planktonic cells. Physiological evaluation of ammonium oxidation and nitrate reduction suggested that biofilms were also less inhibited by Cu-NPs than planktonic cells. Furthermore, functional gene expression for ammonium oxidation (amoA) and nitrite reduction (nirK) showed lower inhibition by NPs in biofilms relative to planktonic-grown cells. These results suggest that biofilms mitigate NP impacts, and that nitrogen-cycling bacteria in wastewater, wetlands, and soils might be more resilient to NPs than planktonic-based assessments suggest. Environ Toxicol Chem 2015;34:887-897. © 2014 SETAC. PMID:25556815

  17. Induced T cell cytokine production is enhanced by engineered nanoparticles

    PubMed Central

    Chen, Weimin; Zhang, Quanxuan; Kaplan, Barbara L. F.; Baker, Gregory L.; Kaminski, Norbert E.

    2014-01-01

    Engineered nanoparticles are widely used in commercial products, and yet due to the paucity of safety information, there are concerns surrounding potential adverse health effects, especially from inhaled nanoparticles and their putative contribution to allergic airway disease. The objective of this study was to investigate whether size or surface chemistry of engineered nanoparticles can influence the immune enhancing properties of these agents on antigen-specific T cell responses. Ovalbumin (OVA)-derived peptides were presented to T cells by either spleen-derived endogenous antigen presenting cells or a mouse dendritic cell (DC) line, DC2.4. In all models, interferon (IFN)-? and interleukin (IL)-2 production by CD8+ or CD4+ T cells in response to peptide OVA257–264 or OVA323–339, respectively, was measured by flow cytometry. To address the study objective, silica nanoparticles (SNPs) were modified with alkyne-terminated surfaces and appended with polyethylene glycol chains via “click” chemistry. These modified SNPs were resistant to agglomerate in in vitro culture media, suggesting that their modulation of T cell responses is the result of true nanoscale-mediated effects. Under conditions of suboptimal T-cell activation, modified SNPs (up to 10 ?g/ml) enhanced the proportion of CD8+, but not CD4+, T cells producing IFN-? and IL-2. Various functional groups (–COOH, –NH2 and –OH) on modified SNPs enhanced IFN-? and IL-2 production to different levels, with –COOH SNPs being the most effective. Furthermore, 51 nm –COOH SNPs exhibited a greater enhancing effect on the CD8+ T cell response than other sized particles. Collectively, our results show that modified SNPs can enhance antigen-specific CD8+ T cell responses, suggesting that certain modified SNPs exhibit potential adjuvant-like properties. PMID:24256152

  18. Encapsulation of plasmid DNA in calcium phosphate nanoparticles: stem cell uptake and gene transfer efficiency

    PubMed Central

    Cao, Xia; Deng, Wenwen; Wei, Yuan; Su, Weiyan; Yang, Yan; Wei, Yawei; Yu, Jiangnan; Xu, Ximing

    2011-01-01

    Background The purpose of this study was to develop calcium phosphate nanocomposite particles encapsulating plasmid DNA (CP-pDNA) nanoparticles as a nonviral vector for gene delivery. Methods CP-pDNA nanoparticles employing plasmid transforming growth factor beta 1 (TGF-?1) were prepared and characterized. The transfection efficiency and cell viability of the CP-pDNA nanoparticles were evaluated in mesenchymal stem cells, which were identified by immunofluorescence staining. Cytotoxicity of plasmid TGF-?1 and calcium phosphate to mesenchymal stem cells were evaluated by MTT assay. Results The integrity of TGF-?1 encapsulated in the CP-pDNA nanoparticles was maintained. The well dispersed CP-pDNA nanoparticles exhibited an ultralow particle size (20–50 nm) and significantly lower cytotoxicity than Lipofectamine™ 2000. Immunofluorescence staining revealed that the cultured cells in this study were probably mesenchymal stem cells. The cellular uptake and transfection efficiency of the CP-pDNA nanoparticles into the mesenchymal stem cells were higher than that of needle-like calcium phosphate nanoparticles and a standard calcium phosphate transfection kit. Furthermore, live cell imaging and confocal laser microscopy vividly showed the transportation process of the CP-pDNA nanoparticles in mesenchymal stem cells. The results of a cytotoxicity assay found that both plasmid TGF-?1 and calcium phosphate were not toxic to mesenchymal stem cells. Conclusion CP-pDNA nanoparticles can be developed into an effective alternative as a nonviral gene delivery system that is highly efficient and has low cytotoxicity. PMID:22229000

  19. Targeted biodegradable nanoparticles for drug delivery to smooth muscle cells.

    PubMed

    Kona, Soujanya; Specht, Danyel; Rahimi, Maham; Shah, Bhavik P; Gilbertson, Timothy A; Nguyen, Kytai T

    2012-01-01

    Targeted delivery of therapeutic agents to prevent smooth muscle cell (SMC) proliferation is important in averting restenosis (a narrowing of blood vessels). Since platelet derived growth factor (PDGF) receptors are over-expressed in proliferating SMCs after injury from cardiovascular interventions, such as angioplasty and stent implantation, our hypothesis is that conjugation of PDGF-BB (platelet-derived growth factor BB (homodimer)) peptides to biodegradable poly (D,L-lactic-co-glycolide) (PLGA) nanoparticles (NPs) would exhibit an increased uptake of these NPs by proliferating SMCs. In this study, poly (D,L-lactide-co-glycolide) (PLGA) nanoparticles containing dexamethasone were formulated and conjugated with PDGF-BB peptides. These NPs were stable, biocompatible, and exhibited a sustained drug release over 14 days. Various particle uptake studies using HASMCs (human aortic smooth muscle cells) demonstrated that PDGF-BB peptide-conjugated nanoparticles significantly increased cellular uptake and decreased proliferation of HASMCs compared to control nanoparticles (without conjugation of PDGF-BB peptides). These NPs were internalized primarily by clathrin-mediated endocytosis and macropinocytosis. Our in vitro results suggest that PDGF-BB peptide-conjugated NPs could represent as an effective targeted, sustained therapeutic delivery system to reduce restenosis and neointimal hyperplasia. PMID:22523971

  20. Stem cell tracking with optically active nanoparticles

    PubMed Central

    Gao, Yu; Cui, Yan; Chan, Jerry KY; Xu, Chenjie

    2013-01-01

    Stem-cell-based therapies hold promise and potential to address many unmet clinical needs. Cell tracking with modern imaging modalities offers insight into the underlying biological process of the stem-cell-based therapies, with the goal to reveal cell survival, migration, homing, engraftment, differentiation, and functions. Adaptability, sensitivity, resolution, and non-invasiveness have contributed to the longstanding use of optical imaging for stem cell tracking and analysis. To identify transplanted stem cells from the host tissue, optically active probes are usually used to label stem cells before the administration. In comparison to the traditional fluorescent probes like fluorescent proteins and dyes, nanoparticle-based probes are advantageous in terms of the photo-stabilities and minimal changes to the cell phenotype. The main focus here is to overview the recent development of optically active nanoparticles for stem cells tracking. The related optical imaging modalities include fluorescence imaging, photoacoustic imaging, Raman and surface enhanced Raman spectroscopy imaging. PMID:23638335

  1. Protamine sulfate-nanodiamond hybrid nanoparticles as a vector for MiR-203 restoration in esophageal carcinoma cells.

    PubMed

    Cao, Minjun; Deng, Xiongwei; Su, Shishuai; Zhang, Fang; Xiao, Xiangqian; Hu, Qin; Fu, Yongwei; Yang, Burton B; Wu, Yan; Sheng, Wang; Zeng, Yi

    2013-12-21

    We report an innovative approach for miRNA-203 delivery in esophageal cancer cells using protamine sulphate (PS)-nanodiamond (ND) nanoparticles. The efficient delivery of miR-203 significantly suppressed the proliferation and migration of cancer cells through targeting Ran and ?Np63, exhibiting a great potential for PS@ND nanoparticles in miRNA-based cancer therapy. PMID:24154605

  2. Interaction of dermatologically relevant nanoparticles with skin cells and skin

    PubMed Central

    Rancan, Fiorenza; Ahlberg, Sebastian; Nazemi, Berouz; Choe, Chun Sik; Darvin, Maxim E; Hadam, Sabrina; Blume-Peytavi, Ulrike; Loza, Kateryna; Diendorf, Jörg; Epple, Matthias; Graf, Christina; Rühl, Eckart; Meinke, Martina C; Lademann, Jürgen

    2014-01-01

    Summary The investigation of nanoparticle interactions with tissues is complex. High levels of standardization, ideally testing of different material types in the same biological model, and combinations of sensitive imaging and detection methods are required. Here, we present our studies on nanoparticle interactions with skin, skin cells, and biological media. Silica, titanium dioxide and silver particles were chosen as representative examples for different types of skin exposure to nanomaterials, e.g., unintended environmental exposure (silica) versus intended exposure through application of sunscreen (titanium dioxide) or antiseptics (silver). Because each particle type exhibits specific physicochemical properties, we were able to apply different combinations of methods to examine skin penetration and cellular uptake, including optical microscopy, electron microscopy, X-ray microscopy on cells and tissue sections, flow cytometry of isolated skin cells as well as Raman microscopy on whole tissue blocks. In order to assess the biological relevance of such findings, cell viability and free radical production were monitored on cells and in whole tissue samples. The combination of technologies and the joint discussion of results enabled us to look at nanoparticle–skin interactions and the biological relevance of our findings from different angles. PMID:25551064

  3. ZnFe2O4 nanoparticles as radiosensitizers in radiotherapy of human prostate cancer cells.

    PubMed

    Meidanchi, Alireza; Akhavan, Omid; Khoei, Samideh; Shokri, Ali A; Hajikarimi, Zahra; Khansari, Nakisa

    2015-01-01

    Nanoparticles of high-Z elements exhibit stronger photoelectric effects than soft tissues under gamma irradiation. Hence, they can be used as effective radiosensitizers for increasing the efficiency of current radiotherapy. In this work, superparamagnetic zinc ferrite spinel (ZnFe2O4) nanoparticles were synthesized by a hydrothermal reaction method and used as radiosensitizers in cancer therapy. The magnetic nanoparticles showed fast separation from solutions (e.g., ~1 min for 2 mg mL(-1) of the nanoparticles in ethanol) by applying an external magnetic field (~1T). The ZnFe2O4 nanoparticles were applied in an in vitro radiotherapy of lymph node carcinoma of prostate cells (as high radioresistant cells) under gamma irradiation of (60)Co source. The nanoparticles exhibited no significant effects on the cancer cells up to the high concentration of 100 ?g mL(-1), in the absence of gamma irradiation. The gamma irradiation alone (2Gy dose) also showed no significant effects on the cells. However, gamma irradiation in the presence of 100 ?g mL(-1) ZnFe2O4 nanoparticles resulted in ~53% inactivation of the cells (~17 times higher than the inactivation that occurred under gamma irradiation alone) after 24h. The higher cell inactivation was assigned to interaction of gamma radiation with nanoparticles (photoelectric effect), resulting in a high level electron release in the media of the radioresistant cells. Our results indicated that ZnFe2O4 nanoparticles not only can be applied in increasing the efficiency of radiotherapy, but also can be easily separated from the cell environment by using an external magnetic field after the radiotherapy. PMID:25492003

  4. Nanoparticle Solar Cell Final Technical Report

    SciTech Connect

    Breeze, Alison, J; Sahoo, Yudhisthira; Reddy, Damoder; Sholin, Veronica; Carter, Sue

    2008-06-17

    The purpose of this work was to demonstrate all-inorganic nanoparticle-based solar cells with photovoltaic performance extending into the near-IR region of the solar spectrum as a pathway towards improving power conversion efficiencies. The field of all-inorganic nanoparticle-based solar cells is very new, with only one literature publication in the prior to our project. Very little is understood regarding how these devices function. Inorganic solar cells with IR performance have previously been fabricated using traditional methods such as physical vapor deposition and sputtering, and solution-processed devices utilizing IR-absorbing organic polymers have been investigated. The solution-based deposition of nanoparticles offers the potential of a low-cost manufacturing process combined with the ability to tune the chemical synthesis and material properties to control the device properties. This work, in collaboration with the Sue Carter research group at the University of California, Santa Cruz, has greatly expanded the knowledge base in this field, exploring multiple material systems and several key areas of device physics including temperature, bandgap and electrode device behavior dependence, material morphological behavior, and the role of buffer layers. One publication has been accepted to Solar Energy Materials and Solar Cells pending minor revision and another two papers are being written now. While device performance in the near-IR did not reach the level anticipated at the beginning of this grant, we did observe one of the highest near-IR efficiencies for a nanoparticle-based solar cell device to date. We also identified several key parameters of importance for improving both near-IR performance and nanoparticle solar cells in general, and demonstrated multiple pathways which showed promise for future commercialization with further research.

  5. Analysis of the Cytotoxicity of Carbon-Based Nanoparticles, Diamond and Graphite, in Human Glioblastoma and Hepatoma Cell Lines

    PubMed Central

    Wierzbicki, Mateusz; Jaworski, S?awomir; Kutwin, Marta; Sawosz, Ewa; Chwalibog, André; Pijanowska, Dorota Genowefa; Pluta, Krzysztof Dariusz

    2015-01-01

    Nanoparticles have attracted a great deal of attention as carriers for drug delivery to cancer cells. However, reports on their potential cytotoxicity raise questions of their safety and this matter needs attentive consideration. In this paper, for the first time, the cytotoxic effects of two carbon based nanoparticles, diamond and graphite, on glioblastoma and hepatoma cells were compared. First, we confirmed previous results that diamond nanoparticles are practically nontoxic. Second, graphite nanoparticles exhibited a negative impact on glioblastoma, but not on hepatoma cells. The studied carbon nanoparticles could be a potentially useful tool for therapeutics delivery to the brain tissue with minimal side effects on the hepatocytes. Furthermore, we showed the influence of the nanoparticles on the stable, fluorescently labeled tumor cell lines and concluded that the labeled cells are suitable for drug cytotoxicity tests. PMID:25816103

  6. Lunar soil simulant and synthesized nanoparticles of magnetite exhibit diverse neurotoxic potential

    NASA Astrophysics Data System (ADS)

    Borisova, Tatiana; Krisanova, Natalia; Sivko, Roman; Nazarova, Anastasiya; Borysov, Arseniy

    Lunar soli simulant can be deleterious to human physiology and the components of lunar soil may be internalized by lung epithelium and may overcome the blood-brain barrier. Nanoparticles of ferric oxide are one of the components of Lunar soil simulants. Neurotoxic potential of lunar soil simulant and synthesized nanoparticles of magnetite was analyzed. The size of particles, their effects on membrane potential, acidification of synaptic vesicles, uptake and ambient level of glutamate, which is the major excitatory neurotransmitter in the CNS, were studied in isolated rat brain nerve terminals (synaptosomes) using photon correlation spectroscopy, spectrofluorimetry, radiolabeled assay, respectively. No significant effect of Lunar soil simulant and synthesized nanoparticles of magnetite on acidification of synaptic vesicles were found in synaptosomes. Also, nanoparticles did not influence the potential of the plasma membrane of synaptosomes. Unspecific binding of L-[14C]glutamate to synaptosomes was not altered by nanoparticles of magnetite, whereas in the presence of Lunar soil simulant this parameter was changed. Thus, it was suggested that Lunar soil simulant might disturb glutamate homeostasis in the mammalian CNS.

  7. Cell toxicity of superparamagnetic iron oxide nanoparticles.

    PubMed

    Mahmoudi, M; Simchi, A; Milani, A S; Stroeve, P

    2009-08-15

    The performance of nanoparticles for biomedical applications is often assessed by their narrow size distribution, suitable magnetic saturation and low toxicity effects. In this work, superparamagnetic iron oxide nanoparticles (SPIONs) with different size, shape and saturation magnetization levels were synthesized via a co-precipitation technique using ferrous salts with a Fe(3+)/Fe(2+) mole ratio equal to 2. A parametric study is conducted, based on a uniform design-of-experiments methodology and a critical polymer/iron mass ratio (r-ratio) for obtaining SPION with narrow size distribution, suitable magnetic saturation, and optimum biocompatibility is identified. Polyvinyl alcohol (PVA) has been used as the nanoparticle coating material, owing to its low toxicity. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay is used to investigate the cell biocompatibility/toxicity effects of the samples. From the MTT assay results, it is observed that the biocompatibility of the nanoparticles, based on cell viabilities, can be enhanced by increasing the r-ratio, regardless of the stirring rate. This effect is mainly due to the growth of the particle hydrodynamic size, causing lower cell toxicity effects. PMID:19476952

  8. Endometriotic mesenchymal stem cells exhibit a distinct immune phenotype.

    PubMed

    Koippallil Gopalakrishnan Nair, Aghila Rani; Pandit, Hrishikesh; Warty, Neeta; Madan, Taruna

    2015-04-01

    Endometriosis is a significant debilitating gynecological problem affecting women of the reproductive age group and post-menopause. Recent reports suggest a role for endometriotic mesenchymal stem cells (ectopic MSCs) in the pathogenesis of endometriosis. To investigate the plausible mechanisms leading to the pathogenic behavior of ectopic MSCs, we compared the immunomodulatory properties of eutopic (healthy) and ectopic MSCs. We analyzed MSC phenotypes, differentiation potential, differential gene expression for an array of pattern recognition receptors (PRRs) and pro-inflammatory cytokine release along with markers of migration and angiogenesis among eutopic and ectopic MSCs. Further, alterations in immunosuppressive functions of eutopic and ectopic MSCs were examined by co-culturing them with mitogen-activated allogeneic PBMCs. Transcripts of PRRs such as all Toll-like receptors (TLR1-10), except TLR8, collectins (CL-L1, CL-P1 and CL-K1), NOD-1 and NOD-2 receptors and secreted pro-inflammatory cytokines like IL-6, IFN-?, vascular endothelial growth factor (VEGF), epidermal growth factor and MCP-1 were significantly up-regulated in ectopic MSCs. The anti-inflammatory cytokine transforming growth factor-? showed significant down-regulation, while IL-10 showed a significant increase in ectopic MSCs. Further, ectopic MSCs showed up-regulated expression for markers of migration and angiogenesis such as matrix metalloproteinase-2 (MMP-2), MMP-3 and MMP-9 and VEGF, respectively. We report here that proliferation of PBMCs was less inhibited upon co-culture with ectopic MSCs compared with eutopic MSCs. The findings suggest that ectopic MSCs with increased levels of TLRs, collectins, pro-inflammatory cytokines and markers of migration and angiogenesis exhibit a distinct immune phenotype compared to eutopic MSCs. This distinct phenotype may be responsible for the reduced immunosuppressive property of ectopic MSCs and may be associated with the pathogenesis of endometriosis. PMID:25416515

  9. Improvement of the separation of tumour cells from peripheral blood cells using magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Schwalbe, M.; Pachmann, K.; Höffken, K.; Clement, J. H.

    2006-09-01

    Circulating tumour cells are a key challenge in tumour therapy. Numerous approaches are on the way to achieving the elimination of these potential sources of metastasis formation. Antibody-directed magnetic cell sorting is supposed to enrich tumour cells with high selectivity, but low efficiency. The short term application of carboxymethyl dextran (CMD) coated magnetit/maghemit nanoparticles allows the discrimination of tumour cells from leukocytes. In the present work we show that the interaction of CMD nanoparticles is cell-type specific and time dependent. The breast cancer cell line MCF-7 and the CML cell line K-562 are characterized by a rapid and high interaction rate, whereas leukocytes exhibit a decelerated behaviour. The addition of carboxymethyl dextran or glucose stimulated the magnetic labelling of leukocytes. The variation of the degree of substitution of dextran with carboxymethyl groups did not affect the labelling profile of leukocytes and MCF-7 cells. In order to verify the in vitro results, whole blood samples from 13 cancer patients were analysed ex vivo. Incubation of the purified leukocyte fraction with CMD nanoparticles in the presence of low amounts of plasma reduced the overall cell content in the positive fraction. In contrast, the absolute number of residual tumour cells in the positive fraction was 90% of the initial amount.

  10. Nanoparticle-based monitoring of cell therapy

    PubMed Central

    Xu, Chenjie; Mu, Luye; Roes, Isaac; Miranda-Nieves, David; Nahrendorf, Matthias; Ankrum, James A; Zhao, Weian; Karp, Jeffrey M

    2012-01-01

    Exogenous cell therapy aims to replace/repair diseased or dysfunctional cells and promises to revolutionize medicine by restoring tissue and organ function. To develop effective cell therapy, the location, distribution and long-term persistence of transplanted cells must be evaluated. Nanoparticle (NP) based imaging technologies have the potential to track transplanted cells non-invasively. Here we summarize the most recent advances in NP-based cell tracking with emphasis on (1) the design criteria for cell tracking NPs, (2) protocols for cell labeling, (3) a comparison of available imaging modalities and their corresponding contrast agents, (4) a summary of preclinical studies on NP-based cell tracking and finally (5) perspectives and future directions. PMID:22101191

  11. Biocompatibility of cerium dioxide and silicon dioxide nanoparticles with endothelial cells.

    PubMed

    Strobel, Claudia; Förster, Martin; Hilger, Ingrid

    2014-01-01

    Cerium dioxide (CeO2) and silicon dioxide (SiO2) nanoparticles are of widespread use in modern life. This means that human beings are markedly exposed to them in their everyday life. Once passing biological barriers, these nanoparticles are expected to interact with endothelial cells, leading to systemic alterations with distinct influences on human health. In the present study we observed the metabolic impact of differently sized CeO2 (8 nm; 35 nm) and SiO2 nanoparticles (117 nm; 315 nm) on immortalized human microvascular (HMEC-1) and primary macrovascular endothelial cells (HUVEC), with particular focus on the CeO2 nanoparticles. The characterization of the CeO2 nanoparticles in cell culture media with varying serum content indicated a steric stabilization of nanoparticles due to interaction with proteins. After cellular uptake, the CeO2 nanoparticles were localized around the nucleus in a ring-shaped manner. The nanoparticles revealed concentration and time, but no size-dependent effects on the cellular adenosine triphosphate levels. HUVEC reacted more sensitively to CeO2 nanoparticle exposure than HMEC-1. This effect was also observed in relation to cytokine release after nanoparticle treatment. The CeO2 nanoparticles exhibited a specific impact on the release of diverse proteins. Namely, a slight trend towards pro-inflammatory effects, a slight pro-thrombotic impact, and an increase of reactive oxygen species after nanoparticle exposure were observed with increasing incubation time. For SiO2 nanoparticles, concentration- and time-dependent effects on the metabolic activity as well as pro-inflammatory reactions were detectable. In general, the effects of the investigated nanoparticles on endothelial cells were rather insignificant, since the alterations on the metabolic cell activity became visible at a nanoparticle concentration that is by far higher than those expected to occur in the in vivo situation (CeO2 nanoparticles: 100 µg/mL; SiO2 nanoparticles: 10 µg/mL). PMID:25383291

  12. Biocompatibility of cerium dioxide and silicon dioxide nanoparticles with endothelial cells

    PubMed Central

    Strobel, Claudia; Förster, Martin

    2014-01-01

    Summary Cerium dioxide (CeO2) and silicon dioxide (SiO2) nanoparticles are of widespread use in modern life. This means that human beings are markedly exposed to them in their everyday life. Once passing biological barriers, these nanoparticles are expected to interact with endothelial cells, leading to systemic alterations with distinct influences on human health. In the present study we observed the metabolic impact of differently sized CeO2 (8 nm; 35 nm) and SiO2 nanoparticles (117 nm; 315 nm) on immortalized human microvascular (HMEC-1) and primary macrovascular endothelial cells (HUVEC), with particular focus on the CeO2 nanoparticles. The characterization of the CeO2 nanoparticles in cell culture media with varying serum content indicated a steric stabilization of nanoparticles due to interaction with proteins. After cellular uptake, the CeO2 nanoparticles were localized around the nucleus in a ring-shaped manner. The nanoparticles revealed concentration and time, but no size-dependent effects on the cellular adenosine triphosphate levels. HUVEC reacted more sensitively to CeO2 nanoparticle exposure than HMEC-1. This effect was also observed in relation to cytokine release after nanoparticle treatment. The CeO2 nanoparticles exhibited a specific impact on the release of diverse proteins. Namely, a slight trend towards pro-inflammatory effects, a slight pro-thrombotic impact, and an increase of reactive oxygen species after nanoparticle exposure were observed with increasing incubation time. For SiO2 nanoparticles, concentration- and time-dependent effects on the metabolic activity as well as pro-inflammatory reactions were detectable. In general, the effects of the investigated nanoparticles on endothelial cells were rather insignificant, since the alterations on the metabolic cell activity became visible at a nanoparticle concentration that is by far higher than those expected to occur in the in vivo situation (CeO2 nanoparticles: 100 µg/mL; SiO2 nanoparticles: 10 µg/mL). PMID:25383291

  13. Surface-charge-dependent cell localization and cytotoxicity of cerium oxide nanoparticles.

    PubMed

    Asati, Atul; Santra, Santimukul; Kaittanis, Charalambos; Perez, J Manuel

    2010-09-28

    Cerium oxide nanoparticles (nanoceria) have shown great potential as antioxidant and radioprotective agents for applications in cancer therapy. Recently, various polymer-coated nanoceria preparations have been developed to improve their aqueous solubility and allow for surface functionalization of these nanoparticles. However, the interaction of polymer-coated nanoceria with cells, their uptake mechanism, and subcellular localization are poorly understood. Herein, we engineered polymer-coated cerium oxide nanoparticles with different surface charges (positive, negative, and neutral) and studied their internalization and toxicity in normal and cancer cell lines. The results showed that nanoceria with a positive or neutral charge enters most of the cell lines studied, while nanoceria with a negative charge internalizes mostly in the cancer cell lines. Moreover, upon entry into the cells, nanoceria is localized to different cell compartments (e.g., cytoplasm and lysosomes) depending on the nanoparticle's surface charge. The internalization and subcellular localization of nanoceria plays a key role in the nanoparticles' cytotoxicity profile, exhibiting significant toxicity when they localize in the lysosomes of the cancer cells. In contrast, minimal toxicity is observed when they localize into the cytoplasm or do not enter the cells. Taken together, these results indicate that the differential surface-charge-dependent localization of nanoceria in normal and cancer cells plays a critical role in the nanoparticles' toxicity profile. PMID:20690607

  14. Magnetic nanoparticles coated with carboxymethylated polysaccharide shells—Interaction with human cells

    NASA Astrophysics Data System (ADS)

    Wotschadlo, Jana; Liebert, Tim; Heinze, Thomas; Wagner, Kerstin; Schnabelrauch, Matthias; Dutz, Silvio; Müller, Robert; Steiniger, Frank; Schwalbe, Manuela; Kroll, Torsten C.; Höffken, Klaus; Buske, Norbert; Clement, Joachim H.

    2009-05-01

    The interaction of magnetic core shell nanoparticles with living cells depends on the structure of the shell. In this paper we demonstrate a strong difference in the cell-nanoparticle interaction depending on the backbone of carboxymethylated polysaccharides used as shell material. Carboxymethyl cellulose with its ?-1?4 linked structure and the carboxymethylated pullulan [?-1?6 linked maltotriose with ?-1?4 linkages] show a constant interaction rate with both, tumor cells and leukocytes. In contrast, carboxymethyl dextran with a ?-1?6 linked backbone exhibits a rapid interaction kinetic with tumor cells that is reduced with leukocytes as target.

  15. Synergistic Targeting of Cell Membrane, Cytoplasm and Nucleus of Cancer Cells using Rod-Shaped Nanoparticles

    PubMed Central

    Barua, Sutapa; Mitragotri, Samir

    2014-01-01

    Design of carriers for effective delivery and targeting of drugs to cellular and sub-cellular compartments is an unmet need in medicine. Here, we report pure drug nanoparticles comprising camptothecin (CPT), trastuzumab (TTZ) and doxorubicin (DOX) to enable cell-specific interactions, subcellular accumulation and growth inhibition of breast cancer cells. CPT is formulated in the form of nanorods which are coated with TTZ. DOX is encapsulated in the TTZ corona around the CPT nanoparticle. Our results show that TTZ/DOX-coated CPT nanorods exhibit cell-specific internalization in BT-474 breast cancer cells, after which TTZ is recycled to the plasma membrane leaving CPT nanorods in the perinuclear region and delivering DOX into the nucleus of the cells. The effects of CPT-TTZ-DOX nanoparticles on growth inhibition are synergistic (combination index = 0.17±0.03) showing 10-10,000 fold lower inhibitory concentrations (IC50) compared to those of individual drugs. The design of antibody-targeted pure drug nanoparticles offers a promising design strategy to facilitate intracellular delivery and therapeutic efficiency of anticancer drugs. PMID:24053162

  16. Nanoparticle labeling identifies slow cycling human endometrial stromal cells

    PubMed Central

    2014-01-01

    Introduction Evidence suggests that the human endometrium contains stem or progenitor cells that are responsible for its remarkable regenerative capability. A common property of somatic stem cells is their quiescent state. It remains unclear whether slow-cycling cells exist in the human endometrium. We hypothesized that the human endometrium contains a subset of slow-cycling cells with somatic stem cell properties. Here, we established an in vitro stem cell assay to isolate human endometrial-derived mesenchymal stem-like cells (eMSC). Methods Single-cell stromal cultures were initially labeled with fluorescent nanoparticles and a small population of fluorescent persistent cells (FPC) remained after culture of 21 days. Two populations of stromal cells, namely FPC and non-FPC were sorted. Results Quantitative analysis of functional assays demonstrated that the FPC had higher colony forming ability, underwent more rounds of self-renewal and had greater enrichment of phenotypically defined prospective eMSC markers: CD146+/CD140b+ and W5C5+ than the non-FPC. They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC. The FPC exhibit low proliferation activities. A proliferation dynamics study revealed that more FPC had a prolonged G1 phase. Conclusions With this study we present an efficient method to label and isolate slow-proliferating cells obtained from human endometrial stromal cultures without genetic modifications. The FPC population could be easily maintained in vitro and are of interest for tissue-repair and engineering perspectives. In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined. PMID:24996487

  17. Rabeprazole exhibits antiproliferative effects on human gastric cancer cell lines

    PubMed Central

    GU, MENGLI; ZHANG, YAN; ZHOU, XINXIN; MA, HAN; YAO, HANGPING; JI, FENG

    2014-01-01

    Intracellular proton extrusion in gastric cancer cells has been reported to promote cancer cell survival under acidic conditions via hydrogen/potassium adenosine triphosphatase (H+/K+-ATPase). Rabeprazole is a frequently used second-generation proton pump inhibitor (PPI) that irreversibly inactivates gastric H+/K+-ATPase. Therefore, we hypothesized that rabeprazole could reduce the viability of gastric cancer cells. In the present study, four human gastric cancer cell lines and one non-cancer gastric cell line were cultured. Cell viability, the ?- and ?-subunits of H+/K+-ATPase and cellular apoptosis were analyzed by dye exclusion assay, reverse transcription-polymerase chain reaction and annexin V-fluorescein isothiocyanate/propidium iodide staining, respectively. The expression level of total extracellular signal-regulated protein kinase 1/2 (ERK 1/2) and phosphorylated-ERK protein was detected by western blot analysis. Gastric cancer cell lines were more tolerant of the acidic culture media than non-cancer cells. Administration of rabeprazole led to a marked decrease in the viability of MKN-28 cells. Exposure to rabeprazole induced significant apoptosis in AGS cells. Rabeprazole completely inhibited the phosphorylation of ERK 1/2 in the MKN-28 cells, whereas the same effect was not observed in either the KATO III or MKN-45 cells. The ERK 1/2 inhibitor, PD98059, attenuated the viability of the AGS cells. A similar antiproliferative effect was observed in the rabeprazole treatment group. In addition, PD98059 and rabeprazole were able to efficaciously inhibit the phosphorylation of ERK 1/2 in the gastric cancer cells. Therefore, it was concluded that rabeprazole can attenuate the cell viability of human gastric cancer cells through inactivation of the ERK1/2 signaling pathway. The results of the present study demonstrate that rabeprazole inhibits the viability of gastric cancer cells in vitro and may serve as a novel antineoplastic agent. PMID:25202402

  18. Rabeprazole exhibits antiproliferative effects on human gastric cancer cell lines.

    PubMed

    Gu, Mengli; Zhang, Yan; Zhou, Xinxin; Ma, Han; Yao, Hangping; Ji, Feng

    2014-10-01

    Intracellular proton extrusion in gastric cancer cells has been reported to promote cancer cell survival under acidic conditions via hydrogen/potassium adenosine triphosphatase (H(+)/K(+)-ATPase). Rabeprazole is a frequently used second-generation proton pump inhibitor (PPI) that irreversibly inactivates gastric H(+)/K(+)-ATPase. Therefore, we hypothesized that rabeprazole could reduce the viability of gastric cancer cells. In the present study, four human gastric cancer cell lines and one non-cancer gastric cell line were cultured. Cell viability, the ?- and ?-subunits of H(+)/K(+)-ATPase and cellular apoptosis were analyzed by dye exclusion assay, reverse transcription-polymerase chain reaction and annexin V-fluorescein isothiocyanate/propidium iodide staining, respectively. The expression level of total extracellular signal-regulated protein kinase 1/2 (ERK 1/2) and phosphorylated-ERK protein was detected by western blot analysis. Gastric cancer cell lines were more tolerant of the acidic culture media than non-cancer cells. Administration of rabeprazole led to a marked decrease in the viability of MKN-28 cells. Exposure to rabeprazole induced significant apoptosis in AGS cells. Rabeprazole completely inhibited the phosphorylation of ERK 1/2 in the MKN-28 cells, whereas the same effect was not observed in either the KATO III or MKN-45 cells. The ERK 1/2 inhibitor, PD98059, attenuated the viability of the AGS cells. A similar antiproliferative effect was observed in the rabeprazole treatment group. In addition, PD98059 and rabeprazole were able to efficaciously inhibit the phosphorylation of ERK 1/2 in the gastric cancer cells. Therefore, it was concluded that rabeprazole can attenuate the cell viability of human gastric cancer cells through inactivation of the ERK1/2 signaling pathway. The results of the present study demonstrate that rabeprazole inhibits the viability of gastric cancer cells in vitro and may serve as a novel antineoplastic agent. PMID:25202402

  19. [Gd@C82(OH)22]n nanoparticles inhibit the migration and adhesion of glioblastoma cells

    PubMed Central

    WANG, JING; GU, FENG; DING, TING; LIU, XIAOLI; XING, GENGMEI; ZHAO, YULIANG; ZHANG, NING; MA, YONGJIE

    2010-01-01

    In our previous study, [Gd@C82(OH)22]n, a fullerene-based nanoparticle, exhibited potent anti-tumor effects in mouse tumor-bearing models without detectable toxicity. The mechanism involved in the anti-tumor effect exerted by [Gd@C82(OH)22]n remains to be elucidated. This study found that glioblastoma cells treated with [Gd@C82(OH)22]n nanoparticles showed a significant impairment in migration and adhesion by cell chemotaxis, scratch and adhesion assays in vitro. Furthermore, our data showed that the key proteins, CD40 and ICAM-1, were involved in the inhibition of adhesion in the [Gd@C82(OH)22]n nanoparticle-treated glioblastoma cells. Thus, our study suggests that the [Gd@C82(OH)22]n nanoparticle is a new potential anti-tumor effector and a therapeutic component for malignant glioblastoma infiltration. PMID:22966378

  20. Carbon and clay nanoparticles induce minimal stress responses in gram negative bacteria and eukaryotic fish cells.

    PubMed

    Taylor, Alicia A; Aron, Gary M; Beall, Gary W; Dharmasiri, Nihal; Zhang, Yixin; McLean, Robert J C

    2014-08-01

    We investigated in vitro the potential mutagenic and toxic effects of two clay-based nanoparticles, Cloisite® Na(+) (Cloisite) and halloysite; and multi-walled carbon nanotubes (MWCNT), commonly used in the polymer composite industry. Using the Ames test, the three nanoparticles did not have a true mutagenic effect, although growth of Salmonella enterica var. Typhimurium (S.typhimurium) was diminished at higher nanoparticle concentrations. We investigated the impact of nanoparticles on Escherichia coli and S. typhimurium including oxyR and rpoS mutants, which are susceptible to oxidative stress. The oxyR mutants were inhibited in the presence of nanoparticles, when grown aerobically with light. Toxicity was not observed in the absence of light or during anaerobic growth. E. coli rpoS mutants exhibited some toxicity when cultured with Cloisite and MWCNT only when grown aerobically with light. There was no effect with other nanoparticles, or with S. typhimurium rpoS mutants. MWCNT exhibited a slight toxic effect against Epithelioma papulosum cyprini (EPC) cells only at the highest concentration tested. There was no discernable toxicity to EPC cells caused by the clay nanoparticles. We conclude that clay-based nanoparticles and MWCNT do not exert a mutagenic effect and do not have a general toxic effect across all bacterial species or between prokaryotic and eukaryotic cells. Modest toxicity was only observed in eukaryotic EPC cells against MWCNT at the highest concentration tested. Limited species-specific toxicity to clay based and MWCNT nanoparticles was seen in bacterial strains primarily due to culture conditions and mutations that exacerbate oxidative stress. PMID:23125163

  1. Gold nanoparticles enhanced electroporation for mammalian cell transfection.

    PubMed

    Zu, Yingbo; Huang, Shuyan; Liao, Wei-Ching; Lu, Yang; Wang, Shengnian

    2014-06-01

    Electroporation figured prominently as an effective nonviral gene delivery approach for its balance on the transfection efficiency and cell viability, no restrictions of probe or cell type, and operation simplicity. The commercial electroporation systems have been widely adopted in the past two decades while still carry drawbacks associated with the high applied electric voltage, unsatisfied delivery efficiency, and/or low cell viability. By adding highly conductive gold nanoparticles (AuNPs) in electroporation solution, we demonstrated enhanced electroporation performance (i.e., better DNA delivery efficiency and higher cell viability) on mammalian cells from two different aspects: the free, naked AuNPs reduce the resistance of the electroporation solution so that the local pulse strength on cells was enhanced; targeting AuNPs (e.g., Tf-AuNPs) were brought to the cell membrane to work as virtual microelectrodes to porate cells with limited area from many different sites. The enhancement was confirmed with leukemia cells in both a commercial batch electroporation system and a home-made flow-through system using pWizGFP plasmid DNA probes. Such enhancement depends on the size, concentration, and the mixing ratio of free AuNPs/Tf-AuNPs. An equivalent mixture of free AuNPs and Tf-AuNPs exhibited the best enhancement with the transfection efficiency increased 2-3 folds at minimum sacrifice of cell viability. This new delivery concept, the combination of nanoparticles and electroporation technologies, may stimulate various in vitro and in vivo biomedical applications which rely on the efficient delivery of nucleic acids, anticancer drugs, or other therapeutic materials. PMID:24749393

  2. pH-sensitive pullulan-based nanoparticle carrier for adriamycin to overcome drug-resistance of cancer cells.

    PubMed

    Guo, Hua; Liu, Yuanyuan; Wang, Yan; Wu, Jing; Yang, Xiaoying; Li, Rongshan; Wang, Yinsong; Zhang, Ning

    2014-10-13

    Urocanic acid was conjugated to pullulan to synthesize O-urocanyl pullulan (URPA) with degree of substitution (DS) of 8.2%. URPA nanoparticles prepared by dialysis method had spherical shapes and a mean diameter of 156.8 ± 16.8 nm. Adriamycin (ADR) was successfully loaded into URPA nanoparticles and exhibited pH-sensitive in vitro release property. MTT assay showed that ADR-loaded URPA (ADR/URPA) nanoparticles had a significant higher toxicity against drug resistant MCF-7/ADR cells than free ADR, and the reversal index reached up to 9.6. The results of flow cytometry and confocal microscopy showed that URPA nanoparticles efficiently enhanced accumulation and retention of ADR in MCF-7/ADR cells and successfully delivered ADR into cell nucleus. The reversal effect of ADR/URPA nanoparticles on the drug resistance of MCF-7/ADR cells was perhaps related with their cell entry and intracellular drug release mechanisms. PMID:25037431

  3. Bio-synthesis of gold nanoparticles by human epithelial cells, in vivo.

    PubMed

    Larios-Rodriguez, E; Rangel-Ayon, C; Castillo, S J; Zavala, G; Herrera-Urbina, R

    2011-09-01

    Healthy epithelial cells, in vivo, have the ability to synthesize gold nanoparticles when aqueous tetrachloroauric acid is made to react with human skin. Neither a reducing agent nor a protecting chemical is needed for this bio-synthesis method. The first indication of gold nanoparticle formation is the staining of the skin, which turns deep purple. Stereoscopic optical micrographs of human skin tissue in contact with aqueous tetrachloroauric acid clearly show the staining of the epithelial cells. The UV-Vis spectrum of these epithelial cells shows an absorption band with a maximum at 553 nm. This absorption peak is within the wavelength region where the surface plasmon resonance (SPR) band of aqueous colloidal gold exhibits a maximum. Transmission electron micrographs show that gold nanoparticles synthesized by epithelial cells have sizes between 1 and 100 nm. The electron diffraction pattern of these nanoparticles reveals a crystalline structure whose interplanar distances correspond to fcc metallic gold. Transmission electron micrographs of ultra-thin (70 nm thick) slices of epithelial cells clearly and undoubtedly demonstrate that gold nanoparticles are inside the cell. According to high resolution transmission electron micrographs of intracellular single gold nanoparticles, they have the shape of a polyhedron. PMID:21817787

  4. Glucose transporter 1-positive endothelial cells in infantile hemangioma exhibit features of facultative stem cells.

    PubMed

    Huang, Lan; Nakayama, Hironao; Klagsbrun, Michael; Mulliken, John B; Bischoff, Joyce

    2015-01-01

    Endothelial glucose transporter 1 (GLUT1) is a definitive and diagnostic marker for infantile hemangioma (IH), a vascular tumor of infancy. To date, GLUT1-positive endothelial cells in IH have not been quantified nor directly isolated and studied. We isolated GLUT1-positive and GLUT1-negative endothelial cells from IH specimens and characterized their proliferation, differentiation, and response to propranolol, a first-line therapy for IH, and to rapamycin, an mTOR pathway inhibitor used to treat an increasingly wide array of proliferative disorders. Although freshly isolated GLUT1-positive cells, selected using anti-GLUT1 magnetic beads, expressed endothelial markers CD31, VE-Cadherin, and vascular endothelial growth factor receptor 2, they converted to a mesenchymal phenotype after 3 weeks in culture. In contrast, GLUT1-negative endothelial cells exhibited a stable endothelial phenotype in vitro. GLUT1-selected cells were clonogenic when plated as single cells and could be induced to redifferentiate into endothelial cells, or into pericytes/smooth muscle cells or into adipocytes, indicating a stem cell-like phenotype. These data demonstrate that, although they appear and function in the tumor as bona fide endothelial cells, the GLUT1-positive endothelial cells display properties of facultative stem cells. Pretreatment with rapamycin for 4 days significantly slowed proliferation of GLUT1-selected cells, whereas propranolol pretreatment had no effect. These results reveal for the first time the facultative nature of GLUT1-positive endothelial cells in IH. PMID:25187207

  5. Nanoparticles

    NASA Astrophysics Data System (ADS)

    Vaez-Zadeh, Mehdi; Mohammadi, Ali

    2014-07-01

    MnFe2O4 nanoparticles of various particle sizes were prepared by co-precipitation, in which different hydroxide concentrations were employed to control particle growth. X-ray diffraction and scanning electron microscopy were used to investigate the nanoparticle structure and morphology (shape and size). The particle size increased with increasing hydroxide concentration. The magnetization and coercivity field were measured by vibrating sample magnetometry. Changes in magnetic behavior were observed in the magnetic hysteresis loop curves of nanoparticles with increasing hydroxide concentration. In the absence of hydroxide, nanoparticles exhibited paramagnetic behavior. Increasing the hydroxide concentration caused a gradual conversion to ferrimagnetic behavior. An increased Néel temperature was observed with increasing hydroxide concentration, and the saturation magnetization exhibited a sharp decrease. Nonuniform hysteresis was observed in the magnetization curve for the sample prepared from hydroxide and ammonium.

  6. Canine osteosarcoma cells exhibit resistance to aurora kinase inhibitors.

    PubMed

    Cannon, C M; Pozniak, J; Scott, M C; Ito, D; Gorden, B H; Graef, A J; Modiano, J F

    2015-03-01

    We evaluated the effect of Aurora kinase inhibitors AZD1152 and VX680 on canine osteosarcoma cells. Cytotoxicity was seen in all four cell lines; however, half-maximal inhibitory concentrations were significantly higher than in human leukaemia and canine lymphoma cells. AZD1152 reduced Aurora kinase B phosphorylation, indicating resistance was not because of failure of target recognition. Efflux mediated by ABCB1 and ABCG2 transporters is one known mechanism of resistance against these drugs and verapamil enhanced AZD1152-induced apoptosis; however, these transporters were only expressed by a small percentage of cells in each line and the effects of verapamil were modest, suggesting other mechanisms contribute to resistance. Our results indicate that canine osteosarcoma cells are resistant to Aurora kinase inhibitors and suggest that these compounds are unlikely to be useful as single agents for this disease. Further investigation of these resistance mechanisms and the potential utility of Aurora kinase inhibitors in multi-agent protocols is warranted. PMID:23410058

  7. Hybrid silver nanoparticle/conjugated polyelectrolyte nanocomposites exhibiting controllable metal-enhanced fluorescence.

    PubMed

    Wang, Xiaoyu; He, Fang; Zhu, Xi; Tang, Fu; Li, Lidong

    2014-01-01

    Metal-enhanced fluorescence of conjugated polyelectrolytes (CPs) is realized using a simple, green hybrid Ag nanocomposite film. Ag nanoparticles (Ag NPs) are pre-prepared by sodium citrate reduction and incorporated into agarose by mixing to form an Ag-containing agarose film (Ag@agarose). Through variation of the amount of Ag NPs in the Ag@agarose film as well as the thickness of the interlayer between CPs and the Ag@agarose film prepared of layer-by-layer assembly of chitosan and sodium alginate, a maximum 8.5-fold increase in the fluorescence of CPs is obtained. After introducing tyrosinase, this system also can be used to detect phenolic compounds with high sensitivity and good visualization under ultraviolet light. PMID:24638208

  8. Controlled self-assembly of multiferroic core-shell nanoparticles exhibiting strong magneto-electric effects

    NASA Astrophysics Data System (ADS)

    Sreenivasulu, Gollapudi; Popov, Maksym; Chavez, Ferman A.; Hamilton, Sean L.; Lehto, Piper R.; Srinivasan, Gopalan

    2014-02-01

    Ferromagnetic-ferroelectric composites show strain mediated coupling between the magnetic and electric sub-systems due to magnetostriction and piezoelectric effects associated with the ferroic phases. We have synthesized core-shell multiferroic nano-composites by functionalizing 10-100 nm barium titanate and nickel ferrite nanoparticles with complementary coupling groups and allowing them to self-assemble in the presence of a catalyst. The core-shell structure was confirmed by electron microscopy and magnetic force microscopy. Evidence for strong strain mediated magneto-electric coupling was obtained by static magnetic field induced variations in the permittivity over 16-18 GHz and polarization and by electric field induced by low-frequency ac magnetic fields.

  9. Hybrid silver nanoparticle/conjugated polyelectrolyte nanocomposites exhibiting controllable metal-enhanced fluorescence

    PubMed Central

    Wang, Xiaoyu; He, Fang; Zhu, Xi; Tang, Fu; Li, Lidong

    2014-01-01

    Metal-enhanced fluorescence of conjugated polyelectrolytes (CPs) is realized using a simple, green hybrid Ag nanocomposite film. Ag nanoparticles (Ag NPs) are pre-prepared by sodium citrate reduction and incorporated into agarose by mixing to form an Ag-containing agarose film (Ag@agarose). Through variation of the amount of Ag NPs in the Ag@agarose film as well as the thickness of the interlayer between CPs and the Ag@agarose film prepared of layer-by-layer assembly of chitosan and sodium alginate, a maximum 8.5-fold increase in the fluorescence of CPs is obtained. After introducing tyrosinase, this system also can be used to detect phenolic compounds with high sensitivity and good visualization under ultraviolet light. PMID:24638208

  10. Controlled self-assembly of multiferroic core-shell nanoparticles exhibiting strong magneto-electric effects

    SciTech Connect

    Sreenivasulu, Gollapudi; Hamilton, Sean L.; Lehto, Piper R.; Srinivasan, Gopalan, E-mail: srinivas@oakland.edu [Physics Department, Oakland University, Rochester, Michigan 48309-4401 (United States); Popov, Maksym [Physics Department, Oakland University, Rochester, Michigan 48309-4401 (United States); Radiophysics Department, Taras Shevchenko National University of Kyiv, Kyiv 01601 (Ukraine); Chavez, Ferman A. [Chemistry Department, Oakland University, Rochester, Michigan 48309-4401 (United States)

    2014-02-03

    Ferromagnetic-ferroelectric composites show strain mediated coupling between the magnetic and electric sub-systems due to magnetostriction and piezoelectric effects associated with the ferroic phases. We have synthesized core-shell multiferroic nano-composites by functionalizing 10–100?nm barium titanate and nickel ferrite nanoparticles with complementary coupling groups and allowing them to self-assemble in the presence of a catalyst. The core-shell structure was confirmed by electron microscopy and magnetic force microscopy. Evidence for strong strain mediated magneto-electric coupling was obtained by static magnetic field induced variations in the permittivity over 16–18?GHz and polarization and by electric field induced by low-frequency ac magnetic fields.

  11. Hybrid silver nanoparticle/conjugated polyelectrolyte nanocomposites exhibiting controllable metal-enhanced fluorescence

    NASA Astrophysics Data System (ADS)

    Wang, Xiaoyu; He, Fang; Zhu, Xi; Tang, Fu; Li, Lidong

    2014-03-01

    Metal-enhanced fluorescence of conjugated polyelectrolytes (CPs) is realized using a simple, green hybrid Ag nanocomposite film. Ag nanoparticles (Ag NPs) are pre-prepared by sodium citrate reduction and incorporated into agarose by mixing to form an Ag-containing agarose film (Ag@agarose). Through variation of the amount of Ag NPs in the Ag@agarose film as well as the thickness of the interlayer between CPs and the Ag@agarose film prepared of layer-by-layer assembly of chitosan and sodium alginate, a maximum 8.5-fold increase in the fluorescence of CPs is obtained. After introducing tyrosinase, this system also can be used to detect phenolic compounds with high sensitivity and good visualization under ultraviolet light.

  12. Cerium fluoride nanoparticles protect cells against oxidative stress.

    PubMed

    Shcherbakov, Alexander B; Zholobak, Nadezhda M; Baranchikov, Alexander E; Ryabova, Anastasia V; Ivanov, Vladimir K

    2015-05-01

    A novel facile method of non-doped and fluorescent terbium-doped cerium fluoride stable aqueous sols synthesis is proposed. Intense green luminescence of CeF3:Tb nanoparticles can be used to visualize these nanoparticles' accumulation in cells using confocal laser scanning microscopy. Cerium fluoride nanoparticles are shown for the first time to protect both organic molecules and living cells from the oxidative action of hydrogen peroxide. Both non-doped and terbium-doped CeF3 nanoparticles are shown to provide noteworthy protection to cells against the vesicular stomatitis virus. PMID:25746257

  13. Aluminum plasmonic nanoparticles enhanced dye sensitized solar cells.

    PubMed

    Xu, Qi; Liu, Fang; Liu, Yuxiang; Meng, Weisi; Cui, Kaiyu; Feng, Xue; Zhang, Wei; Huang, Yidong

    2014-03-10

    We present an investigation on utilizing plasmonic aluminium (Al) nanoparticles (NPs) to enhance the optical absorption of dye-sensitized solar cells (DSCs). The Al NPs exhibit not only the light absorption enhancement in solar cells with localized surface plasmon (LSP) effect but also the chemical stability to iodide/triiodide electrolyte. Besides, the lower work function (~4.06 eV), compared with that of TiO? (~4.6 eV), may suppress the quenching processes, such as charge transfer to metal NPs, to reduce the loss. Thus, high concentration of Al NPs could be incorporated into the TiO? anodes, and the power conversion efficiency (PCE) of DSCs is improved by nearly 13%. Moreover, electrochemical impedance spectroscopy (EIS) characterization also indicates that the plasmonic DSCs with Al NPs present better electrochemical performance than regular ones, which contributes to the improvement of PCE of the device. PMID:24922239

  14. Ocular surface epithelia contain ABCG2-dependent side population cells exhibiting features associated with stem cells

    PubMed Central

    Budak, Murat T.; Alpdogan, Onder S.; Zhou, Mingyuan; Lavker, Robert M.; Akinci, M.A. Murat; Wolosin, J. Mario

    2005-01-01

    Summary When cell populations are incubated with the DNA-binding dye Hoechst 33342 and subjected to flow cytometry analysis for Hoechst 33342 emissions, active efflux of the dye by the ABCG2/BCRP1 transporter causes certain cells to appear as a segregated cohort, known as a side population (SP). Stem cells from several tissues have been shown to possess the SP phenotype. As the lack of specific surface markers has hindered the isolation and subsequent biochemical characterization of epithelial stem cells this study sought to determine the existence of SP cells and expression of ABCG2 in the epithelia of the ocular surface and evaluate whether such SP cells had features associated with epithelial stem cells. Human and rabbit limbal-corneal and conjunctival epithelial cells were incubated with Hoechst 33342, and analyzed and sorted by flow cytometry. Sorted cells were subjected to several tests to determine whether the isolated SP cells displayed features consistent with the stem cell phenotype. Side populations amounting to <1% of total cells, which were sensitive to the ABCG2-inhibitor fumitremorgin C, were found in the conjunctival and limbal epithelia, but were absent from the stem cell-free corneal epithelium. Immunohistochemistry was used to establish the spatial expression pattern of ABCG2. The antigen was detected in clusters of conjunctival and limbal epithelia basal cells but was not present in the corneal epithelium. SP cells were characterized by extremely low light side scattering and contained a high percentage of cells that: showed slow-cycling prior to tissue collection; exhibited an initial delay in proliferation after culturing; and displayed clonogenic capacity and resistance to phorbol-induced differentiation; all features that are consistent with a stem cell phenotype. PMID:15811951

  15. Drug-loaded nanoparticles induce gene expression in human pluripotent stem cell derivatives

    PubMed Central

    Gajbhiye, Virendra; Escalante, Leah; Chen, Guojun; Laperle, Alex; Zheng, Qifeng; Steyer, Benjamin; Gong, Shaoqin; Saha, Krishanu

    2014-01-01

    Tissue engineering and advanced manufacturing of human stem cells requires a suite of tools to control gene expression spatiotemporally in culture. Inducible gene expression systems offer cell-extrinsic control, typically through addition of small molecules, but small molecule inducers typically contain few functional groups for further chemical modification. Doxycycline (DXC), a potent small molecule inducer of tetracycline (Tet) transgene systems, was conjugated to a hyperbranched dendritic polymer (Boltorn H40) and subsequently reacted with polyethylene glycol (PEG). The resulting PEG-H40-DXC nanoparticle exhibited pH-sensitive drug release behavior and successfully controlled gene expression in stem-cell-derived fibroblasts with a Tet-On system. While free DXC inhibited fibroblast proliferation and matrix metalloproteinase (MMP) activity, PEG-H40-DXC nanoparticles maintained higher fibroblast proliferation levels and MMP activity. The results demonstrate that the PEG-H40-DXC nanoparticle system provides an effective tool to controlling gene expression in human stem cell derivatives. PMID:24232694

  16. Micropatterned cell sheets with defined cell and extracellular matrix orientation exhibit anisotropic mechanical properties.

    PubMed

    Isenberg, Brett C; Backman, Daniel E; Kinahan, Michelle E; Jesudason, Rajiv; Suki, Bela; Stone, Phillip J; Davis, Elaine C; Wong, Joyce Y

    2012-03-15

    For an arterial replacement graft to be effective, it must possess the appropriate strength in order to withstand long-term hemodynamic stress without failure, yet be compliant enough that the mismatch between the stiffness of the graft and the native vessel wall is minimized. The native vessel wall is a structurally complex tissue characterized by circumferentially oriented collagen fibers/cells and lamellar elastin. Besides the biochemical composition, the functional properties of the wall, including stiffness, depend critically on the structural organization. Therefore, it will be crucial to develop methods of producing tissues with defined structures in order to more closely mimic the properties of a native vessel. To this end, we sought to generate cell sheets that have specific ECM/cell organization using micropatterned polydimethylsiloxane (PDMS) substrates to guide cell organization and tissue growth. The patterns consisted of large arrays of alternating grooves and ridges. Adult bovine aortic smooth muscle cells cultured on these substrates in the presence of ascorbic acid produced ECM-rich sheets several cell layers thick in which both the cells and ECM exhibited strong alignment in the direction of the micropattern. Moreover, mechanical testing revealed that the sheets exhibited mechanical anisotropy similar to that of native vessels with both the stiffness and strength being significantly larger in the direction of alignment, demonstrating that the microscale control of ECM organization results in functional changes in macroscale material behavior. PMID:22177672

  17. DNA-assembled nanoparticle rings exhibit electric and magnetic resonances at visible frequencies.

    PubMed

    Roller, Eva-Maria; Khorashad, Larousse Khosravi; Fedoruk, Michael; Schreiber, Robert; Govorov, Alexander O; Liedl, Tim

    2015-02-11

    Metallic nanostructures can be used to manipulate light on the subwavelength scale to create tailored optical material properties. Next to electric responses, artificial optical magnetism is of particular interest but difficult to achieve at visible wavelengths. DNA-self-assembly has proved to serve as a viable method to template plasmonic materials with nanometer precision and to produce large quantities of metallic objects with high yields. We present here the fabrication of self-assembled ring-shaped plasmonic metamolecules that are composed of four to eight single metal nanoparticles with full stoichiometric and geometric control. Scattering spectra of single rings as well as absorption spectra of solutions containing the metamolecules are used to examine the unique plasmonic features, which are compared to computational simulations. We demonstrate that the electric and magnetic plasmon resonance modes strongly correlate with the exact shape of the structures. In particular, our computations reveal the magnetic plasmons only for particle rings of broken symmetries, which is consistent with our experimental data. We stress the feasibility of DNA self-assembly as a method to create bulk plasmonic materials and metamolecules that may be applied as building blocks in plasmonic devices. PMID:25611357

  18. Silver polyvinyl pyrrolidone nanoparticles exhibit a capsular polysaccharide influenced bactericidal effect against Streptococcus pneumoniae

    PubMed Central

    Bibbs, Ronda K.; Harris, Rhonda D.; Peoples, Veolanda A.; Barnett, Cleon; Singh, Shree R.; Dennis, Vida A.; Coats, Mamie T.

    2014-01-01

    Streptococcus pneumoniae remains a leading cause of morbidity and mortality worldwide. The highly adaptive nature of S. pneumoniae exemplifies the need for next generation antimicrobials designed to avoid high level resistance. Metal based nanomaterials fit this criterion. Our study examined the antimicrobial activity of gold nanospheres, silver coated polyvinyl pyrrolidone (AgPVP), and titanium dioxide (TiO2) against various serotypes of S. pneumoniae. Twenty nanometer spherical AgPVP demonstrated the highest level of killing among the tested materials. AgPVP (0.6 mg/mL) was able to kill pneumococcal serotypes 2, 3, 4, and 19F within 4 h of exposure. Detailed analysis of cultures during exposure to AgPVP showed that both the metal ions and the solid nanoparticles participate in the killing of the pneumococcus. The bactericidal effect of AgPVP was lessened in the absence of the pneumococcal capsular polysaccharide. Capsule negative strains, JD908 and RX1, were only susceptible to AgPVP at concentrations at least 33% higher than their respective capsule expressing counterparts. These findings suggest that mechanisms of killing used by nanomaterials are not serotype dependent and that the capsular polysaccharide participates in the inhibition. In the near future these mechanisms will be examined as targets for novel antimicrobials. PMID:25520713

  19. Stem cell tracking using iron oxide nanoparticles

    PubMed Central

    Bull, Elizabeth; Madani, Seyed Yazdan; Sheth, Roosey; Seifalian, Amelia; Green, Mark; Seifalian, Alexander M

    2014-01-01

    Superparamagnetic iron oxide nanoparticles (SPIONs) are an exciting advancement in the field of nanotechnology. They expand the possibilities of noninvasive analysis and have many useful properties, making them potential candidates for numerous novel applications. Notably, they have been shown that they can be tracked by magnetic resonance imaging (MRI) and are capable of conjugation with various cell types, including stem cells. In-depth research has been undertaken to establish these benefits, so that a deeper level of understanding of stem cell migratory pathways and differentiation, tumor migration, and improved drug delivery can be achieved. Stem cells have the ability to treat and cure many debilitating diseases with limited side effects, but a main problem that arises is in the noninvasive tracking and analysis of these stem cells. Recently, researchers have acknowledged the use of SPIONs for this purpose and have set out to establish suitable protocols for coating and attachment, so as to bring MRI tracking of SPION-labeled stem cells into common practice. This review paper explains the manner in which SPIONs are produced, conjugated, and tracked using MRI, as well as a discussion on their limitations. A concise summary of recently researched magnetic particle coatings is provided, and the effects of SPIONs on stem cells are evaluated, while animal and human studies investigating the role of SPIONs in stem cell tracking will be explored. PMID:24729700

  20. Trypsinization-dependent cell labeling with fluorescent nanoparticles

    PubMed Central

    2014-01-01

    Trypsin is often used to detach adhered cell subculture from a substrate. However, the proteolytic activity of trypsin may harm cells by cleaving the cell membrane proteins. The present study shows that cellular uptake of fluorescent nanoparticles is remarkably increased within 24 h after trypsinization. These results highlight the trypsin-induced protein digestion, provoking leaky cell plasma membrane which leads to the strongly enhanced cellular uptake of the nanoparticles. To prevent this effect, one should expose cells to the nanoparticle (NP)-based fluorescent labels at least 48 h after trypsinization. PMID:25328505

  1. Highly fluorescent and bioresorbable polymeric nanoparticles with enhanced photostability for cell imaging

    NASA Astrophysics Data System (ADS)

    Huang, Shuo; Liu, Shiying; Wang, Kai; Yang, Cangjie; Luo, Yimin; Zhang, Yingdan; Cao, Bin; Kang, Yuejun; Wang, Mingfeng

    2014-12-01

    We report a facile and general strategy for enhancing the photostability of organic fluorophores for bioimaging applications. As a proof of concept, bright and robust fluorescence was observed in solid states of a well-defined synthetic polymer polycaprolactone consisting of di(thiophene-2-yl)-diketopyrrolopyrrole covalently linked in the middle of the polymer chain as a biocompatible and bioresorbable matrix. The nanoparticles prepared through a nanoprecipitation process of these polymers could be internalized by both tumor cells and stem cells with little cytotoxicity. Moreover, these highly fluorescent nanoparticles exhibited significantly enhanced photostability compared to commercial quantum dots or physical blends of dye/polymer complexes in cell imaging and long-term tracing.We report a facile and general strategy for enhancing the photostability of organic fluorophores for bioimaging applications. As a proof of concept, bright and robust fluorescence was observed in solid states of a well-defined synthetic polymer polycaprolactone consisting of di(thiophene-2-yl)-diketopyrrolopyrrole covalently linked in the middle of the polymer chain as a biocompatible and bioresorbable matrix. The nanoparticles prepared through a nanoprecipitation process of these polymers could be internalized by both tumor cells and stem cells with little cytotoxicity. Moreover, these highly fluorescent nanoparticles exhibited significantly enhanced photostability compared to commercial quantum dots or physical blends of dye/polymer complexes in cell imaging and long-term tracing. Electronic supplementary information (ESI) available: Experimental procedures, absorbance spectra, confocal microscopy characterization. See DOI: 10.1039/c4nr05576d

  2. Plasmonic silver nanoparticles loaded titania nanotube arrays exhibiting enhanced photoelectrochemical and photocatalytic activities

    NASA Astrophysics Data System (ADS)

    Nishanthi, S. T.; Iyyapushpam, S.; Sundarakannan, B.; Subramanian, E.; Pathinettam Padiyan, D.

    2015-01-01

    A combination of electrochemical anodization and photochemical reduction is employed to fabricate highly ordered silver loaded titania nanotubes (Ag/TNT) arrays. The Ag/TNT samples show an extended optical absorbance from UV to visible region owing to the surface plasmon resonance effect of Ag. The photoluminescence intensity of Ag/TNT is significantly lower than that of pure titania revealing a decrease in charge carrier recombination. The photoelectrochemical properties of the prepared samples are studied using linear sweep and transient photocurrent measurements. Compared with pure TNT, the Ag loaded samples show a higher photoelectrochemical activity. The results demonstrate an efficient separation of photogenerated electron-hole pairs and the consequent increase in lifetime of charge carriers by Ag/TNT. The photocatalytic results of methyl orange dye degradation show that the Ag/TNT-3-05 sample exhibits the maximum degradation efficiency of 98.85% with kinetic rate constant of 0.0236(5) min-1 for 180 min light illumination.

  3. Cooperative entry of nanoparticles into the cell

    NASA Astrophysics Data System (ADS)

    Wang, Jiuling; Yao, Haimin; Shi, Xinghua

    2014-12-01

    Interaction of nanoparticles (NPs) with cell membrane is a crucial issue in studying drug delivery, photodynamic therapy system and cytotoxicity. Single NP with relatively small size cannot be fully wrapped by the cell membrane, which prohibits its uptake. One feasible way is cooperative entry, i.e., recruiting and assembling multiple small NPs to form a larger NP cluster to enter into a cell. In this work, we present theoretical analysis about the cooperative entry of multiple NPs. Through free energy calculation we investigate how the NPs' size, shape, interval and NP/cell interfacial binding energy influence the feasibility of entry. Interestingly we find that the cooperative entry of oblate ellipsoidal NPs can get larger energy compensation than individual ones as well as spherical ones. We also propose that soft NPs have preference in cooperative entry of the cell. Our work can be used to actively design and transfer NPs in applications such as drug delivery as well as to understand the shape effect on toxic mechanism of ellipsoidal NPs.

  4. Labeling of macrophage cell using biocompatible magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Min, Ji Hyun; Kim, Sung Tae; Lee, Ji Sung; Kim, Kwanghee; Wu, Jun Hua; Jeong, Jaeho; Song, Ah Young; Lee, Kyung-Mi; Kim, Young Keun

    2011-04-01

    This work investigates the intrinsic cell labeling efficiency of the Fe3O4 nanoparticles prepared by a modified thermal decomposition method using nontoxic precursors and a biocompatible polymer surfactant. This method eliminates the current need for additional step of surface modification. The structural analysis reveals the highly crystalline feature of the nanoparticles, while the magnetic measurement shows their superparamagnetic behavior at room temperature. Fe3O4 nanoparticles were efficiently incorporated into the murine macrophage cells (RAW264.7) without visible cytotoxicity. Cell labeling efficiency was found to be over 90% as measured by magnetically activated cell sorting and physical property measurement system. Therefore, such Fe3O4 nanoparticles could provide a useful magnetic cell labeling tool for macrophage cells using their phagocytic/endocytic activity and further apply to the other relevant biomedical applications.

  5. Fullerene nanoparticles exhibit greater retention in freshwater sediment than in model porous media.

    PubMed

    Zhang, Wei; Isaacson, Carl W; Rattanaudompol, U-sa; Powell, Tremaine B; Bouchard, Dermont

    2012-06-01

    Increasing production and use of fullerene-based nanomaterials underscore the need to determine their mobility in environmental transport pathways and potential ecological exposures. This study investigated the transport of two fullerenes (i.e., aqu/C(60) and water-soluble C(60) pyrrolidine tris-acid [C(60) PTA]) in columns packed with model porous media (Iota quartz and Ottawa sand) and a sediment from Call's creek under saturated and unsaturated steady-state flows. The fullerenes had the least retention in Iota quartz, and the greatest retention in the sediment at near neutral pH, correlating with the degree of grain surface chemical heterogeneity (e.g., amorphous Al hydroxides concentration increasing in the order of Iota quartzexhibited a strong dependency on solution pH that could be explained partly by the pH-dependent surface charge of fullerenes and grain surface, and partly by increased hydrophobicity of C(60) PTA when solution pH approaches its isoelectric point (IEP). Finally, fullerene retention was enhanced in unsaturated media, implying that fullerenes may be more attenuated in the vadose zone than in groundwater. PMID:22445188

  6. Nanosize and surface charge effects of hydroxyapatite nanoparticles on red blood cell suspensions.

    PubMed

    Han, Yingchao; Wang, Xinyu; Dai, Honglian; Li, Shipu

    2012-09-26

    In this paper, the effects of size and surface charge of hydroxyapatite (HAP) particles on a red blood cell (RBC) suspension were studied. Results showed that the HAP particles exhibited nanosize and surface charge effects on the RBC suspension. Differing from HAP microparticles, HAP nanoparticles induced some aggregation of the RBCs in the unstructured agglutinates. HAP nanoparticles were adhered to the surface membrane of the RBCs due to their remarkably higher adsorption capacity than the HAP microparticles, resulting in the formation of a sunken appearance ("caves") on the surface membrane of the RBCs without rupturing the lipid bilayer. In the case of high negatively charged HAP nanoparticles after heparin modification, the aggregation of the RBCs induced by the HAP nanoparticles was inhibited. Such HAP nanoparticle-induced aggregation of the RBCs could be attributed to the bridging force via the electrostatic interaction between the positively charged binding sites on the HAP surface and the negatively charged groups on the surface of the RBCs. The surface charge of the HAP nanoparticles is thus a crucial factor influencing the interaction between the HAP nanoparticles and the RBCs. PMID:22860897

  7. Electrical transport and optical studies of ferromagnetic Cobalt doped ZnO nanoparticles exhibiting a metal-insulator transition

    E-print Network

    M. Naeem; S. K. Hasanain; A. Mumtaz

    2007-12-08

    The observed correlation of oxygen vacancies and room temperature ferromagnetic ordering in Co doped ZnO1-o nanoparticles reported earlier (Naeem et al Nanotechnology 17, 2675-2680) has been further explored by transport and optical measurements. In these particles room temperature ferromagnetic ordering had been observed to occur only after annealing in forming gas. In the current work the optical properties have been studied by diffuse reflection spectroscopy in the UV-Vis region and the band gap of the Co doped compositions has been found to decrease with Co addition. Reflections minima are observed at the energies characteristic of Co+2 d-d (tethrahedral symmetry) crystal field transitions, further establishing the presence of Co in substitutional sites. Electrical transport measurements on palletized samples of the nanoparticles show that the effect of a forming gas is to strongly decrease the resistivity with increasing Co concentration. For the air annealed and non-ferromagnetic samples the variation in the resistivity as a function of Co content are opposite to those observed in the particles prepared in forming gas. The ferromagnetic samples exhibit an apparent change from insulator to metal with increasing temperatures for T>380K and this change becomes more pronounced with increasing Co content. The magnetic and resistive behaviors are correlated by considering the model by Calderon et al [M. J. Calderon and S. D. Sarma, Annals of Physics 2007 (Accepted doi: 10.1016/j.aop.2007.01.010] where the ferromagnetism changes from being mediated by polarons in the low temperature insulating region to being mediated by the carriers released from the weakly bound states in the higher temperature metallic region.

  8. Labeling of macrophage cell using biocompatible magnetic nanoparticles

    Microsoft Academic Search

    Ji Hyun Min; Sung Tae Kim; Ji Sung Lee; Kwanghee Kim; Jun Hua Wu; Jaeho Jeong; Ah Young Song; Kyung-Mi Lee; Young Keun Kim

    2011-01-01

    This work investigates the intrinsic cell labeling efficiency of the Fe3O4 nanoparticles prepared by a modified thermal decomposition method using nontoxic precursors and a biocompatible polymer surfactant. This method eliminates the current need for additional step of surface modification. The structural analysis reveals the highly crystalline feature of the nanoparticles, while the magnetic measurement shows their superparamagnetic behavior at room

  9. Manipulating directional cell motility using intracellular superparamagnetic nanoparticles.

    PubMed

    Bradshaw, Michael; Clemons, Tristan D; Ho, Diwei; Gutiérrez, Lucía; Lázaro, Francisco J; House, Michael J; St Pierre, Timothy G; Fear, Mark W; Wood, Fiona M; Iyer, K Swaminathan

    2015-03-01

    This study investigated the ability for magnetic nanoparticles to influence cellular migration in the presence of an external magnetic field. We found that the direction of migrating keratinocytes can be controlled and the migration speed of fibroblasts can be increased with the internalisation of these nanoparticles in the presence of a magnetic field. The possibility of shepherding cells towards a region of interest through the use of internalized nanoparticles is an attractive prospect for cell tracking, cell therapies, and tissue engineering applications. PMID:25695187

  10. Uptake and cytotoxicity of chitosan nanoparticles in human liver cells

    SciTech Connect

    Loh, Jing Wen [Laboratory for Drug Delivery, Pharmacy, University of Western Australia, 35 Stirling Hwy, Crawley, 6009 (Australia); Yeoh, George [School of Biomedical, Biomolecular and Chemical Sciences, University of Western Australia, 35 Stirling Hwy, Crawley, 6009 (Australia); Centre for Medical Research, Western Australian Institute for Medical Research, Nedlands, WA 6009 (Australia); Saunders, Martin [Centre for Microscopy, Characterisation and Analysis, University of Western Australia, 35 Stirling Hwy, Crawley, 6009 (Australia); Lim, Lee-Yong, E-mail: limly@cyllene.uwa.edu.a [Laboratory for Drug Delivery, Pharmacy, University of Western Australia, 35 Stirling Hwy, Crawley, 6009 (Australia); School of Biomedical, Biomolecular and Chemical Sciences, University of Western Australia, 35 Stirling Hwy, Crawley, 6009 (Australia)

    2010-12-01

    Despite extensive research into the biomedical and pharmaceutical applications of nanoparticles, and the liver being the main detoxifying organ in the human body, there are limited studies which delineate the hepatotoxicity of nanoparticles. This paper reports on the biological interactions between liver cells and chitosan nanoparticles, which have been widely recognised as biocompatible. Using the MTT assay, human liver cells were shown to tolerate up to 4 h of exposure to 0.5% w/v of chitosan nanoparticles (18 {+-} 1 nm, 7.5 {+-} 1.0 mV in culture medium). At nanoparticle concentrations above 0.5% w/v, cell membrane integrity was compromised as evidenced by leakage of alanine transaminase into the extracellular milieu, and there was a dose-dependent increase in CYP3A4 enzyme activity. Uptake of chitosan nanoparticles into the cell nucleus was observed by confocal microscopic analysis after 4 h exposure with 1% w/v of chitosan nanoparticles. Electron micrographs further suggest necrotic or autophagic cell death, possibly caused by cell membrane damage and resultant enzyme leakage.

  11. Dual-responsive magnetic core-shell nanoparticles for nonviral gene delivery and cell separation.

    PubMed

    Majewski, Alexander P; Schallon, Anja; Jérôme, Valérie; Freitag, Ruth; Müller, Axel H E; Schmalz, Holger

    2012-03-12

    We present the synthesis of dual-responsive (pH and temperature) magnetic core-shell nanoparticles utilizing the grafting-from approach. First, oleic acid stabilized superparamagnetic maghemite (?-Fe(2)O(3)) nanoparticles (NPs), prepared by thermal decomposition of iron pentacarbonyl, were surface-functionalized with ATRP initiating sites bearing a dopamine anchor group via ligand exchange. Subsequently, 2-(dimethylamino)ethyl methacrylate (DMAEMA) was polymerized from the surface by ATRP, yielding dual-responsive magnetic core-shell NPs (?-Fe(2)O(3)@PDMAEMA). The attachment of the dopamine anchor group on the nanoparticle's surface is shown to be reversible to a certain extent, resulting in a grafting density of 0.15 chains per nm(2) after purification. Nevertheless, the grafted NPs show excellent long-term stability in water over a wide pH range and exhibit a pH- and temperature-dependent reversible agglomeration, as revealed by turbidimetry. The efficiency of ?-Fe(2)O(3)@PDMAEMA hybrid nanoparticles as a potential transfection agent was explored under standard conditions in CHO-K1 cells. Remarkably, ?-Fe(2)O(3)@PDMAEMA led to a 2-fold increase in the transfection efficiency without increasing the cytotoxicity, as compared to polyethyleneimine (PEI), and yielded on average more than 50% transfected cells. Moreover, after transfection with the hybrid nanoparticles, the cells acquired magnetic properties that could be used for selective isolation of transfected cells. PMID:22296556

  12. Toxicity Study of Silver Nanoparticles Synthesized from Suaeda monoica on Hep-2 Cell Line

    PubMed Central

    Satyavani, Kaliyamurthi; Gurudeeban, Selvaraj; Ramanathan, Thiruganasambandam; Balasubramanian, Thangavel

    2012-01-01

    Recently there has been fabulous excitement in the nano-biotechnological area for the study of nanoparticles synthesis using some natural biological system, which has led the growth advanced nanomaterials. This intention made us to assess the biologically synthesized silver nanoparticles from the leaf of Suaeda monoica (S.monoica) using 1 mM silver nitrate. The leaf extract of S.monoica incubated with 1 mM silver nitrate solution and characterized by UV- spectrometer and AFM. The effect of synthesized silver nanoparticles on Human Epidermoid Larynx Carcinoma cell line was evaluated by the MTT colorimetric technique. As a result we observed gradual change in the colour of extract from greenish to brown. The synthesized silver nanoparticles confirmed by UV at 430 nm and spherical shape identified in the range of 31 nm under AFM. The effect of silver nanoparticles on Human Epidermoid Larynx Carcinoma cell line exhibits a dose-dependent toxicity for the cell tested and the viability of Hep-2 cells decreased to 50 % (IC50) at the concentration of 500 nM. Further findings will be determined the exact mechanisms of this cost effective Nano-treatments. PMID:23407847

  13. Enhanced apoptotic effects of dihydroartemisinin-aggregated gelatin and hyaluronan nanoparticles on human lung cancer cells.

    PubMed

    Sun, Qian; Teong, Benjamin; Chen, I-Fen; Chang, Shwu Jen; Gao, Jimin; Kuo, Shyh-Ming

    2014-04-01

    Recent studies suggest that dihydroartemisinin (DHA), a derivative of artemisinin isolated from the traditional Chinese herb Artemisia annua L., has anticancer properties. Due to poor water solubility, poor oral activity, and a short plasma half-life, large doses of DHA have to be injected to achieve the necessary bioavailability. This study examined increasing DHA bioavailability by encapsulating DHA within gelatin (GEL) or hyaluronan (HA) nanoparticles via an electrostatic field system. Observations from transmission electron microscopy show that DHA in GEL and HA nanoparticles formed GEL/DHA and HA/DHA aggregates that were approximately 30-40 nm in diameter. The entrapment efficiencies for DHA were approximately 13 and 35% for the GEL/DHA and HA/DHA aggregates, respectively. The proliferation of A549 cells was inhibited by the GEL/DHA and HA/DHA aggregates. Fluorescent annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) staining displayed low background staining with annexin V-FITC or PI on DHA-untreated cells. In contrast, annexin V-FITC and PI stains dramatically increased when the cells were incubated with GEL/DHA and HA/DHA aggregates. These results suggest that DHA-aggregated GEL and HA nanoparticles exhibit higher anticancer proliferation activities than DHA alone in A549 cells most likely due to the greater aqueous dispersion after hydrophilic GEL or HA nanoparticles aggregation. These results demonstrate that DHA can aggregate with nanoparticles in an electrostatic field environment to form DHA nanosized aggregates. PMID:24039154

  14. Cationic Surface Modification of PLG Nanoparticles Offers Sustained Gene Delivery to Pulmonary Epithelial Cells

    PubMed Central

    BAOUM, ABDULGADER; DHILLON, NAVNEET; BUCH, SHILPA; BERKLAND, CORY

    2010-01-01

    Biodegradable polymeric nanoparticles are currently being explored as a nonviral gene delivery system; however, many obstacles impede the translation of these nanomaterials. For example, nanoparticles delivered systemically are inherently prone to adsorbing serum proteins and agglomerating as a result of their large surface/volume ratio. What is desired is a simple procedure to prepare nanoparticles that may be delivered locally and exhibit minimal toxicity while improving entry into cells for effectively delivering DNA. The objective of this study was to optimize the formulation of poly(D,L-lactide-co-glycolide) (PLG) nanoparticles for gene delivery performance to a model of the pulmonary epithelium. Using a simple solvent diffusion technique, the chemistry of the particle surface was varied by using different coating materials that adsorb to the particle surface during formation. A variety of cationic coating materials were studied and compared to more conventional surfactants used for PLG nanoparticle fabrication. Nanoparticles (~200 nm) efficiently encapsulated plasmids encoding for luciferase (80–90%) and slowly released the same for 2 weeks. In A549 alveolar lung epithelial cells, high levels of gene expression appeared at day 5 for certain positively charged PLG particles and gene expression was maintained for at least 2 weeks. In contrast, PEI gene expression ended at day 5. PLG particles were also significantly less cytotoxic than PEI suggesting the use of these vehicles for localized, sustained gene delivery to the pulmonary epithelium. PMID:19911425

  15. Cationic surface modification of PLG nanoparticles offers sustained gene delivery to pulmonary epithelial cells.

    PubMed

    Baoum, Abdulgader; Dhillon, Navneet; Buch, Shilpa; Berkland, Cory

    2010-05-01

    Biodegradable polymeric nanoparticles are currently being explored as a nonviral gene delivery system; however, many obstacles impede the translation of these nanomaterials. For example, nanoparticles delivered systemically are inherently prone to adsorbing serum proteins and agglomerating as a result of their large surface/volume ratio. What is desired is a simple procedure to prepare nanoparticles that may be delivered locally and exhibit minimal toxicity while improving entry into cells for effectively delivering DNA. The objective of this study was to optimize the formulation of poly(D,L-lactide-co-glycolide) (PLG) nanoparticles for gene delivery performance to a model of the pulmonary epithelium. Using a simple solvent diffusion technique, the chemistry of the particle surface was varied by using different coating materials that adsorb to the particle surface during formation. A variety of cationic coating materials were studied and compared to more conventional surfactants used for PLG nanoparticle fabrication. Nanoparticles (approximately 200 nm) efficiently encapsulated plasmids encoding for luciferase (80-90%) and slowly released the same for 2 weeks. In A549 alveolar lung epithelial cells, high levels of gene expression appeared at day 5 for certain positively charged PLG particles and gene expression was maintained for at least 2 weeks. In contrast, PEI gene expression ended at day 5. PLG particles were also significantly less cytotoxic than PEI suggesting the use of these vehicles for localized, sustained gene delivery to the pulmonary epithelium. PMID:19911425

  16. Endocytosis and exocytosis of nanoparticles in mammalian cells

    PubMed Central

    Oh, Nuri; Park, Ji-Ho

    2014-01-01

    Engineered nanoparticles that can be injected into the human body hold tremendous potential to detect and treat complex diseases. Understanding of the endocytosis and exocytosis mechanisms of nanoparticles is essential for safe and efficient therapeutic application. In particular, exocytosis is of significance in the removal of nanoparticles with drugs and contrast agents from the body, while endocytosis is of great importance for the targeting of nanoparticles in disease sites. Here, we review the recent research on the endocytosis and exocytosis of functionalized nanoparticles based on various sizes, shapes, and surface chemistries. We believe that this review contributes to the design of safe nanoparticles that can efficiently enter and leave human cells and tissues. PMID:24872703

  17. Aptamer-Conjugated Nanoparticles for Cancer Cell Detection

    PubMed Central

    Medley, Colin D.; Bamrungsap, Suwussa; Tan, Weihong; Smith, Joshua E.

    2011-01-01

    Aptamer-conjugated nanoparticles (ACNPs) have been used for a variety of applications, particularly dual nanoparticles for magnetic extraction and fluorescent labeling. In this type of assay, silica-coated magnetic and fluorophore-doped silica nanoparticles are conjugated to highly selective aptamers to detect and extract targeted cells in a variety of matrices. However, considerable improvements are required in order to increase the selectivity and sensitivity of this two-particle assay to be useful in a clinical setting. To accomplish this, several parameters were investigated, including nanoparticle size, conjugation chemistry, use of multiple aptamer sequences on the nanoparticles, and use of multiple nanoparticles with different aptamer sequences. After identifying the best-performing elements, the improvements made to this assay’s conditional parameters were combined to illustrate the overall enhanced sensitivity and selectivity of the two particle assay using an innovative multiple aptamer approach, signifying a critical feature in the advancement of this technique. PMID:21218774

  18. Core/shell Fe3O4 @SiO2 nanoparticles modified with PAH as a vector for EGFP plasmid DNA delivery into HeLa cells.

    PubMed

    Shi, Mengran; Liu, Yiyao; Xu, Mingming; Yang, Hong; Wu, Chunhui; Miyoshi, Hirokazu

    2011-11-10

    Novel stable core/shell Fe(3)O(4)@SiO(2)/PAH nanoparticles are synthesized using 15 nm Fe(3)O(4) as the template that is modified with PAH. The resulting nanoparticles can absorb plasmid DNA to mediate gene transfer in cultured HeLa cells. An electrophoretic assay suggests that the Fe(3)O(4)@SiO(2)/PAH nanoparticles protect the plasmid DNA from serum and DNase I degradation. A cell viability assay shows that the Fe(3)O(4)@SiO(2)/PAH nanoparticles exhibit a low cytotoxicity toward endothelial cells. Qualitative analysis of transfection in HeLa cells by nanoparticles carrying a plasmid DNA encoding EGFP demonstrates a fairly high expression level, even in the presence of serum. Thus, Fe(3)O(4)@SiO(2)/PAH nanoparticles are biocompatible and suitable for nonviral delivery, and may find applications in cancer therapy. PMID:22052564

  19. Photosensitivity of neurons enabled by cell-targeted gold nanoparticles.

    PubMed

    Carvalho-de-Souza, João L; Treger, Jeremy S; Dang, Bobo; Kent, Stephen B H; Pepperberg, David R; Bezanilla, Francisco

    2015-04-01

    Unmodified neurons can be directly stimulated with light to produce action potentials, but such techniques have lacked localization of the delivered light energy. Here we show that gold nanoparticles can be conjugated to high-avidity ligands for a variety of cellular targets. Once bound to a neuron, these particles transduce millisecond pulses of light into heat, which changes membrane capacitance, depolarizing the cell and eliciting action potentials. Compared to non-functionalized nanoparticles, ligand-conjugated nanoparticles highly resist convective washout and enable photothermal stimulation with lower delivered energy and resulting temperature increase. Ligands targeting three different membrane proteins were tested; all showed similar activity and washout resistance. This suggests that many types of ligands can be bound to nanoparticles, preserving ligand and nanoparticle function, and that many different cell phenotypes can be targeted by appropriate choice of ligand. The findings have applications as an alternative to optogenetics and potentially for therapies involving neuronal photostimulation. VIDEO ABSTRACT. PMID:25772189

  20. Platinum folate nanoparticles toxicity: cancer vs. normal cells.

    PubMed

    Mironava, Tatsiana; Simon, Marcia; Rafailovich, Miriam H; Rigas, Basil

    2013-03-01

    Almost for two decades metallic nanoparticles are successfully used for cancer detection, imaging and treatment. Due to their high electron density they can be easily observed by electron microscopy and used in laser and radiofrequency therapy as energy releasing agents. However, the limitation for this practice is an inability to generate tumor-specific heating in a minimally invasive manner to the healthy tissue. To overcome this restraint we proposed to use folic acid coated metallic nanoparticles and determine whether they preferentially penetrate cancer cells. We developed technique for synthesizing platinum nanoparticles using folic acid as stabilizing agent which produced particles of relatively narrow size distribution, having d=2.3 ± 0.5 nm. High resolution TEM and zeta potential analysis indicated that the particles produced by this method had a high degree of crystalline order with no amorphous outer shell and a high degree of colloidal stability. The keratinocytes and mammary breast cells (cancer and normal) were incubated with platinum folate nanoparticles, and the results showed that the IC50 was significantly higher for the normal cells than the cancer cells in both cases, indicating that these nanoparticles preferentially target the cancer cells. TEM images of thin sections taken from the two types of cells indicated that the number of vacuoles and morphology changes after incubation with nanoparticles was also larger for the cancer cells in both types of tissue studied. No preferential toxicity was observed when folic acid receptors were saturated with free folic acid prior to exposure to nanoparticles. These results confirm our hypothesis regarding the preferential penetration of folic acid coated nanoparticles to cancer cells due to receptor mediated endocytosis. PMID:23318730

  1. A pretargeted nanoparticle system for tumor cell labeling.

    PubMed

    Gunn, Jonathan; Park, Steven I; Veiseh, Omid; Press, Oliver W; Zhang, Miqin

    2011-03-01

    Nanoparticle-based cancer diagnostics and therapeutics can be significantly enhanced by selective tissue localization, but the strategy can be complicated by the requirement of a targeting ligand conjugated on nanoparticles, that is specific to only one or a limited few types of neoplastic cells, necessitating the development of multiple nanoparticle systems for different diseases. Here, we present a new nanoparticle system that capitalizes on a targeting pretreatment strategy, where a circulating fusion protein (FP) selectively prelabels the targeted cellular epitope, and a biotinylated iron oxide nanoparticle serves as a secondary label that binds to the FP on the target cell. This approach enables a single nanoparticle formulation to be used with any one of existing fusion proteins to bind a variety of target cells. We demonstrated this approach with two fusion proteins against two model cancer cell lines: lymphoma (Ramos) and leukemia (Jurkat), which showed 72.2% and 91.1% positive labeling, respectively. Notably, TEM analysis showed that a large nanoparticle population was endocytosed via attachment to the non-internalizing CD20 epitope. PMID:21107453

  2. Quantification of Nanoparticle Dose and Vesicular Inheritance in Proliferating Cells

    PubMed Central

    2013-01-01

    Assessing dose in nanoparticle–cell interactions is inherently difficult due to a complex multiplicity of possible mechanisms and metrics controlling particle uptake. The fundamental unit of nanoparticle dose is the number of particles internalized per cell; we show that this can be obtained for large cell populations that internalize fluorescent nanoparticles by endocytosis, through calibration of cytometry measurements to transmission electron microscopy data. Low-throughput, high-resolution electron imaging of quantum dots in U-2 OS cells is quantified and correlated with high-throughput, low-resolution optical imaging of the nanoparticle-loaded cells. From the correlated data, we obtain probability distribution functions of vesicles per cell and nanoparticles per vesicle. Sampling of these distributions and comparison to fluorescence intensity histograms from flow cytometry provide the calibration factor required to transform the cytometry metric to total particle dose per cell, the mean value of which is 2.4 million. Use of the probability distribution functions to analyze particle partitioning during cell division indicates that, while vesicle inheritance is near symmetric, highly variable vesicle loading leads to a highly asymmetric particle dose within the daughter cells. PMID:23773085

  3. Hybrid superparamagnetic iron oxide nanoparticle-branched polyethylenimine magnetoplexes for gene transfection of vascular endothelial cells

    Microsoft Academic Search

    Ran Namgung; Kaushik Singha; Mi Kyung Yu; Sangyong Jon; Yong Sook Kim; Youngkeun Ahn; In-Kyu Park; Won Jong Kim

    2010-01-01

    The work demonstrated the development of thermally cross-linked superparamagnetic nanomaterial which possessed polyethylene glycol moiety and covalently linked branched polyethylenimine (BPEI), and exhibited highly efficient magnetofection even under serum conditioned media. The study showed its high anti-biofouling, cell viability and serum stability and thus revealed a potential magnetic nanoparticle-mediated targeted gene delivery system. This superparamagnetic particle mediated rapid and efficient

  4. Enhanced magnetic resonance imaging and staining of cancer cells using ferrimagnetic H-ferritin nanoparticles with increasing core size

    PubMed Central

    Cai, Yao; Cao, Changqian; He, Xiaoqing; Yang, Caiyun; Tian, Lanxiang; Zhu, Rixiang; Pan, Yongxin

    2015-01-01

    Purpose This study is to demonstrate the nanoscale size effect of ferrimagnetic H-ferritin (M-HFn) nanoparticles on magnetic properties, relaxivity, enzyme mimetic activities, and application in magnetic resonance imaging (MRI) and immunohistochemical staining of cancer cells. Materials and methods M-HFn nanoparticles with different sizes of magnetite cores in the range of 2.7–5.3 nm were synthesized through loading different amounts of iron into recombinant human H chain ferritin (HFn) shells. Core size, crystallinity, and magnetic properties of those M-HFn nanoparticles were analyzed by transmission electron microscope and low-temperature magnetic measurements. The MDA-MB-231 cancer cells were incubated with synthesized M-HFn nanoparticles for 24 hours in Dulbecco’s Modified Eagle’s Medium. In vitro MRI of cell pellets after M-HFn labeling was performed at 7 T. Iron uptake of cells was analyzed by Prussian blue staining and inductively coupled plasma mass spectrometry. Immunohistochemical staining by using the peroxidase-like activity of M-HFn nanoparticles was carried out on MDA-MB-231 tumor tissue paraffin sections. Results The saturation magnetization (Ms), relaxivity, and peroxidase-like activity of synthesized M-HFn nanoparticles were monotonously increased with the size of ferrimagnetic cores. The M-HFn nanoparticles with the largest core size of 5.3 nm exhibit the strongest saturation magnetization, the highest peroxidase activity in immunohistochemical staining, and the highest r2 of 321 mM?1 s?1, allowing to detect MDA-MB-231 breast cancer cells as low as 104 cells mL?1. Conclusion The magnetic properties, relaxivity, and peroxidase-like activity of M-HFn nanoparticles are size dependent, which indicates that M-HFn nanoparticles with larger magnetite core can significantly enhance performance in MRI and staining of cancer cells. PMID:25878496

  5. Bifunctional magnetic-fluorescent nanoparticles: synthesis, characterization, and cell imaging.

    PubMed

    Lu, Yanjiao; Zheng, Yang; You, Shusen; Wang, Feng; Gao, Zhuo; Shen, Jie; Yang, Wantai; Yin, Meizhen

    2015-03-11

    A new type of bifunctional magnetic-fluorescent Fe3O4@SiO2-PDI-PAA/Ca(2+) nanoparticles has been prepared by coating PDI-cored star polymers (PDI-PAA) onto the surface of Fe3O4@SiO2 core-shell nanostructures. The morphology and properties of the composite nanoparticles are investigated by transmission electron microscopy, ultraviolet-visible spectrometry, fluorescence spectrometry, and vibrating sample magnetometry. The composite nanoparticles display a strong red emission and superparamagnetic behavior at room temperature. The cell viability and uptake assays reveal good biocompatibility of these hybrid nanoparticles. Hence, the composite nanoparticles are of potential to be further explored as therapeutic vector in biomedical field. PMID:25691125

  6. Immobilization of gold nanoparticles on cell culture surfaces for safe and enhanced gold nanoparticle-mediated laser transfection

    NASA Astrophysics Data System (ADS)

    Kalies, Stefan; Heinemann, Dag; Schomaker, Markus; Gentemann, Lara; Meyer, Heiko; Ripken, Tammo

    2014-07-01

    In comparison to standard transfection methods, gold nanoparticle-mediated laser transfection has proven to be a versatile alternative. This is based on its minor influence on cell viability and its high efficiency, especially for the delivery of small molecules like small interfering RNA. However, in order to transfer it to routine usage, a safety aspect is of major concern: The avoidance of nanoparticle uptake by the cells is desired. The immobilization of the gold nanoparticles on cell culture surfaces can address this issue. In this study, we achieved this by silanization of the appropriate surfaces and the binding of gold nanoparticles to them. Comparable perforation efficiencies to the previous approaches of gold nanoparticle-mediated laser transfection with free gold nanoparticles are demonstrated. The uptake of the immobilized particles by the cells is unlikely. Consequently, these investigations offer the possibility of bringing gold nanoparticle-mediated laser transfection closer to routine usage.

  7. Nanoparticle accumulation and transcytosis in brain endothelial cell layers

    NASA Astrophysics Data System (ADS)

    Ye, Dong; Raghnaill, Michelle Nic; Bramini, Mattia; Mahon, Eugene; Åberg, Christoffer; Salvati, Anna; Dawson, Kenneth A.

    2013-10-01

    The blood-brain barrier (BBB) is a selective barrier, which controls and limits access to the central nervous system (CNS). The selectivity of the BBB relies on specialized characteristics of the endothelial cells that line the microvasculature, including the expression of intercellular tight junctions, which limit paracellular permeability. Several reports suggest that nanoparticles have a unique capacity to cross the BBB. However, direct evidence of nanoparticle transcytosis is difficult to obtain, and we found that typical transport studies present several limitations when applied to nanoparticles. In order to investigate the capacity of nanoparticles to access and transport across the BBB, several different nanomaterials, including silica, titania and albumin- or transferrin-conjugated gold nanoparticles of different sizes, were exposed to a human in vitro BBB model of endothelial hCMEC/D3 cells. Extensive transmission electron microscopy imaging was applied in order to describe nanoparticle endocytosis and typical intracellular localisation, as well as to look for evidence of eventual transcytosis. Our results show that all of the nanoparticles were internalised, to different extents, by the BBB model and accumulated along the endo-lysosomal pathway. Rare events suggestive of nanoparticle transcytosis were also observed for several of the tested materials.The blood-brain barrier (BBB) is a selective barrier, which controls and limits access to the central nervous system (CNS). The selectivity of the BBB relies on specialized characteristics of the endothelial cells that line the microvasculature, including the expression of intercellular tight junctions, which limit paracellular permeability. Several reports suggest that nanoparticles have a unique capacity to cross the BBB. However, direct evidence of nanoparticle transcytosis is difficult to obtain, and we found that typical transport studies present several limitations when applied to nanoparticles. In order to investigate the capacity of nanoparticles to access and transport across the BBB, several different nanomaterials, including silica, titania and albumin- or transferrin-conjugated gold nanoparticles of different sizes, were exposed to a human in vitro BBB model of endothelial hCMEC/D3 cells. Extensive transmission electron microscopy imaging was applied in order to describe nanoparticle endocytosis and typical intracellular localisation, as well as to look for evidence of eventual transcytosis. Our results show that all of the nanoparticles were internalised, to different extents, by the BBB model and accumulated along the endo-lysosomal pathway. Rare events suggestive of nanoparticle transcytosis were also observed for several of the tested materials. Electronic supplementary information (ESI) available: Nanoparticle characterization in relevant media by Dynamic Light Scattering and SDS-PAGE. Transport study for silica nanoparticles across the BBB layer. Additional Electron Microscopy images of cells treated with the different nanoparticles investigated and details of the filters of the transwell systems. See DOI: 10.1039/c3nr02905k

  8. Au-ZnO hybrid nanoparticles exhibiting strong charge-transfer-induced SERS for recyclable SERS-active substrates.

    PubMed

    Liu, Liping; Yang, Haitao; Ren, Xiao; Tang, Jin; Li, Yongfeng; Zhang, Xiangqun; Cheng, Zhaohua

    2015-03-12

    Flower-shaped Au-ZnO hybrid nanoparticles have been prepared via seeding growth and subsequent wet-chemical etching of Au-ZnO core-shell nanoparticles. The etched Au-ZnO hybrid nanoparticles have shown a stronger surface-enhanced Raman scattering (SERS) signal of the nontotally symmetric (b2) vibrational modes of PATP molecules than Au nanoparticles alone, which is attributed to the chemical enhancement effect of the ZnO layer which is greatly excited by the localized surface plasmon resonance (LSPR) of Au cores. Further, the mechanism of the LSPR-enhanced charge transfer (CT) effect has been proved by the SERS spectra of PATP molecules excited using different laser sources from 325 to 785 nm. Moreover, the photocatalytic experimental results indicated that Au-ZnO hybrid nanoparticles are promising as biologically compatible and recyclable SERS-active platforms for different molecular species. PMID:25721784

  9. Nanoparticle facilitated extracellular electron transfer in microbial fuel cells.

    PubMed

    Jiang, Xiaocheng; Hu, Jinsong; Lieber, Alexander M; Jackan, Charles S; Biffinger, Justin C; Fitzgerald, Lisa A; Ringeisen, Bradley R; Lieber, Charles M

    2014-11-12

    Microbial fuel cells (MFCs) have been the focus of substantial research interest due to their potential for long-term, renewable electrical power generation via the metabolism of a broad spectrum of organic substrates, although the low power densities have limited their applications to date. Here, we demonstrate the potential to improve the power extraction by exploiting biogenic inorganic nanoparticles to facilitate extracellular electron transfer in MFCs. Simultaneous short-circuit current recording and optical imaging on a nanotechnology-enabled platform showed substantial current increase from Shewanella PV-4 after the formation of cell/iron sulfide nanoparticle aggregates. Detailed characterization of the structure and composition of the cell/nanoparticle interface revealed crystalline iron sulfide nanoparticles in intimate contact with and uniformly coating the cell membrane. In addition, studies designed to address the fundamental mechanisms of charge transport in this hybrid system showed that charge transport only occurred in the presence of live Shewanella, and moreover demonstrated that the enhanced current output can be attributed to improved electron transfer at cell/electrode interface and through the cellular-networks. Our approach of interconnecting and electrically contacting bacterial cells through biogenic nanoparticles represents a unique and promising direction in MFC research and has the potential to not only advance our fundamental knowledge about electron transfer processes in these biological systems but also overcome a key limitation in MFCs by constructing an electrically connected, three-dimensional cell network from the bottom-up. PMID:25310721

  10. Metal nanoparticles amplify photodynamic effect on skin cells in vitro

    NASA Astrophysics Data System (ADS)

    Bauer, Brigitte; Chen, Si; Käll, Mikael; Gunnarsson, Linda; Ericson, Marica B.

    2011-03-01

    We report on an investigation aimed to increase the efficiency of photodynamic therapy (PDT) through the influence of localized surface plasmon resonances (LSPR's) in metal nanoparticles. PDT is based on photosensitizers that generate singlet oxygen at the tumour site upon exposure to visible light. Although PDT is a well-established treatment for skin cancer, a major drawback is the low quantum yield for singlet-oxygen production. This motivates the development of novel methods that enhance singlet oxygen generation during treatment. In this context, we study the photodynamic effect on cultured human skin cells in the presence or absence of gold nanoparticles with well established LSPR and field-enhancement properties. The cultured skin cells were exposed to protoporphyrin IX and gold nanoparticles and subsequently illuminated with red light. We investigated the differences in cell viability by tuning different parameters, such as incubation time and light dose. In order to find optimal parameters for specific targeting of tumour cells, we compared normal human epidermal keratinocytes with a human squamous skin cancer cell line. The study indicates significantly enhanced cell death in the presence of nanoparticles and important differences in treatment efficiency between normal and tumour cells. These results are thus promising and clearly motivate further development of nanoparticle enhanced clinical PDT treatment.

  11. Titanium dioxide nanoparticles induce oxidative stress-mediated apoptosis in human keratinocyte cells.

    PubMed

    Shukla, Ritesh K; Kumar, Ashutosh; Pandey, Alok K; Singh, Shashi S; Dhawan, Alok

    2011-02-01

    Titanium dioxide nanoparticles (TiO2 NPs) are the most commonly used metal oxide NPs in various industrial and commercial products. The present study has demonstrated a significant cellular uptake of TiO2 NPs in the human keratinocyte cells (HaCaT) using transmission electron microscopy and flow cytometry. The data exhibited a significant (p < 0.05) concentration dependent decrease in cell viability and glutathione with concomitant increase in lipid peroxidation and reactive oxygen species. The increased oxidative stress further leads to apoptosis after 48 h of exposure. Our study demonstrates oxidative stress mediated apoptosis in human keratinocyte cells exposed to TiO2 NPs. PMID:21485823

  12. Au-ZnO hybrid nanoparticles exhibiting strong charge-transfer-induced SERS for recyclable SERS-active substrates

    NASA Astrophysics Data System (ADS)

    Liu, Liping; Yang, Haitao; Ren, Xiao; Tang, Jin; Li, Yongfeng; Zhang, Xiangqun; Cheng, Zhaohua

    2015-03-01

    Flower-shaped Au-ZnO hybrid nanoparticles have been prepared via seeding growth and subsequent wet-chemical etching of Au-ZnO core-shell nanoparticles. The etched Au-ZnO hybrid nanoparticles have shown a stronger surface-enhanced Raman scattering (SERS) signal of the nontotally symmetric (b2) vibrational modes of PATP molecules than Au nanoparticles alone, which is attributed to the chemical enhancement effect of the ZnO layer which is greatly excited by the localized surface plasmon resonance (LSPR) of Au cores. Further, the mechanism of the LSPR-enhanced charge transfer (CT) effect has been proved by the SERS spectra of PATP molecules excited using different laser sources from 325 to 785 nm. Moreover, the photocatalytic experimental results indicated that Au-ZnO hybrid nanoparticles are promising as biologically compatible and recyclable SERS-active platforms for different molecular species.Flower-shaped Au-ZnO hybrid nanoparticles have been prepared via seeding growth and subsequent wet-chemical etching of Au-ZnO core-shell nanoparticles. The etched Au-ZnO hybrid nanoparticles have shown a stronger surface-enhanced Raman scattering (SERS) signal of the nontotally symmetric (b2) vibrational modes of PATP molecules than Au nanoparticles alone, which is attributed to the chemical enhancement effect of the ZnO layer which is greatly excited by the localized surface plasmon resonance (LSPR) of Au cores. Further, the mechanism of the LSPR-enhanced charge transfer (CT) effect has been proved by the SERS spectra of PATP molecules excited using different laser sources from 325 to 785 nm. Moreover, the photocatalytic experimental results indicated that Au-ZnO hybrid nanoparticles are promising as biologically compatible and recyclable SERS-active platforms for different molecular species. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr00491h

  13. The Effects of Silica Nanoparticles in Macrophage Cells

    PubMed Central

    Kim, Seungjae; Jang, Jiyoung; Kim, Hyojin; Choi, Hoon; Lee, Kangtaek

    2012-01-01

    Silica nanoparticles, which are applicable in many industrial fields, have been reported to induce cellular changes such as cytotoxicity in various cells and fibrosis in lungs. Because the immune system is the primary targeting organ reacting to internalized exogenous nanoparticles, we tried to figure out the immunostimulatory effect of silica nanoparticles in macrophages using differently sized silica nanoparticles. Using U937 cells we assessed cytotoxicity by CCK-8 assay, ROS generation by CM-H2DCFDA, intracellular Ca++ levels by staining with Fluo4-AM and IL-8 production by ELISA. At non-toxic concentration, the intracellular Ca++ level has increased immediately after exposure to 15 nm particles, not to larger particles. ROS generation was detected significantly in response to 15 nm particles. However, all three different sizes of silica nanoparticles induced IL-8 production. 15 nm silica nanoparticles are more stimulatory than larger particles in cytotoxicity, intracellular Ca++ increase and ROS generation. But IL-8 production was induced to same levels with 50 or 100 nm particles. Therefore, IL-8 production induced by silica nanoparticles may be dependent on other mechanisms rather than intracellular Ca++ increase and ROS generation. PMID:23397001

  14. Laser-targeted photofabrication of gold nanoparticles inside cells

    NASA Astrophysics Data System (ADS)

    Smith, Nicholas I.; Mochizuki, Kentaro; Niioka, Hirohiko; Ichikawa, Satoshi; Pavillon, Nicolas; Hobro, Alison J.; Ando, Jun; Fujita, Katsumasa; Kumagai, Yutaro

    2014-10-01

    Nanoparticle manipulation is of increasing interest, since they can report single molecule-level measurements of the cellular environment. Until now, however, intracellular nanoparticle locations have been essentially uncontrollable. Here we show that by infusing a gold ion solution, focused laser light-induced photoreduction allows in situ fabrication of gold nanoparticles at precise locations. The resulting particles are pure gold nanocrystals, distributed throughout the laser focus at sizes ranging from 2 to 20?nm, and remain in place even after removing the gold solution. We demonstrate the spatial control by scanning a laser beam to write characters in gold inside a cell. Plasmonically enhanced molecular signals could be detected from nanoparticles, allowing their use as nano-chemical probes at targeted locations inside the cell, with intracellular molecular feedback. Such light-based control of the intracellular particle generation reaction also offers avenues for in situ plasmonic device creation in organic targets, and may eventually link optical and electron microscopy.

  15. Nanogel-quantum dot hybrid nanoparticles for live cell imaging

    SciTech Connect

    Hasegawa, Urara [Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10, Kanda-surugadai, Chiyoda-ku, Tokyo 101-0062 (Japan); Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone, Tokyo Medical and Dental University, 2-3-10, Kanda-surugadai, Chiyoda-ku, Tokyo 101-0062 (Japan); Nomura, Shin-ichiro M. [Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10, Kanda-surugadai, Chiyoda-ku, Tokyo 101-0062 (Japan); Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone, Tokyo Medical and Dental University, 2-3-10, Kanda-surugadai, Chiyoda-ku, Tokyo 101-0062 (Japan); Kaul, Sunil C. [National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan); Hirano, Takashi [National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan); Akiyoshi, Kazunari [Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10, Kanda-surugadai, Chiyoda-ku, Tokyo 101-0062 (Japan); Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone, Tokyo Medical and Dental University, 2-3-10, Kanda-surugadai, Chiyoda-ku, Tokyo 101-0062 (Japan); E-mail: akiyoshi.org@tmd.ac.jp

    2005-06-17

    We report here a novel carrier of quantum dots (QDs) for intracellular labeling. Monodisperse hybrid nanoparticles (38 nm in diameter) of QDs were prepared by simple mixing with nanogels of cholesterol-bearing pullulan (CHP) modified with amino groups (CHPNH{sub 2}). The CHPNH{sub 2}-QD nanoparticles were effectively internalized into the various human cells examined. The efficiency of cellular uptake was much higher than that of a conventional carrier, cationic liposome. These hybrid nanoparticles could be a promising fluorescent probe for bioimaging.

  16. In Vitro Cytotoxicity of Nanoparticles in Mammalian Germline Stem Cells

    PubMed Central

    Braydich-Stolle, Laura; Hussain, Saber; Schlager, John J.; Hofmann, Marie-Claude

    2010-01-01

    Gametogenesis is a complex biological process that is particularly sensitive to environmental insults such as chemicals. Many chemicals have a negative impact on the germline, either by directly affecting the germ cells, or indirectly through their action on the somatic nursing cells. Ultimately, these effects can inhibit fertility, and they may have negative consequences for the development of the offspring. Recently, nanomaterials such as nanotubes, nanowires, fullerene derivatives (buckyballs), and quantum dots have received enormous national attention in the creation of new types of analytical tools for biotechnology and the life sciences. Despite the wide application of nanomaterials, there is a serious lack of information concerning their impact on human health and the environment. Thus, there are limited studies available on toxicity of nanoparticles for risk assessment of nanomaterials. The purpose of this study was to assess the suitability of a mouse spermatogonial stem cell line as a model to assess nanotoxicity in the male germline in vitro. The effects of different types of nanoparticles on these cells were evaluated by light microscopy, and by cell proliferation and standard cytotoxicity assays. Our results demonstrate a concentration-dependent toxicity for all types of particles tested, whereas the corresponding soluble salts had no significant effect. Silver nanoparticles were the most toxic while molybdenum trioxide (MoO3) nanoparticles were the least toxic. Our results suggest that this cell line provides a valuable model with which to assess the cytotoxicity of nanoparticles in the germ line in vitro. PMID:16014736

  17. Single Walled Carbon Nanotubes Exhibit Dual-Phase Regulation to Exposed Arabidopsis Mesophyll Cells

    NASA Astrophysics Data System (ADS)

    Yuan, Hengguang; Hu, Shanglian; Huang, Peng; Song, Hua; Wang, Kan; Ruan, Jing; He, Rong; Cui, Daxiang

    2010-12-01

    Herein we are the first to report that single-walled carbon nanotubes (SWCNTs) exhibit dual-phase regulation to Arabidopsis mesophyll cells exposed to different concentration of SWCNTs. The mesophyll protoplasts were prepared by enzyme digestion, and incubated with 15, 25, 50, 100 ?g/ml SWCNTs for 48 h, and then were observed by optical microscopy and transmission electron microscopy, the reactive oxygen species (ROS) generation was measured. Partial protoplasts were stained with propidium iodide and 4'-6- diamidino-2-phenylindole, partial protoplasts were incubated with fluorescein isothiocyanate-labeled SWCNTs, and observed by fluorescence microscopy. Results showed that SWCNTs could traverse both the plant cell wall and cell membrane, with less than or equal to 50 ?g/ml in the culture medium, SWCNTs stimulated plant cells to grow out trichome clusters on their surface, with more than 50 ?g/ml SWCNTs in the culture medium, SWCNTs exhibited obvious toxic effects to the protoplasts such as increasing generation of ROS, inducing changes of protoplast morphology, changing green leaves into yellow, and inducing protoplast cells' necrosis and apoptosis. In conclusion, single walled carbon nanotubes can get through Arabidopsis mesophyll cell wall and membrane, and exhibit dose-dependent dual-phase regulation to Arabidopsis mesophyll protoplasts such as low dose stimulating cell growth, and high dose inducing cells' ROS generation, necrosis or apoptosis.

  18. Biocompatibility Study of Gold Nanoparticles to Human Cells

    Microsoft Academic Search

    J. H. Fan; W. I. Hung; W. T. Li; J. M. Yeh

    Gold nanoparticle (GNP) is one of the most stable and popular nanoparticles, which receives considerable attention due to\\u000a their applications in biomedical imaging and diagnostic tests. However, its cytotoxicity has not been fully investigated.\\u000a Here we report the effects on biocompatibility of water-soluble GNPs with different sizes and concentrations to human bone\\u000a marrow mesenchymal stem cells (hBMSCs) and human hepatoma

  19. Nanoparticle derived contacts for photovoltaic cells

    SciTech Connect

    Ginley, D.S.

    1999-10-20

    Contacts are becoming increasingly important as PV devices move to higher efficiency and lower cost. The authors present an approach to developing contacts using nanoparticle-based precursors. Both elemental, alloy and compound nanoparticles can be employed for contacts. Ink based approaches can be utilized at low temperatures and utilize direct write techniques such as ink jet and screen printing. The ability to control the composition of the nanoparticle allows improved control of the contact metallurgy and the potential for thermodynamically stable interfaces. A key requirement is the ability to control the interface between particles and between particles and the substrate. The authors illustrate some of these principals with recent results on Al, Cu and (Hg,Cu)Te. They show that for the elemental materials control of the surface can prevent oxide formation and act as glue to control the reactivity of the nanoparticles.

  20. Toxicity of Cerium Oxide Nanoparticles in Human Lung Cancer Cells

    SciTech Connect

    Weisheng, Lin; Huang, Yue-wern; Zhou, Xiao Dong; Ma, Yinfa

    2006-12-31

    With the fast development of nanotechnology, the nanomaterials start to cause people's attention for potential toxic effect. In this paper, the cytotoxicity and oxidative stress caused by 20-nm cerium oxide (CeO2) nanoparticles in cultured human lung cancer cells was investigated. The sulforhodamine B method was employed to assess cell viability after exposure to 3.5, 10.5, and 23.3 ?g/ml of CeO2 nanoparticles for 24, 48, and 72 h. Cell viability decreased significantly as a function of nanoparticle dose and exposure time. Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species, glutathione, malondialdehyde, ?-tocopherol, and lactate dehydrogenase, were quantitatively assessed. It is concluded from the results that free radicals generated by exposure to 3.5 to 23.3 ?g/ml CeO2 nanoparticles produce significant oxidative stress in the cells, as reflected by reduced glutathione and ?-tocopherol levels; the toxic effects of CeO2 nanoparticles are dose dependent and time dependent; elevated oxidative stress increases the production of malondialdehyde and lactate dehydrogenase, which are indicators of lipid peroxidation and cell membrane damage, respectively.

  1. Drug-loaded nanoparticles induce gene expression in human pluripotent stem cell derivatives

    NASA Astrophysics Data System (ADS)

    Gajbhiye, Virendra; Escalante, Leah; Chen, Guojun; Laperle, Alex; Zheng, Qifeng; Steyer, Benjamin; Gong, Shaoqin; Saha, Krishanu

    2013-12-01

    Tissue engineering and advanced manufacturing of human stem cells requires a suite of tools to control gene expression spatiotemporally in culture. Inducible gene expression systems offer cell-extrinsic control, typically through addition of small molecules, but small molecule inducers typically contain few functional groups for further chemical modification. Doxycycline (DXC), a potent small molecule inducer of tetracycline (Tet) transgene systems, was conjugated to a hyperbranched dendritic polymer (Boltorn H40) and subsequently reacted with polyethylene glycol (PEG). The resulting PEG-H40-DXC nanoparticle exhibited pH-sensitive drug release behavior and successfully controlled gene expression in stem-cell-derived fibroblasts with a Tet-On system. While free DXC inhibited fibroblast proliferation and matrix metalloproteinase (MMP) activity, PEG-H40-DXC nanoparticles maintained higher fibroblast proliferation levels and MMP activity. The results demonstrate that the PEG-H40-DXC nanoparticle system provides an effective tool to controlling gene expression in human stem cell derivatives.Tissue engineering and advanced manufacturing of human stem cells requires a suite of tools to control gene expression spatiotemporally in culture. Inducible gene expression systems offer cell-extrinsic control, typically through addition of small molecules, but small molecule inducers typically contain few functional groups for further chemical modification. Doxycycline (DXC), a potent small molecule inducer of tetracycline (Tet) transgene systems, was conjugated to a hyperbranched dendritic polymer (Boltorn H40) and subsequently reacted with polyethylene glycol (PEG). The resulting PEG-H40-DXC nanoparticle exhibited pH-sensitive drug release behavior and successfully controlled gene expression in stem-cell-derived fibroblasts with a Tet-On system. While free DXC inhibited fibroblast proliferation and matrix metalloproteinase (MMP) activity, PEG-H40-DXC nanoparticles maintained higher fibroblast proliferation levels and MMP activity. The results demonstrate that the PEG-H40-DXC nanoparticle system provides an effective tool to controlling gene expression in human stem cell derivatives. Electronic supplementary information (ESI) available: ESI containing 1H NMR spectra and additional fibroblast characterization data. See DOI: 10.1039/c3nr04794f

  2. Juglans mandshurica Maxim extracts exhibit antitumor activity on HeLa cells in vitro.

    PubMed

    Xin, Nian; Hasan, Murtaza; Li, Wei; Li, Yan

    2014-04-01

    The present study examined the potential application of Juglans mandshurica Maxim extracts (HT) for cancer therapy by assessing their anti?proliferative activity, reduction of telomerase activity, induction of apoptosis and cell cycle arrest in S phase in HeLa cells. From the perspective of using HT as a herbal medicine, photomicroscopy and florescent microscopy techniques were utilized to characterize the effect of the extracts on telomerase activity and cell morphology. Flow cytometry was employed to study apoptosis and cell cycle of HeLa cells, and DNA laddering was performed. The results showed that HT inhibited cell proliferation and telomerase activity, induced apoptosis and caused S phase arrest of HeLa cells in vitro. HT inhibited HeLa cell proliferation significantly, and the highest inhibition rate was 83.7%. A trap?silver staining assay showed that HT was capable of markedly decreasing telomerase activity of HeLa cells and this inhibition was enhanced in a time? and dose?dependent manner. Results of a Hoechst 33258 staining assay showed that HeLa cells treated by HT induced cell death. Through DNA agarose gel electrophoresis, DNA ladders of HeLa cells treated with HT were observed, indicating apoptosis. In conclusion, the present study demonstrated that HT exhibited anti?tumor effects comprising the inhibition of growth and telomerase activity as well as apoptosis and cell cycle arrest in HeLa cells. PMID:24566804

  3. Chitosan cross-linked docetaxel loaded EGF receptor targeted nanoparticles for lung cancer cells.

    PubMed

    Maya, S; Sarmento, Bruno; Lakshmanan, Vinoth-Kumar; Menon, Deepthy; Seabra, Vitor; Jayakumar, R

    2014-08-01

    Lung cancer, associated with the up-regulated epidermal growth factor receptor (EGFR) led to the development of EGFR targeted anticancer therapeutics. The biopolymeric nanoparticles form an outstanding system for the targeted delivery of therapeutic agents. The present work evaluated the in vitro effects of chitosan cross-linked ?-poly(glutamic acid) (?-PGA) nanoparticles (Nps) loaded with docetaxel (DTXL) and decorated with Cetuximab (CET), targeted to EGFR over-expressing non-small-cell-lung-cancer (NSCLC) cells (A549). CET-DTXL-?-PGA Nps was prepared by ionic gelation and CET conjugation via EDC/NHS chemistry. EGFR specificity of targeted Nps was confirmed by the higher uptake rates of EGFR +ve A549 cells compared to that of EGFR -ve cells (NIH3T3). The cytotoxicity of Nps quantified using cell based (MTT/LDH) and flowcytometry (Cell-cycle analysis, Annexin V/PI and JC-1) assays showed superior antiproliferative activity of CET-DTXL-?-PGA Nps over DTXL-?-PGA Nps. The A549 cells treated with CET-DTXL-?-PGA NPs underwent a G2/M phase cell cycle arrest followed by reduction in mitochondrial membrane potential of A549 cells, inducing apoptosis and necrosis resulting in enhanced cancer cell death. CET-DTXL-?-PGA Nps exhibited enhanced cellular internalization and therapeutic activity, by actively targeting EGFR on NSCLC cells and hence could be an effective alternative to non-specific, conventional chemotherapy by increasing its efficiency by many folds. PMID:24950310

  4. Micropatterned mammalian cells exhibit phenotype-specific left-right asymmetry.

    PubMed

    Wan, Leo Q; Ronaldson, Kacey; Park, Miri; Taylor, Grace; Zhang, Yue; Gimble, Jeffrey M; Vunjak-Novakovic, Gordana

    2011-07-26

    Left-right (LR) asymmetry (handedness, chirality) is a well-conserved biological property of critical importance to normal development. Changes in orientation of the LR axis due to genetic or environmental factors can lead to malformations and disease. While the LR asymmetry of organs and whole organisms has been extensively studied, little is known about the LR asymmetry at cellular and multicellular levels. Here we show that the cultivation of cell populations on micropatterns with defined boundaries reveals intrinsic cell chirality that can be readily determined by image analysis of cell alignment and directional motion. By patterning 11 different types of cells on ring-shaped micropatterns of various sizes, we found that each cell type exhibited definite LR asymmetry (p value down to 10(-185)) that was different between normal and cancer cells of the same type, and not dependent on surface chemistry, protein coating, or the orientation of the gravitational field. Interestingly, drugs interfering with actin but not microtubule function reversed the LR asymmetry in some cell types. Our results show that micropatterned cell populations exhibit phenotype-specific LR asymmetry that is dependent on the functionality of the actin cytoskeleton. We propose that micropatterning could potentially be used as an effective in vitro tool to study the initiation of LR asymmetry in cell populations, to diagnose disease, and to study factors involved with birth defects in laterality. PMID:21709270

  5. Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

    PubMed Central

    Zhang, Xi-Feng; Choi, Yun-Jung; Han, Jae Woong; Kim, Eunsu; Park, Jung Hyun; Gurunathan, Sangiliyandi; Kim, Jin-Hoi

    2015-01-01

    Background Silver nanoparticles (AgNPs) possess unique physical, chemical, and biological properties. AgNPs have been increasingly used as anticancer, antiangiogenic, and antibacterial agents for the treatment of bacterial infections in open wounds as well as in ointments, bandages, and wound dressings. The present study aimed to investigate the effects of two different sizes of AgNPs (10 nm and 20 nm) in male somatic Leydig (TM3) and Sertoli (TM4) cells and spermatogonial stem cells (SSCs). Methods Here, we demonstrate a green and simple method for the synthesis of AgNPs using Bacillus cereus culture supernatants. The synthesized AgNPs were characterized using ultraviolet and visible absorption spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and transmission electron microscopy (TEM). The toxicity of the synthesized AgNPs was evaluated by the effects on cell viability, metabolic activity, oxidative stress, apoptosis, and expression of genes encoding steroidogenic and tight junction proteins. Results AgNPs inhibited the viability and proliferation of TM3 and TM4 cells in a dose- and size-dependent manner by damaging cell membranes and inducing the generation of reactive oxygen species, which in turn affected SSC growth on TM3 and TM4 as feeder cells. Small AgNPs (10 nm) were more cytotoxic than medium-sized nanoparticles (20 nm). TEM revealed the presence of AgNPs in the cell cytoplasm and nucleus, and detected mitochondrial damage and enhanced formation of autosomes and autolysosomes in the AgNP-treated cells. Flow cytometry analysis using Annexin V/propidium iodide staining showed massive cell death by apoptosis or necrosis. Real-time polymerase chain reaction and western blot analyses indicated that in TM3 and TM4 cells, AgNPs activated the p53, p38, and pErk1/2 signaling pathways and significantly downregulated the expression of genes related to testosterone synthesis (TM3) and tight junctions (TM4). Furthermore, the exposure of TM3 and TM4 cells to AgNPs inhibited proliferation and self-renewal of SSCs. Conclusion Our results suggest that AgNPs exhibit size-dependent nanoreprotoxicity in male somatic cells and SSCs, strongly suggesting that applications of AgNPs in commercial products must be carefully evaluated. Further studies of AgNPs-induced nanoreprotoxicity in animal models are required. PMID:25733828

  6. Cellular Nanomedicine Cell selective response to gold nanoparticles

    Microsoft Academic Search

    Hirak K. Patra; Utpal Chaudhuri; Prabir Lahiri

    Gold nanoparticles (GNPs) are considered a potential probe to detect cancer. The present article investigates whether GNPs, even in the absence of any specific functionalization, induce any cell- specific response. We report GNP-induced death response in human carcinoma lung cell line A549. In contrast, the two other cell lines tested, BHK21 (baby hamster kidney) and HepG2 (human hepatocellular liver carcinoma),

  7. Dedifferentiation derived cells exhibit phenotypic and functional characteristics of epidermal stem cells.

    PubMed

    Zhang, Cuiping; Fu, Xiaobing; Chen, Peng; Bao, Xiaoxia; Li, Fu; Sun, Xiaoyan; Lei, Yonghong; Cai, Sa; Sun, Tongzhu; Sheng, Zhiyong

    2010-05-01

    Differentiated epidermal cells can dedifferentiate into stem cells or stem cell-like cells in vivo. In this study, we report the isolation and characterization of dedifferentiation-derived cells. Epidermal sheets eliminated of basal stem cells were transplanted onto the skin wounds in 47 nude athymic (BALB/c-nu/nu) mice. After 5 days, cells negative for CK10 but positive for CK19 and beta1-integrin emerged at the wound-neighbouring side of the epidermal sheets. Furthermore, the percentages of CK19 and beta1-integrin+ cells detected by flow cytometric analysis were increased after grafting (P < 0.01) and CK10+ cells in grafted sheets decreased (P < 0.01). Then we isolated these cells on the basis of rapid adhesion to type IV collagen and found that there were 4.56% adhering cells (dedifferentiation-derived cells) in the grafting group within 10 min. The in vitro phenotypic assays showed that the expressions of CK19, beta1-integrin, Oct4 and Nanog in dedifferentiation-derived cells were remarkably higher than those in the control group (differentiated epidermal cells) (P < 0.01). In addition, the results of the functional investigation of dedifferentiation-derived cells demonstrated: (1) the numbers of colonies consisting of 5-10 cells and greater than 10 cells were increased 5.9-fold and 6.7-fold, respectively, as compared with that in the control (P < 0.01); (2) more cells were in S phase and G2/M phase of the cell cycle (proliferation index values were 21.02% in control group, 45.08% in group of dedifferentiation); (3) the total days of culture (28 days versus 130 days), the passage number of cells (3 passages versus 20 passages) and assumptive total cell output (1 x 10(5) cells versus 1 x 10(12) cells) were all significantly increased and (4) dedifferentiation-derived cells, as well as epidermal stem cells, were capable of regenerating a skin equivalent, but differentiated epidermal cells could not. These results suggested that the characteristics of dedifferentiation-derived cells cultured in vitro were similar to epidermal stem cells. This study may also offer a new approach to yield epidermal stem cells for wound repair and regeneration. PMID:19426155

  8. BTLA exhibits immune memory for ?? T cells in patients with active pulmonary tuberculosis

    PubMed Central

    Zeng, Jin-Cheng; Lin, Dong-Zi; Yi, Lai-Long; Liu, Gan-Bin; Zhang, Hui; Wang, Wan-Dang; Zhang, Jun-Ai; Wu, Xian-Jing; Xiang, Wen-Yu; Kong, Bin; Chen, Zheng W; Wang, Cong-Yi; Xu, Jun-Fa

    2014-01-01

    Despite past extensive studies, the role of B and T lymphocyte attenuator (BTLA) in ?? T cells in patients with active pulmonary tuberculosis (ATB) remains poorly understood. Here we demonstrate that BTLA expression on ?? T cells is decreased in patients with M. tuberculosis (Mtb) infection. Particularly, BTLA expression levels are likely critical for ?? T cells to manifest and maintain an active central memory phenotype with high capacity for secretion of IFN-? and perforin, which are important for immune memory against TB infection. BTLAhigh ?? T cells also exhibited higher capacity in response to Mtb peptide stimulation. In contrast to the role of BTLA played for negative regulation of immune responses, our data in the current studies suggest that BTLA expression on ?? T cells is likely associated with protective immune memory against Mtb infection in the setting of patients with active pulmonary tuberculosis. This previous unappreciated role for BTLA may have implications for prevention and treatment of patients with Mtb infection. PMID:25360214

  9. Nanocomposite of tin sulfide nanoparticles with reduced graphene oxide in high-efficiency dye-sensitized solar cells.

    PubMed

    Yang, Bo; Zuo, Xueqin; Chen, Peng; Zhou, Lei; Yang, Xiao; Zhang, Haijun; Li, Guang; Wu, Mingzai; Ma, Yongqing; Jin, Shaowei; Chen, Xiaoshuang

    2015-01-14

    A nanocomposite of SnS2 nanoparticles with reduced graphene oxide (SnS2@RGO) had been successfully synthesized as a substitute conventional Pt counter electrode (CE) in a dye-sensitized solar cell (DSSC) system. The SnS2 nanoparticles were uniformly dispersed onto graphene sheets, which formed a nanosized composite system. The effectiveness of this nanocomposite exhibited remarkable electrocatalytic properties upon reducing the triiodide, owning to synergistic effects of SnS2 nanoparticles dispersed on graphene sheet and improved conductivity. Consequently, the DSSC equipped with SnS2@RGO nanocomposite CE achieved power conversion efficiency (PCE) of 7.12%, which was higher than those of SnS2 nanoparticles (5.58%) or graphene sheet alone (3.73%) as CEs and also comparable to the value (6.79%) obtained with pure Pt CE as a reference. PMID:25230916

  10. Mammalian cells transfected with the listeriolysin gene exhibit enhanced proliferation and focus formation.

    PubMed Central

    Demuth, A; Chakraborty, T; Krohne, G; Goebel, W

    1994-01-01

    Mouse 3T6 and 3T3 fibroblasts and rat epithelial L2 cells were transfected with recombinant plasmids containing the listeriolysin gene (hly) of Listeria monocytogenes. This bacterial gene (with and without the 5' signal sequence) was cloned under the control of a murine metallothionein promoter, resulting in elevated transcription of both forms of the hly gene after induction with ZnSO4. However, the gene product could be observed only when the listeriolysin gene lacking the 5' signal sequence was used. Intact listeriolysin could not be detected in the cytoplasm or in the supernatant of the hly-transfected cells. 3T6 and L2 cells transfected with the intact hly gene exhibited significantly increased cell proliferation and increased formation of actin microfilaments upon induction of hly expression with ZnSO4. Both cell types are not contact inhibited and formed large piles of spherical cells after transfection with hly. In contrast, contact-inhibited 3T3 cells transfected with the hly gene showed increased proliferation but no formation of such cell aggregates. When 3T6 fibroblasts were transfected with the hly gene without the 5' signal sequence, inhibition of growth, lack of cell layer confluency, and altered (spherical) cell morphology were observed. Images PMID:7927793

  11. Crocin Exhibits Antitumor Effects on Human Leukemia HL-60 Cells In Vitro and In Vivo.

    PubMed

    Sun, Yan; Xu, Hui-Juan; Zhao, Yan-Xia; Wang, Ling-Zhen; Sun, Li-Rong; Wang, Zhi; Sun, Xiu-Fang

    2013-01-01

    Crocin is a carotenoid of the saffron extract that exhibits antitumor activity against many human tumors. However, the effects of crocin on HL-60 cells in vivo have not been evaluated. This study aimed to examine the effects of crocin on HL-60 cells in vitro and in vivo and investigate the underlying mechanisms. HL-60 cells were treated by crocin, and cell proliferation, apoptosis, and cell cycle profiles were examined by MTT assay, AO/EB staining, and flow cytometry, respectively. Furthermore, HL-60 cells were xenografted into nude mice and treated by crocin, the tumor weight and size were calculated, and the expression of Bcl-2 and Bax in xenografts was detected by immunohistochemical staining. The results showed that crocin (0.625-5?mg/mL) inhibited HL-60 cell proliferation and induced apoptosis and cell cycle arrest at G0/G1 phase, in a concentration and time-dependent manner. In addition, crocin (6.25, 25?mg/kg) inhibited the tumor weight and size of HL-60 xenografts in nude mice, inhibited Bcl-2 expression, and increased Bax expression in xenografts. In summary, crocin inhibits the proliferation and tumorigenicity of HL-60 cells, which may be mediated by the induction of apoptosis and cell cycle arrest and the regulation of Bcl-2 and Bax expression. PMID:23573146

  12. Nanoparticles blast cancerous cells with killer drugs | KurzweilAI http://www.kurzweilai.net/nanoparticles-blast-cancerous-cells-with-killer-drugs[5/2/2011 12:14:51 PM

    E-print Network

    Brinker, C. Jeffrey

    Nanoparticles blast cancerous cells with killer drugs | KurzweilAI http://www.kurzweilai.net/nanoparticles-blast-cancerous-cells-with-killer-drugs[5/2/2011 12:14:51 PM] You are here: Home News Nanoparticles blast cancerous cells with killer drugs News ALL NEWS Confirm Like You like Nanoparticles blast cancerous cells with killer drugs | Kurzweil

  13. Nanoparticle PEBBLE sensors in live cells and in vivo

    PubMed Central

    Smith, Ron

    2009-01-01

    Nanoparticle sensors have been developed for imaging and dynamic monitoring, in live cells and in vivo, of the molecular or ionic components, constructs, forces and dynamics, all in real time, during biological/chemical/physical processes. With their biocompatible small size and inert matrix, nanoparticle sensors have been successfully applied for non-invasive real-time measurements of analytes and fields in cells and rodents, with spatial, temporal, physical and chemical resolution. This review describes the diverse designs of nanoparticle sensors for ions and small molecules, physical fields and biological features, as well as the characterization, properties, and applications of these nanosensors to in vitro and in vivo measurements. Their floating as well as localization ability in biological media is captured by the acronym PEBBLE: photonic explorer for bioanalysis with biologically localized embedding. PMID:20098636

  14. Human Myogenic Endothelial Cells Exhibit Chondrogenic and Osteogenic Potentials at the Clonal Level

    PubMed Central

    Zheng, Bo; Li, Guangheng; Chen, William C.W.; Deasy, Bridget M.; Pollett, Jonathan B.; Sun, Bin; Drowley, Lauren; Gharaibeh, Burhan; Usas, Arvydas; Péault, Bruno; Huard, Johnny

    2015-01-01

    We have previously reported the high regenerative potential of murine muscle-derived stem cells (mMDSCs) that are capable of differentiating into multiple mesodermal cell lineages, including myogenic, endothelial, chondrocytic, and osteoblastic cells. Recently, we described a putative human counterpart of mMDSCs, the myogenic endothelial cells (MECs), in adult human skeletal muscle, which efficiently repair/regenerate the injured and dystrophic skeletal muscle as well as the ischemic heart in animal disease models. Nevertheless it remained unclear whether human MECs, at the clonal level, preserve mMDSC-like chondrogenic and osteogenic potentials and classic stem cell characteristics including high proliferation and resistance to stress. Herein, we demonstrated that MECs, sorted from fresh postnatal human skeletal muscle biopsies, can be grown clonally and exhibit robust resistance to oxidative stress with no tumorigeneity. MEC clones were capable of differentiating into chondrocytes and osteoblasts under inductive conditions in vitro and participated in cartilage and bone formation in vivo. Additionally, adipogenic and angiogenic potentials of clonal MECs (cMECs) were observed. Overall, our study showed that cMECs not only display typical properties of adult stem cells but also exhibit chondrogenic and osteogenic capacities in vitro and in vivo, suggesting their potential applications in articular cartilage and bone repair/regeneration. PMID:23553740

  15. The role of surface charge on the uptake and biocompatibility of hydroxyapatite nanoparticles with osteoblast cells

    NASA Astrophysics Data System (ADS)

    Chen, Liang; Mccrate, Joseph M.; C-M Lee, James; Li, Hao

    2011-03-01

    The objective of this study is to evaluate the effect of hydroxyapatite (HAP) nanoparticles with different surface charges on the cellular uptake behavior and in vitro cell viability and proliferation of MC3T3-E1 cell lines (osteoblast). The nanoparticles' surface charge was varied by surface modification with two carboxylic acids: 12-aminododecanoic acid (positive) and dodecanedioic acid (negative). The untreated HAP nanoparticles and dodecanoic acid modified HAP nanoparticles (neutral) were used as the control. X-ray diffraction (XRD) revealed that surface modifications by the three carboxylic acids did not change the crystal structure of HAP nanoparticles; Fourier transform infrared spectroscopy (FT-IR) confirmed the adsorption and binding of the carboxylic acids on the HAP nanoparticles' surfaces; and zeta potential measurement confirmed that the chemicals successfully modified the surface charge of HAP nanoparticles in water based solution. Transmission electron microscopy (TEM) images showed that positively charged, negatively charged and untreated HAP nanoparticles, with similar size and shape, all penetrated into the cells and cells had more uptake of HAP nanoparticles with positive charge compared to those with negative charge, which might be attributed to the attractive or repulsive interaction between the negatively charged cell membrane and positively/negatively charged HAP nanoparticles. The neutral HAP nanoparticles could not penetrate the cell membrane due to their larger size. MTT assay and LDH assay results indicated that as compared with the polystyrene control, greater cell viability and cell proliferation were measured on MC3T3-E1 cells treated with the three kinds of HAP nanoparticles (neutral, positive, and untreated), among which positively charged HAP nanoparticles showed the strongest improvement for cell viability and cell proliferation. In summary, the surface charge of HAP nanoparticles can be modified to influence the cellular uptake of HAP nanoparticles and the different uptake also influences the behavior of cells. These in vitro results may also provide useful information for investigations of HAP nanoparticle applications in gene delivery and intracellular drug delivery.

  16. Particle-Cell Contact Enhances Antibacterial Activity of Silver Nanoparticles

    PubMed Central

    Bondarenko, Olesja; Ivask, Angela; Käkinen, Aleksandr; Kurvet, Imbi; Kahru, Anne

    2013-01-01

    Background It is generally accepted that antibacterial properties of Ag nanoparticles (AgNPs) are dictated by their dissolved fraction. However, dissolution-based concept alone does not fully explain the toxic potency of nanoparticulate silver compared to silver ions. Methodology/Principal Findings Herein, we demonstrated that the direct contact between bacterial cell and AgNPs' surface enhanced the toxicity of nanosilver. More specifically, cell-NP contact increased the cellular uptake of particle-associated Ag ions – the single and ultimate cause of toxicity. To prove that, we evaluated the toxicity of three different AgNPs (uncoated, PVP-coated and protein-coated) to six bacterial strains: Gram-negative Escherichia coli, Pseudomonas fluorescens, P. putida and P. aeruginosa and Gram-positive Bacillus subtilis and Staphylococcus aureus. While the toxicity of AgNO3 to these bacteria varied only slightly (the 4-h EC50 ranged from 0.3 to 1.2 mg Ag/l), the 4-h EC50 values of protein-coated AgNPs for various bacterial strains differed remarkably, from 0.35 to 46 mg Ag/l. By systematically comparing the intracellular and extracellular free Ag+ liberated from AgNPs, we demonstrated that not only extracellular dissolution in the bacterial test environment but also additional dissolution taking place at the particle-cell interface played an essential role in antibacterial action of AgNPs. The role of the NP-cell contact in dictating the antibacterial activity of Ag-NPs was additionally proven by the following observations: (i) separation of bacterial cells from AgNPs by particle-impermeable membrane (cut-off 20 kDa, ?4 nm) significantly reduced the toxicity of AgNPs and (ii) P. aeruginosa cells which tended to attach onto AgNPs, exhibited the highest sensitivity to all forms of nanoparticulate Ag. Conclusions/Significance Our findings provide new insights into the mode of antibacterial action of nanosilver and explain some discrepancies in this field, showing that “Ag-ion” and “particle-specific” mechanisms are not controversial but, rather, are two faces of the same coin. PMID:23737965

  17. Microenvironments and different nanoparticle dynamics in living cells revealed by a standard nanoparticle.

    PubMed

    Pack, Chan Gi; Song, Mi Ryoung; Tae, Eunju Lee; Hiroshima, Michio; Byun, Kyung Hee; Kim, Jun Sung; Sako, Yasushi

    2012-11-10

    For quantitative analysis of nanoparticle diffusions and submicro-environments in living cells, use of newly synthesized silica-based fluorescent nanoparticle (Si-FNP) as a standard nanoprobe is successfully demonstrated. The appropriate characteristics of a standard probe were fully analyzed in vitro by single molecule detection, transmission electron microscopy, and dynamic light scattering. Using fluorescence correlation analysis in single living cells, we quantitatively compared the diffusional properties of the standard Si-FNP with a diameter of 50 nm, peptide coated Si-FNP, streptavidin coated Qdot, and GFP molecule which have different sizes and surface properties. The result demonstrates that the standard Si-FNP without coat is minimally trapped in the vesicles in the process of cellular endocytosis. Interestingly, a large proportion of Si-FNP introduced into the cells by electroporation diffuses freely in the cells during a cell cycle suggesting free diffusing NPs are hardly trapped in the vesicles. The simple but highly sensitive method will provide insight into strategies to understanding the hydrodynamic process of nanoparticle delivery into living cells as well as the cellular microenvironment in the view of submicro-size. PMID:22922061

  18. Mechanisms of nanoparticle-mediated photomechanical cell damage

    PubMed Central

    Peeters, Sara; Kitz, Michael; Preisser, Stefan; Wetterwald, Antoinette; Rothen?Rutishauser, Barbara; Thalmann, George N.; Brandenberger, Christina; Bailey, Arthur; Frenz, Martin

    2012-01-01

    Laser-assisted killing of gold nanoparticle targeted macrophages was investigated. Using pressure transient detection, flash photography and transmission electron microscopy (TEM) imaging, we studied the mechanism of single cell damage by vapor bubble formation around gold nanospheres induced by nanosecond laser pulses. The influence of the number of irradiating laser pulses and of particle size and concentration on the threshold for acute cell damage was determined. While the single pulse damage threshold is independent of the particle size, the threshold decreases with increasing particle size when using trains of pulses. The dependence of the cell damage threshold on the nanoparticle concentration during incubation reveals that particle accumulation and distribution inside the cell plays a key role in tissue imaging or cell damaging. PMID:22435092

  19. Ex vivo expansion of canine cytotoxic large granular lymphocytes exhibiting characteristics of natural killer cells

    PubMed Central

    Shin, Dong-Jun; Park, Ji-Yun; Jang, Youn-Young; Lee, Je-Jung; Lee, Youn-Kyung; Shin, Myung-Geun; Jung, Ji-Youn; Carson, William E.; Cho, Duck; Kim, Sang-Ki

    2013-01-01

    Canine NK cells still are not well-characterized due to the lack of information concerning specific NK cell markers and the fact that NK cells are not an abundant cell population. In this study, we selectively expanded the canine cytotoxic large granular lymphocytes (CLGLs) that exhibit morphologic, genetic, and functional characteristics of NK cells from normal donor PBMCs. The cultured CLGLs were characterized by a high proportion of CD5(dim) expressing cells, of which the majority of cells co-expressed CD3 and CD8, but did not express TCR?? and TCR??. The phenotype of the majority of the CLGLs was CD5(dim)CD3+CD8+ TCR???TCR???CD4?CD21?CD11c+/?CD11d+/?CD44+. The expression of mRNAs for NK cell-associated receptors (NKG2D, NKp30, NKp44, Ly49, perforin, and granzyme B) were highly upregulated in cultured CLGLs. Specifically, NKp46 was remarkably upregulated in the cultured CLGLs compared to PBMCs. The mRNAs for the NKT-associated iTCR? gene in CLGLs was present at a basal level. The cytotoxic activity of the CLGLs against canine NK cell-sensitive CTAC cells was remarkably elevated in a dose-dependent manner, and the CLGLs produced large amounts of IFN-?. The antitumor activity of CLGLs extended to different types of canine tumor cells (CF41.Mg and K9TCC-pu-AXC) without specific antigen recognition. These results are consistent with prior reports, and strongly suggest that the selectively expanded CLGLs represent a population of canine NK cells. The results of this study will contribute to future research on canine NK cells as well as NK cell-based immunotherapy. PMID:23548866

  20. Comparative evaluation of the impact on endothelial cells induced by different nanoparticle structures and functionalization

    PubMed Central

    Müller, Ines; Ernst, Peter; Schäfer, Miriam; Rosman, Christina; Schick, Isabel; Köhler, Oskar; Oehring, Hartmut; Breus, Vladimir V; Basché, Thomas; Sönnichsen, Carsten; Tremel, Wolfgang

    2015-01-01

    Summary In the research field of nanoparticles, many studies demonstrated a high impact of the shape, size and surface charge, which is determined by the functionalization, of nanoparticles on cell viability and internalization into cells. This work focused on the comparison of three different nanoparticle types to give a better insight into general rules determining the biocompatibility of gold, Janus and semiconductor (quantum dot) nanoparticles. Endothelial cells were subject of this study, since blood is the first barrier after intravenous nanoparticle application. In particular, stronger effects on the viability of endothelial cells were found for nanoparticles with an elongated shape in comparison to spherical ones. Furthermore, a positively charged nanoparticle surface (NH2, CyA) leads to the strongest reduction in cell viability, whereas neutral and negatively charged nanoparticles are highly biocompatible to endothelial cells. These findings are attributed to a rapid internalization of the NH2-functionalized nanoparticles in combination with the damage of intracellular membranes. Interestingly, the endocytotic pathway seems to be a size-dependent process whereas nanoparticles with a size of 20 nm are internalized by caveolae-mediated endocytosis and nanoparticles with a size of 40 nm are taken up by clathrin-mediated internalization and macropinocytosis. Our results can be summarized to formulate five general rules, which are further specified in the text and which determine the biocompatibility of nanoparticles on endothelial cells. Our findings will help to design new nanoparticles with optimized properties concerning biocompatibility and uptake behavior with respect to the respective intended application.

  1. Comparative cytotoxicity of Al2O3, CeO2, TiO2 and ZnO nanoparticles to human lung cells.

    PubMed

    Kim, In-Sun; Baek, Miri; Choi, Soo-Jin

    2010-05-01

    The increased applications of nanoparticles in a wide range of industrial fields raise the concern about their potential toxicity to human. The aim of this study was to assess and compare the toxicity of four different oxide nanoparticles (Al2O3, CeO2, TiO2 and ZnO) to human lung epithelial cells, A549 carcinoma cells and L-132 normal cells, in vitro. We focused on the toxicological effects of the present nanoparticles on cell proliferation, cell viability, membrane integrity and oxidative stress. The long-term cytotoxicity of nanoparticles was also evaluated by employing the clonogenic assay. Among four nanoparticles tested, ZnO exhibited the highest cytotoxicity in terms of cell proliferation, cell viability, membrane integrity and colony formation in both cell lines. Al2O3, CeO2 and TiO2 showed little adverse effects on cell proliferation and cell viability. However, TiO2 induced oxidative stress in a concentration- and time-dependent manner. CeO2 caused membrane damage and inhibited colony formation in long-term, but with different degree depending on cell lines. Al2O3 seems to be less toxic than the other nanoparticles even after long time exposure. These results highlight the need for caution during manufacturing process of nanomaterials as well as further investigation on the toxicity mechanism. PMID:20358977

  2. New structural analogues of curcumin exhibit potent growth suppressive activity in human colorectal carcinoma cells

    PubMed Central

    2009-01-01

    Background Colorectal carcinoma is one of the major causes of morbidity and mortality in the Western World. Novel therapeutic approaches are needed for colorectal carcinoma. Curcumin, the active component and yellow pigment of turmeric, has been reported to have several anti-cancer activities including anti-proliferation, anti-invasion, and anti-angiogenesis. Clinical trials have suggested that curcumin may serve as a potential preventive or therapeutic agent for colorectal cancer. Methods We compared the inhibitory effects of curcumin and novel structural analogues, GO-Y030, FLLL-11, and FLLL-12, in three independent human colorectal cancer cell lines, SW480, HT-29, and HCT116. MTT cell viability assay was used to examine the cell viability/proliferation and western blots were used to determine the level of PARP cleavages. Half-Maximal inhibitory concentrations (IC50) were calculated using Sigma Plot 9.0 software. Results Curcumin inhibited cell viability in all three of the human colorectal cancer cell lines studied with IC50 values ranging between 10.26 ?M and 13.31 ?M. GO-Y030, FLLL-11, and FLLL-12 were more potent than curcumin in the inhibition of cell viability in these three human colorectal cancer cell lines with IC50 values ranging between 0.51 ?M and 4.48 ?M. In addition, FLLL-11 and FLLL-12 exhibit low toxicity to WI-38 normal human lung fibroblasts with an IC-50 value greater than 1,000 ?M. GO-Y030, FLLL-11, and FLLL-12 are also more potent than curcumin in the induction of apoptosis, as evidenced by cleaved PARP and cleaved caspase-3 in all three human colorectal cancer cell lines studied. Conclusion The results indicate that the three curcumin analogues studied exhibit more potent inhibitory activity than curcumin in human colorectal cancer cells. Thus, they may have translational potential as chemopreventive or therapeutic agents for colorectal carcinoma. PMID:19331692

  3. Water soluble nanoporous nanoparticle for in vivo targeted drug delivery and controlled release in B cells tumor context

    NASA Astrophysics Data System (ADS)

    de Angelis, F.; Pujia, A.; Falcone, C.; Iaccino, E.; Palmieri, C.; Liberale, C.; Mecarini, F.; Candeloro, P.; Luberto, L.; de Laurentiis, A.; Das, G.; Scala, G.; di Fabrizio, E.

    2010-10-01

    Multitasking nanoparticles are gaining great attention for smart drug delivery systems. The exploration of the nano-scale opens new concrete opportunities for revealing new properties and undiscovered cell-particle interactions. Here we present a biodegradable nanoporous silicon nanoparticle that can be successfully employed for in vivo targeted drug delivery and sustained release. The bare nanoporous nanocarriers can be accurately designed and fabricated with an effective control of porosity, surface chemistry and particle size, up to a few nm. The proposed nanoparticles exhibit several remarkable features including high payload, biodegradability, no toxicity, and multiple loading in water without the need of additional chemical reagents at room temperature. The targeting strategy is based on phage display technology that was successfully used to discover cell surface binding peptide for murine B lymphoma A20 cell line. The peptide used in combination with the nanoporous nanoparticles allows an efficient in vivo targeting, a sustained release and a sensible therapeutic effect.Multitasking nanoparticles are gaining great attention for smart drug delivery systems. The exploration of the nano-scale opens new concrete opportunities for revealing new properties and undiscovered cell-particle interactions. Here we present a biodegradable nanoporous silicon nanoparticle that can be successfully employed for in vivo targeted drug delivery and sustained release. The bare nanoporous nanocarriers can be accurately designed and fabricated with an effective control of porosity, surface chemistry and particle size, up to a few nm. The proposed nanoparticles exhibit several remarkable features including high payload, biodegradability, no toxicity, and multiple loading in water without the need of additional chemical reagents at room temperature. The targeting strategy is based on phage display technology that was successfully used to discover cell surface binding peptide for murine B lymphoma A20 cell line. The peptide used in combination with the nanoporous nanoparticles allows an efficient in vivo targeting, a sustained release and a sensible therapeutic effect. Electronic supplementary information (ESI) available: Nanoparticles fabrication; payload evaluation; dissolution and release profiles; multivalent loading; targeting specifity on A20 Cells; cell cycle analysis; in vitro cytotoxicity assay; in vivo cytotoxicity assay. See DOI: 10.1039/c0nr00161a

  4. Protamine sulfate-nanodiamond hybrid nanoparticles as a vector for MiR-203 restoration in esophageal carcinoma cells

    NASA Astrophysics Data System (ADS)

    Cao, Minjun; Deng, Xiongwei; Su, Shishuai; Zhang, Fang; Xiao, Xiangqian; Hu, Qin; Fu, Yongwei; Yang, Burton B.; Wu, Yan; Sheng, Wang; Zeng, Yi

    2013-11-01

    We report an innovative approach for miRNA-203 delivery in esophageal cancer cells using protamine sulphate (PS)-nanodiamond (ND) nanoparticles. The efficient delivery of miR-203 significantly suppressed the proliferation and migration of cancer cells through targeting Ran and ?Np63, exhibiting a great potential for PS@ND nanoparticles in miRNA-based cancer therapy.We report an innovative approach for miRNA-203 delivery in esophageal cancer cells using protamine sulphate (PS)-nanodiamond (ND) nanoparticles. The efficient delivery of miR-203 significantly suppressed the proliferation and migration of cancer cells through targeting Ran and ?Np63, exhibiting a great potential for PS@ND nanoparticles in miRNA-based cancer therapy. Electronic supplementary information (ESI) available: (1) Experimental section; (2) Results: serum stability of miR-203/PS@NDs and miR-203 release curve (Fig. S1). Cytotoxicity assay of PS@NDs to Ec-109 cells (Fig. S2); confocal image and FACS analysis of intracellular uptake of cy3-labeled miR-203 (Fig. S3 and S4); real-time PCR analysis of miR-203 restoration (Fig. S5); Ran and ?Np63 expression (Fig. S6); the sizes and zeta potentials of miRNA/PS@NDs (Table S1); the sequences of the microRNA mimics and primers (Table S2, S3 and S4). See DOI: 10.1039/c3nr04056a

  5. Magnetic CoPt nanoparticles as MRI contrast agent for transplanted neural stem cells detection

    NASA Astrophysics Data System (ADS)

    Meng, Xiaoting; Seton, Hugh C.; Lu, Le T.; Prior, Ian A.; Thanh, Nguyen T. K.; Song, Bing

    2011-03-01

    Neural stem cells (NSCs) exhibit features that make them suitable candidates for stem cell replacement therapy and spinal cord reconstruction. Magnetic resonance imaging (MRI) offers the potential to track cells in vivo using innovative approaches to cell labeling and image acquisition. In this study, experiments were carried out to optimize the loading condition of magnetic CoPt hollow nanoparticles (CoPt NPs) into neural stem cells and to define appropriate MRI parameters. Both cell viability and multipotency analysis showed that CoPt NPs at a concentration of 16 µg ml-1 reduced T2 relaxation times in labeled rat NSCs, producing greater contrast on spin echo acquisitions at 4.7 T, yet did not affect cell viability and in vitro differentiation potential compared to controls. After optimizing nanoparticle loading concentrations and labeled cell numbers for MRI detection, CoPt-loaded NSCs were transplanted into organotypic spinal cord slices. The results showed that MRI could efficiently detect low numbers of CoPt-labeled NSCs with the enhanced image contrast. Our study demonstrated that MRI of grafted NSCs labeled with CoPt NPs is a useful tool to evaluate organotypic spinal cord slice models and has potential applications in other biological systems.

  6. Mice with Reduced Levels of p53 Protein Exhibits the Testicular Giant-Cell Degenerative Syndrome

    Microsoft Academic Search

    Varda Rotter; Dov Schwartz; Einat Almon; Naomi Goldfinger; Ahuva Kapon; Asher Meshorer; Larry A. Donehower; Arnold J. Levine

    1993-01-01

    Transgenic mice which carry hybrid p53 promoter-chloramphenicol acetyltransferase (CAT) transgenes were found to express CAT enzymatic activity predominantly in the testes. Endogenous levels of p53 mRNA and protein were lower than in the nontransgenic control mice. The various p53 promoter-CAT transgenic mice exhibited in their testes multinucleated giant cells, a degenerative syndrome resulting presumably from the inability of the tetraploid

  7. Targeting myeloid cells using nanoparticles to improve cancer immunotherapy.

    PubMed

    Amoozgar, Zohreh; Goldberg, Michael S

    2014-10-01

    While nanoparticles have traditionally been used to deliver cytotoxic drugs directly to tumors to induce cancer cell death, emerging data suggest that nanoparticles are likely to generate a larger impact on oncology through the delivery of agents that can stimulate antitumor immunity. Tumor-targeted nanocarriers have generally been used to localize chemotherapeutics to tumors and thus decrease off-target toxicity while enhancing efficacy. Challengingly, tumor heterogeneity and evolution render tumor-intrinsic approaches likely to succumb to relapse. The immune system offers exquisite specificity, cytocidal potency, and long-term activity that leverage an adaptive memory response. For this reason, the ability to manipulate immune cell specificity and function would be desirable, and nanoparticles represent an exciting means by which to perform such manipulation. Dendritic cells and tumor-associated macrophages are cells of the myeloid lineage that function as natural phagocytes, so they naturally take up nanoparticles. Dendritic cells direct the specificity and potency of cellular immune responses that can be targeted for cancer vaccines. Herein, we discuss the specific criteria needed for efficient vaccine design, including but not limited to the route of administration, size, morphology, surface charge, targeting ligands, and nanoparticle composition. In contrast, tumor-associated macrophages are critical mediators of immunosuppression whose trans-migratory abilities can be exploited to localize therapeutics to the tumor core and which can be directly targeted for elimination or for repolarization to a tumor suppressive phenotype. It is likely that a combination of targeting dendritic cells to stimulate antitumor immunity and tumor-associated macrophages to reduce immune suppression will impart significant benefits and result in durable antitumor responses. PMID:25280471

  8. Biogenic-Production of SnO2 Nanoparticles and Its Cytotoxic Effect Against Hepatocellular Carcinoma Cell Line (HepG2).

    PubMed

    Roopan, Selvaraj Mohana; Kumar, Subramanian Hari Subbish; Madhumitha, Gunabalan; Suthindhiran, Krishnamurthy

    2015-02-01

    In this paper, we have established for the first time, the terrific efficiency of aqueous extract of agricultural waste dried peel of sugar apple (Annona squamosa) in the rapid synthesis of stable SnO2 nanoparticles. In topical years, the deployment of secondary metabolites from plant extract has emerged as a novel technology for the synthesis of various nanoparticles. In this paper, we have studied the potential of SnO2 nanoparticles assembly using agricultural waste source for the first time. The synthesized nanoparticles were characterized and confirmed as SnO2 nanoparticles by using UV-visible spectroscopy, XRD, and TEM analysis. The motivation of this study was to examine cytotoxicity study of SnO2 nanoparticles against hepatocellular carcinoma cell line (HepG2). SnO2 nanoparticles inhibited the cell proliferation in a dose- and time-dependent manner with an IC50 value of 148 ?g/mL. The treated cells showed an altered morphology with increasing concentrations of SnO2 nanoparticles. Our result shows that the SnO2 nanoparticles exhibit moderate cytotoxicity towards the hepatocellular carcinoma (HepG2) at tested concentrations. PMID:25410804

  9. Monitoring the endocytosis of magnetic nanoparticles by cells using permanent micro-flux sources.

    PubMed

    Osman, O; Zanini, L F; Frénéa-Robin, M; Dumas-Bouchiat, F; Dempsey, N M; Reyne, G; Buret, F; Haddour, N

    2012-10-01

    Trapping of cells is essential to perform basic handling operations in cell-based microsystems, such as media exchange, concentration, cell isolation and cell sorting. Cell trapping by magnetophoresis typically requires cell labeling with magnetic nanoparticles. Here we report on endocytotic uptake of 100 nm magnetic nanoparticles by Human Embryonic Kidney 293 cells. The attraction of labeled cells by micro-magnet arrays characterised by very high magnetic field gradients (?10? T/m) was studied as a function of labeling conditions (nanoparticle concentration in the extracellular medium, incubation time). The threshold incubation conditions for effective magnetophoretic trapping were established. This simple technique may be exploited to minimise the quantity of magnetic nanoparticles needed for efficient cell trapping, thus reducing stress or nanoparticle-mediated toxicity. Nanoparticle internalization into cells was confirmed using both confocal and Transmission Electron Microscopy (TEM). PMID:22773161

  10. Immunosuppressive Compounds Exhibit Particular Effects on Functional Properties of Human Anti-Aspergillus TH1 Cells

    PubMed Central

    Tramsen, Lars; Schmidt, Stanislaw; Roeger, Frauke; Schubert, Ralf; Salzmann-Manrique, Emilia; Latgé, Jean-Paul; Klingebiel, Thomas

    2014-01-01

    Allogeneic hematopoietic stem cell transplant (HSCT) recipients are at high risk for invasive aspergillosis. Whereas adoptive immunotherapy transferring donor-derived anti-Aspergillus TH1 cells has been shown to be beneficial for HSCT recipients suffering from invasive aspergillosis, little is known about the impact of commonly used immunosuppressants on the functional properties of anti-Aspergillus TH1 cells. Anti-Aspergillus TH1 cells were coincubated with different concentrations of methylprednisolone, cyclosporine (CsA), mycophenolic acid (MPA), the active component of mycophenolate mofetil, and rapamycin. Immunosuppressants were tested in concentrations reflecting common target levels in serum and in significantly lower and higher concentrations. Apoptosis of anti-Aspergillus TH1 cells, as well as proliferation and production of gamma interferon (IFN-?) and CD154 upon restimulation, was evaluated in the presence and absence of immunosuppressive compounds. All dosages of CsA, MPA, and methylprednisolone significantly decreased the number of viable anti-Aspergillus TH1 cells in the cell culture, which was due partly to an impaired proliferative capacity of the cells and partly to an increased rate of apoptosis. In addition, CsA significantly decreased the number of IFN-?-producing cells and had the highest impact of all immunosuppressants on IFN-? levels in the supernatant. CsA also significantly decreased the expression of CD154 by anti-Aspergillus TH1 cells. Variant dosages of immunosuppressants exhibit particular effects on essential functional properties of anti-Aspergillus TH1 cells. Our findings may have an important impact on the design of clinical trials evaluating the therapeutic benefit of anti-Aspergillus TH1 cells in allogeneic HSCT recipients suffering from invasive aspergillosis. PMID:24711569

  11. Rare somatic cells from human breast tissue exhibit extensive lineage plasticity

    PubMed Central

    Roy, Somdutta; Gascard, Philippe; Dumont, Nancy; Zhao, Jianxin; Pan, Deng; Petrie, Sarah; Margeta, Marta; Tlsty, Thea D.

    2013-01-01

    We identified cell surface markers associated with repression of p16INK4a/cyclin-dependent kinase inhibitor 2A(CDKN2A), a critical determinant in the acquisition of a plastic state. These cell surface markers allowed direct isolation of rare cells from healthy human breast tissue that exhibit extensive lineage plasticity. This subpopulation is poised to transcribe plasticity markers, OCT3/4, SOX2, and NANOG, at levels similar to those measured in human embryonic stem cells and to acquire a plastic state sensitive to environmental programming. In vitro, in vivo, and teratoma assays demonstrated that either a directly sorted (uncultured) or a single-cell (clonogenic) cell population from primary tissue can differentiate into functional derivatives of each germ layer, ectodermal, endodermal, and mesodermal. In contrast to other cells that express OCT3/4, SOX2, and NANOG, these human endogenous plastic somatic cells are mortal, express low telomerase activity, expand for an extensive but finite number of population doublings, and maintain a diploid karyotype before arresting in G1. PMID:23487770

  12. Myocytes and fibroblasts exhibit functional synergism in mixed cultures of neonatal rat heart cells.

    PubMed

    Schroedl, N A; Hartzell, C R

    1983-12-01

    Cardiac cells obtained from neonatal rat heart contain a mixed population of cell types that can be enriched in culture in either myocytes or fibroblast-like cells. A metabolic comparison of mixed heart cell cultures with enriched cultures of the same age-in-culture and initial cell density showed that mixed cultures used glucose more rapidly than either enriched myocytes or fibroblasts. Mixed cultures were shown to respond to deprivation of insulin and of serum with decreases in the rate of glucose usage and decreases in the protein content of cells, whereas enriched cultures did not respond in the expected manner to insulin deprivation. Mixed, 11-day-old cells also exhibited greater increases in cellular protein and greater resistance to the stress of starvation than enriched cultures. Palmitate usage, however, was similar in all cultures examined. We conclude that mixed cultures may serve as a better model system to study cardiac metabolism and to monitor the effects of drugs and hormones on the neonatal myocardium. In addition, it is clear from our results that myocytes and fibroblastic-like cells coexist in a metabolically functional synergism. PMID:6361043

  13. In ViWo Tumor Cell Targeting with "Click" Nanoparticles Geoffrey von Maltzahn,,

    E-print Network

    Bhatia, Sangeeta

    In ViWo Tumor Cell Targeting with "Click" Nanoparticles Geoffrey von Maltzahn,, Yin Ren,, Ji of inorganic nanoparticles to tumors. We find that "click" chemistry allows cyclic LyP-1 targeting peptides to be specifically linked to azido-nanoparticles and to direct their binding to p32-expressing tumor cells in Vitro

  14. T cells enhance gold nanoparticle delivery to tumors in vivo

    PubMed Central

    2011-01-01

    Gold nanoparticle-mediated photothermal therapy (PTT) has shown great potential for the treatment of cancer in mouse studies and is now being evaluated in clinical trials. For this therapy, gold nanoparticles (AuNPs) are injected intravenously and are allowed to accumulate within the tumor via the enhanced permeability and retention (EPR) effect. The tumor is then irradiated with a near infrared laser, whose energy is absorbed by the AuNPs and translated into heat. While reliance on the EPR effect for tumor targeting has proven adequate for vascularized tumors in small animal models, the efficiency and specificity of tumor delivery in vivo, particularly in tumors with poor blood supply, has proven challenging. In this study, we examine whether human T cells can be used as cellular delivery vehicles for AuNP transport into tumors. We first demonstrate that T cells can be efficiently loaded with 45 nm gold colloid nanoparticles without affecting viability or function (e.g. migration and cytokine production). Using a human tumor xenograft mouse model, we next demonstrate that AuNP-loaded T cells retain their capacity to migrate to tumor sites in vivo. In addition, the efficiency of AuNP delivery to tumors in vivo is increased by more than four-fold compared to injection of free PEGylated AuNPs and the use of the T cell delivery system also dramatically alters the overall nanoparticle biodistribution. Thus, the use of T cell chaperones for AuNP delivery could enhance the efficacy of nanoparticle-based therapies and imaging applications by increasing AuNP tumor accumulation. PMID:21711861

  15. T cells enhance gold nanoparticle delivery to tumors in vivo

    NASA Astrophysics Data System (ADS)

    Kennedy, Laura C.; Bear, Adham S.; Young, Joseph K.; Lewinski, Nastassja A.; Kim, Jean; Foster, Aaron E.; Drezek, Rebekah A.

    2011-12-01

    Gold nanoparticle-mediated photothermal therapy (PTT) has shown great potential for the treatment of cancer in mouse studies and is now being evaluated in clinical trials. For this therapy, gold nanoparticles (AuNPs) are injected intravenously and are allowed to accumulate within the tumor via the enhanced permeability and retention (EPR) effect. The tumor is then irradiated with a near infrared laser, whose energy is absorbed by the AuNPs and translated into heat. While reliance on the EPR effect for tumor targeting has proven adequate for vascularized tumors in small animal models, the efficiency and specificity of tumor delivery in vivo, particularly in tumors with poor blood supply, has proven challenging. In this study, we examine whether human T cells can be used as cellular delivery vehicles for AuNP transport into tumors. We first demonstrate that T cells can be efficiently loaded with 45 nm gold colloid nanoparticles without affecting viability or function (e.g. migration and cytokine production). Using a human tumor xenograft mouse model, we next demonstrate that AuNP-loaded T cells retain their capacity to migrate to tumor sites in vivo. In addition, the efficiency of AuNP delivery to tumors in vivo is increased by more than four-fold compared to injection of free PEGylated AuNPs and the use of the T cell delivery system also dramatically alters the overall nanoparticle biodistribution. Thus, the use of T cell chaperones for AuNP delivery could enhance the efficacy of nanoparticle-based therapies and imaging applications by increasing AuNP tumor accumulation.

  16. Anionic polymers and 10 nm Fe?O?@UA wound dressings support human foetal stem cells normal development and exhibit great antimicrobial properties.

    PubMed

    Grumezescu, Alexandru Mihai; Holban, Alina Maria; Andronescu, Ecaterina; Mogo?anu, George Dan; Vasile, Bogdan Stefan; Chifiriuc, Mariana Carmen; Lazar, Veronica; Andrei, Eugen; Constantinescu, Andrei; Maniu, Horia

    2014-03-25

    The aims of this study were the development, characterization and bioevaluation of a novel biocompatible, resorbable and bio-active wound dressing prototype, based on anionic polymers (sodium alginate--AlgNa, carboximethylcellulose--CMC) and magnetic nanoparticles loaded with usnic acid (Fe?O?@UA). The antimicrobial activity was tested against Staphylococcus aureus grown in biofilms. The biocompatibility testing model included an endothelial cell line from human umbilical vein and human foetal progenitor cells derived from the amniotic fluid, that express a wide spectrum of surface molecules involved in different vascular functions and inflammatory response, and may be used as skin regenerative support. The obtained results demonstrated that CMC/Fe?O?@UA and AlgNa/Fe?O?@UA are exhibiting structural and functional properties that recommend them for further applications in the biomedical field. They could be used alone or coated with different bio-active compounds, such as Fe?O?@UA, for the development of novel, multifunctional porous materials used in tissues regeneration, as antimicrobial substances releasing devices, providing also a mechanical support for the eukaryotic cells adhesion, and exhibiting the advantage of low cytotoxicity on human progenitor cells. The great antimicrobial properties exhibited by the newly synthesized nano-bioactive coatings are recommending them as successful candidates for improving the implanted devices surfaces used in regenerative medicine. PMID:23994366

  17. Helicobacter pylori Protein JHP0290 Exhibits Proliferative and Anti-Apoptotic Effects in Gastric Epithelial Cells

    PubMed Central

    Tavares, Raquel; Pathak, Sushil Kumar

    2015-01-01

    The influence of Helicobacter pylori infection on gastric epithelial cell proliferation, apoptosis and signaling pathways contributes to the development of infection-associated diseases. Here we report that JHP0290, which is a poorly functionally characterized protein from H. pylori, regulates multiple responses in human gastric epithelial cells. The differential expression and release of JHP0290 homologues was observed among H. pylori strains. JHP0290 existed in monomeric and dimeric forms in H. pylori cell extracts and culture broth. Recombinant purified JHP0290 (rJHP0290) also showed monomeric and dimeric forms, whereas the rJHP0290 C162A mutant exhibited only a monomeric form. The dimeric form of the protein was found to bind more efficiently to gastric epithelial cells than the monomeric form. The exposure of gastric epithelial cells to rJHP0290 induced proliferation in a dose-dependent manner. Faster progression into the cell cycle was observed in rJHP0290-challenged gastric epithelial cells. Furthermore, we detected an anti-apoptotic effect of rJHP0290 in gastric epithelial cells when the cells were treated with rJHP0290 in combination with Camptothecin (CPT), which is an inducer of apoptosis. CPT-induced caspase 3 activation was significantly reduced in the presence of rJHP0290. In addition, the activation of ERK MAPK and the transcription factor NF?B was observed in rJHP0290-challenged gastric epithelial cells lines. Our results suggest that JHP0290 may affect H. pylori-induced gastric diseases via the regulation of gastric epithelial cell proliferation and anti-apoptotic pathways. PMID:25879227

  18. Nanoparticles Based Stem Cell Tracking in Regenerative Medicine

    PubMed Central

    Edmundson, Matthew; Thanh, Nguyen TK; Song, Bing

    2013-01-01

    Stem cell therapies offer great potentials in the treatment for a wide range of diseases and conditions. With so many stem cell replacement therapies going through clinical trials currently, there is a great need to understand the mechanisms behind a successful therapy, and one of the critical points of discovering them is to track stem cell migration, proliferation and differentiation in vivo. To be of most use tracking methods should ideally be non-invasive, high resolution and allow tracking in three dimensions. Magnetic resonance imaging (MRI) is one of the ideal methods, but requires a suitable contrast agent to be loaded to the cells to be tracked, and one of the most wide-spread in stem cell tracking is a group of agents known as magnetic nanoparticles. This review will explore the current use of magnetic nanoparticles in developing and performing stem cell therapies, and will investigate their potential limitations and the future directions magnetic nanoparticle tracking is heading in. PMID:23946823

  19. Enhanced reactive oxygen species overexpression by CuO nanoparticles in poorly differentiated hepatocellular carcinoma cells

    NASA Astrophysics Data System (ADS)

    Kung, Mei-Lang; Hsieh, Shu-Ling; Wu, Chih-Chung; Chu, Tian-Huei; Lin, Yu-Chun; Yeh, Bi-Wen; Hsieh, Shuchen

    2015-01-01

    Copper oxide nanoparticles (CuO NPs) are known to exhibit toxic effects on a variety of cell types and organs. To determine the oxidative impact of CuO NPs on hepatocellular carcinoma (HCC) cells, well-differentiated (HepG2) and poorly differentiated (SK-Hep-1) cells were exposed to CuO NPs. Cell viability assay showed that the median inhibition concentration (IC50) for SK-Hep-1 and HepG2 cells was 25 ?g ml-1 and 85 ?g ml-1, respectively. Cellular fluorescence intensity using DCFH-DA staining analysis revealed significant intracellular reactive oxygen species (ROS) generation of up to 242% in SK-Hep-1 cells, compared with 86% in HepG2 cells. HPLC analysis demonstrated that a CuO NP treatment caused cellular GSH depletion of 58% and a GSH/GSSG ratio decrease to ~0.1 in SK-Hep-1 cells. The oxidative stress caused by enhanced superoxide anion production was observed in both HepG2 (146%) and SK-Hep-1 (192%) cells. The Griess assay verified that CuO NPs induced NO production (170%) in SK-Hep-1 cells. Comet assay and western blot further demonstrated that CuO NPs induced severe DNA strand breakage (70%) in SK-Hep-1 cells and caused DNA damage via increased ?-H2AX levels. These results suggest that well-differentiated HepG2 cells possess a robust antioxidant defense system against CuO NP-induced ROS stress and exhibit more tolerance to oxidative stress. Conversely, poorly differentiated SK-Hep-1 cells exhibited a deregulated antioxidant defense system that allowed accumulation of CuO NP-induced ROS and resulted in severe cytotoxicity.Copper oxide nanoparticles (CuO NPs) are known to exhibit toxic effects on a variety of cell types and organs. To determine the oxidative impact of CuO NPs on hepatocellular carcinoma (HCC) cells, well-differentiated (HepG2) and poorly differentiated (SK-Hep-1) cells were exposed to CuO NPs. Cell viability assay showed that the median inhibition concentration (IC50) for SK-Hep-1 and HepG2 cells was 25 ?g ml-1 and 85 ?g ml-1, respectively. Cellular fluorescence intensity using DCFH-DA staining analysis revealed significant intracellular reactive oxygen species (ROS) generation of up to 242% in SK-Hep-1 cells, compared with 86% in HepG2 cells. HPLC analysis demonstrated that a CuO NP treatment caused cellular GSH depletion of 58% and a GSH/GSSG ratio decrease to ~0.1 in SK-Hep-1 cells. The oxidative stress caused by enhanced superoxide anion production was observed in both HepG2 (146%) and SK-Hep-1 (192%) cells. The Griess assay verified that CuO NPs induced NO production (170%) in SK-Hep-1 cells. Comet assay and western blot further demonstrated that CuO NPs induced severe DNA strand breakage (70%) in SK-Hep-1 cells and caused DNA damage via increased ?-H2AX levels. These results suggest that well-differentiated HepG2 cells possess a robust antioxidant defense system against CuO NP-induced ROS stress and exhibit more tolerance to oxidative stress. Conversely, poorly differentiated SK-Hep-1 cells exhibited a deregulated antioxidant defense system that allowed accumulation of CuO NP-induced ROS and resulted in severe cytotoxicity. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr05843g

  20. Olfactory ensheathing glia and Schwann cells exhibit a distinct interaction behavior with meningeal cells.

    PubMed

    Franssen, Elske H P; Roet, Kasper C D; de Bree, Freddy M; Verhaagen, Joost

    2009-05-15

    Schwann cells (SCs) and olfactory ensheathing glia (OEG) have both been used as cellular transplants to promote spinal cord repair. Both cell types support axonal regeneration and have beneficial effects on functional recovery. A significant difference between SCs and OEG is the effect of these cell types on astrocytes (ACs) present in the neural scar. In contrast to OEG, which associate and intermingle with ACs, SCs and ACs form separate cellular territories. Here, we show that OEG and SCs also interact differently with meningeal cells (MCs), another major cellular component of the neural scar. Whereas OEG intermingle with MCs in cocultures, SCs aggregate into well-defined cell clusters. Our data suggest that (a) soluble factor(s) as well as direct cellular contact are involved in the MC-induced SC clustering. Furthermore, the cluster formation of SCs in coculture with MCs is different from the previously reported segregation of SCs and ACs in coculture. The present results help to understand the differential behavior of both cell types after transplantation in the injured spinal cord and will be important to either determine which cell has optimal capacities to render the scar more permissive for regeneration, or to exploit the transplantation of both cell types in combination. PMID:19140223

  1. Two-photon excited fluorescence of silica nanoparticles loaded with a fluorene-based monomer and its cross-conjugated polymer: their application to cell imaging

    NASA Astrophysics Data System (ADS)

    Aparicio-Ixta, Laura; Ramos-Ortiz, Gabriel; Pichardo-Molina, Juan L.; Maldonado, José Luis; Rodríguez, Mario; Tellez-Lopez, Víctor M.; Martinez-Fong, Daniel; Zolotukhin, Mikhail G.; Fomine, Serguei; Meneses-Nava, Marco. A.; Barbosa-García, Oracio

    2012-11-01

    In this work the two-photon activity of nanoparticles obtained from a fluorene monomer (M1) and its cross-conjugated polymer (P1) is reported. Aqueous suspensions of M1 and P1 nanoparticles prepared through the reprecipitation method exhibited maximum two-photon absorption (TPA) cross-sections of 84 and 9860 GM (1 GM = 10-50 cm4 s) at 740 nm, respectively, and a fluorescence quantum yield of ~1. Such a two-photon activity was practically equal with respect to that for molecular solutions of M1 and P1. These materials were then successfully encapsulated into silica nanoparticles to provide bio-compatibly. A lung cancer cell line (A549) and a human cervical cancer cell line (HeLa cells) were incubated with our fluorescent silica nanoparticles to carry out two-photon imaging. By means of these studies we demonstrate that optimized nonlinear optical polymers loaded in silica nanoparticles can be used as efficient probes with low cytotoxicity and good photostability for two-photon fluorescence microscopy. To the best of our knowledge, studies concerning polymer-doped silica nanoparticles exhibiting large two-photon activity have not been reported in the literature.In this work the two-photon activity of nanoparticles obtained from a fluorene monomer (M1) and its cross-conjugated polymer (P1) is reported. Aqueous suspensions of M1 and P1 nanoparticles prepared through the reprecipitation method exhibited maximum two-photon absorption (TPA) cross-sections of 84 and 9860 GM (1 GM = 10-50 cm4 s) at 740 nm, respectively, and a fluorescence quantum yield of ~1. Such a two-photon activity was practically equal with respect to that for molecular solutions of M1 and P1. These materials were then successfully encapsulated into silica nanoparticles to provide bio-compatibly. A lung cancer cell line (A549) and a human cervical cancer cell line (HeLa cells) were incubated with our fluorescent silica nanoparticles to carry out two-photon imaging. By means of these studies we demonstrate that optimized nonlinear optical polymers loaded in silica nanoparticles can be used as efficient probes with low cytotoxicity and good photostability for two-photon fluorescence microscopy. To the best of our knowledge, studies concerning polymer-doped silica nanoparticles exhibiting large two-photon activity have not been reported in the literature. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr31925j

  2. Epirubicin loaded to pre-polymerized poly(butyl cyanoacrylate) nanoparticles: preparation and in vitro evaluation in human lung adenocarcinoma cells.

    PubMed

    Yordanov, Georgi; Evangelatov, Alexander; Skrobanska, Ralica

    2013-07-01

    This article describes the preparation of epirubicin-loaded nanoparticles, prepared by loading of the drug in pre-polymerized poly(butyl cyanoacrylate) nanoparticles, their physicochemical characterization and in vitro evaluation on human lung adenocarcinoma (A549) cells. Nanoparticles were also coated in aqueous dispersions with two different non-ionic surfactants (Pluronic F68 and Polysorbate 80). All particles were spherical in shape, with monomodal size distributions. The zeta-potentials at pH 7.4 increased with augmentation of the particle drug content. The increased drug content was found to correlate with the initial concentration of the drug, used for the particle preparation. In vitro studies on A549 cells showed that the drug-loaded nanoparticles, as well as the combinations of free drug and empty nanoparticles, exhibited higher cytotoxicity than the free drug alone. The presence of surfactants also resulted in increased cytotoxicity. Fluorescent imaging of epirubicin internalization by the adenocarcinoma cells revealed that the free drug was predominantly localized in the cell nucleus, while a cytoplasmic localization was observed for the nanoparticle-bound drug formulations, suggesting the probability of nanoparticle endocytosis. Thus the hereby presented results could be useful for development of nanoparticle-based anthracycline formulations for treatment of lung adenocarcinoma. PMID:23466549

  3. Influence of surface-modified maghemite nanoparticles on in vitro survival of human stem cells

    PubMed Central

    Babi?, Michal; Lukash, Lyubov L; Ruban, Tetiana A; Kolomiets, Yurii N; Shpylova, Svitlana P; Grypych, Oksana A

    2014-01-01

    Summary Surface-modified maghemite (?-Fe2O3) nanoparticles were obtained by using a conventional precipitation method and coated with D-mannose and poly(N,N-dimethylacrylamide). Both the initial and the modified particles were characterized by transmission electron microscopy and dynamic light scattering with regard to morphology, particle size and polydispersity. In vitro survival of human stem cells was then investigated by using the methyl thiazolyl tetrazolium (MTT) assay, which showed that D-mannose- and poly(N,N-dimethylacrylamide)-coated ?-Fe2O3 particles exhibit much lower level of cytotoxicity than the non-coated ?-Fe2O3. PMID:25383284

  4. Synthesis of nanoparticles of P3HT and PCBM for optimizing morphology in polymeric solar cells

    NASA Astrophysics Data System (ADS)

    Satapathi, Soumitra; Gill, Hardeep Singh; Li, Lian; Samuelson, Lynne; Kumar, Jayant; Mosurkal, Ravi

    2014-12-01

    Nanoparticles (NPs) with opposite charges of a donor polymer, poly(3-hexyl thiophene) (P3HT) and an acceptor molecule, phenyl-C61-butyric acid methyl ester (PCBM) were synthesized using simple mini-emulsion technique. The NPs were examined by dynamic light scattering, atomic force microscopy and scanning electron microscopy to confirm the formation of the NPs. The NPs were assembled into thin films by spin-coating of a blend of the NPs dispersion. The built-up of a five layered film was monitored by UV-vis absorption spectroscopy. Our preliminary study demonstrated that a solar cell made from an annealed NP film can exhibit photovoltaic response.

  5. Enhanced reactive oxygen species overexpression by CuO nanoparticles in poorly differentiated hepatocellular carcinoma cells.

    PubMed

    Kung, Mei-Lang; Hsieh, Shu-Ling; Wu, Chih-Chung; Chu, Tian-Huei; Lin, Yu-Chun; Yeh, Bi-Wen; Hsieh, Shuchen

    2015-02-01

    Copper oxide nanoparticles (CuO NPs) are known to exhibit toxic effects on a variety of cell types and organs. To determine the oxidative impact of CuO NPs on hepatocellular carcinoma (HCC) cells, well-differentiated (HepG2) and poorly differentiated (SK-Hep-1) cells were exposed to CuO NPs. Cell viability assay showed that the median inhibition concentration (IC50) for SK-Hep-1 and HepG2 cells was 25 ?g ml(-1) and 85 ?g ml(-1), respectively. Cellular fluorescence intensity using DCFH-DA staining analysis revealed significant intracellular reactive oxygen species (ROS) generation of up to 242% in SK-Hep-1 cells, compared with 86% in HepG2 cells. HPLC analysis demonstrated that a CuO NP treatment caused cellular GSH depletion of 58% and a GSH/GSSG ratio decrease to ?0.1 in SK-Hep-1 cells. The oxidative stress caused by enhanced superoxide anion production was observed in both HepG2 (146%) and SK-Hep-1 (192%) cells. The Griess assay verified that CuO NPs induced NO production (170%) in SK-Hep-1 cells. Comet assay and western blot further demonstrated that CuO NPs induced severe DNA strand breakage (70%) in SK-Hep-1 cells and caused DNA damage via increased ?-H2AX levels. These results suggest that well-differentiated HepG2 cells possess a robust antioxidant defense system against CuO NP-induced ROS stress and exhibit more tolerance to oxidative stress. Conversely, poorly differentiated SK-Hep-1 cells exhibited a deregulated antioxidant defense system that allowed accumulation of CuO NP-induced ROS and resulted in severe cytotoxicity. PMID:25521936

  6. Sensitization of ovarian cancer cells to cisplatin by gold nanoparticles

    PubMed Central

    Saha, Sounik; Robertson, David J.; McMeekin, Scott; Bhattacharya, Resham; Mukherjee, Priyabrata

    2014-01-01

    Recently we reported that gold nanoparticles (AuNPs) inhibit ovarian tumor growth and metastasis in mice by reversing epithelial-mesenchymal transition (EMT). Since EMT is known to confer drug resistance to cancer cells, we wanted to investigate whether anti-EMT property of AuNP could be utilized to sensitize ovarian cancer cells to cisplatin. Herein, we report that AuNPs prevent cisplatin-induced acquired chemoresistance and stemness in ovarian cancer cells and sensitize them to cisplatin. AuNPs inhibit cisplatin induced EMT, decrease the side population cells and key stem cell markers such as ALDH1, CD44, CD133, Sox2, MDR1 and ABCG2 in ovarian cancer cells. Mechanistically, AuNPs prevent cisplatin-induced activation of Akt and NF-?B signaling axis in ovarian cancer cells that are critical for EMT, stem cell maintenance and drug resistance. In vivo, AuNPs sensitize orthotopically implanted ovarian tumor to a low dose of cisplatin and significantly inhibit tumor growth via facilitated delivery of both AuNP and cisplatin. These findings suggest that by depleting stem cell pools and inhibiting key molecular pathways gold nanoparticles sensitize ovarian cancer cells to cisplatin and may be used in combination to inhibit tumor growth and metastasis in ovarian cancer. PMID:25071019

  7. Casearin X exhibits cytotoxic effects in leukemia cells triggered by apoptosis.

    PubMed

    Ferreira, Paulo M Pinheiro; Santos, André G; Tininis, Aristeu G; Costa, Patricia M; Cavalheiro, Alberto J; Bolzani, Vanderlan S; Moraes, Manoel O; Costa-Lotufo, Letícia V; Montenegro, Raquel C; Pessoa, Cláudia

    2010-12-01

    Clerodane diterpenes have demonstrated cytotoxic, antiplasmodial and anti-ulcer properties. In the present work, we determined the cytotoxic effect of casearin L (Cas L), O (Cas O) and X (Cas X) and (-)-hardwickiic acid isolated from Casearia sylvestris leaves, and investigated the underlying mechanisms involved in in vitro cell death induced by Cas X in HL-60 leukemia cells (0.7, 1.5 and 3.0?M). Cytotoxicity tests demonstrated that Cas X was the most active compound studied, showing greater cytotoxic effects against CEM and HL-60 lines (IC(50) of 0.4?M) and human peripheral blood mononuclear cells (PBMC, IC(50) of 1.2?M). After 24h exposure, Cas X caused a decrease in 5-bromo-20-deoxyuridine (BrdU) incorporation (36.6 and 24.5% labeling at 0.7 and 1.5?M, respectively), reduction in viability, and increase in apoptotic and necrotic leukemia cells in a dose-dependent manner evidenced by the trypan blue and AO/EB (acridine orange/ethidium bromide) assays. Moreover, Cas X-treated cells exhibited nuclear fragmentation and cytoplasmic vacuolization depending on the concentration tested. These characteristics of apoptosis or secondary necrosis were confirmed by flow cytometry which revealed DNA fragmentation, phosphatidylserine externalization, activation of the effector caspases 3/7 and mitochondrial depolarization. We then found evidence that Cas X causes cell death via apoptotic pathways, corroborating the potential of casearins as compounds with promising antitumor-related properties. PMID:20816779

  8. Mice deficient in heparanase exhibit impaired dendritic cell migration and reduced airway inflammation.

    PubMed

    Poon, Ivan K H; Goodall, Katharine J; Phipps, Simon; Chow, Jenny D Y; Pagler, Eloisa B; Andrews, Daniel M; Conlan, Carly L; Ryan, Gemma F; White, Julie A; Wong, Michael K L; Horan, Catherine; Matthaei, Klaus I; Smyth, Mark J; Hulett, Mark D

    2014-04-01

    Heparanase is a ?-d-endoglucuronidase that cleaves heparan sulphate, a key component of the ECM and basement membrane. The remodelling of the ECM by heparanase has been proposed to regulate both normal physiological and pathological processes, including wound healing, inflammation, tumour angiogenesis and cell migration. Heparanase is also known to exhibit non-enzymatic functions by regulating cell adhesion, cell signalling and differentiation. In this study, constitutive heparanase-deficient (Hpse(-/-) ) mice were generated on a C57BL/6 background using the Cre/loxP recombination system, with a complete lack of heparanase mRNA, protein and activity. Although heparanase has been implicated in embryogenesis and development, Hpse(-/-) mice are anatomically normal and fertile. Interestingly, consistent with the suggested function of heparanase in cell migration, the trafficking of dendritic cells from the skin to the draining lymph nodes was markedly reduced in Hpse(-/-) mice. Furthermore, the ability of Hpse(-/-) mice to generate an allergic inflammatory response in the airways, a process that requires dendritic cell migration, was also impaired. These findings establish an important role for heparanase in immunity and identify the enzyme as a potential target for regulation of an immune response. PMID:24532362

  9. Human T-cell leukemia virus types I and II exhibit different DNase I protection patterns.

    PubMed Central

    Altman, R; Harrich, D; Garcia, J A; Gaynor, R B

    1988-01-01

    Human T-cell leukemia virus types I (HTLV-I) and II (HTLV-II) are human retroviruses which normally infect T-lymphoid cells. HTLV-I infection is associated with adult T-cell leukemia-lymphoma, and HTLV-II is associated with an indolent form of hairy-cell leukemia. To identify potential transcriptional regulatory elements of these two related human retroviruses, we performed DNase I footprinting of both the HTLV-I and HTLV-II long terminal repeats (LTRs) by using extracts prepared from uninfected T cells, HTLV-I and HTLV-II transformed T cells, and HeLa cells. Five regions of the HTLV-I LTR and three regions of the HTLV-II LTR showed protection by DNase I footprinting. All three of the 21-base-pair repeats previously shown to be important in HTLV transcriptional regulation were protected in the HTLV-I LTR, whereas only one of these repeats was protected in the HTLV-II LTR. Several regions exhibited altered protection in extracts prepared from lymphoid cells as compared with HeLa cells, but there were minimal differences in the protection patterns between HTLV-infected and uninfected lymphoid extracts. A number of HTLV-I and HTLV-II LTR fragments which contained regions showing protection in DNase I footprinting were able to function as inducible enhancer elements in transient CAT gene expression assays in the presence of the HTLV-II tat protein. The alterations in the pattern of the cellular proteins which bind to the HTLV-I and HTLV-II LTRs may in part be responsible for differences in the transcriptional regulation of these two related viruses. Images PMID:2831395

  10. Mice lacking TrkB in parvalbumin-positive cells exhibit sexually dimorphic behavioral phenotypes.

    PubMed

    Lucas, Elizabeth K; Jegarl, Anita; Clem, Roger L

    2014-11-01

    Activity-dependent brain-derived neurotrophic factor (BDNF) signaling through receptor tyrosine kinase B (TrkB) is required for cued fear memory consolidation and extinction. Although BDNF is primarily secreted from glutamatergic neurons, TrkB is expressed by other genetically defined cells whose contributions to the behavioral effects of BDNF remain poorly understood. Parvalbumin (PV)-positive interneurons, which are highly enriched in TrkB, are emerging as key regulators of fear memory expression. We therefore hypothesized that activity-dependent BDNF signaling in PV-interneurons may modulate emotional learning. To test this hypothesis, we utilized the LoxP/Cre system for conditional deletion of TrkB in PV-positive cells to examine the impact of cell-autonomous BDNF signaling on Pavlovian fear conditioning and extinction. However, behavioral abnormalities indicative of vestibular dysfunction precluded the use of homozygous conditional knockouts in tests of higher cognitive functioning. While vestibular dysfunction was apparent in both sexes, female conditional knockouts exhibited an exacerbated phenotype, including extreme motor hyperactivity and circling behavior, compared to their male littermates. Heterozygous conditional knockouts were spared of vestibular dysfunction. While fear memory consolidation was unaffected in heterozygotes of both sexes, males exhibited impaired extinction consolidation compared to their littermate controls. Our findings complement evidence from human and rodent studies suggesting that BDNF signaling promotes consolidation of extinction and point to PV-positive neurons as a discrete population that mediates these effects in a sex-specific manner. PMID:25127683

  11. Intracellular Uptake and Trafficking of Difluoroboron Dibenzoylmethane-Poly(lactic acid) Nanoparticles in HeLa Cells

    PubMed Central

    Contreras, Janette; Xie, Jiansong; Chen, Yin Jie; Pei, Hua; Zhang, Guoqing; Fraser, Cassandra L.; Hamm-Alvarez, Sarah F.

    2010-01-01

    In this study, nanoparticles based on difluoroboron dibenzoylmethane-poly(lactic acid) (BF2dbmPLA) are prepared. Polylactic acid or polylactide is a commonly used degradable polymer, while the boron dye possesses a large extinction coefficient, high emission quantum yield, 2-photon absorption, and sensitivity to the surrounding environment. BF2dbmPLA exhibits molecular weight-dependent emission properties, and can be formulated as stable nanoparticles, suggesting that its unique optical properties may be useful in multiple contexts for probing intracellular environments. Here we show that BF2dbmPLA nanoparticles are internalized into cultured HeLa cells by endocytosis, and that within the cellular milieu they retain their fluorescence properties. BF2dbmPLA nanoparticles are photostable, resisting laser-induced photobleaching under conditions that destroy the fluorescence of a common photostable probe, LysoTracker™ blue. Their endocytosis is also lipid raft-dependent, as evidenced by their significant co-localization with cholera toxin B subunit in membrane compartments after uptake, and their sensitivity of uptake to methyl-?-cyclodextrin. Additionally, BF2dbmPLA nanoparticle endocytosis utilizes microtubules and actin filaments. Internalized BF2dbmPLA nanoparticles do not accumulate in acidic late endosomes and lysosomes, but within a perinuclear non-lysosomal compartment. These findings demonstrate the feasibility of using novel BF2dbmPLA nanoparticles exhibiting diverse emission properties for in situ, live cell imaging, and suggest that their endogenous uptake occurs through a lipid-raft dependent endocytosis mechanism. PMID:20420413

  12. In Vivo Tumor Cell Targeting with “Click” Nanoparticles

    PubMed Central

    von Maltzahn, Geoffrey; Ren, Yin; Park, Ji-Ho; Min, Dal-Hee; Kotamraju, Venkata Ramana; Jayakumar, Jayanthi; Fogel, Valentina; Sailor, Michael J.; Ruoslahti, Erkki; Bhatia, Sangeeta N.

    2008-01-01

    The in vivo fate of nanomaterials strongly determines their biomedical efficacy. Accordingly, much effort has been invested into the development of library screening methods to select targeting ligands for a diversity of sites in vivo. Still, broad application of chemical and biological screens to the in vivo targeting of nanomaterials requires ligand attachment chemistries that are generalizable, efficient, covalent, orthogonal to diverse biochemical libraries, applicable under aqueous conditions, and stable in in vivo environments. To date, the copper(I)-catalyzed Huisgen 1,3-dipolar cycloaddition or “click” reaction has shown considerable promise as a method for developing targeted nanomaterials in vitro. Here, we investigate the utility of “click” chemistry for the in vivo targeting of inorganic nanoparticles to tumors. We find that “click” chemistry allows cyclic LyP-1 targeting peptides to be specifically linked to azido-nanoparticles and to direct their binding to p32-expressing tumor cells in vitro. Moreover, “click” nanoparticles are able to stably circulate for hours in vivo following intravenous administration (>5h circulation time), extravasate into tumors, and penetrate the tumor interstitium to specifically bind p32-expressing cells in tumors. In the future, in vivo use of “click” nanomaterials should expedite the progression from ligand discovery to in vivo evaluation and diversify approaches toward multifunctional nanoparticle development. PMID:18611045

  13. T cells from CLL patients exhibit features of T-cell exhaustion but retain capacity for cytokine production

    PubMed Central

    Davies, Jeffrey K.; McClanahan, Fabienne; Fatah, Rewas; Iqbal, Sameena; Agrawal, Samir; Ramsay, Alan G.; Gribben, John G.

    2013-01-01

    T-cell exhaustion, originally described in chronic viral infections, was recently reported in solid and hematologic cancers. It is not defined whether exhaustion contributes to T-cell dysfunction observed in chronic lymphocytic leukemia (CLL). We investigated the phenotype and function of T cells from CLL patients and age-matched controls. CD8+ and CD4+ T cells from CLL patients had increased expression of exhaustion markers CD244, CD160, and PD1, with expansion of a PD1+BLIMP1HI subset. These molecules were most highly expressed in the expanded population of effector T cells in CLL. CLL CD8+ T cells showed functional defects in proliferation and cytotoxicity, with the cytolytic defect caused by impaired granzyme packaging into vesicles and nonpolarized degranulation. In contrast to virally induced exhaustion, CLL T cells showed increased production of interferon-? and TNF? and increased expression of TBET, and normal IL2 production. These defects were not restricted to expanded populations of cytomegalovirus (CMV)–specific cells, although CMV seropositivity modulated the distribution of lymphocyte subsets, the functional defects were present irrespective of CMV serostatus. Therefore, although CLL CD8+ T cells exhibit features of T-cell exhaustion, they retain the ability to produce cytokines. These findings also exclude CMV as the sole cause of T-cell defects in CLL. PMID:23247726

  14. Normal and cancer breast epithelial cells endocytosis study of nanoparticles by combined AFM and NSOM microscopy

    Microsoft Academic Search

    Yu Zhang; Vahid Yazdanpanah; Mo Yang; Mihrimah Ozkan; Cengiz S. Ozkan

    2007-01-01

    A substantial understanding of nanoparticles recognition and uptake by biological cells and tissues is very important for reaching their fullest potential in biomedical research. In this work, we investigated the cell endocytosis of iron oxide nanoparticles by both normal breast epithelial cells (MCF10A) and cancer breast epithelial cells (MCF7) using a combination of atomic force microscope (AFM) and nearfield scanning

  15. Selective reduction of the interaction of magnetic nanoparticles with leukocytes and tumor cells by human plasma

    NASA Astrophysics Data System (ADS)

    Schwalbe, Manuela; Jörke, Cornelia; Buske, Norbert; Höffken, Klaus; Pachmann, Katharina; Clement, Joachim H.

    2005-05-01

    Carboxymethyl-dextran coated magnetic nanoparticles can interact with viable human cells. The interaction of the nanoparticles is cell-type specific. The addition of human plasma led to a dramatic reduction of magnetically separable leukocytes in comparison to tumor cells. We conclude that low plasma concentrations might support an efficient enrichment of circulating epithelial cells from the peripheral blood of tumor patients.

  16. RECQL5 and BLM exhibit divergent functions in cells defective for the Fanconi anemia pathway

    PubMed Central

    Kim, Tae Moon; Son, Mi Young; Dodds, Sherry; Hu, Lingchuan; Luo, Guangbin; Hasty, Paul

    2015-01-01

    Fanconi anemia (FA) patients exhibit bone marrow failure, developmental defects and cancer. The FA pathway maintains chromosomal stability in concert with replication fork maintenance and DNA double strand break (DSB) repair pathways including RAD51-mediated homologous recombination (HR). RAD51 is a recombinase that maintains replication forks and repairs DSBs, but also rearranges chromosomes. Two RecQ helicases, RECQL5 and Bloom syndrome mutated (BLM) suppress HR through nonredundant mechanisms. Here we test the impact deletion of RECQL5 and BLM has on mouse embryonic stem (ES) cells deleted for FANCB, a member of the FA core complex. We show that RECQL5, but not BLM, conferred resistance to mitomycin C (MMC, an interstrand crosslinker) and camptothecin (CPT, a type 1 topoisomerase inhibitor) in FANCB-defective cells. RECQL5 suppressed, while BLM caused, breaks and radials in FANCB-deleted cells exposed to CPT or MMC, respectively. RECQL5 protected the nascent replication strand from MRE11-mediated degradation and restarted stressed replication forks in a manner additive to FANCB. By contrast BLM restarted, but did not protect, replication forks in a manner epistatic to FANCB. RECQL5 also lowered RAD51 levels in FANCB-deleted cells at stressed replication sites implicating a rearrangement avoidance mechanism. Thus, RECQL5 and BLM impact FANCB-defective cells differently in response to replication stress with relevance to chemotherapeutic regimes. PMID:25520194

  17. RECQL5 and BLM exhibit divergent functions in cells defective for the Fanconi anemia pathway.

    PubMed

    Kim, Tae Moon; Son, Mi Young; Dodds, Sherry; Hu, Lingchuan; Luo, Guangbin; Hasty, Paul

    2015-01-01

    Fanconi anemia (FA) patients exhibit bone marrow failure, developmental defects and cancer. The FA pathway maintains chromosomal stability in concert with replication fork maintenance and DNA double strand break (DSB) repair pathways including RAD51-mediated homologous recombination (HR). RAD51 is a recombinase that maintains replication forks and repairs DSBs, but also rearranges chromosomes. Two RecQ helicases, RECQL5 and Bloom syndrome mutated (BLM) suppress HR through nonredundant mechanisms. Here we test the impact deletion of RECQL5 and BLM has on mouse embryonic stem (ES) cells deleted for FANCB, a member of the FA core complex. We show that RECQL5, but not BLM, conferred resistance to mitomycin C (MMC, an interstrand crosslinker) and camptothecin (CPT, a type 1 topoisomerase inhibitor) in FANCB-defective cells. RECQL5 suppressed, while BLM caused, breaks and radials in FANCB-deleted cells exposed to CPT or MMC, respectively. RECQL5 protected the nascent replication strand from MRE11-mediated degradation and restarted stressed replication forks in a manner additive to FANCB. By contrast BLM restarted, but did not protect, replication forks in a manner epistatic to FANCB. RECQL5 also lowered RAD51 levels in FANCB-deleted cells at stressed replication sites implicating a rearrangement avoidance mechanism. Thus, RECQL5 and BLM impact FANCB-defective cells differently in response to replication stress with relevance to chemotherapeutic regimes. PMID:25520194

  18. Firefly Luciferase and Rluc8 Exhibit Differential Sensitivity to Oxidative Stress in Apoptotic Cells

    PubMed Central

    Czupryna, Julie; Tsourkas, Andrew

    2011-01-01

    Over the past decade, firefly Luciferase (fLuc) has been used in a wide range of biological assays, providing insight into gene regulation, protein-protein interactions, cell proliferation, and cell migration. However, it has also been well established that fLuc activity can be highly sensitive to its surrounding environment. In this study, we found that when various cancer cell lines (HeLa, MCF-7, and 293T) stably expressing fLuc were treated with staurosporine (STS), there was a rapid loss in bioluminescence. In contrast, a stable variant of Renilla luciferase (RLuc), RLuc8, exhibited significantly prolonged functionality under the same conditions. To identify the specific underlying mechanism(s) responsible for the disparate sensitivity of RLuc8 and fLuc to cellular stress, we conducted a series of inhibition studies that targeted known intracellular protein degradation/modification pathways associated with cell death. Interestingly, these studies suggested that reactive oxygen species, particularly hydrogen peroxide (H2O2), was responsible for the diminution of fLuc activity. Consistent with these findings, the direct application of H2O2 to HeLa cells also led to a reduction in fLuc bioluminescence, while H2O2 scavengers stabilized fLuc activity. Comparatively, RLuc8 was far less sensitive to ROS. These observations suggest that fLuc activity can be substantially altered in studies where ROS levels become elevated and can potentially lead to ambiguous or misleading findings. PMID:21603648

  19. Hepatitis B virus X protein mutants exhibit distinct biological activities in hepatoma Huh7 cells

    SciTech Connect

    Liu Xiaohong; Zhang Shuhui; Lin Jing; Zhang Shunmin [Department of Pathology, Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai 200433 (China); Feitelson, Mark A. [Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122 (United States); Gao Hengjun [National Engineering Center for Biochip at Shanghai, Shanghai 201203 (China); Zhu Minghua [Department of Pathology, Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai 200433 (China)], E-mail: mhzhu2000@hotmail.com

    2008-09-05

    The role of the hepatitis B virus X protein (HBx) in hepatocarcinogenesis remains controversial. To investigate the biological impact of hepatitis B virus x gene (HBx) mutation on hepatoma cells, plasmids expressing the full-length HBx or HBx deletion mutants were constructed. The biological activities in these transfectants were analyzed by a series of assays. Results showed that HBx3'-20 and HBx3'-40 amino acid deletion mutants exhibited an increase in cellular proliferation, focus formation, tumorigenicity, and invasive growth and metastasis through promotion of the cell cycle from G0/G1 to the S phase, when compared with the full-length HBx. In contrast, HBx3'-30 amino acid deletion mutant repressed cell proliferation by blocking in G1 phase. The expression of P53, p21{sup WAF1}, p14{sup ARF}, and MDM2 proteins was regulated by expression of HBx mutants. In conclusions, HBx variants showed different effects and functions on cell proliferation and invasion by regulation of the cell cycle progression and its associated proteins expression.

  20. Effect of nonendocytic uptake of nanoparticles on human bronchial epithelial cells.

    PubMed

    Zhao, Xi; Wu, Yun; Gallego-Perez, Daniel; Kwak, Kwang Joo; Gupta, Cherry; Ouyang, Xilian; Lee, L James

    2015-03-17

    The toxicity of artificial nanoparticles is a major concern in industrial applications. Cellular uptake of hard nanoparticles could follow either endocytic or nonendocytic pathways, leading to different stimuli to the cells. Yet the cellular responses to nanoparticles following different pathways have not been compared due to the lack of an independent nonendocytic delivery method. We applied a unique delivery method, nanochannel electroporation (NEP), to produce predominantly nonendocytic uptakes of quantum dots (Q-dots) and multiwalled carbon nanotubes (MWCNTs) with different chemical modifications. NEP delivery bypassed endocytosis by electrophoretic injection of nanoparticles into human bronchial epithelial (BEAS-2B) cells at different dosages. Conventional exposure by direct nanoparticle suspending in cell culture medium was also performed as control. The dosage-dependent responses to nanoparticles under different uptake pathways were compared. Fluorescence colocalization demonstrated that nanoparticles followed both endocytic and nonendocytic pathways for cell entry in contact exposure, whereas NEP delivery of nanoparticles bypassed endocytosis. Nonendocytic entry resulted in much higher oxidation stress and, for MWCNTs, more cell death in BEAS-2B cells. Despite the observation that most nanoparticles were taken up by cells through endocytosis, the minor nonendocytic entry of nanoparticles seemed to dominate the overall cellular response in conventional contact exposure. Our finding suggests that prevention against nonendocytic uptake could help reduce the toxicity of hard nanoparticles. PMID:25671340

  1. Zebrafish kidney marrow contains ABCG2-dependent side population cells exhibiting hematopoietic stem cell properties

    Microsoft Academic Search

    Oleg Tsinkalovsky; Audun Osland Vik-Mo; Sara Ferreira; Ole Didrik Laerum; Anders Fjose

    2007-01-01

    Zebrafish (Danio rerio) has emerged as a powerful genetic model for the study of vertebrate hematopoiesis. However, methods for detection and isolation of hematopoietic stem cells (HSCs) have not yet been reported. In mammals, the combination of Hoechst 33342 staining with flow cytometry can be used for separation of a bone marrow side population (SP), which is highly enriched for

  2. PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells

    NASA Astrophysics Data System (ADS)

    Feng Liang, Gao; Zhu, Yan Liang; Sun, Bo; Hu, Fei Hu; Tian, Tian; Li, Shu Chun; Xiao, Zhong Dang

    2011-07-01

    The biggest challenge in the field of gene therapy is how to effectively deliver target genes to special cells. This study aimed to develop a new type of poly( D, L-lactide-co-glycolide) (PLGA)-based nanoparticles for gene delivery, which are capable of overcoming the disadvantages of polyethylenimine (PEI)- or cationic liposome-based gene carrier, such as the cytotoxicity induced by excess positive charge, as well as the aggregation on the cell surface. The PLGA-based nanoparticles presented in this study were synthesized by emulsion evaporation method and characterized by transmission electron microscopy, dynamic light scattering, and energy dispersive spectroscopy. The size of PLGA/PEI nanoparticles in phosphate-buffered saline (PBS) was about 60 nm at the optimal charge ratio. Without observable aggregation, the nanoparticles showed a better monodispersity. The PLGA-based nanoparticles were used as vector carrier for miRNA transfection in HepG2 cells. It exhibited a higher transfection efficiency and lower cytotoxicity in HepG2 cells compared to the PEI/DNA complex. The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes. Moreover, nanocomplex shows better serum compatibility than commercial liposome. PLGA nanocomplexes obviously accumulate in tumor cells after transfection, which indicate that the complexes contribute to cellular uptake of pDNA and pronouncedly enhance the treatment effect of miR-26a by inducing cell cycle arrest. Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

  3. Simvastatin exhibits antiproliferative effects on spheres derived from canine mammary carcinoma cells.

    PubMed

    Torres, Cristian G; Olivares, Araceli; Stoore, Caroll

    2015-05-01

    Mammary cancer is the most frequent type of tumor in the female canine. Treatments are mainly limited to surgery and chemotherapy; however, these tumors may develop clinical recurrence, metastasis and chemoresistance. The existence of a subpopulation of cancer cells with stemness features called cancer stem-like cells, may explain in part these characteristics of tumor progression. The statins, potent blockers of cholesterol synthesis, have also shown antitumor effects on cancer mammary cells, changes mediated by a decrease in the isoprenylation of specific proteins. Few studies have shown that simvastatin, a lipophilic statin, sensitizes cancer stem-like cells eliminating drug resistance. The aim of the present study was to evaluate the effects of simvastatin on spheres derived from CF41.Mg canine mammary tumor cells, which were characterized by phenotypic and functional analyses. Spheres exhibited characteristics of stemness, primarily expressing a CD44+/CD24-/low phenotype, displaying auto-renewal and relative chemoresistance. Exposure to simvastatin induced a decrease in the sphere-forming capacity and cell viability, accompanied by a concentration- and time-dependent increase in caspase-3/7 activity. In addition, modulation of ?-catenin and p53 expression was observed. Simvastatin triggered a synergistic effect with doxorubicin, sensitizing the spheres to the cytotoxic effect exerted by the drug. Invasiveness of spheres was decreased in response to simvastatin and this effect was counteracted by the presence of geranylgeranyl pyrophosphate. Our results suggest that simvastatin targets canine mammary cancer stem-like cells, supporting its therapeutical application as a novel agent to treat canine mammary cancer. PMID:25778435

  4. Glial cells, but not neurons, exhibit a controllable response to a localized inflammatory microenvironment in vitro

    PubMed Central

    Sommakia, Salah; Rickus, Jenna L.; Otto, Kevin J.

    2014-01-01

    The ability to design long-lasting intracortical implants hinges on understanding the factors leading to the loss of neuronal density and the formation of the glial scar. In this study, we modify a common in vitro mixed cortical culture model using lipopolysaccharide (LPS) to examine the responses of microglia, astrocytes, and neurons to microwire segments. We also use dip-coated polyethylene glycol (PEG), which we have previously shown can modulate impedance changes to neural microelectrodes, to control the cellular responses. We find that microglia, as expected, exhibit an elevated response to LPS-coated microwire for distances of up to 150 ?m, and that this elevated response can be mitigated by co-depositing PEG with LPS. Astrocytes exhibit a more complex, distance-dependent response, whereas neurons do not appear to be affected by the type or magnitude of glial response within this in vitro model. The discrepancy between our in vitro responses and typically observed in vivo responses suggest the importance of using a systems approach to understand the responses of the various brain cell types in a chronic in vivo setting, as well as the necessity of studying the roles of cell types not native to the brain. Our results further indicate that the loss of neuronal density observed in vivo is not a necessary consequence of elevated glial activation. PMID:25452724

  5. Self-crack-filled graphene films by metallic nanoparticles for high-performance graphene heterojunction solar cells.

    PubMed

    Ho, Po-Hsun; Liou, Yi-Ting; Chuang, Chien-Hsun; Lin, Shih-Wei; Tseng, Chi-Yang; Wang, Di-Yan; Chen, Chia-Chun; Hung, Wen-Yi; Wen, Cheng-Yen; Chen, Chun-Wei

    2015-03-01

    Graphene, with cracks filled with gold nanoparticles, is grown by chemical vapor deposition on a Cu substrate. The crack-filled graphene not only exhibits superior electrical properties but also forms a better junction with other semiconductors. A high-quality crack-filled graphene/Si Schottky junction solar cell is achieved, demonstrating the highest fill factor (0.79) and best efficiency (12.3%). PMID:25619427

  6. Amyloid fibrils enhance transport of metal nanoparticles in living cells and induced cytotoxicity.

    PubMed

    Bolisetty, Sreenath; Boddupalli, Chandra Sekhar; Handschin, Stephan; Chaitanya, Krishna; Adamcik, Jozef; Saito, Yasuyuki; Manz, Markus G; Mezzenga, Raffaele

    2014-07-14

    Amyloid protein fibrils occur in vivo as pathological agents, in the case of neurodegenerative diseases, or as functional amyloids, when playing biologically vital roles. Here we show how amyloid fibrils generated from a food protein, ?-lactoglobulin, can be used as nanoreactors for the synthesis of metal nanoparticles and demonstrate that the resulting hybrids can play a central role in the internalization of nanoparticles into living cells, with up to 3-fold-enhanced transport properties over pristine nanoparticles. We conjugate gold, silver, and palladium nanoparticles onto amyloid fibrils by chemical reduction, and we study their effect on dendritic and MCF7 breast cancer cells. Transmission electron microscopy indicates localization of nanoparticles inside vesicles of the cells. Flow cytometry reveals that silver nanoparticle-amyloid hybrids are cytotoxic, while gold and palladium nanoparticle-amyloid hybrids produce no notable effect on cell viability and activation status. PMID:24941321

  7. Visualizing DNA Nanoparticle Motion under Graphene Liquid Cell TEM

    NASA Astrophysics Data System (ADS)

    Chen, Qian; Smith, Jessica; Park, Jungwon; Lee, Somin; Zettl, Alex; Alivisatos, Paul

    2013-03-01

    We think of a simple colloidal nanocrystal as one type of artificial atoms. They mutually interact, cluster into artificial molecules, and further arrange into macroscopically functional artificial solids. The ``atomic'' resolution dynamics of this bottom-up strategy in materials design is studied here in a system of artificial molecules composed of DNA and nanoparticle. The observation of dynamics in their liquid environment is recently enabled by graphene liquid cell transmission electron microscopy (TEM). In comparison to conventional TEM, wherein the assembled 3D artificial structures are dried out during sample preparation and thus are collapsed, this graphene liquid cell introduces a special local liquid structure that retains the conformations as well as the dynamics of the assemblies. In situ imaging of correlated motions of DNA and nanoparticle provides insights into the design principles of artificial nanocrystal molecules and solids linked by DNA.

  8. BDNF+/- mice exhibit deficits in oligodendrocyte lineage cells of the basal forebrain.

    PubMed

    Vondran, Melissa W; Clinton-Luke, Patricia; Honeywell, Jean Z; Dreyfus, Cheryl F

    2010-05-01

    Previous work indicated that brain-derived neurotrophic factor (BDNF), through the trkB receptor, increases DNA synthesis in oligodendrocyte (OLG) progenitor cells (OPCs) and differentiation of postmitotic OLGs of the basal forebrain (BF). In the present studies, BDNF knockout animals were used to investigate BDNF's effects on OLG lineage cells (OLCs) in vivo. OLCs of the BF were found to express the trkB receptor, suggesting they are responsive to BDNF. Immunohistochemistry using NG2 and CC1 antibodies was utilized to examine the numbers of NG2+ OPCs and CC1+ postmitotic BF OLGs. At embryonic day 17 (E17), BDNF-/- animals display reduced NG2+ cells. This reduction was also observed in BDNF+/- mice at E17 and at postnatal day 1 (P1), P14, and adult stage, suggesting that BDNF plays a role in OPC development. BDNF+/- mice do not exhibit deficits in numbers of CC1+ OLGs. However, myelin basic protein, myelin associated glycoprotein, and proteolipid protein are reduced in BDNF+/- mice, suggesting that BDNF plays a role in differentiation. These data indicate that progenitor cells and myelin proteins may be affected in vivo by a decrease in BDNF. PMID:20091777

  9. Silver Nanoparticle Enhanced Freestanding Thin-Film Silicon Solar Cells

    NASA Astrophysics Data System (ADS)

    Winans, Joshua David

    As the supply of fossil fuels diminishes in quantity the demand for alternative energy sources will consistently increase. Solar cells are an environmentally friendly and proven technology that suffer in sales due to a large upfront cost. In order to help facilitate the transition from fossil fuels to photovoltaics, module costs must be reduced to prices well below $1/Watt. Thin-film solar cells are more affordable because of the reduced materials costs, but lower in efficiency because less light is absorbed before passing through the cell. Silver nanoparticles placed at the front surface of the solar cell absorb and reradiate the energy of the light in ways such that more of the light ends being captured by the silicon. Silver nanoparticles can do this because they have free electron clouds that can take on the energy of an incident photon through collective action. This bulk action of the electrons is called a plasmon. This work begins by discussing the economics driving the need for reduced material use, and the pros and cons of taking this step. Next, the fundamental theory of light-matter interaction is briefly described followed by an introduction to the study of plasmonics. Following that we discuss a traditional method of silver nanoparticle formation and the initial experimental studies of their effects on the ability of thin-film silicon to absorb light. Then, Finite-Difference Time-Domain simulation software is used to simulate the effects of nanoparticle morphology and size on the scattering of light at the surface of the thin-film.

  10. Phagocytosis independent extracellular nanoparticle clearance by human immune cells.

    PubMed

    Bartneck, Matthias; Keul, Heidrun A; Zwadlo-Klarwasser, Gabriele; Groll, Jürgen

    2010-01-01

    It has recently been discovered that human immune cells, especially neutrophil granulocytes, form neutrophil extracellular traps (NETs) that abolish pathogens. Our study provides evidence that extracellular traps formed by neutrophils, monocytes and macrophages act as physical barriers for nanoparticles, thus presenting a new nanomaterial clearance mechanism of the human immune system. While particle shape is of minor importance, positive charges significantly enhance particle trapping. PMID:19994869

  11. Amorphous silica nanoparticles enhance cross-presentation in murine dendritic cells

    SciTech Connect

    Hirai, Toshiro [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan)] [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Yoshioka, Yasuo, E-mail: yasuo@phs.osaka-u.ac.jp [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan)] [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Takahashi, Hideki; Ichihashi, Ko-ichi; Yoshida, Tokuyuki; Tochigi, Saeko [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan)] [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Nagano, Kazuya [Laboratory of Biopharmaceutical Research, National Institute of Biomedical Innovation, 7-6-8 Saitoasagi, Ibaraki, Osaka 567-0085 (Japan)] [Laboratory of Biopharmaceutical Research, National Institute of Biomedical Innovation, 7-6-8 Saitoasagi, Ibaraki, Osaka 567-0085 (Japan); Abe, Yasuhiro [Cancer Biology Research Center, Sanford Research/USD, 2301 E. 60th Street N, Sioux Falls, SD 57104 (United States)] [Cancer Biology Research Center, Sanford Research/USD, 2301 E. 60th Street N, Sioux Falls, SD 57104 (United States); Kamada, Haruhiko; Tsunoda, Shin-ichi [Laboratory of Biopharmaceutical Research, National Institute of Biomedical Innovation, 7-6-8 Saitoasagi, Ibaraki, Osaka 567-0085 (Japan) [Laboratory of Biopharmaceutical Research, National Institute of Biomedical Innovation, 7-6-8 Saitoasagi, Ibaraki, Osaka 567-0085 (Japan); The Center for Advanced Medical Engineering and Informatics, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Nabeshi, Hiromi [Division of Foods, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan)] [Division of Foods, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan); Yoshikawa, Tomoaki [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan)] [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Tsutsumi, Yasuo, E-mail: ytsutsumi@phs.osaka-u.ac.jp [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan) [Laboratory of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Laboratory of Biopharmaceutical Research, National Institute of Biomedical Innovation, 7-6-8 Saitoasagi, Ibaraki, Osaka 567-0085 (Japan); The Center for Advanced Medical Engineering and Informatics, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan)

    2012-10-26

    Highlights: Black-Right-Pointing-Pointer Silica nanoparticles enhanced cross-presentation. Black-Right-Pointing-Pointer Silica nanoparticles induced endosomal release of exogenous antigens. Black-Right-Pointing-Pointer Silica nanoparticle-induced cross-presentation was mediated by scavenger receptors. Black-Right-Pointing-Pointer Surface-modification may enable the manufacture of safer silica nanoparticles. -- Abstract: Nanomaterials (NMs) exhibit unique physicochemical properties and innovative functions, and they are increasingly being used in a wide variety of fields. Ensuring the safety of NMs is now an urgent task. Recently, we reported that amorphous silica nanoparticles (nSPs), one of the most widely used NMs, enhance antigen-specific cellular immune responses and may therefore aggravate immune diseases. Thus, to ensure the design of safer nSPs, investigations into the effect of nSPs on antigen presentation in dendritic cells, which are central orchestrators of the adaptive immune response, are now needed. Here, we show that nSPs with diameters of 70 and 100 nm enhanced exogenous antigen entry into the cytosol from endosomes and induced cross-presentation, whereas submicron-sized silica particles (>100 nm) did not. Furthermore, we show that surface modification of nSPs suppressed cross-presentation. Although further studies are required to investigate whether surface-modified nSPs suppress immune-modulating effects in vivo, the current results indicate that appropriate regulation of the characteristics of nSPs, such as size and surface properties, will be critical for the design of safer nSPs.

  12. Biomedical Applications of Magnetic Nanoparticles: Delivering Genes and Remote Control of Cells

    NASA Astrophysics Data System (ADS)

    Dobson, Jon

    2013-03-01

    The use of magnetic micro- and nanoparticles for biomedical applications was first proposed in the 1920s as a way to measure the rehological properties of the cell's cytoplasm. Since that time, magnetic micro- and nanoparticle synthesis, coating and bio-functionalization have advanced significantly, as have the applications for these particles. Magnetic micro- and nanoparticles are now used in a variety of biomedical techniques such as targeted drug delivery, MRI contrast enhancement, gene transfection, immno-assay and cell sorting. More recently, magnetic micro- and nanoparticles have been used to investigate and manipulate cellular processes both in vitro and in vivo. This talk will focus on magnetic nanoparticle targeting to and actuation of cell surface receptors to control cell signaling cascades to control cell behavior. This technology has applications in disease therapy, cell engineering and regenerative medicine. The use of magnetic nanoparticles and oscillating magnet arrays for enhanced gene delivery will also be discussed.

  13. Iron oxide nanoparticles as drug delivery agents in MIA PaCa-2 pancreatic cells

    NASA Astrophysics Data System (ADS)

    Perry, Christopher; Randriamahefa, Alexandrine; Lokko, Carl; Evans, Whitney; Watkins, Julian; Carrell, Holly; King, Natalie; Patel, Darayas

    2007-02-01

    Oleic acid (OA)-Pluronic-coated iron oxide nanoparticles were synthesized and characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and Atomic Force Microscopy (AFM). FT-IR confirmed the bonding of oleic acid and Pluronic (surfactant) to the nanoparticles. AFM measurements on these nanoparticles indicated a root mean square (RMS) roughness, a measure of nanoparticle size of (50 +/- 20) nm. The efficiency of these functionalized nanoparticles was investigated by loading with 5-Fluorouracil (5-FU) in aqueous solution. AFM measurements were used to characterize modified iron oxide nanoparticles and pancreatic MIA PaCa-2 cells, including size distribution, stability and cellular uptake. Nanoparticles were added to MIA PaCa-2 cells and assayed for their cytotoxic effects after 24 and 48 hours. The outcome of this study demonstrated the effectiveness of oleic acid (OA)-Pluronic-coated iron oxide nanoparticles as a non-toxic drug delivery agent for pancreatic cancer.

  14. Targeting Cancer Stem Cells with Nanoparticle-Enabled Therapies

    PubMed Central

    Burke, Andrew R.; Singh, Ravi N.; Carroll, David L.; Torti, Frank M.; Torti, Suzy V.

    2013-01-01

    Emerging evidence suggests that multiple tumor types are sustained by a small population of transformed stem-like cells that have the ability to both self-renew and give rise to non-tumorigenic daughter cells that constitute the bulk of a tumor. These cells, which generally constitute a minority of the overall cancer cell population, are highly resistant to conventional therapies and persist following treatment, leading to disease relapse and the formation of distant metastases. Therapies that disrupt the maintenance and survival of cancer stem cells are the subject of active current investigation. This review discusses recent approaches to the application of nanomedicine to the targeting and elimination of cancer stem cells. Specifically, recent publications in the areas of nanoparticle-enabled drug and nucleic acid delivery and photothermal therapy are addressed. PMID:24383043

  15. Cellular uptake and toxicity effects of silver nanoparticles in mammalian kidney cells.

    PubMed

    Mili?, Mirta; Leitinger, Gerd; Pavi?i?, Ivan; Zebi? Avdi?evi?, Maja; Dobrovi?, Slaven; Goessler, Walter; Vinkovi? Vr?ek, Ivana

    2014-10-28

    The rapid progress and early commercial acceptance of silver-based nanomaterials is owed to their biocidal activity. Besides embracing the antimicrobial potential of silver nanoparticles (AgNPs), it is imperative to give special attention to the potential adverse health effects of nanoparticles owing to prolonged exposure. Here, we report a detailed study on the in vitro interactions of citrate-coated AgNPs with porcine kidney (Pk15) cells. As uncertainty remains whether biological/cellular responses to AgNPs are solely as a result of the release of silver ions or whether the AgNPs themselves have toxic effects, we investigated the effects of Ag(+) on Pk15 cells for comparison. Next, we investigated the cellular uptake of both AgNPs and Ag(+) in Pk15 cells at various concentrations applied. The detected Ag contents in cells exposed to 50?mg?l(-1) AgNPs and 50?mg?l(-1) Ag(+) were 209 and 25?µg of Ag per 10(6) cells, respectively. Transmission electron microscopy (TEM) images indicated that the Pk15 cells internalized AgNPs by endocytosis. Both forms of silver, nano and ionic, decreased the number of viable Pk15 cells after 24?h in a dose-dependent manner. In spite of a significant uptake into the cells, AgNPs had only insignificant toxicity at concentrations lower than 25?mg?l(-1) , whereas Ag(+) exhibited a significant decrease in cell viability at one-fifth of this concentration. The Comet assay suggested that a rather high concentration of AgNP (above 25?mg?l(-1) ) is able to induce genotoxicity in Pk15 cells. Further studies must seek deeper understanding of AgNP behavior in biological media and their interactions with cellular membranes. Copyright © 2014 John Wiley & Sons, Ltd. PMID:25352480

  16. Genotoxic responses to titanium dioxide nanoparticles and fullerene in gpt delta transgenic MEF cells

    Microsoft Academic Search

    An Xu; Yunfei Chai; Takehiko Nohmi; Tom K Hei

    2009-01-01

    BACKGROUND: Titanium dioxide (TiO2) nanoparticles and fullerene (C60) are two attractive manufactured nanoparticles with great promise in industrial and medical applications. However, little is known about the genotoxic response of TiO2 nanoparticles and C60 in mammalian cells. In the present study, we determined the mutation fractions induced by either TiO2 nanoparticles or C60 in gpt delta transgenic mouse primary embryo

  17. Carbon black and titanium dioxide nanoparticles elicit distinct apoptotic pathways in bronchial epithelial cells

    PubMed Central

    2010-01-01

    Background Increasing environmental and occupational exposures to nanoparticles (NPs) warrant deeper insight into the toxicological mechanisms induced by these materials. The present study was designed to characterize the cell death induced by carbon black (CB) and titanium dioxide (TiO2) NPs in bronchial epithelial cells (16HBE14o- cell line and primary cells) and to investigate the implicated molecular pathways. Results Detailed time course studies revealed that both CB (13 nm) and TiO2(15 nm) NP exposed cells exhibit typical morphological (decreased cell size, membrane blebbing, peripheral chromatin condensation, apoptotic body formation) and biochemical (caspase activation and DNA fragmentation) features of apoptotic cell death. A decrease in mitochondrial membrane potential, activation of Bax and release of cytochrome c from mitochondria were only observed in case of CB NPs whereas lipid peroxidation, lysosomal membrane destabilization and cathepsin B release were observed during the apoptotic process induced by TiO2 NPs. Furthermore, ROS production was observed after exposure to CB and TiO2 but hydrogen peroxide (H2O2) production was only involved in apoptosis induction by CB NPs. Conclusions Both CB and TiO2 NPs induce apoptotic cell death in bronchial epithelial cells. CB NPs induce apoptosis by a ROS dependent mitochondrial pathway whereas TiO2 NPs induce cell death through lysosomal membrane destabilization and lipid peroxidation. Although the final outcome is similar (apoptosis), the molecular pathways activated by NPs differ depending upon the chemical nature of the NPs. PMID:20398356

  18. Biocompatible fluorescent nanoparticles for in vivo stem cell tracking

    NASA Astrophysics Data System (ADS)

    Cova, Lidia; Bigini, Paolo; Diana, Valentina; Sitia, Leopoldo; Ferrari, Raffaele; Pesce, Ruggiero Maria; Khalaf, Rushd; Bossolasco, Patrizia; Ubezio, Paolo; Lupi, Monica; Tortarolo, Massimo; Colombo, Laura; Giardino, Daniela; Silani, Vincenzo; Morbidelli, Massimo; Salmona, Mario; Moscatelli, Davide

    2013-06-01

    Efficient application of stem cells to the treatment of neurodegenerative diseases requires safe cell tracking to follow stem cell fate over time in the host environment after transplantation. In this work, for the first time, fluorescent and biocompatible methyl methacrylate (MMA)-based nanoparticles (fluoNPs) were synthesized through a free-radical co-polymerization process with a fluorescent macromonomer obtained by linking Rhodamine B and hydroxyethyl methacrylate. We demonstrate that the fluoNPs produced by polymerization of MMA-Rhodamine complexes (1) were efficient for the labeling and tracking of multipotent human amniotic fluid cells (hAFCs); (2) did not alter the main biological features of hAFCs (such as viability, cell growth and metabolic activity); (3) enabled us to determine the longitudinal bio-distribution of hAFCs in different brain areas after graft in the brain ventricles of healthy mice by a direct fluorescence-based technique. The reliability of our approach was furthermore confirmed by magnetic resonance imaging analyses, carried out by incubating hAFCs with both superparamagnetic iron oxide nanoparticles and fluoNPs. Our data suggest that these finely tunable and biocompatible fluoNPs can be exploited for the longitudinal tracking of stem cells.

  19. The novel protein PTPIP51 exhibits tissue- and cell-specific expression.

    PubMed

    Stenzinger, Albrecht; Kajosch, Tobias; Tag, Claudia; Porsche, Alexandra; Welte, Inka; Hofer, Hans Werner; Steger, Klaus; Wimmer, Monika

    2005-01-01

    The expression patterns of both mRNA and protein of the novel protein tyrosine phosphatase interacting protein 51 (PTPIP51) were studied in various organs by in situ hybridization, immunoblotting, and immunocytochemistry. The protein was found in all mammalian species investigated: guinea pig, rat, mouse, pig, and human. The presence of the protein was, however, restricted to specific organs. High levels of PTPIP51 were found in epidermis and seminiferous epithelium. The expression appears to be associated with distinct stages of differentiation. While basal cells in the epidermis and spermatogonia showed no perceptible amount of PTPIP51, keratinocytes of suprabasal layers and differentiating first-order spermatocytes up to spermatids exhibited high expression. In skeletal muscle, the presence of PTPIP51 was restricted to fibers of the fast twitch type. In surface epithelia containing ciliated cells, the protein was associated with the microtubular structures responsible for ciliary movement. Furthermore, specific structures of the central nervous system, for example, neurons of the hippocampal region, ganglion cells of the autonomic nervous system, and axons of the peripheral nervous system showed a distinct staining pattern with the antibody to PTPIP51. Our data suggest that PTPIP51 might be involved in the regulation of cellular processes associated with differentiation, movement, or cytoskeletal organization. PMID:15609043

  20. Damnacanthal, a noni component, exhibits antitumorigenic activity in human colorectal cancer cells.

    PubMed

    Nualsanit, Thararat; Rojanapanthu, Pleumchitt; Gritsanapan, Wandee; Lee, Seong-Ho; Lawson, Darunee; Baek, Seung Joon

    2012-08-01

    Damnacanthal, an anthraquinone compound, is isolated from the roots of Morinda citrifolia L. (noni), which has been used for traditional therapy in several chronic diseases including cancer. Although noni has been consumed for a long time in Asian and Polynesian countries, the molecular mechanisms by which it exerts several benefits are starting to emerge. In this report, we examined systematic approaches on the cancer-suppressing capability of damnacanthal in colorectal tumorigenesis. Damnacanthal exhibits cell growth arrest as well as caspase activity induction in colorectal cancer cells. We also examined several potential target proteins and found that the proapoptotic protein nonsteroidal anti-inflammatory activated gene-1 (NAG-1) is highly induced. Subsequently, we have found that damnacanthal also enhances transcription factor CCAAT/enhancer binding protein ? (C/EBP?), which controls NAG-1 transcriptional activity. Blocking of C/EBP? by shRNA results in the reduction of NAG-1 expression as well as caspase activity in the presence of damnacanthal. Taken together, these results indicate that damnacanthal increases antitumorigenic activity in human colorectal cancer cells and that C/EBP? plays a role in damnacanthal-induced NAG-1 expression. PMID:21852088

  1. Damnacanthal, a Noni component, exhibits anti-tumorigenic activity in human colorectal cancer cells

    PubMed Central

    Nualsanit, Thararat; Rojanapanthu, Pleumchitt; Gritsanapan, Wandee; Lee, Seong-Ho; Lawson, Darunee; Baek, Seung Joon

    2011-01-01

    Damnacanthal, an anthraquinone compound, is isolated from the roots of Morinda citrifolia L. (noni), which has been used for traditional therapy in several chronic diseases including cancer. Although noni has been consumed for a long time in Asian and Polynesian countries, the molecular mechanisms by which it exerts several benefits are starting to emerge. In this report, we examined systematic approaches on the cancer suppressing capability of damnacanthal in colorectal tumorigenesis. Damnacanthal exhibits cell growth arrest as well as caspase activity induction in colorectal cancer cells. We also examined several potential target proteins and found that the pro-apoptotic protein Nonsteroidal anti-inflammatory activated gene-1 (NAG-1) is highly induced. Subsequently, we have found that damnacanthal also enhances transcription factor C/EBP?, which controls NAG-1 transcriptional activity. Blocking of C/EBP? by shRNA results in the reduction of NAG-1 expression as well as caspase activity in the presence of damnacanthal. Taken together, these results indicate that damnacanthal increases anti-tumorigenic activity in human colorectal cancer cells, and C/EBP? plays a role in damnacanthal-induced NAG-1 expression. PMID:21852088

  2. Congenital amegakaryocytic thrombocytopenia iPS cells exhibit defective MPL-mediated signaling

    PubMed Central

    Hirata, Shinji; Takayama, Naoya; Jono-Ohnishi, Ryoko; Endo, Hiroshi; Nakamura, Sou; Dohda, Takeaki; Nishi, Masanori; Hamazaki, Yuhei; Ishii, Ei-ichi; Kaneko, Shin; Otsu, Makoto; Nakauchi, Hiromitsu; Kunishima, Shinji; Eto, Koji

    2013-01-01

    Congenital amegakaryocytic thrombocytopenia (CAMT) is caused by the loss of thrombopoietin receptor–mediated (MPL-mediated) signaling, which causes severe pancytopenia leading to bone marrow failure with onset of thrombocytopenia and anemia prior to leukopenia. Because Mpl–/– mice do not exhibit the human disease phenotype, we used an in vitro disease tracing system with induced pluripotent stem cells (iPSCs) derived from a CAMT patient (CAMT iPSCs) and normal iPSCs to investigate the role of MPL signaling in hematopoiesis. We found that MPL signaling is essential for maintenance of the CD34+ multipotent hematopoietic progenitor (MPP) population and development of the CD41+GPA+ megakaryocyte-erythrocyte progenitor (MEP) population, and its role in the fate decision leading differentiation toward megakaryopoiesis or erythropoiesis differs considerably between normal and CAMT cells. Surprisingly, complimentary transduction of MPL into normal or CAMT iPSCs using a retroviral vector showed that MPL overexpression promoted erythropoiesis in normal CD34+ hematopoietic progenitor cells (HPCs), but impaired erythropoiesis and increased aberrant megakaryocyte production in CAMT iPSC–derived CD34+ HPCs, reflecting a difference in the expression of the transcription factor FLI1. These results demonstrate that impaired transcriptional regulation of the MPL signaling that normally governs megakaryopoiesis and erythropoiesis underlies CAMT. PMID:23908116

  3. Labeling of immune cells for in vivo imaging using magnetofluorescent nanoparticles

    Microsoft Academic Search

    Filip K Swirski; Frederick Reynolds; Lee Josephson; Ralph Weissleder; Mikael J Pittet

    2006-01-01

    Observation of immune and stem cells in their native microenvironments requires the development of imaging agents to allow their in vivo tracking. We describe here the synthesis of magnetofluorescent nanoparticles for cell labeling in vitro and for multimodality imaging of administered cells in vivo. MION-47, a prototype monocrystalline iron oxide nanoparticle, was first converted to an intermediate bearing a fluorochrome

  4. Novel cell lines isolated from mouse embryonic stem cells exhibiting de novo methylation of the E-cadherin promoter.

    PubMed

    Hawkins, Kate; Keramari, Maria; Soncin, Francesca; Segal, Joe M; Mohamet, Lisa; Miazga, Natalie; Ritson, Sarah; Bobola, Nicoletta; Merry, Catherine L R; Ward, Christopher M

    2014-11-01

    Mouse embryonic stem cells (mESCs) and epiblast stem cells represent the naïve and primed pluripotent states, respectively. These cells self-renew via distinct signaling pathways and can transition between the two states in the presence of appropriate growth factors. Manipulation of signaling pathways has therefore allowed the isolation of novel pluripotent cell types such as Fibroblast growth factor, Activin and BIO-derived stem cells and IESCs. However, the effect of cell seeding density on pluripotency remains unexplored. In this study, we have examined whether mESCs can epigenetically regulate E-cadherin to enter a primed-like state in response to low cell seeding density. We show that low density seeding in the absence of leukaemia inhibitory factor (LIF) induces decreased apoptosis and maintenance of pluripotency via Activin/Nodal, concomitant with loss of E-cadherin, Signal transducer and activator of transcription phosphorylation, and chimera-forming ability. These cells, E-cadherin negative proliferating stem cells (ENPSCs) can be reverted to a naïve phenotype by addition of LIF or forced E-cadherin expression. However, prolonged culture of ENPSCs without LIF leads to methylation of the E-cadherin promoter (ENPSC(M)), which cannot be reversed by LIF supplementation, and increased histone H3K27 and decreased H3K4 trimethylation. Transcript analysis of ENPSC(M) revealed a primed-like phenotype and their differentiation leads to enrichment of neuroectoderm cells. The generation of ENPSCs is similar to tumorigenesis as ENPSCs exhibit transcript alterations associated with neoplasia, hyperplasia, carcinoma, and metastasis. We therefore describe a novel cell model to elucidate the role of E-cadherin in pluripotency and to investigate epigenetic regulation of this gene during mESC differentiation and tumor metastasis. PMID:25074424

  5. Nanoparticle assisted photothermal deformation of individual neuronal organelles and cells

    PubMed Central

    Romero, V. H.; Kereselidze, Z.; Egido, W.; Michaelides, E. A.; Santamaria, F.; Peralta, X. G.

    2014-01-01

    Stimulation of the localized surface plasmon of metallic nanoparticles has been shown to be an effective mechanism to induce photothermal damage in biological tissues. However, few studies have focused on single cell or subcellular ablation. Our results show that, upon incubation, gold nanostars are internalized by neurons of acute mouse cerebellar brain slices, clustering inside or close to the nucleus. By stimulating the nanostars’ surface plasmon using a femtosecond laser, we show deformation of single nuclei and single cells. Given its precision and extremely localized effect, this is a promising technique for photothermal therapy in areas sensitive to collateral thermal damage such as the nervous system. PMID:25426326

  6. Toxicity of gold nanoparticles on somatic and reproductive cells.

    PubMed

    Taylor, U; Barchanski, A; Garrels, W; Klein, S; Kues, W; Barcikowski, S; Rath, D

    2012-01-01

    Along with the number of potential applications for gold nanoparticles (AuNP) especially for medical and scientific purposes, the interest in possible toxic effects of such particles is rising. The general perception views nanosized gold colloids as relatively inert towards biological systems. However, a closer analysis of pertinent studies reveals a more complex picture. While the chemical compound of which the nanoparticles consists plays an important role, further biocompatibility determining aspects have been made out. The vast majority of trials concerning AuNP-toxicity were performed using somatic cell culture lines. The results show a considerable dependency of toxic effects on size, zeta potential and surface functionalisation. In vivo studies on this subject are still rare. Based on the existing data it can be assumed, that a dosage of under <400 µg Au/kg showed no untoward effects. If higher amounts were applied toxicity depended on route of administration and particle size. Since nanoparticles have been shown to cross reproduction-relevant biological barriers such as the blood-testicle and the placental barrier the question of their reprotoxicity arises. Yet data concerning this subject is far from adequate. Regarding gametes, recent experiments showed a dose-dependent sensitivity of spermatozoa towards AuNP. Oocytes have not yet been tested in that respect. Interestingly, so far no effects were detected on embryos after gold nanoparticle exposure. In conclusion, the biocompatibility of gold nanoparticles depends on a range of particle specific aspects as well as the choice of target tissue. Further clarification of such matters are subject to ongoing research. PMID:22101718

  7. Commercial Nanoparticles for Stem Cell Labeling and Tracking

    PubMed Central

    Wang, Yaqi; Xu, Chenjie; Ow, Hooisweng

    2013-01-01

    Stem cell therapy provides promising solutions for diseases and injuries that conventional medicines and therapies cannot effectively treat. To achieve its full therapeutic potentials, the homing process, survival, differentiation, and engraftment of stem cells post transplantation must be clearly understood. To address this need, non-invasive imaging technologies based on nanoparticles (NPs) have been developed to track transplanted stem cells. Here we summarize existing commercial NPs which can act as contrast agents of three commonly used imaging modalities, including fluorescence imaging, magnetic resonance imaging and photoacoustic imaging, for stem cell labeling and tracking. Specifically, we go through their technologies, industry distributors, applications and existing concerns in stem cell research. Finally, we provide an industry perspective on the potential challenges and future for the development of new NP products. PMID:23946821

  8. Comparative evaluation of novel biodegradable nanoparticles for the drug targeting to breast cancer cells.

    PubMed

    Mattu, C; Pabari, R M; Boffito, M; Sartori, S; Ciardelli, G; Ramtoola, Z

    2013-11-01

    Nanomedicine formulations such as biodegradable nanoparticles (nps) and liposomes offer several advantages over traditional routes of administration: due to their small size, nanocarriers are able to selectively accumulate inside tumours or inflammatory tissues, resulting in improved drug efficacy and reduced side effects. To further augment targeting ability of nanoparticles towards tumour cells, specific ligands or antibodies that selectively recognise biomarkers over-expressed on cancer cells, can be attached to the surface either by chemical bond or by hydrophilic/hydrophobic interactions. In the present work, Herceptin (HER), a monoclonal antibody (mAb) able to selectively recognise HER-2 over-expressing tumour cells (such as breast and ovarian cancer cells), was absorbed on the surface of nanoparticles through hydrophilic/hydrophobic interactions. Nps were prepared by a modified single emulsion solvent evaporation method with five different polymers: three commercial polyesters (poly(?-caprolactone) (PCL), poly (D,L-lactide) (PLA) and poly (D,L-lactide-co-.glycolide) (PLGA)) and two novel biodegradable polyesterurethanes (PURs) based on Poly(?-caprolactone) blocks, synthesised with different chain extenders (1,4-cyclohexane dimethanol (CDM) and N-Boc-serinol). Polyurethanes were introduced as matrix-forming materials for nanoparticles due to their high chemical versatility, which allows tailoring of the materials final properties by properly selecting the reagents. All nps exhibited a small size and negative surface charge, suitable for surface functionalisation with mAb through hydrophilic/hydrophobic interactions. The extent of cellular internalisation was tested on two different cell lines: MCF-7 and SK-BR-3 breast cancer cells showing a normal and a high expression of the HER-2 receptor, respectively. Paclitaxel, a model anti-neoplastic drug, was encapsulated inside all nps, and release profiles and cytotoxicity on SK-BR-3 cells were also assessed. Interestingly, PUR nps were superior to commercial polyester-based nps in terms of higher cellular internalisation and cytotoxic activity on the tested cell lines. Results obtained warrants further investigation on the application of these PUR nps for controlled drug delivery and targeting. PMID:23916461

  9. Cell-specific expression of artificial microRNAs targeting essential genes exhibit potent antitumor effect on hepatocellular carcinoma cells.

    PubMed

    Mao, Chenyu; Liu, Hao; Chen, Ping; Ye, Jingjia; Teng, Lisong; Jia, Zhenyu; Cao, Jiang

    2015-03-20

    To achieve specific and potent antitumor effect of hepatocyte carcinoma cells, replication defective adenoviral vectors, namely rAd/AFP-amiRG, rAd/AFP-amiRE and rAd/AFP-amiRP, were constructed which were armed with artificial microRNAs (amiRs) targeting essential functional genes glyceraldehyde-3-phosphate dehydrogenase, eukaryotic translation initiation factor 4E and DNA polymerase ? respectively under the control of a recombinant promoter comprised of human ?-fetoprotein enhancer and basal promoter. The AFP enhancer/promoter showed specific high transcription activity in AFP-positive HCC cells Hep3B, HepG2 and SMMC7721, while low in AFP-negative cell Bcap37. All artificial microRNAs exhibited efficient knockdown of target genes. Decreased ATP production and protein synthesis was observed in rAd/AFP-amiRG and rAd/AFP-amiRE treated HCC cells. All three recombinant adenoviruses showed efficient blockage of cell cycle progression and significant suppression of HCC cells in vitro. In nude mice model bearing Hep3B xenograft, administration of rAd/AFP-amiRG showed potent antitumor effect. The strategy of tumor-specific knockdown of genes essential for cell survival and proliferation may suggest a novel promising approach for HCC gene therapy. PMID:25691059

  10. Do cancer cells in human and meristematic cells in plant exhibit similar responses toward plant extracts with cytotoxic activities?

    PubMed

    Khalifa, Noha S; Barakat, Hoda S; Elhallouty, Salwa; Salem, Dina

    2015-01-01

    We examined the effect of water extracts of Persea americana fruit, and of the leaves of Tabernamontana divericata, Nerium oleander and Annona cherimolia (positive control) on Vicia faba root cells. We had confirmed in our previously published data the cytotoxicity of these plant extracts on four human cancer cell lines: liver (HepG-2), lung (A549), colon (HT-29) and breast (MCF-7). Vicia faba roots were soaked in plant extracts at dilutions of 100, 1,250, 2,500, 5,000, 10,000, 20,000 ppm for 4 and 24 h. All treatments resulted in a significant reduction in the mitotic index in a dose dependant manner. Root cells treated with T. divericata, N. oleander and A. cherimolia exhibited a decrease in prophase cell percentage, increase in micronuclei and chromosomal abnormalities as concentration increased. The P. americana treatment showed the highest cytotoxic effect on cancer cells, prophase cell percentage increased linearly with the applied concentration and no micronuclei were detected. This study shows that root tip assay of beans can be used in initial screening for new plant extracts to validate their use as candidates for containing active cytotoxic agents against malignant cells. This will greatly help in exploring new plant extracts as drugs for cancer treatment. PMID:24705601

  11. Single-molecule imaging in live cell using gold nanoparticles.

    PubMed

    Leduc, Cécile; Si, Satyabrata; Gautier, Jérémie J; Gao, Zhenghong; Shibu, Edakkattuparambil S; Gautreau, Alexis; Giannone, Grégory; Cognet, Laurent; Lounis, Brahim

    2015-01-01

    Optimal single particle tracking experiments in live cells requires small and photostable probes, which do not modify the behavior of the molecule of interest. Current fluorescence-based microscopy of single molecules and nanoparticles is often limited by bleaching and blinking or by the probe size. As an alternative, we present in this chapter the synthesis of a small and highly specific gold nanoprobe whose detection is based on its absorption properties. We first present a protocol to synthesize 5-nm-diameter gold nanoparticles and functionalize them with a nanobody, a single-domain antibody from camelid, targeting the widespread green fluorescent protein (GFP)-tagged proteins with a high affinity. Then we describe how to detect and track these individual gold nanoparticles in live cell using photothermal imaging microscopy. The combination of a probe with small size, perfect photostability, high specificity, and versatility through the vast existing library of GFP-proteins, with a highly sensitive detection technique enables long-term tracking of proteins with minimal hindrance in confined and crowded environments such as intracellular space. PMID:25640421

  12. Mast cells contribute to altered vascular reactivity and ischemia-reperfusion injury following cerium oxide nanoparticle instillation

    PubMed Central

    WINGARD, CHRISTOPHER J.; WALTERS, DIANNE M.; CATHEY, BROOK L.; HILDERBRAND, SUSANA C.; KATWA, PRANITA; LIN, SIJIE; KE, PU CHUN; PODILA, RAMAKRISHNA; RAO, APPARAO; LUST, ROBERT M.; BROWN, JARED M.

    2011-01-01

    Cerium oxide (CeO2) represents an important nanomaterial with wide ranging applications. However, little is known regarding how CeO2 exposure may influence pulmonary or systemic inflammation. Furthermore, how mast cells would influence inflammatory responses to a nanoparticle exposure is unknown. We thus compared pulmonary and cardiovascular responses between C57BL/6 and B6.Cg-KitW-sh mast cell deficient mice following CeO2 nanoparticle instillation. C57BL/6 mice instilled with CeO2 exhibited mild pulmonary inflammation. However, B6.Cg-KitW-sh mice did not display a similar degree of inflammation following CeO2 instillation. Moreover, C57BL/6 mice instilled with CeO2 exhibited altered aortic vascular responses to adenosine and an increase in myocardial ischemia/reperfusion injury which was absent in B6.Cg-KitW-sh mice. In vitro CeO2 exposure resulted in increased production of PGD2, TNF-?, IL-6 and osteopontin by cultured mast cells. These findings demonstrate that CeO2 nanoparticles activate mast cells contributing to pulmonary inflammation, impairment of vascular relaxation and exacerbation of myocardial ischemia/reperfusion injury. PMID:21043986

  13. Proper design of silica nanoparticles combines high brightness, lack of cytotoxicity and efficient cell endocytosis.

    PubMed

    Rampazzo, Enrico; Voltan, Rebecca; Petrizza, Luca; Zaccheroni, Nelsi; Prodi, Luca; Casciano, Fabio; Zauli, Giorgio; Secchiero, Paola

    2013-09-01

    Silica-based luminescent nanoparticles (SiNPs) show promising prospects in nanomedicine in light of their chemical properties and versatility. In this study, we have characterized silica core-PEG shell SiNPs derivatized with PEG moieties (NP-PEG), with external amino- (NP-PEG-amino) or carboxy-groups (NP-PEG-carbo), both in cell cultures as well as in animal models. By using different techniques, we could demonstrate that these SiNPs were safe and did not exhibit appreciable cytotoxicity in different relevant cell models, of normal or cancer cell types, growing either in suspension (JVM-2 leukemic cell line and primary normal peripheral blood mononuclear cells) or in adherence (human hepatocarcinoma Huh7 and umbilical vein endothelial cells). Moreover, by multiparametric flow cytometry, we could demonstrate that the highest efficiency of cell uptake and entry was observed with NP-PEG-amino, with a stable persistence of the fluorescence signal associated with SiNPs in the loaded cell populations both in vitro and in vivo settings suggesting this as an innovative method for cell traceability and detection in whole organisms. Finally, experiments performed with the endocytosis inhibitor Genistein clearly suggested the involvement of a caveolae-mediated pathway in SiNP endocytosis. Overall, these data support the safe use of these SiNPs for diagnostic and therapeutic applications. PMID:23851463

  14. Graphene nanosheets inserted by silver nanoparticles as zero-dimensional nanospacers for dye sensitized solar cells.

    PubMed

    Chang, Quanhong; Wang, Zhenping; Wang, Jinzhong; Yan, Yuan; Ma, Zhoujing; Zhu, Jianxiao; Shi, Wangzhou; Chen, Qi; Yu, Qingjiang; Huang, Lei

    2014-05-21

    Three-dimensional Ag nanoparticle/GNs (Ag/GNs) hybrids as highly efficient counter electrode (CE) materials for dye sensitized solar cells (DSSCs) is described, highlighting the Ag nanoparticles as zero-dimensional nanospacers inserting into GNs to lift the interspacing layer between individual GNs. It is demonstrated that, when the hybrids are used as CE materials for DSSCs, compared to their pure GNs, Ag/GNs hybrids without agglomerates have a significant improvement in their electrochemical properties such as high current density, narrow peak-to-peak separation (Epp) and low charge transfer resistance (RCT). The enhancement of electrochemical performance can be attributed to the increased electrode conductivity, an extended interlayer distance and the reduction of the restacking of graphene sheets due to the insertion of metallic Ag nanoparticles into GNs. The DSSC with this hybrid CE exhibited an energy conversion efficiency (?) of 7.72% with an open circuit voltage (VOC), short circuit photocurrent density (JSC), and fill factor (FF) of 732 mV, 14.67 mA cm(-2), and 71.8%, respectively. PMID:24710127

  15. Manganese nanoparticle activates mitochondrial dependent apoptotic signaling and autophagy in dopaminergic neuronal cells

    SciTech Connect

    Afeseh Ngwa, Hilary; Kanthasamy, Arthi [Department of Biomedical Sciences, Iowa Center for Advanced Neurotoxicology, Iowa State University, Ames, IA 50011 (United States)] [Department of Biomedical Sciences, Iowa Center for Advanced Neurotoxicology, Iowa State University, Ames, IA 50011 (United States); Gu, Yan; Fang, Ning [Department of Chemistry, Iowa State University, Ames, IA 50011 (United States)] [Department of Chemistry, Iowa State University, Ames, IA 50011 (United States); Anantharam, Vellareddy [Department of Biomedical Sciences, Iowa Center for Advanced Neurotoxicology, Iowa State University, Ames, IA 50011 (United States)] [Department of Biomedical Sciences, Iowa Center for Advanced Neurotoxicology, Iowa State University, Ames, IA 50011 (United States); Kanthasamy, Anumantha G., E-mail: akanthas@iastate.edu [Department of Biomedical Sciences, Iowa Center for Advanced Neurotoxicology, Iowa State University, Ames, IA 50011 (United States)

    2011-11-15

    The production of man-made nanoparticles for various modern applications has increased exponentially in recent years, but the potential health effects of most nanoparticles are not well characterized. Unfortunately, in vitro nanoparticle toxicity studies are extremely limited by yet unresolved problems relating to dosimetry. In the present study, we systematically characterized manganese (Mn) nanoparticle sizes and examined the nanoparticle-induced oxidative signaling in dopaminergic neuronal cells. Differential interference contrast (DIC) microscopy and transmission electron microscopy (TEM) studies revealed that Mn nanoparticles range in size from single nanoparticles ({approx} 25 nM) to larger agglomerates when in treatment media. Manganese nanoparticles were effectively internalized in N27 dopaminergic neuronal cells, and they induced a time-dependent upregulation of the transporter protein transferrin. Exposure to 25-400 {mu}g/mL Mn nanoparticles induced cell death in a time- and dose-dependent manner. Mn nanoparticles also significantly increased ROS, accompanied by a caspase-mediated proteolytic cleavage of proapoptotic protein kinase C{delta} (PKC{delta}), as well as activation loop phosphorylation. Blocking Mn nanoparticle-induced ROS failed to protect against the neurotoxic effects, suggesting the involvement of other pathways. Further mechanistic studies revealed changes in Beclin 1 and LC3, indicating that Mn nanoparticles induce autophagy. Primary mesencephalic neuron exposure to Mn nanoparticles induced loss of TH positive dopaminergic neurons and neuronal processes. Collectively, our results suggest that Mn nanoparticles effectively enter dopaminergic neuronal cells and exert neurotoxic effects by activating an apoptotic signaling pathway and autophagy, emphasizing the need for assessing possible health risks associated with an increased use of Mn nanoparticles in modern applications. -- Highlights: Black-Right-Pointing-Pointer Mn nanoparticles activate mitochondrial cell death signaling in dopaminergic neuron. Black-Right-Pointing-Pointer Mn nanoparticles activate caspase-mediated proteolytic cleavage of PKC{delta} cascade. Black-Right-Pointing-Pointer Mn nanoparticles induce autophagy in dopaminergic neuronal cells. Black-Right-Pointing-Pointer Mn nanoparticles induce loss of TH{sup +} neurons in primary mesencephalic cultures. Black-Right-Pointing-Pointer Study emphasizes neurotoxic risks of Mn nanoparticles to nigral dopaminergic system.

  16. Comparative cytotoxicity studies of carbon-encapsulated iron nanoparticles in murine glioma cells.

    PubMed

    Grudzinski, Ireneusz P; Bystrzejewski, Michal; Cywinska, Monika A; Kosmider, Anita; Poplawska, Magdalena; Cieszanowski, Andrzej; Fijalek, Zbigniew; Ostrowska, Agnieszka

    2014-05-01

    Carbon-encapsulated iron nanoparticles (CEINs) have recently emerged as a new class of magnetic nanomaterials with a great potential for an increasing number of biomedical applications. To address the current deficient knowledge of cellular responses due to CEIN exposures, we focused on the investigation of internalization profile and resulting cytotoxic effects of CEINs (0.0001-100 ?g/ml) in murine glioma cells (GL261) in vitro. The studied CEIN samples were characterized (TEM, FT-IR, Zeta potential, Boehm titration) and examined as raw and purified nanomaterials with various surface chemistry composition. Of the four type CEINs (the mean diameter 47-56 nm) studied here, the as-synthesized raw nanoparticles (Fe@C/Fe) exhibited high cytotoxic effects on the plasma cell membrane (LDH, Calcein AM/PI) and mitochondria (MTT, JC-1) causing some pro-apoptotic evens (Annexin V/PI) in glioma cells. The effects of the purified (Fe@C) and surface-modified (Fe@C-COOH and Fe@C-(CH2)2COOH) CEINs were found in quite similar patterns; however, most of these cytotoxic events were slightly diminished compared to those induced by Fe@C/Fe. The study showed that the surface-functionalized CEINs affected the cell cycle progression in both S and G2/M phases to a greater extent compared to that of the rest of nanoparticles studied to data. Taken all together, the present results highlight the importance of the rational design of CEINs as their physicochemical features such as morphology, hydrodynamic size, impurity profiles, and especially surface characteristics are critical determinants of different cytotoxic responses. PMID:24632386

  17. Engineered nanoparticles interacting with cells: size matters

    PubMed Central

    2014-01-01

    With the rapid advancement of nanoscience and nanotechnology, detailed knowledge of interactions between engineered nanomaterials and cells, tissues and organisms has become increasingly important, especially in regard to possible hazards to human health. This review intends to give an overview of current research on nano-bio interactions, with a focus on the effects of NP size on their interactions with live cells. We summarize common techniques to characterize NP size, highlight recent work on the impact of NP size on active and passive cellular internalization and intracellular localization. Cytotoxic effects are also discussed. PMID:24491160

  18. Cytotoxicity of silica nanoparticles on HaCaT cells.

    PubMed

    Liang, Hao; Jin, Chan; Tang, Ying; Wang, Fude; Ma, Chunwang; Yang, Yongji

    2014-04-01

    Despite the widespread use of silica nanoparticles (SiO2 NPs) in biological and medical fields, their adverse effects have not been clearly elucidated. In this study, spherical SiO2 NPs with a 50 nm diameter were used to study their interaction with HaCaT cells. SiO2 NPs were found to be readily internalized into HaCaT cells and localized in the cytoplasm, lysosomes and autophagosomes. Decreased cell viability and damaged cell membrane integrity showed the cytotoxicity of SiO2 NPs. Significant glutathione depletion and reactive oxygen species generation, which reduced the cellular antioxidant level, could be the major factor of cytotoxicity induced by SiO2 NPs. PMID:24155225

  19. Titanium dioxide nanoparticles cause genotoxicity in human lung epithelial cells

    EPA Science Inventory

    The use of engineered nanoparticles in consumer products is steadily increasing. However, the health effects of exposure to these nanoparticles are not thoroughly understood. This study investigated the genotoxicity of six titanium dioxide and two cerium oxide nanoparticles of va...

  20. Enhanced penetration into 3D cell culture using two and three layered gold nanoparticles

    PubMed Central

    England, Christopher G; Priest, Thomas; Zhang, Guandong; Sun, Xinghua; Patel, Dhruvinkumar N; McNally, Lacey R; van Berkel, Victor; Gobin, André M; Frieboes, Hermann B

    2013-01-01

    Nano-scale particles sized 10–400 nm administered systemically preferentially extravasate from tumor vasculature due to the enhanced permeability and retention effect. Therapeutic success remains elusive, however, because of inhomogeneous particle distribution within tumor tissue. Insufficient tumor vascularization limits particle transport and also results in avascular hypoxic regions with non-proliferating cells, which can regenerate tissue after nanoparticle-delivered cytotoxicity or thermal ablation. Nanoparticle surface modifications provide for increasing tumor targeting and uptake while decreasing immunogenicity and toxicity. Herein, we created novel two layer gold-nanoshell particles coated with alkanethiol and phosphatidylcholine, and three layer nanoshells additionally coated with high-density-lipoprotein. We hypothesize that these particles have enhanced penetration into 3-dimensional cell cultures modeling avascular tissue when compared to standard poly(ethylene glycol) (PEG)-coated nanoshells. Particle uptake and distribution in liver, lung, and pancreatic tumor cell cultures were evaluated using silver-enhancement staining and hyperspectral imaging with dark field microscopy. Two layer nanoshells exhibited significantly higher uptake compared to PEGylated nanoshells. This multilayer formulation may help overcome transport barriers presented by tumor vasculature, and could be further investigated in vivo as a platform for targeted cancer therapies. PMID:24124360

  1. Micro-Raman Spectroscopy of Silver Nanoparticle Induced Stress on Optically-Trapped Stem Cells

    PubMed Central

    Bankapur, Aseefhali; Krishnamurthy, R. Sagar; Zachariah, Elsa; Santhosh, Chidangil; Chougule, Basavaraj; Praveen, Bhavishna; Valiathan, Manna; Mathur, Deepak

    2012-01-01

    We report here results of a single-cell Raman spectroscopy study of stress effects induced by silver nanoparticles in human mesenchymal stem cells (hMSCs). A high-sensitivity, high-resolution Raman Tweezers set-up has been used to monitor nanoparticle-induced biochemical changes in optically-trapped single cells. Our micro-Raman spectroscopic study reveals that hMSCs treated with silver nanoparticles undergo oxidative stress at doping levels in excess of 2 µg/ml, with results of a statistical analysis of Raman spectra suggesting that the induced stress becomes more dominant at nanoparticle concentration levels above 3 µg/ml. PMID:22514708

  2. Effect of ZnO nanoparticles on nasopharyngeal cancer cells viability and respiration

    NASA Astrophysics Data System (ADS)

    Prasanth, R.; Gopinath, D.

    2013-03-01

    Development of a therapeutic drugs based on nanoparticles requires a better understanding of the mechanism of selective cyto-toxic effects of nanopaticles over cancer cells. Scanning electrochemical microscopy provides opportunity to measure the real time chemical process at cell proximity in the presence of nanoparticle. Herein, the respiration process in nasopharyngeal cancer cells is investigated with the help of scanning electrochemical microscopy. The cell viability has been tested with MTT assay. The results show that ZnO nanoparticles have time and dose dependent effect in nasopharyngeal cancer cells and the cell respiration rate decreases with time.

  3. Luminescent solar concentrators and all-inorganic nanoparticle solar cells for solar energy harvesting

    NASA Astrophysics Data System (ADS)

    Sholin, Veronica

    Increasing energy demand and the parallel increase of greenhouse gas emissions are challenging researchers to find new and cleaner energy sources. Solar energy harvesting is arguably the most promising candidate for replacing fossil-fuel power generation. Photovoltaics are the most direct way of collecting solar energy; cost continues to hinder large-scale implementation of photovoltaics, however. Therefore, alternative technologies that will allow the extraction of solar power, while maintaining the overall costs of fabrication, installation, collection, and distribution low, must be explored. This thesis focuses on the fabrication and testing of two types of devices that step up to this challenge: the luminescent solar concentrator (LSC) and all-inorganic nanoparticle solar cells. In these devices I make use of novel materials, semiconducting polymers and inorganic nanoparticles, both of which have lower costs than the crystalline materials used in the fabrication of traditional photovoltaics. Furthermore, the cost of manufacturing LSCs and the nanoparticle solar cells is lower than the manufacturing cost of traditional optics-based concentrators and crystalline solar cells. An LSC is essentially a slab of luminescent material that acts as a planar light pipe. The LSC absorbs incoming photons and channels fluoresced photons toward appropriately located solar cells, which perform the photovoltaic conversion. By covering large areas with relatively inexpensive fluorescing organic dyes or semiconducting polymers, the area of solar cell needed is greatly reduced. Because semiconducting polymers and quantum dots may have small absorption/emission band overlaps, tunable absorption, and longer lifetimes, they are good candidates for LSC fabrication, promising improvement with respect to laser dyes traditionally used to fabricate LSCs. Here the efficiency of LSCs consisting of liquid solutions of semiconducting polymers encased in glass was measured and compared to the efficiency of LSCs based on small molecule laser dyes and on quantum dots. Factors affecting the optical efficiency of the system such as the luminescing properties of the fluorophors were examined. The experimental results were compared to Monte-Carlo simulations. Our results suggest that commercially available quantum dots cannot serve as viable LSC dyes because of large absorption/emission band overlap and relatively low quantum yield. Materials such as Red F demonstrate that semi-conducting polymers with high quantum yield and small absorption/emission band overlap are good candidates for LSCs. Recently, a solar cell system based purely on CdSe and Cite nanoparticles as the absorbing materials was proposed ans it was suggested that its operational mechanism was that of polymer donor/acceptor systems. Here we present solar cells consisting of a sintered active bilayer of CdSe and PbSe nanoparticles in the structure ITO/CdSe/interlayer/PbSe/Al, where an interlayer of LiF or Al2O3 was found necessary to prevent low shunt resistance from suppressing the photovoltaic behavior. We fabricated unoptimized solar cells with a short-circuit current of 6 mA/cm2, an open-circuit voltage of 0.18 V, and a fill factor of 41%. External quantum efficiency spectra revealed that photons from the infrared portion of the spectrum were not collected, suggesting that the low bandgap PbSe film did not contribute to the photocurrent of the structure despite exhibiting photoconductivity. Other measurements, however, showed that the PbSe film was indeed necessary to produce a photovoltage and transport electrons. Through sintering, the nanoparticle films acquired bandgaps similar to those of the corresponding bulk materials and became more conductive. Because the PbSe films were found to be considerably more conductive than the CdSe ones, we suggest that the PbSe layer is effectively behaving like a low conductivity electrical contact. Therefore, in contrast to the photovoltaics presented in the seminal research on CdSe/Cite solar cells, the CdSe/PbSe solar cell system presented here d

  4. Biodegradable nanoparticles for targeted ultrasound imaging of breast cancer cells in vitro

    NASA Astrophysics Data System (ADS)

    Liu, Jun; Li, Jie; Rosol, Thomas J.; Pan, Xueliang; Voorhees, Jeffrey L.

    2007-08-01

    Disease-specific enhanced imaging through a targeted agent promises to improve the specificity of medical ultrasound. Nanoparticles may provide unique advantages for targeted ultrasound imaging due to their novel physical and surface properties. In this study, we examined a nanoparticle agent developed from a biodegradable polymer, polylactic acid (PLA). The nanoparticles (mean diameter = 250 nm) were surface conjugated to an anti-Her2 antibody (i.e., Herceptin) for specific binding to breast cancer cells that overexpress Her2 receptors. We examined the targeting specificity and the resultant ultrasound enhancement in Her2-positive and negative cells. Flow cytometry and confocal imaging were used to assess the nanoparticle-cell binding. Her2-positive cells demonstrated substantial staining after incubation with nanoparticle/antibody conjugates, while minimal staining was found in Her2-negative cells, indicating receptor-specific binding of the conjugated PLA nanoparticles. In high-resolution ultrasound B-mode images, the average gray scale of the Her2-positive cells was consistently and significantly higher after nanoparticle treatment (133 ± 4 in treated cells versus 109 ± 4 in control, p < 0.001, n = 5), while no difference was detected in the cells that did not overexpress the receptors (117 ± 3 in treated cells versus 118 ± 5 in control). In conclusion, the feasibility of using targeted nanoparticles to enhance ultrasonic images was demonstrated in vitro. This may be a promising approach to target cancer biomarkers for site-specific ultrasound imaging.

  5. nanoparticles

    NASA Astrophysics Data System (ADS)

    Olive-Méndez, Sion F.; Santillán-Rodríguez, Carlos R.; González-Valenzuela, Ricardo A.; Espinosa-Magaña, Francisco; Matutes-Aquino, José A.

    2014-04-01

    In this work, we present the role of vanadium ions (V+5 and V+3), oxygen vacancies (VO), and interstitial zinc (Zni) to the contribution of specific magnetization for a mixture of ZnO-V2O5 nanoparticles (NPs). Samples were obtained by mechanical milling of dry powders and ethanol-assisted milling for 1 h with a fixed atomic ratio V/Zn?=?5% at. For comparison, pure ZnO samples were also prepared. All samples exhibit a room temperature magnetization ranging from 1.18?×?10-3 to 3.5?×?10-3 emu/gr. Pure ZnO powders (1.34?×?10-3 emu/gr) milled with ethanol exhibit slight increase in magnetization attributed to formation of Zni, while dry milled ZnO powders exhibit a decrease of magnetization due to a reduction of VO concentration. For the ZnO-V2O5 system, dry milled and thermally treated samples under reducing atmosphere exhibit a large paramagnetic component associated to the formation of V2O3 and secondary phases containing V+3 ions; at the same time, an increase of VO is observed with an abrupt fall of magnetization to ??~?0.7?×?10-3 emu/gr due to segregation of V oxides and formation of secondary phases. As mechanical milling is an aggressive synthesis method, high disorder is induced at the surface of the ZnO NPs, including VO and Zni depending on the chemical environment. Thermal treatment restores partially structural order at the surface of the NPs, thus reducing the amount of Zni at the same time that V2O5 NPs segregate reducing the direct contact with the surface of ZnO NPs. Additional samples were milled for longer time up to 24 h to study the effect of milling on the magnetization; 1-h milled samples have the highest magnetizations. Structural characterization was carried out using X-ray diffraction and transmission electron microscopy. Identification of VO and Zni was carried out with Raman spectra, and energy-dispersive X-ray spectroscopy was used to verify that V did not diffuse into ZnO NPs as well to quantify O/Zn ratios.

  6. Detection of fluorescent nanoparticle interactions with primary immune cell subpopulations by flow cytometry.

    PubMed

    Gamucci, Olimpia; Bertero, Alice; Malvindi, Maria Ada; Sabella, Stefania; Pompa, Pier Paolo; Mazzolai, Barbara; Bardi, Giuseppe

    2014-01-01

    Engineered nanoparticles are endowed with very promising properties for therapeutic and diagnostic purposes. This work describes a fast and reliable method of analysis by flow cytometry to study nanoparticle interaction with immune cells. Primary immune cells can be easily purified from human or mouse tissues by antibody-mediated magnetic isolation. In the first instance, the different cell populations running in a flow cytometer can be distinguished by the forward-scattered light (FSC), which is proportional to cell size, and the side-scattered light (SSC), related to cell internal complexity. Furthermore, fluorescently labeled antibodies against specific cell surface receptors permit the identification of several subpopulations within the same sample. Often, all these features vary when cells are boosted by external stimuli that change their physiological and morphological state. Here, 50 nm FITC-SiO2 nanoparticles are used as a model to identify the internalization of nanostructured materials in human blood immune cells. The cell fluorescence and side-scattered light increase after incubation with nanoparticles allowed us to define time and concentration dependence of nanoparticle-cell interaction. Moreover, such protocol can be extended to investigate Rhodamine-SiO2 nanoparticle interaction with primary microglia, the central nervous system resident immune cells, isolated from mutant mice that specifically express the Green Fluorescent Protein (GFP) in the monocyte/macrophage lineage. Finally, flow cytometry data related to nanoparticle internalization into the cells have been confirmed by confocal microscopy. PMID:24747480

  7. Detection of Fluorescent Nanoparticle Interactions with Primary Immune Cell Subpopulations by Flow Cytometry

    PubMed Central

    Gamucci, Olimpia; Bertero, Alice; Malvindi, Maria Ada; Sabella, Stefania; Pompa, Pier Paolo; Mazzolai, Barbara; Bardi, Giuseppe

    2014-01-01

    Engineered nanoparticles are endowed with very promising properties for therapeutic and diagnostic purposes. This work describes a fast and reliable method of analysis by flow cytometry to study nanoparticle interaction with immune cells. Primary immune cells can be easily purified from human or mouse tissues by antibody-mediated magnetic isolation. In the first instance, the different cell populations running in a flow cytometer can be distinguished by the forward-scattered light (FSC), which is proportional to cell size, and the side-scattered light (SSC), related to cell internal complexity. Furthermore, fluorescently labeled antibodies against specific cell surface receptors permit the identification of several subpopulations within the same sample. Often, all these features vary when cells are boosted by external stimuli that change their physiological and morphological state. Here, 50 nm FITC-SiO2 nanoparticles are used as a model to identify the internalization of nanostructured materials in human blood immune cells. The cell fluorescence and side-scattered light increase after incubation with nanoparticles allowed us to define time and concentration dependence of nanoparticle-cell interaction. Moreover, such protocol can be extended to investigate Rhodamine-SiO2 nanoparticle interaction with primary microglia, the central nervous system resident immune cells, isolated from mutant mice that specifically express the Green Fluorescent Protein (GFP) in the monocyte/macrophage lineage. Finally, flow cytometry data related to nanoparticle internalization into the cells have been confirmed by confocal microscopy. PMID:24747480

  8. Scaffold-independent Patterning of Cells using Magnetic Nanoparticles

    NASA Astrophysics Data System (ADS)

    Ghosh, Suvojit; Biswas, Moanaro; Elankumaran, Subbiah; Puri, Ishwar

    2013-03-01

    Spatial patterning of cells in vitro relies on direct contact of cells on to solid surfaces. Scaffold independent patterning of cells has never been achieved so far. Patterning of cells has wide applications including stem cell biology, tissue architecture and regenerative medicine besides fundamental biology. Magnetized cells in a suspension can be manipulated using an externally applied magnetic field enabling directed patterning. We magnetized mammalian cells by internalization of superparamagnetic nanoparticles coated with bovine serum albumin (BSA). A magnetic field is then used to arrange cells in a desired pattern on a substrate or in suspension. The control strategy is derived from the self-assembly of magnetic colloids in a liquid considering magnetostatic interactions. The range of achievable structural features promise novel experimental methods investigating the influence of tissue shape and size on cell population dynamics wherein Fickian diffusion of autocrine growth signals are known to play a significant role. By eliminating the need for a scaffold, intercellular adhesion mechanics and the effects of temporally regulated signals can be investigated. The findings can be applied to novel tissue engineering methods.

  9. Cardenolide glycosides from the seeds of Digitalis purpurea exhibit carcinoma-specific cytotoxicity toward renal adenocarcinoma and hepatocellular carcinoma cells.

    PubMed

    Fujino, Tomofumi; Kuroda, Minpei; Matsuo, Yukiko; Kubo, Satoshi; Tamura, Chikako; Sakamoto, Nami; Mimaki, Yoshihiro; Hayakawa, Makio

    2015-02-01

    Four cardenolide glycosides, glucodigifucoside (2), 3'-O-acetylglucoevatromonoside (9), digitoxigenin 3-O-?-D-glucopyranosyl-(1?4)-?-D-glucopyranosyl-(1?4)-3-O-acetyl-?-D-digitoxopyranoside (11), and purpureaglycoside A (12), isolated from the seeds of Digitalis purpurea, exhibited potent cytotoxicity against human renal adenocarcinoma cell line ACHN. These compounds exhibited significantly lower IC50 values against ACHN than that against normal human renal proximal tubule-derived cell line HK-2. In particular, 2 exhibited the most potent and carcinoma-specific cytotoxicity, with a sixfold lower IC50 value against ACHN than that against HK-2. Measurement of cyclin-dependent kinase inhibitor levels revealed that upregulation of p21/Cip1 expression was involved in the carcinoma-specific cytotoxicity of 2. Further, compound 2 also exhibited the carcinoma-specific cytotoxicity toward hepatocellular carcinoma cell line. PMID:25345317

  10. Copper nanoparticle incorporated plasmonic organic bulk-heterojunction solar cells

    NASA Astrophysics Data System (ADS)

    Liu, Zhihai; Lee, Seung Yong; Lee, Eun-Cheol

    2014-12-01

    By embedding copper nanoparticles into poly(3,4-thylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) layers, the power conversion efficiency of organic bulk-heterojunction solar cell using poly(3-hexylthiophene) (P3HT) was increased from 3.58% to 3.96%, and that of the device based on poly[[4,8-bis[(2-ethylhexyl)oxy]benzo[1,2-b:4,5-b']dithiophene-2,6-diyl][3-fluoro-2-[(2-ethylhexyl)carbonyl]thieno[3,4-b]thiophenediyl

  11. Manganese-impregnated mesoporous silica nanoparticles for signal enhancement in MRI cell labelling studies.

    PubMed

    Guillet-Nicolas, Rémy; Laprise-Pelletier, Myriam; Nair, Mahesh M; Chevallier, Pascale; Lagueux, Jean; Gossuin, Yves; Laurent, Sophie; Kleitz, Freddy; Fortin, Marc-André

    2013-12-01

    Mesoporous silica nanoparticles (MSNs) are used in drug delivery and cell tracking applications. As Mn(2+) is already implemented as a "positive" cell contrast agent in preclinical imaging procedures (in the form of MnCl2 for neurological studies), the introduction of Mn in the porous network of MSNs would allow labelling cells and tracking them using MRI. These particles are in general internalized in endosomes, an acidic environment with high saline concentration. In addition, the available MSN porosity could also serve as a carrier to deliver medical/therapeutic substances through the labelled cells. In the present study, manganese oxide was introduced in the porous network of MCM-48 silica nanoparticles (Mn-M48SNs). The particles exhibit a narrow size distribution (~140 nm diam.) and high porosity (~60% vol.), which was validated after insertion of Mn. The resulting Mn-M48SNs were characterized by TEM, N2 physisorption, and XRD. Evidence was found with H2-TPR, and XPS characterization, that Mn(II) is the main oxidation state of the paramagnetic species after suspension in water, most probably in the form of Mn-OOH. The colloidal stability as a function of time was confirmed by DLS in water, acetate buffer and cell culture medium. In NMR data, no significant evidence of Mn(2+) leaching was found in Mn-M48SNs in acidic water (pH 6), up to 96 hours after suspension. High longitudinal relaxivity values of r1 = 8.4 mM(-1) s(-1) were measured at 60 MHz and 37 °C, with the lowest relaxometric ratios (r2/r1 = 2) reported to date for a Mn-MSN system. Leukaemia cells (P388) were labelled with Mn-M48SNs and nanoparticle cell internalization was confirmed by TEM. Finally, MRI contrast enhancement provided by cell labelling with escalated incubation concentrations of Mn-M48SNs was quantified at 1 T. This study confirmed the possibility of efficiently confining Mn into M48SNs using incipient wetness, while maintaining an open porosity and relatively high pore volume. Because these Mn-labelled M48SNs express strong "positive" contrast media properties at low concentrations, they are potentially applicable for cell tracking and drug delivery methodologies. PMID:24178890

  12. Comparison of intracellular accumulation and cytotoxicity of free mTHPC and mTHPC-loaded PLGA nanoparticles in human colon carcinoma cells

    NASA Astrophysics Data System (ADS)

    Löw, Karin; Knobloch, Thomas; Wagner, Sylvia; Wiehe, Arno; Engel, Andrea; Langer, Klaus; von Briesen, Hagen

    2011-06-01

    The second generation photosensitizer mTHPC was approved by the European Medicines Agency (EMA) for the palliative treatment of advanced head and neck cancer in October 2001. It is known that mTHPC possesses a significant phototoxicity against a variety of human cancer cells in vitro but also exhibits dark toxicity and can cause adverse effects (especially skin photosensitization). Due to its poor water solubility, the administration of hydrophobic photosensitizer still presents several difficulties. To overcome the administration problems, the use of nanoparticles as drug carrier systems is much investigated. Nanoparticles based on poly(lactic-co-glycolic acid) (PLGA) have been extensively studied as delivery systems into tumours due to their biocompatibility and biodegradability. The goal of this study was the comparison of free mTHPC and mTHPC-loaded PLGA nanoparticles concerning cytotoxicity and intracellular accumulation in human colon carcinoma cells (HT29). The nanoparticles delivered the photosensitizer to the colon carcinoma cells and enabled drug release without losing its activity. The cytotoxicity assays showed a time- and concentration-dependent decrease in cell proliferation and viability after illumination. However, first and foremost mTHPC lost its dark toxic effects using the PLGA nanoparticles as a drug carrier system. Therefore, PLGA nanoparticles are a promising drug carrier system for the hydrophobic photosensitizer mTHPC.

  13. Comparison of intracellular accumulation and cytotoxicity of free mTHPC and mTHPC-loaded PLGA nanoparticles in human colon carcinoma cells.

    PubMed

    Löw, Karin; Knobloch, Thomas; Wagner, Sylvia; Wiehe, Arno; Engel, Andrea; Langer, Klaus; von Briesen, Hagen

    2011-06-17

    The second generation photosensitizer mTHPC was approved by the European Medicines Agency (EMA) for the palliative treatment of advanced head and neck cancer in October 2001. It is known that mTHPC possesses a significant phototoxicity against a variety of human cancer cells in vitro but also exhibits dark toxicity and can cause adverse effects (especially skin photosensitization). Due to its poor water solubility, the administration of hydrophobic photosensitizer still presents several difficulties. To overcome the administration problems, the use of nanoparticles as drug carrier systems is much investigated. Nanoparticles based on poly(lactic-co-glycolic acid) (PLGA) have been extensively studied as delivery systems into tumours due to their biocompatibility and biodegradability. The goal of this study was the comparison of free mTHPC and mTHPC-loaded PLGA nanoparticles concerning cytotoxicity and intracellular accumulation in human colon carcinoma cells (HT29). The nanoparticles delivered the photosensitizer to the colon carcinoma cells and enabled drug release without losing its activity. The cytotoxicity assays showed a time- and concentration-dependent decrease in cell proliferation and viability after illumination. However, first and foremost mTHPC lost its dark toxic effects using the PLGA nanoparticles as a drug carrier system. Therefore, PLGA nanoparticles are a promising drug carrier system for the hydrophobic photosensitizer mTHPC. PMID:21508461

  14. Casein-coated Iron Oxide Nanoparticles for High MRI Contrast Enhancement and Efficient Cell Targeting

    PubMed Central

    Huang, Jing; Wang, Liya; Lin, Run; Wang, Andrew Y.; Yang, Lily; Kuang, Min; Qian, Weiping; Mao, Hui

    2013-01-01

    Surface properties, as well as inherent physicochemical properties, of the engineered nanomaterials play important roles in their interactions with the biological systems, which eventually affect their efficiency in diagnostic and therapeutic applications. Here we report a new class MRI contrast agent based on milk casein protein coated iron oxide nanoparticles (CNIOs) with a core size of 15 nm and hydrodynamic diameter ~30 nm. These CNIOs exhibited excellent water-solubility, colloidal stability, and biocompatibility. Importantly, CNIOs exhibited prominent T2 enhancing capability with a transverse relaxivity r2 of 273 mM?1s?1 at 3 Tesla. The transverse relaxivity is ~2.5-fold higher than that of iron oxide nanoparticles with the same core but an amphiphilic polymer coating. CNIOs showed pH-responsive properties, formed loose and soluble aggregates near the pI (pH~4.0). The aggregates could be dissociated reversibly when the solution pH was adjusted away from the pI. The transverse relaxation property and MRI contrast enhancing effect of CNIOs remained unchanged in the pH range of 2.0 to 8.0. Further functionalization of CNIOs can be achieved via surface modification of the protein coating. Bio-affinitive ligands, such as a single chain fragment from the antibody of epidermal growth factor receptor (ScFvEGFR), could be readily conjugated onto the protein coating, enabling specific targeting to MDA-MB-231 breast cancer cells over-expressing EGFR. T2-weighted MRI of mice intravenously administered with CNIOs demonstrated strong contrast enhancement in the liver and spleen. These favorable properties suggest CNIOs as a class of biomarker targeted magnetic nanoparticles for MRI contrast enhancement and related biomedical applications. PMID:23633522

  15. Laser nanothermolysis of human leukemia cells using functionalized plasmonic nanoparticles

    PubMed Central

    Liopo, Anton V.; Conjusteau, André; Konopleva, Marina; Andreeff, Michael; Oraevsky, Alexander A.

    2012-01-01

    In the present work, we present the use of gold nanorods as plasmonic nanoparticles for selective photothermal therapy of human acute (HL-60) and chronicle (K-562) leukemia cells using a near-infrared laser. We improved a published methodology of gold nanorods conjugation to generate high yields of narrow band gold nanorods with an optical absorption centered at 760 nm. The manufactured nanorods were pegylated and conjugated with monoclonal antibody to become non-toxic as biocompatible nanothermolysis agent. Gold nanorods are synthesized and conjugated to CD33 monoclonal antibody. After pegylation, or conjugation with CD33 antibody, gold nanorods were non-toxic to acute and chronic leukemia cells. Our modified gold nanorods CD33 conjugates shown high level of accumulation for both leukemia cell lines, and successful used for nanothermolysis of human leukemia cells in vitro. Each sample was illuminated with 1 or 3 laser shots as for low and for high laser fluence. The radiation was provided by a Quanta Systems q-switched titanium sapphire laser, and the system was designed for maximum sample coverage using non-focused illumination. HL-60 and K-562 cells were treated for 45 min with gold nanorods CD33 conjugated, or with pegylated gold nanorods. The effect of pulsed-laser nanothermolysis for acute and chronic leukemia cells were investigated with cell counting for number of living cells, percentage of cell death and functional parameters such as damage of cell membrane and metabolic activity. Gold nanorods CD33 conjugates significantly increase cell damage for low fluence laser and completely destroyed cancer cells after 3 pulses for low fluence (acute leukemia) and for high fluence laser as for HL-60 (acute) and for K-562 (chronicle) leukemia cells. PMID:22720194

  16. Laser nanothermolysis of human leukemia cells using functionalized plasmonic nanoparticles.

    PubMed

    Liopo, Anton V; Conjusteau, André; Konopleva, Marina; Andreeff, Michael; Oraevsky, Alexander A

    2012-01-01

    In the present work, we present the use of gold nanorods as plasmonic nanoparticles for selective photothermal therapy of human acute (HL-60) and chronicle (K-562) leukemia cells using a near-infrared laser. We improved a published methodology of gold nanorods conjugation to generate high yields of narrow band gold nanorods with an optical absorption centered at 760 nm. The manufactured nanorods were pegylated and conjugated with monoclonal antibody to become non-toxic as biocompatible nanothermolysis agent. Gold nanorods are synthesized and conjugated to CD33 monoclonal antibody. After pegylation, or conjugation with CD33 antibody, gold nanorods were non-toxic to acute and chronic leukemia cells. Our modified gold nanorods CD33 conjugates shown high level of accumulation for both leukemia cell lines, and successful used for nanothermolysis of human leukemia cells in vitro. Each sample was illuminated with 1 or 3 laser shots as for low and for high laser fluence. The radiation was provided by a Quanta Systems q-switched titanium sapphire laser, and the system was designed for maximum sample coverage using non-focused illumination. HL-60 and K-562 cells were treated for 45 min with gold nanorods CD33 conjugated, or with pegylated gold nanorods. The effect of pulsed-laser nanothermolysis for acute and chronic leukemia cells were investigated with cell counting for number of living cells, percentage of cell death and functional parameters such as damage of cell membrane and metabolic activity. Gold nanorods CD33 conjugates significantly increase cell damage for low fluence laser and completely destroyed cancer cells after 3 pulses for low fluence (acute leukemia) and for high fluence laser as for HL-60 (acute) and for K-562 (chronicle) leukemia cells. PMID:22720194

  17. Morphological effect of oscillating magnetic nanoparticles in killing tumor cells

    NASA Astrophysics Data System (ADS)

    Cheng, Dengfeng; Li, Xiao; Zhang, Guoxin; Shi, Hongcheng

    2014-04-01

    Forced oscillation of spherical and rod-shaped iron oxide magnetic nanoparticles (MNPs) via low-power and low-frequency alternating magnetic field (AMF) was firstly used to kill cancer cells in vitro. After being loaded by human cervical cancer cells line (HeLa) and then exposed to a 35-kHz AMF, MNPs mechanically damaged cell membranes and cytoplasm, decreasing the cell viability. It was found that the concentration and morphology of the MNPs significantly influenced the cell-killing efficiency of oscillating MNPs. In this preliminary study, when HeLa cells were pre-incubated with 100 ?g/mL rod-shaped MNPs (rMNP, length of 200 ± 50 nm and diameter of 50 to 120 nm) for 20 h, MTT assay proved that the cell viability decreased by 30.9% after being exposed to AMF for 2 h, while the cell viability decreased by 11.7% if spherical MNPs (sMNP, diameter of 200 ± 50 nm) were used for investigation. Furthermore, the morphological effect of MNPs on cell viability was confirmed by trypan blue assay: 39.5% rMNP-loaded cells and 15.1% sMNP-loaded cells were stained after being exposed to AMF for 2 h. It was also interesting to find that killing tumor cells at either higher (500 ?g/mL) or lower (20 ?g/mL) concentration of MNPs was less efficient than that achieved at 100 ?g/mL concentration. In conclusion, the relatively asymmetric morphological rod-shaped MNPs can kill cancer cells more effectively than spherical MNPs when being exposed to AMF by virtue of their mechanical oscillations.

  18. Coating Solid Lipid Nanoparticles with Hyaluronic Acid Enhances Antitumor Activity against Melanoma Stem-like Cells

    PubMed Central

    Shen, Hongxin; Shi, Sanjun; Zhang, Zhirong; Gong, Tao; Sun, Xun

    2015-01-01

    Successful anticancer chemotherapy requires targeting tumors efficiently and further potential to eliminate cancer stem cell (CSC) subpopulations. Since CD44 is present on many types of CSCs, and it binds specially to hyaluronic acid (HA), we tested whether coating solid lipid nanoparticles with hyaluronan (HA-SLNs)would allow targeted delivery of paclitaxel (PTX) to CD44-overexpressing B16F10 melanoma cells. First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44+) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44-) did not. This phenotype included sphere and colony formation, higher proportion of side population cells, expression of CSC-related markers (ALDH, CD133, Oct-4) and tumorigenicity in vivo. Next we showed that administering PTX-loaded HA-SLNs led to efficient intracellular delivery of PTX and induced substantial apoptosis in CD44+ cells in vitro. In the B16F10-CD44+ lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events. These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

  19. IsomiR expression profiles in human lymphoblastoid cell lines exhibit population and gender dependencies.

    PubMed

    Loher, Phillipe; Londin, Eric R; Rigoutsos, Isidore

    2014-09-30

    For many years it was believed that each mature microRNA (miRNA) existed as a single entity with fixed endpoints and a 'static' and unchangeable primary sequence. However, recent evidence suggests that mature miRNAs are more 'dynamic' and that each miRNA precursor arm gives rise to multiple isoforms, the isomiRs. Here we report on our identification of numerous and abundant isomiRs in the lymphoblastoid cell lines (LCLs) of 452 men and women from five different population groups. Unexpectedly, we find that these isomiRs exhibit an expression profile that is population-dependent and gender-dependent. This is important as it indicates that the LCLs of each gender/population combination have their own unique collection of mature miRNA transcripts. Moreover, each identified isomiR has its own characteristic abundance that remains consistent across biological replicates indicating that these are not degradation products. The primary sequences of identified isomiRs differ from the known miRBase miRNA either at their 5´-endpoint (leads to a different 'seed' sequence and suggests a different targetome), their 3´-endpoint, or both simultaneously. Our analysis of Argonaute PAR-CLIP data from LCLs supports the association of many of these newly identified isomiRs with the Argonaute silencing complex and thus their functional roles through participation in the RNA interference pathway. PMID:25229428

  20. IsomiR expression profiles in human lymphoblastoid cell lines exhibit population and gender dependencies

    PubMed Central

    Loher, Phillipe; Londin, Eric R.; Rigoutsos, Isidore

    2014-01-01

    For many years it was believed that each mature microRNA (miRNA) existed as a single entity with fixed endpoints and a ‘static’ and unchangeable primary sequence. However, recent evidence suggests that mature miRNAs are more ‘dynamic’ and that each miRNA precursor arm gives rise to multiple isoforms, the isomiRs. Here we report on our identification of numerous and abundant isomiRs in the lymphoblastoid cell lines (LCLs) of 452 men and women from five different population groups. Unexpectedly, we find that these isomiRs exhibit an expression profile that is population-dependent and gender-dependent. This is important as it indicates that the LCLs of each gender/population combination have their own unique collection of mature miRNA transcripts. Moreover, each identified isomiR has its own characteristic abundance that remains consistent across biological replicates indicating that these are not degradation products. The primary sequences of identified isomiRs differ from the known miRBase miRNA either at their 5´-endpoint (leads to a different ‘seed’ sequence and suggests a different targetome), their 3´-endpoint, or both simultaneously. Our analysis of Argonaute PAR-CLIP data from LCLs supports the association of many of these newly identified isomiRs with the Argonaute silencing complex and thus their functional roles through participation in the RNA interference pathway. PMID:25229428

  1. Nup50 is required for cell differentiation and exhibits transcription-dependent dynamics

    PubMed Central

    Buchwalter, Abigail L.; Liang, Yun; Hetzer, Martin W.

    2014-01-01

    The nuclear pore complex (NPC) plays a critical role in gene expression by mediating import of transcription regulators into the nucleus and export of RNA transcripts to the cytoplasm. Emerging evidence suggests that in addition to mediating transport, a subset of nucleoporins (Nups) engage in transcriptional activation and elongation at genomic loci that are not associated with NPCs. The underlying mechanism and regulation of Nup mobility on and off nuclear pores remain unclear. Here we show that Nup50 is a mobile Nup with a pronounced presence both at the NPC and in the nucleoplasm that can move between these different localizations. Strikingly, the dynamic behavior of Nup50 in both locations is dependent on active transcription by RNA polymerase II and requires the N-terminal half of the protein, which contains importin ?– and Nup153-binding domains. However, Nup50 dynamics are independent of importin ?, Nup153, and Nup98, even though the latter two proteins also exhibit transcription-dependent mobility. Of interest, depletion of Nup50 from C2C12 myoblasts does not affect cell proliferation but inhibits differentiation into myotubes. Taken together, our results suggest a transport-independent role for Nup50 in chromatin biology that occurs away from the NPC. PMID:24943837

  2. Quantum dots incorporated magnetic nanoparticles for imaging colon carcinoma cells

    PubMed Central

    2013-01-01

    Background Engineered multifunctional nanoparticles (NPs) have made a tremendous impact on the biomedical sciences, with advances in imaging, sensing and bioseparation. In particular, the combination of optical and magnetic responses through a single particle system allows us to serve as novel multimodal molecular imaging contrast agents in clinical settings. Despite of essential medical imaging modalities and of significant clinical application, only few nanocomposites have been developed with dual imaging contrast. A new method for preparing quantum dots (QDs) incorporated magnetic nanoparticles (MNPs) based on layer-by-layer (LbL) self-assembly techniques have developed and used for cancer cells imaging. Methods Here, citrate - capped negatively charged Fe3O4 NPs were prepared and coated with positively - charged hexadecyltrimethyl ammonium bromide (CTAB). Then, thiol - capped negatively charged CdTe QDs were electrostatically bound with CTAB. Morphological, optical and magnetic properties of the fluorescent magnetic nanoparticles (FMNPs) were characterized. Prepared FMNPs were additionally conjugated with hCC49 antibodies fragment antigen binding (Fab) having binding affinity to sialylated sugar chain of TAG-72 region of LS174T cancer cells, which was prepared silkworm expression system, and then were used for imaging colon carcinoma cells. Results The prepared nanocomposites were magnetically responsive and fluorescent, simultaneously that are useful for efficient cellular imaging, optical sensing and magnetic separation. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) revealed that the particle size is around 50 nm in diameter with inner magnetic core and outer CdTe QDs core-shell structure. Cytotoxicity test of prepared FMNPs indicates high viability in Vero cells. NPs conjugated with anti cancer antibodies were successfully labeled on colon carcinoma cells (LS174) in vitro and showed significant specificity to target cells. Conclusion The present report demonstrates a simple synthesis of CdTe QDs-Fe3O4 NPs. The surface of the prepared FMNPs was enabled simple conjugation to monoclonal antibodies by electrostatic interaction. This property further extended their in vitro applications as cellular imaging contrast agents. Such labeling of cells with new fluorescent-magneto nanoprobes for living detection is of interest to various biomedical applications and has demonstrated the potential for future medical use. PMID:23957878

  3. Cell death pathway induced by resveratrol-bovine serum albumin nanoparticles in a human ovarian cell line

    PubMed Central

    GUO, LIYUAN; PENG, YAN; LI, YULIAN; YAO, JINGPING; ZHANG, GUANGMEI; CHEN, JIE; WANG, JING; SUI, LIHUA

    2015-01-01

    Resveratrol-bovine serum albumin nanoparticles (RES-BSANP) exhibit chemotherapeutic properties, which trigger apoptosis. The aim of the present study was to investigate the caspase-independent cell death pathway induced by RES-BSANP in human ovarian cancer SKOV3 cells and to analyze its mechanism. Morphological changes were observed by apoptotic body/cell nucleus DNA staining using inverted and fluorescence microscopy. The cell death pathway was determined by phosphatidylserine translocation. Western blot analysis was conducted to detect the activation of apoptosis-inducing factor (AIF), cytochrome c (Cyto c) and B-cell lymphoma 2-associated X protein (Bax). Apoptotic body and nuclear condensation and fragmentation were observed simultaneously following treatment with RES-BSANP. RES-BSANP induced apoptosis in a dose-dependent manner in the human ovarian cancer SKOV3 cells. The translocation of AIF from the mitochondria to the cytoplasm occurred earlier than that of Cyto c. In addition, Bax binding to the mitochondria was required for the release of AIF and Cyto c from the mitochondria. The AIF apoptosis pathway may present an alternative caspase-dependent apoptosis pathway in human ovarian cell death induced by RES-BSANP. Elucidation of this pathway may be critical for the treatment of cancer using high doses of RES-BSANP. PMID:25663913

  4. Modulating Gold Nanoparticle in vivo Delivery for Photothermal Therapy Applications Using a T Cell Delivery System

    NASA Astrophysics Data System (ADS)

    Kennedy, Laura Carpin

    This thesis reports new gold nanoparticle-based methods to treat chemotherapy-resistant and metastatic tumors that frequently evade conventional cancer therapies. Gold nanoparticles represent an innovative generation of diagnostic and treatment agents due to the ease with which they can be tuned to scatter or absorb a chosen wavelength of light. One area of intensive investigation in recent years is gold nanoparticle photothermal therapy (PTT), in which gold nanoparticles are used to heat and destroy cancer. This work demonstrates the utility of gold nanoparticle PTT against two categories of cancer that are currently a clinical challenge: trastuzumab-resistant breast cancer and metastatic cancer. In addition, this thesis presents a new method of gold nanoparticle delivery using T cells that increases gold nanoparticle tumor accumulation efficiency, a current challenge in the field of PTT. I ablated trastuzumab-resistant breast cancer in vitro for the first time using anti-HER2 labeled silica-gold nanoshells, demonstrating the potential utility of PTT against chemotherapy-resistant cancers. I next established for the first time the use of T cells as gold nanoparticle vehicles in vivo. When incubated with gold nanoparticles in culture, T cells can internalize up to 15000 nanoparticles per cell with no detrimental effects to T cell viability or function (e.g. migration and cytokine secretion). These AuNP-T cells can be systemically administered to tumor-bearing mice and deliver gold nanoparticles four times more efficiently than by injecting free nanoparticles. In addition, the biodistribution of AuNP-T cells correlates with the normal biodistribution of T cell carrier, suggesting the gold nanoparticle biodistribution can be modulated through the choice of nanoparticle vehicle. Finally, I apply gold nanoparticle PTT as an adjuvant treatment for T cell adoptive transfer immunotherapy (Hyperthermia-Enhanced Immunotherapy or HIT) of distant tumors in a melanoma mouse model. The results presented in this thesis expand the potential of gold nanoparticle PTT from only chemotherapy-sensitive or localized cancers to chemotherapy-resistant non-localized cancers that currently defy conventional therapies.

  5. Infrared light-absorbing gold/gold sulfide nanoparticles induce cell death in esophageal adenocarcinoma

    PubMed Central

    Li, Yan; Gobin, Andre M; Dryden, Gerald W; Kang, Xinqin; Xiao, Deyi; Li, Su Ping; Zhang, Guandong; Martin, Robert CG

    2013-01-01

    Gold nanoparticles and near infrared-absorbing light are each innocuous to tissue but when combined can destroy malignant tissue while leaving healthy tissue unharmed. This study investigated the feasibility of photothermal ablation therapy for esophageal adenocarcinoma using chitosan-coated gold/gold sulfide (CS-GGS) nanoparticles. A rat esophagoduodenal anastomosis model was used for the in vivo ablation study, and three human esophageal cell lines were used to study the response of cancer cells and benign cells to near infrared light after treatment with CS-GGS. The results indicate that both cancerous tissue and cancer cells took up more gold nanoparticles and were completely ablated after exposure to near infrared light. The benign tissue and noncancerous cells showed less uptake of these nanoparticles, and remained viable after exposure to near infrared light. CS-GGS nanoparticles could provide an optimal endoluminal therapeutic option for near infrared light ablation of esophageal cancer. PMID:23818775

  6. Delivery of a transforming growth factor ?-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles

    PubMed Central

    Deng, Wen Wen; Cao, Xia; Wang, Miao; Qu, Rui; Su, Wei Yan; Yang, Yan; Wei, Ya Wei; Xu, Xi Ming; Yu, Jiang Nan

    2012-01-01

    The objective of this study was to investigate the use of cationized Pleurotus eryngii polysaccharide (CPEPS) as a nonviral gene delivery vehicle to transfer plasmid DNA encoding transforming growth factor beta-1 (pTGF-?1) into mesenchymal stem cells (MSCs) in vitro. Crude P. eryngii polysaccharide was purified, and then cationized by grafting spermine onto the backbone of the polysaccharide. Agarose gel electrophoresis, transmission electron microscopy, and a Nano Sense Zetasizer (Malvern Instruments, Malvern, UK) were used to characterize the CPEPS-pTGF-?1 nanoparticles. The findings of cytotoxicity analysis showed that when the nanoparticles were formulated with a CPEPS/pTGF-?1 weight ratio ? 10:1, a greater gel retardation effect was observed during agarose gel electrophoresis. The CPEPS-pTGF-?1 nanoparticles with a weight ratio of 20:1, respectively, possessed an average particle size of 80.8 nm in diameter and a zeta potential of +17.4 ± 0.1 mV. Significantly, these CPEPS-pTGF-?1 nanoparticles showed lower cytotoxicity and higher transfection efficiency than both polyethylenimine (25 kDa) (P = 0.006, Student’s t-test) and LipofectamineTM 2000 (P = 0.002, Student’s t-test). Additionally, the messenger RNA expression level of TGF-?1 in MSCs transfected with CPEPS-pTGF-?1 nanoparticles was significantly higher than that of free plasmid DNA-transfected MSCs and slightly elevated compared with that of Lipofectamine 2000-transfected MSCs. Flow cytometry analysis demonstrated that 92.38% of MSCs were arrested in the G1 phase after being transfected with CPEPS-pTGF-?1 nanoparticles, indicating a tendency toward differentiation. In summary, the findings of this study suggest that the CPEPS-pTGF-?1 nanoparticles prepared in this work exhibited excellent transfection efficiency and low toxicity. Therefore, they could be developed into a promising nonviral vector for gene delivery in vitro. PMID:22457592

  7. The stoichiometry of single nanoparticles of copper zinc tin selenide.

    PubMed

    Haas, Wernfried; Rath, Thomas; Pein, Andreas; Rattenberger, Johannes; Trimmel, Gregor; Hofer, Ferdinand

    2011-02-21

    Cu(2)ZnSnSe(4) nanoparticles have high potential to be used as ink for printable solar cells. Using transmission electron microscopy we show that these nanoparticles exhibit a broad range of chemical heterogeneity. These results are contrary to the interpretation of previous experimental work and will have considerable impact on the development of these nanoparticles. PMID:21184006

  8. Cerium Oxide Nanoparticles Induce Oxidative Stress and Genotoxicity in Human Skin Melanoma Cells.

    PubMed

    Ali, Daoud; Alarifi, Saud; Alkahtani, Saad; AlKahtane, Abdullah A; Almalik, Abdulaziz

    2014-11-14

    Extensive applications of cerium oxide (CeO2) nanoparticles require a better understanding of their possible effects on human health. However, data demonstrating the effect of CeO2 nanoparticles on the human skin melanoma cell remain scanty. In the current study, we determined the mechanism through which CeO2 nanoparticles (APS <25 nm) induce toxicity in human skin melanoma cells (A375). The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and neutral red uptake assays showed concentration and time-dependent cytotoxicity of CeO2 nanoparticles in A375 cells. CeO2 nanoparticles significantly induced the generation reactive oxygen species (ROS) and malondialdehyde, superoxide dismutase, and decreased glutathione levels in A375 cells. It was also observed that the CeO2 nanoparticles induced chromosomal condensation and caspase-3 activity. CeO2 nanoparticles exposed cells revealed the formation of DNA double-strand breakage as measured by percent tail DNA and olive tail moment through comet assay. The decline of cell viability, production of ROS, and DNA damage in A375 cells specifies that CeO2 nanoparticles have less capable to induce cyto and genotoxicity. PMID:25395198

  9. Nanoparticle interactions with live cells: Quantitative fluorescence microscopy of nanoparticle size effects

    PubMed Central

    Shang, Li; Nienhaus, Karin; Jiang, Xiue; Yang, Linxiao; Landfester, Katharina; Mailänder, Volker; Simmet, Thomas

    2014-01-01

    Summary Engineered nanomaterials are known to enter human cells, often via active endocytosis. Mechanistic details of the interactions between nanoparticles (NPs) with cells are still not well enough understood. NP size is a key parameter that controls the endocytic mechanism and affects the cellular uptake yield. Therefore, we have systematically analyzed the cellular uptake of fluorescent NPs in the size range of 3.3–100 nm (diameter) by live cells. By using spinning disk confocal microscopy in combination with quantitative image analysis, we studied the time courses of NP association with the cell membrane and subsequent internalization. NPs with diameters of less than 10 nm were observed to accumulate at the plasma membrane before being internalized by the cells. In contrast, larger NPs (100 nm) were directly internalized without prior accumulation at the plasma membrane, regardless of their surface charges. We attribute this distinct size dependence to the requirement of a sufficiently strong local interaction of the NPs with the endocytic machinery in order to trigger the subsequent internalization. PMID:25551067

  10. Augmented cellular uptake of nanoparticles using tea catechins: effect of surface modification on nanoparticle-cell interaction

    NASA Astrophysics Data System (ADS)

    Lu, Yi-Ching; Luo, Pei-Chun; Huang, Chun-Wan; Leu, Yann-Lii; Wang, Tzu-Hao; Wei, Kuo-Chen; Wang, Hsin-Ell; Ma, Yunn-Hwa

    2014-08-01

    Nanoparticles may serve as carriers in targeted therapeutics; interaction of the nanoparticles with a biological system may determine their targeting effects and therapeutic efficacy. Epigallocatechin-3-gallate (EGCG), a major component of tea catechins, has been conjugated with nanoparticles and tested as an anticancer agent. We investigated whether EGCG may enhance nanoparticle uptake by tumor cells. Cellular uptake of a dextran-coated magnetic nanoparticle (MNP) was determined by confocal microscopy, flow cytometry or a potassium thiocyanate colorimetric method. We demonstrated that EGCG greatly enhanced interaction and/or internalization of MNPs (with or without polyethylene glycol) by glioma cells, but not vascular endothelial cells. The enhancing effects are both time- and concentration-dependent. Such effects may be induced by a simple mix of MNPs with EGCG at a concentration as low as 1-3 ?M, which increased MNP uptake 2- to 7-fold. In addition, application of magnetic force further potentiated MNP uptake, suggesting a synergetic effect of EGCG and magnetic force. Because the effects of EGCG were preserved at 4 °C, but not when EGCG was removed from the culture medium prior to addition of MNPs, a direct interaction of EGCG and MNPs was implicated. Use of an MNP-EGCG composite produced by adsorption of EGCG and magnetic separation also led to an enhanced uptake. The results reveal a novel interaction of a food component and nanocarrier system, which may be potentially amenable to magnetofection, cell labeling/tracing, and targeted therapeutics.

  11. Augmented cellular uptake of nanoparticles using tea catechins: effect of surface modification on nanoparticle-cell interaction.

    PubMed

    Lu, Yi-Ching; Luo, Pei-Chun; Huang, Chun-Wan; Leu, Yann-Lii; Wang, Tzu-Hao; Wei, Kuo-Chen; Wang, Hsin-Ell; Ma, Yunn-Hwa

    2014-09-01

    Nanoparticles may serve as carriers in targeted therapeutics; interaction of the nanoparticles with a biological system may determine their targeting effects and therapeutic efficacy. Epigallocatechin-3-gallate (EGCG), a major component of tea catechins, has been conjugated with nanoparticles and tested as an anticancer agent. We investigated whether EGCG may enhance nanoparticle uptake by tumor cells. Cellular uptake of a dextran-coated magnetic nanoparticle (MNP) was determined by confocal microscopy, flow cytometry or a potassium thiocyanate colorimetric method. We demonstrated that EGCG greatly enhanced interaction and/or internalization of MNPs (with or without polyethylene glycol) by glioma cells, but not vascular endothelial cells. The enhancing effects are both time- and concentration-dependent. Such effects may be induced by a simple mix of MNPs with EGCG at a concentration as low as 1-3 ?M, which increased MNP uptake 2- to 7-fold. In addition, application of magnetic force further potentiated MNP uptake, suggesting a synergetic effect of EGCG and magnetic force. Because the effects of EGCG were preserved at 4 °C, but not when EGCG was removed from the culture medium prior to addition of MNPs, a direct interaction of EGCG and MNPs was implicated. Use of an MNP-EGCG composite produced by adsorption of EGCG and magnetic separation also led to an enhanced uptake. The results reveal a novel interaction of a food component and nanocarrier system, which may be potentially amenable to magnetofection, cell labeling/tracing, and targeted therapeutics. PMID:25069428

  12. Targeting polymeric fluorescent nanodiamond-gold/silver multi-functional nanoparticles as a light-transforming hyperthermia reagent for cancer cells

    NASA Astrophysics Data System (ADS)

    Cheng, Liang-Chien; Chen, Hao Ming; Lai, Tsung-Ching; Chan, Yung-Chieh; Liu, Ru-Shi; Sung, James C.; Hsiao, Michael; Chen, Chung-Hsuan; Her, Li-Jane; Tsai, Din Ping

    2013-04-01

    This work demonstrates a simple route for synthesizing multi-functional fluorescent nanodiamond-gold/silver nanoparticles. The fluorescent nanodiamond is formed by the surface passivation of poly(ethylene glycol) bis(3-aminopropyl) terminated. Urchin-like gold/silver nanoparticles can be obtained via one-pot synthesis, and combined with each other via further thiolation of nanodiamond. The morphology of the nanodiamond-gold/silver nanoparticles thus formed was identified herein by high-resolution transmission electron microscopy, and clarified using diffraction patterns. Fourier transform infrared spectroscopy clearly revealed the surface functionalization of the nanoparticles. The fluorescence of the materials with high photo stability was examined by high power laser irradiation and long-term storage at room temperature. To develop the bio-recognition of fluorescent nanodiamond-gold/silver nanoparticles, pre-modified transferrin was conjugated with the gold/silver nanoparticles, and the specificity and activity were confirmed in vitro using human hepatoma cell line (J5). The cellular uptake analysis that was conducted using flow cytometry and inductively coupled plasma mass spectrometry exhibited that twice as many transferrin-modified nanoparticles as bare nanoparticles were engulfed, revealing the targeting and ease of internalization of the human hepatoma cell. Additionally, the in situ monitoring of photothermal therapeutic behavior reveals that the nanodiamond-gold/silver nanoparticles conjugated with transferrin was more therapeutic than the bare nanodiamond-gold/silver materials, even when exposed to a less energetic laser source. Ultimately, this multi-functional material has great potential for application in simple synthesis. It is non-cytotoxic, supports long-term tracing and can be used in highly efficient photothermal therapy against cancer cells.This work demonstrates a simple route for synthesizing multi-functional fluorescent nanodiamond-gold/silver nanoparticles. The fluorescent nanodiamond is formed by the surface passivation of poly(ethylene glycol) bis(3-aminopropyl) terminated. Urchin-like gold/silver nanoparticles can be obtained via one-pot synthesis, and combined with each other via further thiolation of nanodiamond. The morphology of the nanodiamond-gold/silver nanoparticles thus formed was identified herein by high-resolution transmission electron microscopy, and clarified using diffraction patterns. Fourier transform infrared spectroscopy clearly revealed the surface functionalization of the nanoparticles. The fluorescence of the materials with high photo stability was examined by high power laser irradiation and long-term storage at room temperature. To develop the bio-recognition of fluorescent nanodiamond-gold/silver nanoparticles, pre-modified transferrin was conjugated with the gold/silver nanoparticles, and the specificity and activity were confirmed in vitro using human hepatoma cell line (J5). The cellular uptake analysis that was conducted using flow cytometry and inductively coupled plasma mass spectrometry exhibited that twice as many transferrin-modified nanoparticles as bare nanoparticles were engulfed, revealing the targeting and ease of internalization of the human hepatoma cell. Additionally, the in situ monitoring of photothermal therapeutic behavior reveals that the nanodiamond-gold/silver nanoparticles conjugated with transferrin was more therapeutic than the bare nanodiamond-gold/silver materials, even when exposed to a less energetic laser source. Ultimately, this multi-functional material has great potential for application in simple synthesis. It is non-cytotoxic, supports long-term tracing and can be used in highly efficient photothermal therapy against cancer cells. Electronic supplementary information (ESI) available: The TEM images of ND and FND-Au/Ag. The photo-stability of FND which is exposures at 561 nm laser for 3 h and stores at room temperature for 1 month. See DOI: 10.1039/c3nr34091k

  13. Carbon nanoparticles for gene transfection in eukaryotic cell lines.

    PubMed

    Zanin, H; Hollanda, L M; Ceragioli, H J; Ferreira, M S; Machado, D; Lancellotti, M; Catharino, R R; Baranauskas, V; Lobo, A O

    2014-06-01

    For the first time, oxygen terminated cellulose carbon nanoparticles (CCN) was synthesised and applied in gene transfection of pIRES plasmid. The CCN was prepared from catalytic of polyaniline by chemical vapour deposition techniques. This plasmid contains one gene that encodes the green fluorescent protein (GFP) in eukaryotic cells, making them fluorescent. This new nanomaterial and pIRES plasmid formed ?-stacking when dispersed in water by magnetic stirring. The frequencies shift in zeta potential confirmed the plasmid strongly connects to the nanomaterial. In vitro tests found that this conjugation was phagocytised by NG97, NIH-3T3 and A549 cell lines making them fluorescent, which was visualised by fluorescent microscopy. Before the transfection test, we studied CCN in cell viability. Both MTT and Neutral Red uptake tests were carried out using NG97, NIH-3T3 and A549 cell lines. Further, we use metabolomics to verify if small amounts of nanomaterial would be enough to cause some cellular damage in NG97 cells. We showed two mechanisms of action by CCN-DNA complex, producing an exogenous protein by the transfected cell and metabolomic changes that contributed by better understanding of glioblastoma, being the major finding of this work. Our results suggested that this nanomaterial has great potential as a gene carrier agent in non-viral based therapy, with low cytotoxicity, good transfection efficiency, and low cell damage in small amounts of nanomaterials in metabolomic tests. PMID:24863237

  14. Chitosan-coated triangular silver nanoparticles as a novel class of biocompatible, highly effective photothermal transducers for in vitro cancer cell therapy.

    PubMed

    Boca, Sanda C; Potara, Monica; Gabudean, Ana-Maria; Juhem, Aurelie; Baldeck, Patrice L; Astilean, Simion

    2011-12-01

    One of the relevant directions that nanotechnology is taking nowadays is connected with nanomedicine and specifically related to the use of light and nanoparticles in early diagnosis and effective therapeutics of cancer. Noble-metal nanoparticles can act under laser irradiation as effective photothermal transducers for triggering localized hyperthermia of tumors. In this work we report the performance of newly synthesized chitosan-coated silver nanotriangles (Chit-AgNTs) with strong resonances in near-infrared (NIR) to operate as photothermal agents against a line of human non-small lung cancer cells (NCI-H460). The hyperthermia experiments were conducted by excitation of nanoparticles-loaded cells at 800 nm wavelength from a Ti:Sapphire laser. We found that the rate of cell mortality in the presence of Chit-AgNTs is higher than in the presence of thiolated poly(ethylene) glycol capped gold nanorods (PEG-AuNRs) - a common hyperthermia agent used as reference-, while no destructive effects were noticed on the control sample (cells without nanoparticles) under identical irradiation conditions. Additionally, we conducted cytotoxicity assays and found Chit-AgNTs to be efficiently uptaken by the cells while exhibiting good biocompatibility for healthy human embryonic cells (HEK), which is essential for any in vivo applications. Our results reveal a novel class of biocompatible plasmonic nanoparticles with high potential to be implemented as effective phototherapeutic agents in the battle against cancer. PMID:21840122

  15. Anticancer studies of the synthesized gold nanoparticles against MCF 7 breast cancer cell lines

    NASA Astrophysics Data System (ADS)

    Kamala Priya, M. R.; Iyer, Priya R.

    2014-09-01

    It has been previously stated that gold nanoparticles have been successfully synthesized using various green extracts of plants. The synthesized gold nanoparticles were characterized under scanning electron microscopy and EDX to identify the size of the nanoparticles. It was found that the nanoparticles were around 30 nm in size, which is a commendable nano dimension achieved through a plant mediated synthesis. The nanoparticles were further studied for their various applications. In the current study, we have made attempts to exploit the anticancer ability of the gold nano particles. The nanoparticles were studied against MCF 7 breast cancer cell lines. The results obtained from the studies of anticancer activity showed that gold nanoparticles gave an equivalent good results, in par with the standard drugs against cancer. The AuNP's proved to be efficient even from the minimum concentrations of 2 ?g/ml, and as the concentration increased the anticancer efficacy as well increased.

  16. Anticancer studies of the synthesized gold nanoparticles against MCF 7 breast cancer cell lines

    NASA Astrophysics Data System (ADS)

    Kamala Priya, M. R.; Iyer, Priya R.

    2015-04-01

    It has been previously stated that gold nanoparticles have been successfully synthesized using various green extracts of plants. The synthesized gold nanoparticles were characterized under scanning electron microscopy and EDX to identify the size of the nanoparticles. It was found that the nanoparticles were around 30 nm in size, which is a commendable nano dimension achieved through a plant mediated synthesis. The nanoparticles were further studied for their various applications. In the current study, we have made attempts to exploit the anticancer ability of the gold nano particles. The nanoparticles were studied against MCF 7 breast cancer cell lines. The results obtained from the studies of anticancer activity showed that gold nanoparticles gave an equivalent good results, in par with the standard drugs against cancer. The AuNP's proved to be efficient even from the minimum concentrations of 2 ?g/ml, and as the concentration increased the anticancer efficacy as well increased.

  17. Toxicological mode of action of ZnO nanoparticles: Impact on immune cells.

    PubMed

    Roy, Ruchi; Das, Mukul; Dwivedi, Premendra D

    2015-02-01

    The use of nanoscale materials is growing exponentially as concerns rise about the human hazards to it. It is assumed that living beings are coevolved with nanoparticles ever since the origin of life on earth and therefore, they must have developed the defense and toxicity mitigating mechanisms for nanoparticles. Although having peculiar properties these new materials also present new health risks upon interacting with biological systems. Zinc oxide is the most widely used nanoparticles among various nanomaterials. Recently, these nanoparticles have been shown to specifically kill cancerous cells; therefore, it is believed that these nanoparticles may be used as an alternative anti-tumor agent. However, it is also known that these nanoparticles pose several deleterious effects to living beings. It is therefore critical to understand the nature and origin of the toxicity imposed by these nanomaterials. Keeping these points in mind the present review provides updated information on various aspects of toxicities induced by these engineered nanoparticles. PMID:25193324

  18. Bridged polysilsesquioxanes: Hybrid organic-inorganic materials as fuel cell polyelectrolyte membranes and functional nanoparticles

    NASA Astrophysics Data System (ADS)

    Khiterer, Mariya

    2007-05-01

    This dissertation describes the design, fabrication, and characterization of organic-inorganic hybrid materials. Several classes of bridged polysilsesquioxanes are presented. The first class is a membrane material suitable for fuel cell technology as a proton conducting polyelectrolyte. The second class includes hybrid nanoparticles for display device applications and chromatographic media. Chapter 1 is an introduction to hybrid organic-inorganic materials. Sol-gel chemistry is discussed, followed by a survey of prominent examples of silica hybrids. Examples of physical organic-silica blends and covalent organo-silicas, including ORMOCERSRTM, polyhedral oligomeric silsesquioxanes, and bridged polysilsesquioxanes are discussed. Bridged polysilsesquioxanes are described in great detail. Monomer synthesis, sol-gel chemistry, processing, characterization, and physical properties are included. Chapter 2 describes the design of polyelectrolyte bridged polysilsesquioxane membranes. The materials contain covalently bound sulfonic acid groups originating from the corresponding disulfides. These organic-inorganic hybrid materials integrate a network supporting component which is systematically changed to fine-tune their physical properties. The membranes are characterized as PEM fuel cell electrolytes, where proton conductivities of 4-6 mS cm-1 were measured. In Chapter 3 techniques for the preparation of bridged polysilsesquioxane nanoparticles are described. An inverse water-in-oil microemulsion polymerization method is developed to prepare cationic nanoparticles, including viologen-bridged materials with applications in electrochromic display devices. An aqueous ammonia system is used to prepare neutral nanoparticles containing hydrocarbon bridging groups, which have potential applications as chromatographic media. Chapter 4 describes electrochromic devices developed in collaboration with the Heflin group of Virginia Tech, which incorporate viologen bridged nanoparticles described in Chapter 3. The devices are prepared via the layer-by-layer deposition technique and characterized by voltammetry and transmission spectroscopy. Contrast ratios between yellow and violet states were 45-50% with switching times of 3-3.5 seconds. Finally, Appendix I describes the resolution of racemic 3,3.3',3'-Tetramethyl-1,1"-spirobisindane-5,5',6,6'-tetrol by diastereomeric complex formation with (8S,9R)-(-)-N-benzylcinchonidinium chloride. Enantiomerically pure bisspirocatechol is used to prepare a chiral polymer, which exhibits differences in solid state packing from polymer made with the racemic monomer. Preliminary results on the use of the chiral polymer in enantioselective membrane separations technology are described.

  19. Targeting polymeric fluorescent nanodiamond-gold/silver multi-functional nanoparticles as a light-transforming hyperthermia reagent for cancer cells.

    PubMed

    Cheng, Liang-Chien; Chen, Hao Ming; Lai, Tsung-Ching; Chan, Yung-Chieh; Liu, Ru-Shi; Sung, James C; Hsiao, Michael; Chen, Chung-Hsuan; Her, Li-Jane; Tsai, Din Ping

    2013-05-01

    This work demonstrates a simple route for synthesizing multi-functional fluorescent nanodiamond-gold/silver nanoparticles. The fluorescent nanodiamond is formed by the surface passivation of poly(ethylene glycol) bis(3-aminopropyl) terminated. Urchin-like gold/silver nanoparticles can be obtained via one-pot synthesis, and combined with each other via further thiolation of nanodiamond. The morphology of the nanodiamond-gold/silver nanoparticles thus formed was identified herein by high-resolution transmission electron microscopy, and clarified using diffraction patterns. Fourier transform infrared spectroscopy clearly revealed the surface functionalization of the nanoparticles. The fluorescence of the materials with high photo stability was examined by high power laser irradiation and long-term storage at room temperature. To develop the bio-recognition of fluorescent nanodiamond-gold/silver nanoparticles, pre-modified transferrin was conjugated with the gold/silver nanoparticles, and the specificity and activity were confirmed in vitro using human hepatoma cell line (J5). The cellular uptake analysis that was conducted using flow cytometry and inductively coupled plasma mass spectrometry exhibited that twice as many transferrin-modified nanoparticles as bare nanoparticles were engulfed, revealing the targeting and ease of internalization of the human hepatoma cell. Additionally, the in situ monitoring of photothermal therapeutic behavior reveals that the nanodiamond-gold/silver nanoparticles conjugated with transferrin was more therapeutic than the bare nanodiamond-gold/silver materials, even when exposed to a less energetic laser source. Ultimately, this multi-functional material has great potential for application in simple synthesis. It is non-cytotoxic, supports long-term tracing and can be used in highly efficient photothermal therapy against cancer cells. PMID:23536050

  20. Use of titanium dioxide nanoparticles biosynthesized by Bacillus mycoides in quantum dot sensitized solar cells

    PubMed Central

    2014-01-01

    Background One of the major challenges of nanotechnology during the last decade has been the development of new procedures to synthesize nanoparticles. In this context, biosynthetic methods have taken hold since they are simple, safe and eco-friendly. Results In this study, we report the biosynthesis of TiO2 nanoparticles by an environmental isolate of Bacillus mycoides, a poorly described Gram-positive bacterium able to form colonies with novel morphologies. This isolate was able to produce TiO2 nanoparticles at 37°C in the presence of titanyl hydroxide. Biosynthesized nanoparticles have anatase polymorphic structure, spherical morphology, polydisperse size (40–60 nm) and an organic shell as determined by UV–vis spectroscopy, TEM, DLS and FTIR, respectively. Also, conversely to chemically produced nanoparticles, biosynthesized TiO2 do not display phototoxicity. In order to design less expensive and greener solar cells, biosynthesized nanoparticles were evaluated in Quantum Dot Sensitized Solar Cells (QDSSCs) and compared with chemically produced TiO2 nanoparticles. Solar cell parameters such as short circuit current density (ISC) and open circuit voltage (VOC) revealed that biosynthesized TiO2 nanoparticles can mobilize electrons in QDSSCs similarly than chemically produced TiO2. Conclusions Our results indicate that bacterial extracellular production of TiO2 nanoparticles at low temperatures represents a novel alternative for the construction of green solar cells. PMID:25027643

  1. Slow intracellular trafficking of catalase nanoparticles targeted to ICAM-1 protects endothelial cells from oxidative stress.

    PubMed

    Muro, Silvia; Cui, Xiumin; Gajewski, Christine; Murciano, Juan-Carlos; Muzykantov, Vladimir R; Koval, Michael

    2003-11-01

    Nanotechnologies promise new means for drug delivery. ICAM-1 is a good target for vascular immunotargeting of nanoparticles to the perturbed endothelium, although endothelial cells do not internalize monomeric anti-ICAM-1 antibodies. However, coupling ICAM-1 antibodies to nanoparticles creates multivalent ligands that enter cells via an amiloride-sensitive endocytic pathway that does not require clathrin or caveolin. Fluorescence microscopy revealed that internalized anti-ICAM nanoparticles are retained in a stable form in early endosomes for an unusually long time (1-2 h) and subsequently were degraded following slow transport to lysosomes. Inhibition of lysosome acidification by chloroquine delayed degradation without affecting anti-ICAM trafficking. Also, the microtubule disrupting agent nocodazole delayed degradation by inhibiting anti-ICAM nanoparticle trafficking to lysosomes. Addition of catalase to create anti-ICAM nanoparticles with antioxidant activity did not affect the mechanisms of nanoparticle uptake or trafficking. Intracellular anti-ICAM/catalase nanoparticles were active, because endothelial cells were resistant to H2O2-induced oxidative injury for 1-2 h after nanoparticle uptake. Chloroquine and nocodazole increased the duration of antioxidant protection by decreasing the extent of anti-ICAM/catalase degradation. Therefore, the unique trafficking pathway followed by internalized anti-ICAM nanoparticles seems well suited for targeted delivery of therapeutic enzymes to endothelial cells and may provide a basis for treatment of acute vascular oxidative stress. PMID:12878488

  2. CdS/CdSe quantum dot-sensitized solar cells based on ZnO nanoparticle/nanorod composite electrodes

    NASA Astrophysics Data System (ADS)

    Kim, Soo-Kyoung; Raj, C. Justin; Kim, Hee-Je

    2014-11-01

    Zinc oxide (ZnO) films were deposited on fluorine-doped tin oxide (FTO) glass substrates with the application of polysulfide redox reactions and a CuS counter electrode to fabricate CdS/CdSe quantum dot-sensitized solar cells (QDSCs). In the present study, ZnO nanoparticles were deposited in the interstices of the ZnO nanorods. The performance of the QDSCs was improved by the ZnO nanoparticle/ nanorod composite structure because the ZnO nanorods exhibit high electron transport, and while the ZnO nanoparticles have a large surface area for QD deposition. The ZnO nanoparticle/nanorod composite films represent a promising achievement for enhancing the conversion efficiency of QDSCs. [Figure not available: see fulltext.

  3. Dual targeted polymeric nanoparticles based on tumor endothelium and tumor cells for enhanced antitumor drug delivery.

    PubMed

    Gupta, Madhu; Chashoo, Gousia; Sharma, Parduman Raj; Saxena, Ajit Kumar; Gupta, Prem Narayan; Agrawal, Govind Prasad; Vyas, Suresh Prasad

    2014-03-01

    Some specific types of tumor cells and tumor endothelial cells represented CD13 proteins and act as receptors for Asn-Gly-Arg (NGR) motifs containing peptide. These CD13 receptors can be specifically recognized and bind through the specific sequence of cyclic NGR (cNGR) peptide and presented more affinity and specificity toward them. The cNGR peptide was conjugated to the poly(ethylene glycol) (PEG) terminal end in the poly(lactic-co-glycolic) acid PLGA-PEG block copolymer. Then, the ligand conjugated nanoparticles (cNGR-DNB-NPs) encapsulating docetaxel (DTX) were synthesized from preformed block copolymer by the emulsion/solvent evaporation method and characterized for different parameters. The various studies such as in vitro cytotoxicity, cell apoptosis, and cell cycle analysis presented the enhanced therapeutic potential of cNGR-DNB-NPs. The higher cellular uptake was also found in cNGR peptide anchored NPs into HUVEC and HT-1080 cells. However, free cNGR could inhibit receptor mediated intracellular uptake of NPs into both types of cells at 37 and 4 °C temperatures, revealing the involvement of receptor-mediated endocytosis. The in vivo biodistribution and antitumor efficacy studies indicated that targeted NPs have a higher therapeutic efficacy through targeting the tumor-specific site. Therefore, the study exhibited that cNGR-functionalized PEG-PLGA-NPs could be a promising approach for therapeutic applications to efficient antitumor drug delivery. PMID:24512060

  4. Silver nanoparticles induce p53-mediated apoptosis in human bronchial epithelial (BEAS-2B) cells.

    PubMed

    Kim, Ha Ryong; Shin, Da Young; Park, Yong Joo; Park, Chang We; Oh, Seung Min; Chung, Kyu Hyuck

    2014-06-01

    Deregulated apoptosis has been associated with many lung diseases. Although many studies have reported the apoptotic effects exhibited by silver nanoparticles (Ag-NPs) in various circumstances, the apoptosis mechanism of Ag-NPs is unclear. We investigated oxidative stress and apoptosis in human normal bronchial epithelial (BEAS-2B) cells to elucidate the role of p53 in apoptosis by Ag-NPs. First, dispersion and stability of Ag-NPs improved using bronchial epithelial cell growth medium with 5% fetal bovine serum. Then, we observed oxidative stress in BEAS-2B cells exposed to Ag-NPs. Second, we carried out a cell death assay to measure DNA fragmentation as a biomarker of apoptosis. BEAS-2B cells were treated with p53-specific short interfering RNA (siRNA) or p53 inhibitor (pifithrin-?) to investigate whether p53 is involved in apoptosis by Ag-NPs. As a result, Ag-NPs significantly enhanced DNA fragmentation dose-dependently and treatment with p53 siRNA or pifithrin-? prevented DNA fragmentation. We also found that apoptosis-related genes (caspase-3, Bax, and Bcl-2) were regulated by Ag-NPs, which was detected by mRNA and protein levels. These results suggest that Ag-NPs induced p53-mediated apoptosis in BEAS-2B cells. Our findings may contribute to understanding the potential roles of Ag-NPs in pulmonary disease. PMID:24849675

  5. Influences of Au nanoparticle density on the performance of GaAs solar cells

    NASA Astrophysics Data System (ADS)

    Lam, Nguyen Dinh; Kim, Youngjo; Kim, Kangho; Lee, Jaejin

    2014-03-01

    The influence of the Au nanoparticle density on the performance of single-junction GaAs solar cells has been investigated. The single-junction GaAs solar cells are grown on (100) GaAs substrates by using a low-pressure metalorganic chemical vapor deposition (MOCVD) system. Au nanoparticles are dispersed on the surfaces of the cells by micro-pipetted casting and the density of the dispersed nanoparticles varies from 4.7 × 109 to 4.3 × 1010 cm-2. The conversion efficiencies of the GaAs solar cells are observed to depend strongly on the Au nanoparticle densities. For Au nanoparticle densities lower than 2.8 × 1010 cm-2, the conversion efficiency improvement of the GaAs solar cells mainly originates from the antireflection coating effect. In contrast, for the sample with a Au nanoparticle density of 2.8 × 1010 cm-2, the antireflection effect decreases due to the shadow effect and the local field enhancement effect due to the Au nanoparticles starts to be effective. The GaAs solar cell with a Au nanoparticle density of 1.9 × 1010 cm-2 shows a maximum conversion enhancement compared to that of the reference sample.

  6. Enhancing light trapping properties of thin film solar cells by plasmonic effect of silver nanoparticles.

    PubMed

    Jung, Junhee; Ha, Kyungyeon; Cho, Jaehyun; Ahn, Shihyun; Park, Hyeongsik; Hussain, Shahzada Qamar; Choi, Mansoo; Yi, Junsin

    2013-12-01

    The preparation of thin film silicon solar cells containing Ag nanoparticles is reported in this article. Ag nanoparticles were deposited on fluorine doped tin oxide coated glass substrates by the evaporation and condensation method. a-Si:H solar cells were deposited on these substrates by cluster type plasma enhanced chemical vapor deposition. We discuss the double textured surface effect with respect to both the surface morphology of the substrate and the plasmonic effect of the Ag nanoparticles. Ag nanoparticles of various sizes from 10 to 100 nm were deposited. The haze values of the Ag embedded samples increased with increasing particle size whereas the optical transmittance decreased at the same conditions. The solar cell with the 30 nm size Ag nanoparticles showed a short circuit current density of 12.97 mA/cm2, which is 0.53 mA/cm2 higher than that of the reference solar cell without Ag nanoparticles, and the highest quantum efficiency for wavelengths from 550 to 800 nm. When 30 nm size nanoparticles were employed, the conversion efficiency of the solar cell was increased from 6.195% to 6.696%. This study reports the application of the scattering effect of Ag nanoparticles for the improvement of the conversion efficiency of amorphous silicon solar cells. PMID:24266153

  7. Human iNKT and MAIT cells exhibit a PLZF-dependent proapoptotic propensity that is counterbalanced by XIAP

    PubMed Central

    Gérart, Stéphane; Sibéril, Sophie; Martin, Emmanuel; Lenoir, Christelle; Aguilar, Claire; Picard, Capucine; Lantz, Olivier; Fischer, Alain; Latour, Sylvain

    2013-01-01

    Invariant natural killer (iNKT) T cells and mucosal-associated invariant T (MAIT) cells represent peculiar T-lymphocyte subpopulations with innate-like properties that differ from conventional T cells. iNKT are reduced in the primary immunodeficiency caused by mutations in the X-linked inhibitor of apoptosis (XIAP). By studying the mechanism of this depletion, we herein report that iNKT cells exhibit a high susceptibility to apoptosis that is not observed with conventional T cells. Elevated expression of caspases 3 and 7 accounts for the proapoptotic phenotype of iNKT cells, which is inhibited by XIAP although it exerts a moderate effect in conventional T cells. Similarly, MAIT cells exhibit a proapoptotic propensity with elevated expression of activated caspases and are decreased in XIAP-deficient individuals. Knockdown of the transcription factor PLZF/ZBTB-16, which is involved in the effector program of iNKT cells, diminishes their proapoptotic phenotype. Conversely, overexpression of PLZF/ZBTB-16 in conventional T cells leads to a proapoptotic phenotype. Our findings identify a previously unknown pathway of regulation of innate-like T-cell homeostasis depending on XIAP and PLZF. The proapoptotic feature of iNKT cells also gives a reliable explanation of their exhaustion observed in different human conditions including the XIAP immunodeficiency. PMID:23223428

  8. Recognition and transmembrane delivery of bioconjugated Fe2O3@Au nanoparticles with living cells

    NASA Astrophysics Data System (ADS)

    Sun, Linlin; Wang, Jine; Wang, Zhenxin

    2010-02-01

    Here, we describe the synthesis of peptide- and/or protein-functionalized Fe2O3 core-Au shell (Fe2O3@Au) nanoparticles for imaging and targeting of living cells. When functionalized with the transmembrane peptide RRRRRRRR (R8), the Fe2O3@Au nanoparticles (R8-Fe2O3@Au) are able to serve as cellular trafficking agents with excellent biocompatibility. The internalization mechanism and delivery efficiency of the R8-Fe2O3@Au nanoparticles have been characterized with dark-field microscopy and fluorescence confocal scanning laser microcopy. Experimental result suggests that the R8-Fe2O3@Au nanoparticles are internalized initially by binding with the membrane-associated proteoglycans on cell surfaces, especially heparan sulfate proteoglycans (HSPGs), following an energy-dependent endocytosis process to enter into living cells. After conjugation with the epidermal growth factor receptor antibody (anti-EGFR), these nanoparticles can also be used for the recognition of cell membrane antigens to specifically label tumor cells.Here, we describe the synthesis of peptide- and/or protein-functionalized Fe2O3 core-Au shell (Fe2O3@Au) nanoparticles for imaging and targeting of living cells. When functionalized with the transmembrane peptide RRRRRRRR (R8), the Fe2O3@Au nanoparticles (R8-Fe2O3@Au) are able to serve as cellular trafficking agents with excellent biocompatibility. The internalization mechanism and delivery efficiency of the R8-Fe2O3@Au nanoparticles have been characterized with dark-field microscopy and fluorescence confocal scanning laser microcopy. Experimental result suggests that the R8-Fe2O3@Au nanoparticles are internalized initially by binding with the membrane-associated proteoglycans on cell surfaces, especially heparan sulfate proteoglycans (HSPGs), following an energy-dependent endocytosis process to enter into living cells. After conjugation with the epidermal growth factor receptor antibody (anti-EGFR), these nanoparticles can also be used for the recognition of cell membrane antigens to specifically label tumor cells. Electronic supplementary information (ESI) available: TEM images of Fe2O3 nanoparticles, stability of peptide conjugated Fe2O3@Au nanoparticles; Inhibition of cellular proliferation by nanoparticles and biotin-R8, ICP-MS analysis of HeLa cells incubated with R8-Fe2O3@Au, F-R8-Fe2O3@Au, or F-Fe2O3@Au nanoparticles as a function of incubation time and anti-EGFR blocking study. See DOI: 10.1039/b9nr00152b

  9. Mice Deficient of Glutamatergic Signaling from Intrinsically Photosensitive Retinal Ganglion Cells Exhibit Abnormal Circadian Photoentrainment

    PubMed Central

    Purrier, Nicole; Engeland, William C.; Kofuji, Paulo

    2014-01-01

    Several aspects of behavior and physiology, such as sleep and wakefulness, blood pressure, body temperature, and hormone secretion exhibit daily oscillations known as circadian rhythms. These circadian rhythms are orchestrated by an intrinsic biological clock in the suprachiasmatic nuclei (SCN) of the hypothalamus which is adjusted to the daily environmental cycles of day and night by the process of photoentrainment. In mammals, the neuronal signal for photoentrainment arises from a small subset of intrinsically photosensitive retinal ganglion cells (ipRGCs) that send a direct projection to the SCN. ipRGCs also mediate other non-image-forming (NIF) visual responses such as negative masking of locomotor activity by light, and the pupillary light reflex (PLR) via co-release of neurotransmitters glutamate and pituitary adenylate cyclase-activating peptide (PACAP) from their synaptic terminals. The relative contribution of each neurotransmitter system for the circadian photoentrainment and other NIF visual responses is still unresolved. We investigated the role of glutamatergic neurotransmission for circadian photoentrainment and NIF behaviors by selective ablation of ipRGC glutamatergic synaptic transmission in mice. Mutant mice displayed delayed re-entrainment to a 6 h phase shift (advance or delay) in the light cycle and incomplete photoentrainment in a symmetrical skeleton photoperiod regimen (1 h light pulses between 11 h dark periods). Circadian rhythmicity in constant darkness also was reduced in some mutant mice. Other NIF responses such as the PLR and negative masking responses to light were also partially attenuated. Overall, these results suggest that glutamate from ipRGCs drives circadian photoentrainment and negative masking responses to light. PMID:25357191

  10. Toxicity of silver nanoparticles in mouse embryonic stem cells and chemical based reprogramming of somatic cells to sphere cells

    NASA Astrophysics Data System (ADS)

    Rajanahalli Krishnamurthy, Pavan

    Abstract 1: Silver nanoparticles (Ag Np's) have an interesting surface chemistry and unique plasmonic properties. They are used in a wide variety of applications ranging from consumer products like socks, medical dressing, computer chips and it is also shown to have antimicrobial, anti bacterial activity and wound healing. Ag Np toxicity studies have been limited to date which needs to be critically addressed due to its wide applications. Mouse embryonic stem (MES) cells represent a unique cell population with the ability to undergo both self renewal and differentiation. They exhibit very stringent and tightly regulated mechanisms to circumvent DNA damage and stress response. We used 10 nm coated (polysaccharide) and uncoated Ag Np's to test its toxic effects on MES cells. MES cells and embryoid bodies (EB's) were treated with two concentrations of Ag Np's: 5 microg/ml and 50 ug/ml and exposed for 24, 48 and 72 hours. Increased cell death, ROS production and loss of mitochondrial membrane potential and alkaline phosphatase (AP) occur in a time and a concentration dependant manner. Due to increased cell death, there is a progressive increase in Annexin V (apoptosis) and Propidium Iodide (PI) staining (necrosis). Oct4 and Nanog undergo ubiquitination and dephosphorylation post-translational modifications in MES cells thereby altering gene expression of pluripotency factors and differentiation of EB's into all the three embryonic germ layers with specific growth factors were also inhibited after Ag Np exposure. Flow cytometry analysis revealed Ag Np's treated cells had altered cell cycle phases correlating with altered self renewal capacity. Our results suggest that Ag Np's effect MES cell self renewal, pluripotency and differentiation and serves as a perfect model system for studying toxicity induced by engineered Ag Np's. Abstract 2: The reprogramming of fibroblasts to pluripotent stem cells and the direct conversion of fibroblasts to functional neurons has been successfully manipulated by ectopic expression of defined factors. We demonstrate that mouse fibroblasts can be converted into sphere cells by detaching fibroblast cells by proteases and then using AlbuMAX I-containing culture medium without genetic alteration. AlbuMAX I is a lipid-rich albumin. Albumin-associated lipids arachidonic acid (AA) and pluronic F-68 were responsible for this effect. The converted colonies were positive for both alkaline phosphatase and stage specific embryonic antigen-1 (SSEA-1) staining. Global gene expression analysis indicated that the sphere cells were in an intermediate state compared with MES cells and MEF cells. The sphere cells were able to differentiate into tissues representing all three embryonic germ layers following retinoic acid treatment, and also differentiated into smooth muscle cells following treatment with vascular endothelial growth factor (VEGF). The study presented a potential novel approach to transdifferentiate mouse fibroblast cells into other cell lineages mediated by AlbuMAX I-containing culture medium.

  11. Drug permeation across intestinal epithelial cells using porous silicon nanoparticles.

    PubMed

    Bimbo, Luis M; Mäkilä, Ermei; Laaksonen, Timo; Lehto, Vesa-Pekka; Salonen, Jarno; Hirvonen, Jouni; Santos, Hélder A

    2011-04-01

    Mesoporous silicon particles hold great potential in improving the solubility of otherwise poorly soluble drugs. To effectively translate this feature into the clinic, especially via oral or parenteral administration, a thorough understanding of the interactions of the micro- and nanosized material with the physiological environment during the delivery process is required. In the present study, the behaviour of thermally oxidized porous silicon particles of different sizes interacting with Caco-2 cells (both non-differentiated and polarized monolayers) was investigated in order to establish their fate in a model of intestinal epithelial cell barrier. Particle interactions and TNF-? were measured in RAW 264.7 macrophages, while cell viabilities, reactive oxygen species and nitric oxide levels, together with transmission electron microscope images of the polarized monolayers, were assessed with both the Caco-2 cells and RAW 264.7 macrophages. The results showed a concentration and size dependent influence on cell viability and ROS-, NO- and TNF-? levels. There was no evidence of the porous nanoparticles crossing the Caco-2 cell monolayers, yet increased permeation of the loaded poorly soluble drug, griseofulvin, was shown. PMID:21194747

  12. Cell uptake and oral absorption of titanium dioxide nanoparticles.

    PubMed

    Janer, G; Mas del Molino, E; Fernández-Rosas, E; Fernández, A; Vázquez-Campos, S

    2014-07-15

    Large efforts are invested on the development of in vitro tests to evaluate nanomaterial (NM) toxicity. In order to assess the relevance of the adverse effects identified in in vitro toxicity tests a thorough understanding of the biokinetics of NMs is critical. We used different in vitro and in vivo test methods to evaluate cell uptake and oral absorption of titanium dioxide nanoparticles (TiO2 NPs). These NPs were readily uptaken by A549 cells (carcinomic human alveolar basal epithelial cells) in vitro. Such rapid uptake contrasted with a very low oral absorption in a differentiated Caco-2 monolayer system (human epithelial colorectal adenocarcinoma cells) and after oral gavage administration to rats. In this oral study, no significant increase in the levels of titanium was recorded by ICP-MS in any of the tissues evaluated (including among other: small intestine, Peyer's patches, mesenteric lymph nodes, liver, and spleen). No NPs were observed by TEM in sections of the small intestine, except for several particles in the cytoplasm of a cell from a Peyer's Patch area. The observation of NPs in Peyer's Patch suggests that the Caco-2 monolayer system is likely to underestimate the potential for oral absorption of NPs and that the model could be improved by including M-cells in co-culture. PMID:24793716

  13. Polymeric nanoparticles for targeted radiosensitization of prostate cancer cells.

    PubMed

    Menon, Jyothi U; Tumati, Vasu; Hsieh, Jer-Tsong; Nguyen, Kytai T; Saha, Debabrata

    2015-05-01

    One of the many issues of using radiosensitizers in a clinical setting is timing daily radiation treatments to coincide with peak drug concentration in target tissue. To overcome this deficit, we have synthesized a novel nanoparticle (NP) system consisting of poly (lactic-co-glycolic acid) (PLGA) NPs conjugated with prostate cancer cell penetrating peptide-R11 and encapsulated with a potent radio-sensitizer 8-dibenzothiophen-4-yl-2-morpholin-4-yl-chromen-4-one (NU7441) to allow prostate cancer-specific targeting and sustained delivery over 3 weeks. Preliminary characterization studies showed that the R11-conjugated NPs (R11-NU7441 NPs) had an average size of about 274?±?80 nm and were stable for up to 5 days in deionized water and serum. The NPs were cytocompatible with immortalized prostate cells (PZ-HPV-7). Further, the particles showed a bi-phasic release of encapsulated NU7441 and were taken up by PC3 prostate cancer cells in a dose- and magnetic field-dependent manner while not being taken up in nonprostate cancer cell lines. In addition, R11-NU7441 NPs were effective radiation sensitizers of prostate cancer cell lines in vitro. These results thus demonstrate the potential of R11-conjugated PLGA NPs as novel platforms for targeted radiosensitization of prostate cancer cells. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 1632-1639, 2015. PMID:25088162

  14. LaB6 nanoparticles with carbon-doped silica coating for fluorescence imaging and near-IR photothermal therapy of cancer cells.

    PubMed

    Lai, B-H; Chen, D-H

    2013-07-01

    In this study, LaB6 nanoparticles are used as a novel nanomaterial for near-infrared (NIR) photothermal therapy because they are cheaper than nanostructured gold, are easy to prepare and have an excellent NIR photothermal conversion property. Furthermore, the surface of LaB6 nanoparticles is coated with a carbon-doped silica (C-SiO2) shell to introduce a fluorescent property and improve stability and biocompatibility. The resulting LaB6@C-SiO2 nanoparticles retain the excellent NIR photothermal conversion property and exhibit a bright blue emission under UV irradiation or a green emission under visible irradiation. Using a HeLa cancer cell line, it is demonstrated that LaB6@C-SiO2 nanoparticles have no significant cytotoxicity, but their presence leads to remarkable cell death after NIR irradiation. In addition, from the observation of cellular uptake, the fluorescence labeling function of LaB6@SiO2 (LaB6 core/SiO2 shell) nanoparticles is also confirmed. These results suggest that LaB6@C-SiO2 nanoparticles may potentially serve as an efficient multifunctional nano-platform for simultaneous fluorescent imaging and NIR-triggered photothermal therapy of cancer cells. PMID:23542555

  15. Effects of poly-(lactide-co-glycolide) nanoparticles on electrophysiological properties of enteroendocrine cells.

    PubMed

    Shah, Bhavik; Kona, Soujanya; Gilbertson, Timothy A; Nguyen, Kytai T

    2011-04-01

    PLGA nanoparticles are widely used to deliver pharmacological compounds and genes to a variety of cell types. Despite the fact that many of these cells types depend critically on ion channel activity to function normally, there have been no studies on the effect of nanoparticles on the ion channel activity. To this end, we have investigated the effect of nanoparticles on cholecystokinin (CCK)-releasing enteroendocrine cell (EEC) line STC-1. It has been shown that regulation of CCK release from STC-1 cells in response to food depends on the normal electrogenic properties of these cells, including the activity of voltage-gated calcium and potassium channels. Due to the importance of voltage-gated ion channels in the normal physiological responses of STC-1 cells, we performed electrophysiological (patch clamp) experiments to assess the effects of PLGA nanoparticles on the voltage-gated calcium and potassium channels. Whole-cell patch clamp recordings on STC-1 cells containing 100 nm nanoparticles show no macroscopic differences in calcium and potassium channel activity. Additional experiments determined that the activation, inactivation, and use-dependent inactivation of these voltage-gated ion channels did not have any significant effect of nanoparticles on these basic biophysical properties. Lastly, we have examined the effects of PLGA nanoparticles on stimulus-induced rise in intracellular calcium concentration in STC-1 cells, which is necessary for release of CCK. Our data demonstrate that the use of PLGA nanoparticles did not alter the electrophysiological properties of STC-1 cells and supports the use of PLGA nanoparticles as an attractive option for delivering pharmaceuticals/genes to cells of the digestive system that might eventually prove useful for reducing appetite/food intake and in treatment of various gastrointestinal illnesses. PMID:21776734

  16. Oxidative stress mediated cytotoxicity of biologically synthesized silver nanoparticles in human lung epithelial adenocarcinoma cell line

    PubMed Central

    2014-01-01

    The goal of the present study was to investigate the toxicity of biologically prepared small size of silver nanoparticles in human lung epithelial adenocarcinoma cells A549. Herein, we describe a facile method for the synthesis of silver nanoparticles by treating the supernatant from a culture of Escherichia coli with silver nitrate. The formation of silver nanoparticles was characterized using various analytical techniques. The results from UV-visible (UV-vis) spectroscopy and X-ray diffraction analysis show a characteristic strong resonance centered at 420 nm and a single crystalline nature, respectively. Fourier transform infrared spectroscopy confirmed the possible bio-molecules responsible for the reduction of silver from silver nitrate into nanoparticles. The particle size analyzer and transmission electron microscopy results suggest that silver nanoparticles are spherical in shape with an average diameter of 15 nm. The results derived from in vitro studies showed a concentration-dependent decrease in cell viability when A549 cells were exposed to silver nanoparticles. This decrease in cell viability corresponded to increased leakage of lactate dehydrogenase (LDH), increased intracellular reactive oxygen species generation (ROS), and decreased mitochondrial transmembrane potential (MTP). Furthermore, uptake and intracellular localization of silver nanoparticles were observed and were accompanied by accumulation of autophagosomes and autolysosomes in A549 cells. The results indicate that silver nanoparticles play a significant role in apoptosis. Interestingly, biologically synthesized silver nanoparticles showed more potent cytotoxicity at the concentrations tested compared to that shown by chemically synthesized silver nanoparticles. Therefore, our results demonstrated that human lung epithelial A549 cells could provide a valuable model to assess the cytotoxicity of silver nanoparticles. PMID:25242904

  17. Adult Cardiac Progenitor Cell Aggregates Exhibit Survival Benefit Both In Vitro and In Vivo

    E-print Network

    Bauer, Michael

    Background: A major hurdle in the use of exogenous stems cells for therapeutic regeneration of injured myocardium remains the poor survival of implanted cells. To date, the delivery of stem cells into myocardium has largely ...

  18. Efficient gene transfer into rat mesenchymal stem cells with cationized Lycium barbarum polysaccharides nanoparticles

    Microsoft Academic Search

    Miao Wang; Wenwen Deng; Min Fu; Xia Cao; Yan Yang; Weiyan Su; Jiangnan Yu; Ximing Xu

    2011-01-01

    A new gene vector, cationized Lycium barbarum polysaccharides (cLBP) nanoparticles, prepared with different amine compounds for non-viral delivery of plasmid DNA encoding for transforming growth factor beta 1 (TGF?-1) was developed and the efficiency of these nanoparticles to transfect rat mesenchymal stem cells in vitro was determined. The spherical cLBP-plasmid TGF?-1 nanoparticles were 160–330nm in diameter and can retard the

  19. One-pot synthesis of CdS nanoparticles exhibiting quantum size effect prepared within a sol-gel derived ureasilicate matrix

    NASA Astrophysics Data System (ADS)

    Gonçalves, Luis F. F. F.; Kanodarwala, Fehmida K.; Stride, John A.; Silva, Carlos J. R.; Gomes, Maria J. M.

    2013-12-01

    This paper describes a novel single-pot synthesis process based on sol-gel for the production of a highly transparent hybrid matrix containing CdS nanoparticles (NPs). The reaction between cadmium and sulphide ions in the presence of 3-mercaptopropyltrimethoxysilane (MPTMS) that originates the formation of quantum confined CdS NPs occurs simultaneously with the condensation and polymerization of the gel precursors that evolves to a macromolecular hybrid organic-inorganic network. The obtained xerogel matrix is based on the reaction of organically modified alkoxysilane (3-isocyanatepropyltriethoxysilane) and a di-amine functionalized oligopolyoxyethylene (Jeffamine ED-600). The final material is characterized as highly transparent, homogeneous and flexible xerogel incorporating stabilized and high crystalline CdS NPs that exhibit size-dependent optical properties due to quantum confinement of photogenerated e-h pairs as observed from UV-visible and photoluminescence spectroscopy and HRTEM microscopy measurements. The developed approach has obvious advantages comparatively to the alternative and more complex routes of production of composite materials with embedded semiconductor NPs because of the simplified one-pot preparative procedure used. The developed sol-gel process allows the control of the optical characteristics of the obtained CdS NPs embedded within the network by adjusting the molar ratio between cadmium ion and MPTMS and between cadmium and sulphide ions.

  20. In vitro toxicity of silica nanoparticles in human lung cancer cells

    SciTech Connect

    Lin Weisheng [Department of Chemistry and Environmental Research Center, University of Missouri-Rolla, Rolla, MO 65409 (United States); Huang Yuewern [Department of Biological Sciences and Environmental Research Center, University of Missouri-Rolla, Rolla, MO 65409 (United States); Zhou Xiaodong [Pacific Northwest National Laboratory, Richland, WA 99352 (United States); Ma Yinfa [Department of Chemistry and Environmental Research Center, University of Missouri-Rolla, Rolla, MO 65409 (United States)]. E-mail: yinfa@umr.edu

    2006-12-15

    The cytotoxicity of 15-nm and 46-nm silica nanoparticles was investigated by using crystalline silica (Min-U-Sil 5) as a positive control in cultured human bronchoalveolar carcinoma-derived cells. Exposure to 15-nm or 46-nm SiO{sub 2} nanoparticles for 48 h at dosage levels between 10 and 100 {mu}g/ml decreased cell viability in a dose-dependent manner. Both SiO{sub 2} nanoparticles were more cytotoxic than Min-U-Sil 5; however, the cytotoxicities of 15-nm and 46-nm silica nanoparticles were not significantly different. The 15-nm SiO{sub 2} nanoparticles were used to determine time-dependent cytotoxicity and oxidative stress responses. Cell viability decreased significantly as a function of both nanoparticle dosage (10-100 {mu}g/ml) and exposure time (24 h, 48 h, and 72 h). Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species (ROS), glutathione, malondialdehyde, and lactate dehydrogenase, were quantitatively assessed. Exposure to SiO{sub 2} nanoparticles increased ROS levels and reduced glutathione levels. The increased production of malondialdehyde and lactate dehydrogenase release from the cells indicated lipid peroxidation and membrane damage. In summary, exposure to SiO{sub 2} nanoparticles results in a dose-dependent cytotoxicity in cultural human bronchoalveolar carcinoma-derived cells that is closely correlated to increased oxidative stress.

  1. Orthopaedic applications of nanoparticle-based stem cell therapies

    PubMed Central

    2012-01-01

    Stem cells have tremendous applications in the field of regenerative medicine and tissue engineering. These are pioneering fields that aim to create new treatments for disease that currently have limited therapies or cures. A particularly popular avenue of research has been the regeneration of bone and cartilage to combat various orthopaedic diseases. Magnetic nanoparticles (MNPs) have been applied to aid the development and translation of these therapies from research to the clinic. This review highlights contemporary research for the applications of iron-oxide-based MNPs for the therapeutic implementation of stem cells in orthopaedics. These MNPs comprise of an iron oxide core, coated with a choice of biological polymers that can facilitate the uptake of MNPs by cells through improving endocytic activity. The combined use of these oxides and the biological polymer coatings meet biological requirements, effectively encouraging the use of MNPs in regenerative medicine. The association of MNPs with stem cells can be achieved via the process of endocytosis resulting in the internalisation of these particles or the attachment to cell surface receptors. This allows for the investigation of migratory patterns through various tracking studies, the targeting of particle-labelled cells to desired locations via the application of an external magnetic field and, finally, for activation stem cells to initiate various cellular responses to induce the differentiation. Characterisation of cell localisation and associated tissue regeneration can therefore be enhanced, particularly for in vivo applications. MNPs have been shown to have the potential to stimulate differentiation of stem cells for orthopaedic applications, without limiting proliferation. However, careful consideration of the use of active agents associated with the MNP is suggested, for differentiation towards specific lineages. This review aims to broaden the knowledge of current applications, paving the way to translate the in vitro and in vivo work into further orthopaedic clinical studies. PMID:22520594

  2. Breaking tolerance to self, circulating natural killer cells expressing inhibitory KIR for non-self HLA exhibit effector function after T cell–depleted allogeneic hematopoietic cell transplantation

    PubMed Central

    Yu, Junli; Venstrom, Jeffrey M.; Liu, Xiao-Rong; Pring, James; Hasan, Reenat S.; O'Reilly, Richard J.

    2009-01-01

    Alloreactive natural killer (NK) cells are an important influence on hematopoietic stem cell transplantation (HSCT) outcome. In HLA-mismatched HSCT, alloreactivity occurs when licensed donor NK cells expressing inhibitory killer Ig-like receptors (KIR) for donor MHC class I ligands recognize the lack of the class I ligands in the mismatched recipient (“missing self”). Studies in HLA-matched HSCT, however, have also demonstrated improved outcome in patients lacking class I ligands for donor inhibitory KIR (“missing ligand”), indicating that classically nonlicensed donor NK cells expressing KIR for non-self MHC class I ligands may exhibit functional competence in HSCT. We examined NK function in 16 recipients of T cell–depleted allografts from HLA-identical or KIR-ligand matched donors after myeloablative therapy. After HSCT, nonlicensed NK cells expressing inhibitory KIR for non-self class I exhibit robust intracellular IFN-? and cytotoxic response to target cells lacking cognate ligand, gradually becoming tolerized to self by day 100. These findings could not be correlated with cytokine environment or phenotypic markers of NK development, nor could they be attributed to non-KIR receptors such as CD94/NKG2A. These findings confirm that NK alloreactivity can occur in HLA-matched HSCT, where tolerance to self is either acquired by the stem cell–derived NK cell after exiting the bone marrow or where tolerance to self can be temporarily overcome. PMID:19179302

  3. Internalized Chitosan Nanoparticles Persist for Long Time in Cultured Cells

    PubMed Central

    Malatesta, M.; Grecchi, S.; Chiesa, E.; Cisterna, B.; Costanzo, M.; Zancanaro, C.

    2015-01-01

    Chitosan-based nanoparticles (chiNPs) are considered to be potentially good carriers for the sustained intracellular delivery of specific molecules. However, scarce attention has been paid to the long-lasting permanence of these NPs in the intracellular milieu, as well as to their intracellular fate (i.e., distribution, interaction with cell organelles, and degradation) in the long term. In the present study, the presence and subcellular location of FITC-labelled chiNPs were monitored in HeLa cells up to 14 days post-administration using multicolorfluorescence confocal microscopy and diaminobenzidine photo-oxidation at transmission electron microscopy. The main result of the present study is the demonstration that internalized chiNPs persist inside the cell up to two weeks, occurring in both the cytoplasm and nucleus; accordingly, chiNPs are able to pass from mother to daughter cells through several mitotic cycles. The cells did not show increased mortality or structural damage up to 14 days after chiNP exposure. PMID:25820565

  4. Ligand modified nanoparticles increases cell uptake, alters endocytosis and elevates glioma distribution and internalization

    PubMed Central

    Gao, Huile; Yang, Zhi; Zhang, Shuang; Cao, Shijie; Shen, Shun; Pang, Zhiqing; Jiang, Xinguo

    2013-01-01

    Nanoparticles (NPs) were widely used in drugs/probes delivery for improved disease diagnosis and/or treatment. Targeted delivery to cancer cells is a highly attractive application of NPs. However, few studies have been performed on the targeting mechanisms of these ligand-modified delivery systems. Additional studies are needed to understand the transport of nanoparticles in the cancer site, the interactions between nanoparticles and cancer cells, the intracellular trafficking of nanoparticles within the cancer cells and the subcellular destiny and potential toxicity. Interleukin 13 (IL-13) peptide can specifically bind IL-13R?2, a receptor that is highly expressed on glioma cells but is expressed at low levels on other normal cells. It was shown that the nanoparticels modification with the IL-13 peptide could improve glioma treatment by selectively increasing cellular uptake, facilitating cell internalization, altering the uptake pathway and increasing glioma localization. PMID:23982586

  5. Human circulating CD14+ monocytes as a source of progenitors that exhibit mesenchymal cell differentiation

    Microsoft Academic Search

    Masataka Kuwana; Yuka Okazaki; Hiroaki Kodama; Keisuke Izumi; Hidekata Yasuoka; Yoko Ogawa; Yutaka Kawakami; Yasuo Ikeda

    2003-01-01

    Circulating CD14 monocytes are pre- cursors of phagocytes, such as macrophages and dendritic cells. Here we report primitive cells with a fibroblast-like morphology derived from human peripheral blood CD14 monocytes that can dif- ferentiate into several distinct mesenchymal cell lineages. We named this cell population monocyte- derived mesenchymal progenitor (MOMP). MOMPs were obtained in vitro from human peripheral blood mononuclear

  6. Development and characterization of multifunctional nanoparticles for drug delivery to cancer cells

    NASA Astrophysics Data System (ADS)

    Nahire, Rahul Rajaram

    Lipid and polymeric nanoparticles, although proven to be effective drug delivery systems compared to free drugs, have shown considerable limitations pertaining to their uptake and release at tumor sites. Spatial and temporal control over the delivery of anticancer drugs has always been challenge to drug delivery scientists. Here, we have developed and characterized multifunctional nanoparticles (liposomes and polymersomes) which are targeted specifically to cancer cells, and release their contents with tumor specific internal triggers. To enable these nanoparticles to be tracked in blood circulation, we have imparted them with echogenic characteristic. Echogenicity of nanoparticles is evaluated using ultrasound scattering and imaging experiments. Nanoparticles demonstrated effective release with internal triggers such as elevated levels of MMP-9 enzyme found in the extracellular matrix of tumor cells, decreased pH of lysosome, and differential concentration of reducing agents in cytosol of cancer cells. We have also successfully demonstrated the sensitivity of these particles towards ultrasound to further enhance the release with internal triggers. To ensure the selective uptake by folate receptor- overexpressing cancer cells, we decorated these nanoparticles with folic acid on their surface. Fluorescence microscopic images showed significantly higher uptake of folate-targeted nanoparticles by MCF-7 (breast cancer) and PANC-1 (pancreatic cancer) cells compared to particles without any targeting ligand on their surface. To demonstrate the effectiveness of these nanoparticles to carry the drugs inside and kill cancer cells, we encapsulated doxorubicin and/or gemcitabine employing the pH gradient method. Drug loaded nanoparticles showed significantly higher killing of the cancer cells compared to their non-targeted counterparts and free drugs. With further development, these nanoparticles certainly have potential to be used as a multifunctional nanocarriers for image guided, targeted delivery of anticancer drugs.

  7. Hydroxyapatite nanoparticles inhibit the growth of human glioma cells in vitro and in vivo.

    PubMed

    Chu, Sheng-Hua; Feng, Dong-Fu; Ma, Yan-Bin; Li, Zhi-Qiang

    2012-01-01

    Hydroxyapatite nanoparticles (nano-HAPs) have been reported to exhibit antitumor effects on various human cancers, but the effects of nano-HAPs on human glioma cells remain unclear. The aim of this study was to explore the inhibitory effect of nano-HAPs on the growth of human glioma U251 and SHG44 cells in vitro and in vivo. Nano-HAPs could inhibit the growth of U251 and SHG44 cells in a dose- and time-dependent manner, according to methyl thiazoletetrazolium assay and flow cytometry. Treated with 120 mg/L and 240 mg/L nano-HAPs for 48 hours, typical apoptotic morphological changes were noted under Hoechst staining and transmission electron microscopy. The tumor growth of cells was inhibited after the injection in vivo, and the related side effects significantly decreased in the nano-HAP-and-drug combination group. Because of the function of nano-HAPs, the expression of c-Met, SATB1, Ki-67, and bcl-2 protein decreased, and the expression of SLC22A18 and caspase-3 protein decreased noticeably. The findings indicate that nano-HAPs have an evident inhibitory action and induce apoptosis of human glioma cells in vitro and in vivo. In a drug combination, they can significantly reduce the adverse reaction related to the chemotherapeutic drug 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). PMID:22888225

  8. Hyaluronic acid modified mesoporous silica nanoparticles for targeted drug delivery to CD44-overexpressing cancer cells.

    PubMed

    Yu, Meihua; Jambhrunkar, Siddharth; Thorn, Peter; Chen, Jiezhong; Gu, Wenyi; Yu, Chengzhong

    2013-01-01

    In this paper, a targeted drug delivery system has been developed based on hyaluronic acid (HA) modified mesoporous silica nanoparticles (MSNs). HA-MSNs possess a specific affinity to CD44 over-expressed on the surface of a specific cancer cell line, HCT-116 (human colon cancer cells). The cellular uptake performance of fluorescently labelled MSNs with and without HA modification has been evaluated by confocal microscopy and fluorescence-activated cell sorter (FACS) analysis. Compared to bare MSNs, HA-MSNs exhibit a higher cellular uptake via HA receptor mediated endocytosis. An anticancer drug, doxorubicin hydrochloride (Dox), has been loaded into MSNs and HA-MSNs as drug delivery vehicles. Dox loaded HA-MSNs show greater cytotoxicity to HCT-116 cells than free Dox and Dox-MSNs due to the enhanced cell internalization behavior of HA-MSNs. It is expected that HA-MSNs have a great potential in targeted delivery of anticancer drugs to CD44 over-expressing tumors. PMID:23076766

  9. Influence of nanoparticle shape on charge transport and recombination in polymer/nanocrystal solar cells.

    PubMed

    Li, Zhe; Wang, Weiyuan; Greenham, Neil C; McNeill, Christopher R

    2014-12-21

    A key consideration for the efficient operation of hybrid solar cells based upon conjugated polymers and inorganic semiconductor nanocrystals is charge transport in the nanocrystal phase. Here we report the results of a study into the charge transport kinetics of polymer/nanocrystal solar cells based on blends poly(3-hexylthiophene) (P3HT) with either CdSe nano-dots or CdSe nano-tetrapods. Transient photocurrent measurements reveal significant differences in the charge transport kinetics of nano-dot and nano-tetrapod hybrid cells, with the charge collection of the P3HT/CdSe nano-dot device severely limited by charge trapping. In comparison the nano-tetrapod cell exhibits significantly reduced charge trapping compared to the nano-dot cell accounting for the improved fill-factor and overall device efficiency. Transient photovoltage measurements have also been employed that demonstrate slower recombination rates in the P3HT/CdSe tetrapod device compared to the P3HT/CdSe dot device. These observations directly identify nanoparticle shape as a critical factor influencing the charge transport and hence recombination in this benchmark hybrid system, confirming the hypothesis that the use of tetrapods improves device performance through an improvement in electron transport in the nanocrystal phase. PMID:24781139

  10. Fabrication, performance and atmospheric stability of inverted ZnO nanoparticle/polymer solar cell

    NASA Astrophysics Data System (ADS)

    Yuan, Zhaolin

    2015-01-01

    Zinc oxide (ZnO) nanoparticles (NPs, ~5 nm) were first synthesized by a simple wet chemical method. A mixture of poly(3-hexylthiophene-2,5-diyl):(6,6)-phenyl C61 butyric acid methyl ester (P3HT:PCBM) was used as the photoactive layer, and an inverted solar cell with a structure of ITO/ZnO NPs/P3HT:PCBM/MoO3/Ag was fabricated. Its performance and stability in the ambient atmosphere were investigated in detail. The results showed that the fabricated solar cell under 100 mW/cm2 AM1.5 illumination exhibited a power conversion efficiency (PCE) of 0.28 %. In addition, illumination intensity had significant effect on open circuit voltage ( V oc), short circuit current ( J sc), fill factor (FF), and PCE of the fabricated solar cell. The dark storability (darkness, room temperature, and 50-60 % relative humidity) was shown to exceed 4,416 h without notable loss in PCE. The fabricated solar cell with excellent long-term stability was achieved in an ambient atmosphere; also, the stable mechanism of the solar cell in the ambient atmosphere was illuminated.

  11. Hyaluronic acid modified mesoporous silica nanoparticles for targeted drug delivery to CD44-overexpressing cancer cells

    NASA Astrophysics Data System (ADS)

    Yu, Meihua; Jambhrunkar, Siddharth; Thorn, Peter; Chen, Jiezhong; Gu, Wenyi; Yu, Chengzhong

    2012-12-01

    In this paper, a targeted drug delivery system has been developed based on hyaluronic acid (HA) modified mesoporous silica nanoparticles (MSNs). HA-MSNs possess a specific affinity to CD44 over-expressed on the surface of a specific cancer cell line, HCT-116 (human colon cancer cells). The cellular uptake performance of fluorescently labelled MSNs with and without HA modification has been evaluated by confocal microscopy and fluorescence-activated cell sorter (FACS) analysis. Compared to bare MSNs, HA-MSNs exhibit a higher cellular uptake via HA receptor mediated endocytosis. An anticancer drug, doxorubicin hydrochloride (Dox), has been loaded into MSNs and HA-MSNs as drug delivery vehicles. Dox loaded HA-MSNs show greater cytotoxicity to HCT-116 cells than free Dox and Dox-MSNs due to the enhanced cell internalization behavior of HA-MSNs. It is expected that HA-MSNs have a great potential in targeted delivery of anticancer drugs to CD44 over-expressing tumors.

  12. Development of a dose-controlled multiculture cell exposure chamber for efficient delivery of airborne and engineered nanoparticles

    NASA Astrophysics Data System (ADS)

    Asimakopoulou, Akrivi; Daskalos, Emmanouil; Lewinski, Nastassja; Riediker, Michael; Papaioannou, Eleni; Konstandopoulos, Athanasios G.

    2013-04-01

    In order to study the various health influencing parameters related to engineered nanoparticles as well as to soot emitted by Diesel engines, there is an urgent need for appropriate sampling devices and methods for cell exposure studies that simulate the respiratory system and facilitate associated biological and toxicological tests. The objective of the present work was the further advancement of a Multiculture Exposure Chamber (MEC) into a dose-controlled system for efficient delivery of nanoparticles to cells. It was validated with various types of nanoparticles (Diesel engine soot aggregates, engineered nanoparticles for various applications) and with state-of-the-art nanoparticle measurement instrumentation to assess the local deposition of nanoparticles on the cell cultures. The dose of nanoparticles to which cell cultures are being exposed was evaluated in the normal operation of the in vitro cell culture exposure chamber based on measurements of the size specific nanoparticle collection efficiency of a cell free device. The average efficiency in delivering nanoparticles in the MEC was approximately 82%. The nanoparticle deposition was demonstrated by Transmission Electron Microscopy (TEM). Analysis and design of the MEC employs Computational Fluid Dynamics (CFD) and true to geometry representations of nanoparticles with the aim to assess the uniformity of nanoparticle deposition among the culture wells. Final testing of the dose-controlled cell exposure system was performed by exposing A549 lung cell cultures to fluorescently labeled nanoparticles. Delivery of aerosolized nanoparticles was demonstrated by visualization of the nanoparticle fluorescence in the cell cultures following exposure. Also monitored was the potential of the aerosolized nanoparticles to generate reactive oxygen species (ROS) (e.g. free radicals and peroxides generation), thus expressing the oxidative stress of the cells which can cause extensive cellular damage or damage on DNA.

  13. Distinct novel quinazolinone exhibits selective inhibition in MGC-803 cancer cells by dictating mutant p53 function.

    PubMed

    Zhang, Guo-Hai; Xue, Wen-Bin; An, Yun-Feng; Yuan, Jing-Mei; Qin, Jiang-Ke; Pan, Cheng-Xue; Su, Gui-Fa

    2015-05-01

    The mutant p53 proteins and their corresponding cellular response can be manipulated by novel quinazolinone derivatives 4-8 (a-i) in p53 mutant cancer cells. Of the two most potent compounds, 4a exhibited promising broad-spectrum anti-cancer effects, whereas 6c showed selective and exclusive inhibition activity in p53 mutant cancer cell lines but low toxicity to wild-type p53 cancer cell A375 and normal lung fibroblast WI-38 cells. Furthermore, 6c exhibited a more sophisticated mechanism for cell-destructive response by causing S/G2 phase arrest effect and cell size reduction. Compared with the cellular response of 6b and genetic background of cell lines studied, p53 mutation was found to be the key factor and main target for 6c evoked cell-destructive response. Molecular mechanism studies indicated that p53 phosphorylation and acetylation dual-targeting inhibitor 6c exerted anti-cancer activities with a special mechanism in evoking cell apoptosis by arresting mutant p53 function to trigger the deregulation of Cdk2 caused Bim-mediated apoptosis. To the best of our knowledge, 6c is the first quinazolinone derivative to dictate mutant p53 function for apoptotic cell death. PMID:25828929

  14. Mechanodelivery of nanoparticles to the cytoplasm of living cells

    NASA Astrophysics Data System (ADS)

    Emerson, Nyssa T.; Hsia, Chih-Hao; Rafalska-Metcalf, Ilona U.; Yang, Haw

    2014-04-01

    Nanotechnology has opened up the opportunity to probe, sense, and manipulate the chemical environment of biological systems with an unprecedented level of spatiotemporal control. A major obstacle to the full realization of these novel technologies is the lack of a general, robust, and simple method for the delivery of arbitrary nanostructures to the cytoplasm of intact live cells. Here, we identify a new delivery modality, based on mechanical disruption of the plasma membrane, which efficiently mediates the delivery of nanoparticles to the cytoplasm of mammalian cells. We use two distinct execution modes, two adherent cell lines, and three sizes of semiconducting nanocrystals, or quantum dots, to demonstrate its applicability and effectiveness. As the underlying mechanism is purely physical, we anticipate that such ``mechanodelivery'' can be generalized to other modes of execution as well as to the cytoplasmic introduction of a structurally diverse array of functional nanomaterials.Nanotechnology has opened up the opportunity to probe, sense, and manipulate the chemical environment of biological systems with an unprecedented level of spatiotemporal control. A major obstacle to the full realization of these novel technologies is the lack of a general, robust, and simple method for the delivery of arbitrary nanostructures to the cytoplasm of intact live cells. Here, we identify a new delivery modality, based on mechanical disruption of the plasma membrane, which efficiently mediates the delivery of nanoparticles to the cytoplasm of mammalian cells. We use two distinct execution modes, two adherent cell lines, and three sizes of semiconducting nanocrystals, or quantum dots, to demonstrate its applicability and effectiveness. As the underlying mechanism is purely physical, we anticipate that such ``mechanodelivery'' can be generalized to other modes of execution as well as to the cytoplasmic introduction of a structurally diverse array of functional nanomaterials. Electronic supplementary information (ESI) available: Characterization of the QD diameter, passivation of QDs, electroporation protocol, flow cytometric data analysis, and additional epifluorescence images of QD labeled cells, including Table S1 and Fig. S1-S15. See DOI: 10.1039/c3nr06468a

  15. Manganese-impregnated mesoporous silica nanoparticles for signal enhancement in MRI cell labelling studies

    NASA Astrophysics Data System (ADS)

    Guillet-Nicolas, Rémy; Laprise-Pelletier, Myriam; Nair, Mahesh M.; Chevallier, Pascale; Lagueux, Jean; Gossuin, Yves; Laurent, Sophie; Kleitz, Freddy; Fortin, Marc-André

    2013-11-01

    Mesoporous silica nanoparticles (MSNs) are used in drug delivery and cell tracking applications. As Mn2+ is already implemented as a ``positive'' cell contrast agent in preclinical imaging procedures (in the form of MnCl2 for neurological studies), the introduction of Mn in the porous network of MSNs would allow labelling cells and tracking them using MRI. These particles are in general internalized in endosomes, an acidic environment with high saline concentration. In addition, the available MSN porosity could also serve as a carrier to deliver medical/therapeutic substances through the labelled cells. In the present study, manganese oxide was introduced in the porous network of MCM-48 silica nanoparticles (Mn-M48SNs). The particles exhibit a narrow size distribution (~140 nm diam.) and high porosity (~60% vol.), which was validated after insertion of Mn. The resulting Mn-M48SNs were characterized by TEM, N2 physisorption, and XRD. Evidence was found with H2-TPR, and XPS characterization, that Mn(ii) is the main oxidation state of the paramagnetic species after suspension in water, most probably in the form of Mn-OOH. The colloidal stability as a function of time was confirmed by DLS in water, acetate buffer and cell culture medium. In NMR data, no significant evidence of Mn2+ leaching was found in Mn-M48SNs in acidic water (pH 6), up to 96 hours after suspension. High longitudinal relaxivity values of r1 = 8.4 mM-1 s-1 were measured at 60 MHz and 37 °C, with the lowest relaxometric ratios (r2/r1 = 2) reported to date for a Mn-MSN system. Leukaemia cells (P388) were labelled with Mn-M48SNs and nanoparticle cell internalization was confirmed by TEM. Finally, MRI contrast enhancement provided by cell labelling with escalated incubation concentrations of Mn-M48SNs was quantified at 1 T. This study confirmed the possibility of efficiently confining Mn into M48SNs using incipient wetness, while maintaining an open porosity and relatively high pore volume. Because these Mn-labelled M48SNs express strong ``positive'' contrast media properties at low concentrations, they are potentially applicable for cell tracking and drug delivery methodologies.Mesoporous silica nanoparticles (MSNs) are used in drug delivery and cell tracking applications. As Mn2+ is already implemented as a ``positive'' cell contrast agent in preclinical imaging procedures (in the form of MnCl2 for neurological studies), the introduction of Mn in the porous network of MSNs would allow labelling cells and tracking them using MRI. These particles are in general internalized in endosomes, an acidic environment with high saline concentration. In addition, the available MSN porosity could also serve as a carrier to deliver medical/therapeutic substances through the labelled cells. In the present study, manganese oxide was introduced in the porous network of MCM-48 silica nanoparticles (Mn-M48SNs). The particles exhibit a narrow size distribution (~140 nm diam.) and high porosity (~60% vol.), which was validated after insertion of Mn. The resulting Mn-M48SNs were characterized by TEM, N2 physisorption, and XRD. Evidence was found with H2-TPR, and XPS characterization, that Mn(ii) is the main oxidation state of the paramagnetic species after suspension in water, most probably in the form of Mn-OOH. The colloidal stability as a function of time was confirmed by DLS in water, acetate buffer and cell culture medium. In NMR data, no significant evidence of Mn2+ leaching was found in Mn-M48SNs in acidic water (pH 6), up to 96 hours after suspension. High longitudinal relaxivity values of r1 = 8.4 mM-1 s-1 were measured at 60 MHz and 37 °C, with the lowest relaxometric ratios (r2/r1 = 2) reported to date for a Mn-MSN system. Leukaemia cells (P388) were labelled with Mn-M48SNs and nanoparticle cell internalization was confirmed by TEM. Finally, MRI contrast enhancement provided by cell labelling with escalated incubation concentrations of Mn-M48SNs was quantified at 1 T. This study confirmed the possibility of efficiently confining Mn into M48SNs using incipient wetness

  16. Memory CD8+ T cells exhibit increased antigen threshold requirements for recall proliferation.

    PubMed

    Mehlhop-Williams, Erin R; Bevan, Michael J

    2014-02-10

    A hallmark of immunological memory is the ability of previously primed T cells to undergo rapid recall responses upon antigen reencounter. Classic work has suggested that memory T cells proliferate in response to lower doses of antigen than naive T cells and with reduced requirements for co-stimulation. In contrast to this premise, we observed that naive but not memory T cells proliferate in vivo in response to limited antigen presentation. To reconcile these observations, we tested the antigen threshold requirement for cell cycle entry in naive and central memory CD8(+) T cells. Although both naive and memory T cells detect low dose antigen, only naive T cells activate cell cycle effectors. Direct comparison of TCR signaling on a single cell basis indicated that central memory T cells do not activate Zap70, induce cMyc expression, or degrade p27 in response to antigen levels that activate these functions in naive T cells. The reduced sensitivity of memory T cells may result from both decreased surface TCR expression and increased expression of protein tyrosine phosphatases as compared with naive T cells. Our data describe a novel aspect of memory T cell antigen threshold sensitivity that may critically regulate recall expansion. PMID:24493801

  17. Interactions of lipid-based liquid crystalline nanoparticles with model and cell membranes.

    PubMed

    Barauskas, Justas; Cervin, Camilla; Jankunec, Marija; Spandyreva, Marija; Ribokaite, Kristina; Tiberg, Fredrik; Johnsson, Markus

    2010-05-31

    Lipid-based liquid crystalline nanoparticles (LCNPs) are interesting candidates for drug delivery applications, for instance as solubilizing or encapsulating carriers for intravenous (i.v.) drugs. Here it is important that the carriers are safe and tolerable and do not have, e.g. hemolytic activity. In the present study we have studied LCNP particles of different compositions with respect to their mixing behavior and membrane destabilizing effects in model and cell membrane systems. Different types of non-lamellar LCNPs were studied including cubic phase nanoparticles (Cubosome) based on glycerol monooleate (GMO), hexagonal phase nanoparticles (Hexosome) based on diglycerol monooleate (DGMO) and glycerol dioleate (GDO), sponge phase nanoparticles based on DGMO/GDO/polysorbate 80 (P80) and non-lamellar nanoparticles based on soy phosphatidylcholine (SPC)/GDO. Importantly, the LCNPs based on the long-chain monoacyl lipid, GMO, were shown to display a very fast and complete lipid mixing with model membranes composed of multilamellar SPC liposomes as assessed by a fluorescence energy transfer (FRET) assay. The result correlated well with pronounced hemolytic properties observed when the GMO-based LCNPs were mixed with rat whole blood. In sharp contrast, LCNPs based on mixtures of the long-chain diacyl lipids, SPC and GDO, were found to be practically inert towards both hemolysis in rat whole blood as well as lipid mixing with SPC model membranes. The LCNP dispersions based on a mixture of long-chain monoacyl and diacyl lipids, DGMO/GDO, displayed an intermediate behavior compared to the GMO and SPC/GDO-based systems with respect to both hemolysis and lipid mixing. It is concluded that GMO-based LCNPs are unsuitable for parenteral drug delivery applications (e.g. i.v. administration) while the SPC/GDO-based LCNPs exhibit good properties with limited lipid mixing and hemolytic activity. The correlation between results from lipid mixing or FRET experiments and the in vitro hemolysis data indicates that FRET assays can be one useful screening tool for parenteral drug delivery systems. It is argued that the hemolytic potential is correlated with chemical activity of the monomers in the mixtures. PMID:20214966

  18. Enhanced relative biological effectiveness of proton radiotherapy in tumor cells with internalized gold nanoparticles

    SciTech Connect

    Polf, Jerimy C.; Gillin, Michael [Department of Radiation Physics, University of Texas MD Anderson Cancer Center, Houston, Texas 77030 (United States); Bronk, Lawrence F. [Department of Radiation Physics, University of Texas MD Anderson Cancer Center, Houston, Texas 77030 (United States); David H. Koch Center, University of Texas MD Anderson Cancer Center, Houston, Texas 77030 (United States); Driessen, Wouter H. P.; Arap, Wadih; Pasqualini, Renata [David H. Koch Center, University of Texas MD Anderson Cancer Center, Houston, Texas 77030 (United States)

    2011-05-09

    The development and use of sensitizing agents to improve the effectiveness of radiotherapy have long been sought to improve our ability to treat cancer. In this letter, we have studied the relative biological effectiveness of proton beam radiotherapy on prostate tumor cells with and without internalized gold nanoparticles. The effectiveness of proton radiotherapy for the killing of prostate tumor cells was increased by approximately 15%-20% for those cells containing internalized gold nanoparticles.

  19. Oxidative stress induced by cerium oxide nanoparticles in cultured BEAS-2B cells

    Microsoft Academic Search

    Eun-Jung Park; Jinhee Choi; Young-Kwon Park

    2008-01-01

    Cerium oxide nanoparticles of different sizes (15, 25, 30, 45nm) were prepared by the supercritical synthesis method, and cytotoxicity was evaluated using cultured human lung epithelial cells (BEAS-2B). Exposure of the cultured cells to nanoparticles (5, 10, 20, 40?g\\/ml) led to cell death, ROS increase, GSH decrease, and the inductions of oxidative stress-related genes such as heme oxygenase-1, catalase, glutathione

  20. Hyperthermia HeLa cell treatment with silica coated manganese oxide nanoparticles

    E-print Network

    Villanueva, A; Alonso, JM; Rueda, T; Martínez, A; Crespo, P; Morales, MP; Fernandez, MA Gonzalez; Valdes, J; Rivero, G

    2009-01-01

    HeLa tumour cells incubated with ferromagnetic nanoparticles of manganese oxide perovskite La0.56(SrCa)0.22MnO3 were treated with a high frequency alternating magnetic field. The particles were previously coated with silica to improve their biocompatibility. The control assays made with HeLa tumour cells showed that cell survival and growth rate were not affected by the particle internalization in cells, or by the electromagnetic field on cells without nanoparticles. The application of an alternating electromagnetic field to cells incubated with this silica coated manganese oxide induced a significant cellular damage that finally lead to cell death by an apoptotic mechanism.

  1. Antibody-mediated targeting of iron oxide nanoparticles to the folate receptor alpha increases tumor cell association in vitro and in vivo

    PubMed Central

    Ndong, Christian; Toraya-Brown, Seiko; Kekalo, Katsiaryna; Baker, Ian; Gerngross, Tillman U; Fiering, Steven N; Griswold, Karl E

    2015-01-01

    Active molecular targeting has become an important aspect of nanoparticle development for oncology indications. Here, we describe molecular targeting of iron oxide nanoparticles (IONPs) to the folate receptor alpha (FOLR?) using an engineered antibody fragment (Ffab). Compared to control nanoparticles targeting the non-relevant botulinum toxin, the Ffab-IONP constructs selectively accumulated on FOLR?-overexpressing cancer cells in vitro, where they exhibited the capacity to internalize into intracellular vesicles. Similarly, Ffab-IONPs homed to FOLR?-positive tumors upon intraperitoneal administration in an orthotopic murine xenograft model of ovarian cancer, whereas negative control particles showed no detectable tumor accumulation. Interestingly, Ffab-IONPs built with custom 120 nm nanoparticles exhibited lower in vitro targeting efficiency when compared to those built with commercially sourced 180 nm nanoparticles. In vivo, however, the two Ffab-IONP platforms achieved equivalent tumor homing, although the smaller 120 nm IONPs were more prone to liver sequestration. Overall, the results show that Ffab-mediated targeting of IONPs yields specific, high-level accumulation within cancer cells, and this fact suggests that Ffab-IONPs could have future utility in ovarian cancer diagnostics and therapy.

  2. Proper design of silica nanoparticles combines high brightness, lack of cytotoxicity and efficient cell endocytosis

    NASA Astrophysics Data System (ADS)

    Rampazzo, Enrico; Voltan, Rebecca; Petrizza, Luca; Zaccheroni, Nelsi; Prodi, Luca; Casciano, Fabio; Zauli, Giorgio; Secchiero, Paola

    2013-08-01

    Silica-based luminescent nanoparticles (SiNPs) show promising prospects in nanomedicine in light of their chemical properties and versatility. In this study, we have characterized silica core-PEG shell SiNPs derivatized with PEG moieties (NP-PEG), with external amino- (NP-PEG-amino) or carboxy-groups (NP-PEG-carbo), both in cell cultures as well as in animal models. By using different techniques, we could demonstrate that these SiNPs were safe and did not exhibit appreciable cytotoxicity in different relevant cell models, of normal or cancer cell types, growing either in suspension (JVM-2 leukemic cell line and primary normal peripheral blood mononuclear cells) or in adherence (human hepatocarcinoma Huh7 and umbilical vein endothelial cells). Moreover, by multiparametric flow cytometry, we could demonstrate that the highest efficiency of cell uptake and entry was observed with NP-PEG-amino, with a stable persistence of the fluorescence signal associated with SiNPs in the loaded cell populations both in vitro and in vivo settings suggesting this as an innovative method for cell traceability and detection in whole organisms. Finally, experiments performed with the endocytosis inhibitor Genistein clearly suggested the involvement of a caveolae-mediated pathway in SiNP endocytosis. Overall, these data support the safe use of these SiNPs for diagnostic and therapeutic applications.Silica-based luminescent nanoparticles (SiNPs) show promising prospects in nanomedicine in light of their chemical properties and versatility. In this study, we have characterized silica core-PEG shell SiNPs derivatized with PEG moieties (NP-PEG), with external amino- (NP-PEG-amino) or carboxy-groups (NP-PEG-carbo), both in cell cultures as well as in animal models. By using different techniques, we could demonstrate that these SiNPs were safe and did not exhibit appreciable cytotoxicity in different relevant cell models, of normal or cancer cell types, growing either in suspension (JVM-2 leukemic cell line and primary normal peripheral blood mononuclear cells) or in adherence (human hepatocarcinoma Huh7 and umbilical vein endothelial cells). Moreover, by multiparametric flow cytometry, we could demonstrate that the highest efficiency of cell uptake and entry was observed with NP-PEG-amino, with a stable persistence of the fluorescence signal associated with SiNPs in the loaded cell populations both in vitro and in vivo settings suggesting this as an innovative method for cell traceability and detection in whole organisms. Finally, experiments performed with the endocytosis inhibitor Genistein clearly suggested the involvement of a caveolae-mediated pathway in SiNP endocytosis. Overall, these data support the safe use of these SiNPs for diagnostic and therapeutic applications. Electronic supplementary information (ESI) available: Synthetic procedures, 1H and 13C NMR spectra, TEM and DLS measurements, and absorption and emission spectra. See DOI: 10.1039/c3nr02563b

  3. Laser-ablated titania nanoparticles for aqueous processed hybrid solar cells.

    PubMed

    Körstgens, V; Pröller, S; Buchmann, T; Moseguí González, D; Song, L; Yao, Y; Wang, W; Werhahn, J; Santoro, G; Roth, S V; Iglev, H; Kienberger, R; Müller-Buschbaum, P

    2015-02-01

    Titania nanoparticles are produced by laser ablation in liquid in order to initiate functionalization of titania with the polymer for the active layer. By combining these titania nanoparticles and water-soluble poly[3-(potassium-6-hexanoate)thiophene-2,5-diyl] (P3P6T) hybrid solar cells are realized. PMID:25623567

  4. Biofunctionalized nanoparticles with pH-responsive and cell penetrating blocks for gene delivery

    NASA Astrophysics Data System (ADS)

    Gaspar, V. M.; Marques, J. G.; Sousa, F.; Louro, R. O.; Queiroz, J. A.; Correia, I. J.

    2013-07-01

    Bridging the gap between nanoparticulate delivery systems and translational gene therapy is a long sought after requirement in nanomedicine-based applications. However, recent developments regarding nanoparticle functionalization have brought forward the ability to synthesize materials with biofunctional moieties that mimic the evolved features of viral particles. Herein we report the versatile conjugation of both cell penetrating arginine and pH-responsive histidine moieties into the chitosan polymeric backbone, to improve the physicochemical characteristics of the native material. Amino acid coupling was confirmed by 2D TOCSY NMR and Fourier transform infrared spectroscopy. The synthesized chitosan-histidine-arginine (CH-H-R) polymer complexed plasmid DNA biopharmaceuticals, and spontaneously assembled into stable 105 nm nanoparticles with spherical morphology and positive surface charge. The functionalized delivery systems were efficiently internalized into the intracellular compartment, and exhibited remarkably higher transfection efficiency than unmodified chitosan without causing any cytotoxic effect. Additional findings regarding intracellular trafficking events reveal their preferential escape from degradative lysosomal pathways and nuclear localization. Overall, this assembly of nanocarriers with bioinspired moieties provides the foundations for the design of efficient and customizable materials for cancer gene therapy.

  5. Nanotoxicity of silver nanoparticles to red blood cells: size dependent adsorption, uptake, and hemolytic activity.

    PubMed

    Chen, Li Qiang; Fang, Li; Ling, Jian; Ding, Cheng Zhi; Kang, Bin; Huang, Cheng Zhi

    2015-03-16

    Silver nanoparticles (AgNPs) are increasingly being used as antimicrobial agents and drug carriers in biomedical fields. However, toxicological information on their effects on red blood cells (RBCs) and the mechanisms involved remain sparse. In this article, we examined the size dependent nanotoxicity of AgNPs using three different characteristic sizes of 15 nm (AgNPs15), 50 nm (AgNPs50), and 100 nm (AgNPs100) against fish RBCs. Optical microscopy and transmission electron microscopy observations showed that AgNPs exhibited a size effect on their adsorption and uptake by RBCs. The middle sized AgNPs50, compared with the smaller or bigger ones, showed the highest level of adsorption and uptake by the RBCs, suggesting an optimal size of ?50 nm for passive uptake by RBCs. The toxic effects determined based on the hemolysis, membrane injury, lipid peroxidation, and antioxidant enzyme production were fairly size and dose dependent. In particular, the smallest sized AgNPs15 displayed a greater ability to induce hemolysis and membrane damage than AgNPs50 and AgNPs100. Such cytotoxicity induced by AgNPs should be attributed to the direct interaction of the nanoparticle with the RBCs, resulting in the production of oxidative stress, membrane injury, and subsequently hemolysis. Overall, the results suggest that particle size is a critical factor influencing the interaction between AgNPs and the RBCs. PMID:25602487

  6. Beauty is Skin Deep: A Surface Monolayer Perspective on Nanoparticle Interactions with Cells and Biomacromolecules**

    PubMed Central

    Saha, Krishnendu; Bajaj, Avinash; Duncan, Bradley; Rotello, Vincent M.

    2012-01-01

    Surface recognition of biosystems is a critical component in the development of novel biosensors, delivery vehicles and for the therapeutic regulation of biological processes. Monolayer-protected nanoparticles present a highly versatile scaffold for selective interaction with biomacromolecules and cells. Through engineering of the monolayer surface, nanoparticles can be tailored for surface recognition of biomolecules and cells. This review highlights recent progress in nanoparticle-biomacromolecule/cellular interactions, emphasizing the effect of the surface monolayer structure on the interactions with proteins, DNA and cell surfaces. The extension of these tailored interactions to hybrid nanomaterials, biosensing platforms and delivery vehicles is also discussed. PMID:21671432

  7. Different cell responses induced by exposure to maghemite nanoparticles

    NASA Astrophysics Data System (ADS)

    Luengo, Yurena; Nardecchia, Stefania; Morales, María Puerto; Serrano, M. Concepción

    2013-11-01

    Recent advances in nanotechnology have permitted the development of a wide repertoire of inorganic magnetic nanoparticles (NPs) with extensive promise for biomedical applications. Despite this remarkable potential, many questions still arise concerning the biocompatible nature of NPs when in contact with biological systems. Herein, we have investigated how controlled changes in the physicochemical properties of iron oxide NPs at their surface (i.e., surface charge and hydrodynamic size) affect, first, their interaction with cell media components and, subsequently, cell responses to NP exposure. For that purpose, we have prepared iron oxide NPs with three different coatings (i.e., dimercaptosuccinic acid - DMSA, (3-aminopropyl)triethoxysilane - APS and dextran) and explored the response of two different cell types, murine L929 fibroblasts and human Saos-2 osteoblasts, to their exposure. Interestingly, different cell responses were found depending on the NP concentration, surface charge and cell type. In this sense, neutral NPs, as those coated with dextran, induced negligible cell damage, as their cellular internalization was significantly reduced. In contrast, surface-charged NPs (i.e., those coated with DMSA and APS) caused significant cellular changes in viability, morphology and cell cycle under certain culture conditions, as a result of a more active cellular internalization. These results also revealed a particular cellular ability to detect and remember the original physicochemical properties of the NPs, despite the formation of a protein corona when incubated in culture media. Overall, conclusions from these studies are of crucial interest for future biomedical applications of iron oxide NPs.Recent advances in nanotechnology have permitted the development of a wide repertoire of inorganic magnetic nanoparticles (NPs) with extensive promise for biomedical applications. Despite this remarkable potential, many questions still arise concerning the biocompatible nature of NPs when in contact with biological systems. Herein, we have investigated how controlled changes in the physicochemical properties of iron oxide NPs at their surface (i.e., surface charge and hydrodynamic size) affect, first, their interaction with cell media components and, subsequently, cell responses to NP exposure. For that purpose, we have prepared iron oxide NPs with three different coatings (i.e., dimercaptosuccinic acid - DMSA, (3-aminopropyl)triethoxysilane - APS and dextran) and explored the response of two different cell types, murine L929 fibroblasts and human Saos-2 osteoblasts, to their exposure. Interestingly, different cell responses were found depending on the NP concentration, surface charge and cell type. In this sense, neutral NPs, as those coated with dextran, induced negligible cell damage, as their cellular internalization was significantly reduced. In contrast, surface-charged NPs (i.e., those coated with DMSA and APS) caused significant cellular changes in viability, morphology and cell cycle under certain culture conditions, as a result of a more active cellular internalization. These results also revealed a particular cellular ability to detect and remember the original physicochemical properties of the NPs, despite the formation of a protein corona when incubated in culture media. Overall, conclusions from these studies are of crucial interest for future biomedical applications of iron oxide NPs. Electronic supplementary information (ESI) available: Additional details about NP characterization and viability controls of Saos-2 osteoblasts. See DOI: 10.1039/c3nr02148c

  8. Nonendocytic Delivery of Functional Engineered Nanoparticles into the Cytoplasm of Live Cells Using a Novel, High-Throughput Microfluidic Device

    E-print Network

    Lee, Jungmin

    The ability to straightforwardly deliver engineered nanoparticles into the cell cytosol with high viability will vastly expand the range of biological applications. Nanoparticles could potentially be used as delivery ...

  9. Optimum surface condition for plasmonic Ag nanoparticles in polycrystalline silicon thin film solar cells

    NASA Astrophysics Data System (ADS)

    Park, Jongsung; Park, Nochang; Varlamov, Sergey

    2014-01-01

    Excitation of surface plasmons in silver nanoparticles is a promising method for increasing the light absorption in solar cells and hence, the cell photocurrent. The optical environment is an important key factor to consider when designing plasmonic solar cells because it affects the surface plasmon characteristics. In this paper, we applied the silver nanoparticles on the rear side of polycrystalline silicon thin film solar cells and systematically investigated the optimum surface condition for maximising plasmonic enhanced light absorption in the cells. Three different environments, thermal silicon dioxide (SiO2), native SiO2, and oxide-free silicon surface were investigated. We found that the existence of the SiO2 layer between Si and nanoparticles has a major effect on Qscat and therefore, the absorption in the cells. We also found that nanoparticles on the thermal SiO2 layer showed that the peak of Qscat is located at the visible light wavelengths <700 nm, nanoparticles on the native SiO2 layer and directly on Si sample showed that their peaks of Qscat are positioned at the longer wavelengths >700 nm. The sample with nanoparticles on the native SiO2 layer showed the highest potential short circuit current density (Jsc) enhancement, 62.5%, and absolute Jsc enhancement, 32.3%. On the other hand, the sample with nanoparticles on the thermal SiO2 layer indicated 19.6% enhancement of Jsc.

  10. The Fate of ZnO Nanoparticles Administered to Human Bronchial Epithelial Cells

    PubMed Central

    Gilbert, Benjamin; Fakra, Sirine C.; Xia, Tian; Pokhrel, Suman; Mädler, Lutz; Nel, André E.

    2014-01-01

    A particular challenge for nanotoxicology is the evaluation of the biological fate and toxicity of nanomaterials that dissolve in aqueous fluids. Zinc oxide nanomaterials are of particular concern because dissolution leads to release of the toxic divalent zinc ion. Although dissolved zinc ions have been implicated in ZnO cytotoxicity, direct identification of the chemical form of zinc taken up by cells exposed to ZnO nanoparticles, and its intracellular fate, has not yet been achieved. We combined high resolution X-ray spectromicroscopy and high elemental sensitivity X-ray microprobe analyses to determine the fate of ZnO and less soluble iron-doped ZnO nanoparticles following exposure to cultures of human bronchial epithelial cells, BEAS-2B. We complemented two-dimensional X-ray imaging methods with atomic force microscopy of cell surfaces to distinguish between nanoparticles that were transported inside the cells from those that adhered to the cell exterior. The data suggest cellular uptake of ZnO nanoparticles is a mechanism of zinc accumulation in cells. Following uptake, ZnO nanoparticles dissolved completely generating intracellular Zn2+ complexed by molecular ligands. These results corroborate a model for ZnO nanoparticle toxicity that is based on nanoparticle uptake followed by intracellular dissolution. PMID:22646753

  11. The Role of Dextran Coatings on the Cytotoxicity Properties of Ceria Nanoparticles Toward Bone Cancer Cells

    NASA Astrophysics Data System (ADS)

    Yazici, Hilal; Alpaslan, Ece; Webster, Thomas J.

    2015-03-01

    Cerium oxide nanoparticles have demonstrated great potential as antioxidant and radioprotective agents for nanomedicine applications especially for cancer therapy. The surface chemistry of nanoparticles is an important property that has a significant effect on their performance in biological applications including cancer diagnosis, cancer treatment, and bacterial infection. Recently, various nanosized cerium oxide particles with different types of polymer coatings have been developed to improve aqueous solubility and allow for surface functionalization for distinct applications. In this study, the role of ceria nanoparticles coated with dextran on the cytotoxicity properties of bone cancer cells was shown. Specifically, 0.1 M and 0.01 M dextran-coated, <5-nm ceria nanoparticles, were synthesized. The cytotoxicity of 0.1 M and 0.01 M dextran-coated ceria nanoparticles was evaluated against osteosarcoma cells. A change in cell viability was observed when treating osteosarcoma cells with 0.1 M dextran-coated ceria nanoparticles in the 250-1000 ?g/mL concentration range. In contrast, minimal toxicity to bone cancer cells was observed for the 0.01 M dextran coating after 3 days compared with the 0.1 M dextran coating. These results indicated that surface dextran functionalization had a positive impact on the cytotoxicity of cerium oxide nanoparticles against osteosarcoma cells.

  12. Gold nanoparticles cellular toxicity and recovery: adipose Derived Stromal cells.

    PubMed

    Mironava, Tatsiana; Hadjiargyrou, Michael; Simon, Marcia; Rafailovich, Miriam H

    2014-03-01

    Gold nanoparticles (AuNPs) are currently used in numerous medical applications. Herein, we describe their in vitro impact on human adipose-derived stromal cells (ADSCs) using 13 nm and 45 nm citrate-coated AuNPs. In their non-differentiated state, ADSCs were penetrated by the AuNPs and stored in vacuoles. The presence of the AuNPs in ADSCs resulted in increased population doubling times, decreased cell motility and cell-mediated collagen contraction. The degree to which the cells were impacted was a function of particle concentration, where the smaller particles required a sevenfold higher concentration to have the same effect as the larger ones. Furthermore, AuNPs reduced adipogenesis as measured by lipid droplet accumulation and adiponectin secretion. These effects correlated with transient increases in DLK1 and with relative reductions in fibronectin. Upon removal of exogenous AuNPs, cellular NP levels decreased and normal ADSC functions were restored. As adiponectin helps regulate energy metabolism, local fluctuations triggered by AuNPs can lead to systemic changes. Hence, careful choice of size, concentration and clinical application duration of AuNPs is warranted. PMID:23330784

  13. Single cells from human primary colorectal tumors exhibit polyfunctional heterogeneity in secretions of ELR+ CXC chemokines

    E-print Network

    Adalsteinsson, Viktor A.

    Cancer is an inflammatory disease of tissue that is largely influenced by the interactions between multiple cell types, secreted factors, and signal transduction pathways. While single-cell sequencing continues to refine ...

  14. Synthesis and characterization of folate conjugated chitosan and cellular uptake of its nanoparticles in HT-29 cells.

    PubMed

    Li, Puwang; Wang, Yichao; Zeng, Fanbo; Chen, Lijue; Peng, Zheng; Kong, Ling Xue

    2011-05-01

    Folate-chitosan (FA-CS) conjugates synthesized by coupling FA with CS render new and improved functions because the original properties of CS are maintained and the targeting ligand of FA is incorporated. In this work, FA-CS conjugates were synthesized based on chemical linking of carboxylic group of FA with amino group of CS as confirmed by Fourier transform spectroscopy (FTIR) and nuclear magnetic resonance ((1)H NMR). FA-CS conjugates displayed less crystal nature when compared to CS. The FA-CS nanoparticles (NPs) were prepared by crosslinking FA-CS conjugates with sodium tripolyphosphate (STPP). Positively charged FA-CS nanoparticles were spherical in shape with a particle size of about 100 nm. Cellular uptake of CS or FA-CS nanoparticles was assayed by fluorescent microscopy using calcein as fluorescent marker in colon cancer cells (HT-29). The FA-CS nanoparticles exhibited improved uptake of HT-29 and could become a potential targeted drug delivery system for colorectal cancer. PMID:21397214

  15. In vitro toxicity of iron oxide nanoparticle: oxidative damages on Hep G2 cells.

    PubMed

    Sadeghi, Leila; Tanwir, Farzeen; Yousefi Babadi, Vahid

    2015-02-01

    During the past years many studies have been done highlighting the great need for a more thorough understanding of cell-iron oxide nanoparticle interactions. To improve our knowledge in this field, there is a great need for standardized protocols that would allow to comparing the cytotoxic potential of any Fe2O3-NP type with previously studied particles. Several approaches are reported that several parameters which are of great importance for Fe2O3 nanoparticle induced toxicity. Nanoparticles because of their very small size can pass through the cell membrane and can make oxidative damage in all parts of the cells such as mitochondria, membrane, DNA due to high surface area. This study focuses on acute cytotoxicity of reactive oxygen species and DNA damaging effects of mentioned nanoparticles. Results showed increase of the oxidative damage leads cells to the apoptosis, therefore reduced cell viability. It is interesting that all of the results are concentration and time dependent. PMID:25497787

  16. Study of gold nanoparticles and live cells interactions by using planar evanescent wave excitation

    NASA Astrophysics Data System (ADS)

    Lee, Chia-Wei; Lin, En-Hong; Cheng, Ji-Yen; Wei, Pei-Kuen

    2009-03-01

    We present a planar evanescent wave (PEW) technique combined with phase contrast optical microscopy to study the interactions between cells and gold nanoparticles (AuNPs). The PEW method employs a dual-fiber-line guide to couple light into a thin glass slide. It produces a uniform and long evanescent wave near the glass surface, as verified by the optical near-field measurement. High-contrast AuNP images are obtained by the PEW illumination. At the same time, cells are observed only by using the phase contrast microscopy. The nanoparticles and cell images indicated that unmodified AuNPs had no interactions with cells, possibly due to the negative surface charges on both cells and nanoparticles. The electrostatic concept was further verified by coating AuNPs with positively charged poly (L-lysine). DNA aptamers for surface mucin glycoprotein were coated on AuNPs to demonstrate the application for single nanoparticle tracking.

  17. Metallofullerene nanoparticles promote osteogenic differentiation of bone marrow stromal cells through BMP signaling pathway.

    PubMed

    Yang, Kangning; Cao, Weipeng; Hao, Xiaohong; Xue, Xue; Zhao, Jing; Liu, Juan; Zhao, Yuliang; Meng, Jie; Sun, Baoyun; Zhang, Jinchao; Liang, Xing-Jie

    2013-02-01

    Although endohedral metallofullerenol [Gd@C(82)(OH)(22)](n) nanoparticles have anti-tumor efficiency and mostly deposit in the bones of mice, how these nanoparticles act in bone marrow stromal cells (MSCs) remains largely unknown. Herein, we observed that [Gd@C(82)(OH)(22)](n) nanoparticles facilitated the differentiation of MSCs toward osteoblasts, as evidenced by the enhancement of alkaline phosphatase (ALP) activity and mineralized nodule formation upon [Gd@C(82)(OH)(22)](n) nanoparticle treatment. Mechanistically, the effect of [Gd@C(82)(OH)(22)](n) nanoparticles on ALP activity was inhibited by the addition of noggin as an inhibitor of the BMP signaling pathway. Moreover, the in vivo results of the ovariectomized rats further indicated that [Gd@C(82)(OH)(22)](n) nanoparticles effectively improved bone density and prevented osteoporosis. PMID:23299786

  18. Cells From Long-Lived Mutant Mice Exhibit Enhanced Repair of Ultraviolet Lesions

    Microsoft Academic Search

    Adam B. Salmon; Mats Ljungman; Richard A. Miller

    2008-01-01

    Fibroblasts isolated from long-lived hypopituitary dwarf mice are resistant to many cell stresses, including ultraviolet (UV) light and methyl methane sulfonate (MMS), which induce cell death by producing DNA damage. Here we report that cells from Snell dwarf mice recover more rapidly than controls from the inhibition of RNA synthesis induced by UV damage. Recovery of messenger RNA (mRNA) synthesis

  19. Superparamagnetic MFe2O 4 (M = Ni, Co, Zn, Mn) nanoparticles: synthesis, characterization, induction heating and cell viability studies for cancer hyperthermia applications.

    PubMed

    Sabale, Sandip; Jadhav, Vidhya; Khot, Vishwajeet; Zhu, Xiaoli; Xin, Meiling; Chen, Hongxia

    2015-03-01

    Superparamagnetic nanoferrites are prepared by simple and one step refluxing in polyol synthesis. The ferrite nanoparticles prepared by this method exhibit particle sizes below 10 nm and high degree of crystallinity. These ferrite nanoparticles are compared by means of their magnetic properties, induction heating and cell viability studies for its application in magnetic fluid hyperthermia. Out of all studied nanoparticles in present work, only ZnFe2O4 and CoFe2O4 MNPs are able to produce threshold hyperthermia temperature. This rise in temperature is discussed in detail in view of their magneto-structural properties. Therefore ZnFe2O4 and CoFe2O4 MNPs with improved stability, magnetic induction heating and cell viability are suitable candidates for magnetic hyperthermia. PMID:25690622

  20. CYP26A1 knockout embryonic stem cells exhibit reduced differentiation and growth arrest in response to retinoic acid.

    PubMed

    Langton, Simne; Gudas, Lorraine J

    2008-03-15

    CYP26A1, a cytochrome P450 enzyme, metabolizes all-trans-retinoic acid (RA) into polar metabolites, e.g. 4-oxo-RA and 4-OH-RA. To determine if altering RA metabolism affects embryonic stem (ES) cell differentiation, we disrupted both alleles of Cyp26a1 by homologous recombination. CYP26a1(-/-) ES cells had a 11.0+/-3.2-fold higher intracellular RA concentration than Wt ES cells after RA treatment for 48 h. RA-treated CYP26A1(-/-) ES cells exhibited 2-3 fold higher mRNA levels of Hoxa1, a primary RA target gene, than Wt ES cells. Despite increased intracellular RA levels, CYP26a1(-/-) ES cells were more resistant than Wt ES cells to RA-induced proliferation arrest. Transcripts for parietal endodermal differentiation markers, including laminin, J6(Hsp 47), and J31(SPARC, osteonectin) were expressed at lower levels in RA-treated CYP26a1(-/-) ES cells, indicating that the lack of CYP26A1 activity inhibits RA-associated differentiation. Microarray analyses revealed that RA-treated CYP26A1(-/-) ES cells exhibited lower mRNA levels than Wt ES cells for genes involved in differentiation, particularly in neural (Epha4, Pmp22, Nrp1, Gap43, Ndn) and smooth muscle differentiation (Madh3, Nrp1, Tagln Calponin, Caldesmon1). In contrast, genes involved in the stress response (e.g. Tlr2, Stk2, Fcgr2b, Bnip3, Pdk1) were expressed at higher levels in CYP26A1(-/-) than in Wt ES cells without RA. Collectively, our results show that CYP26A1 activity regulates intracellular RA levels, cell proliferation, transcriptional regulation of primary RA target genes, and ES cell differentiation to parietal endoderm. PMID:18241852

  1. Release of magnetic nanoparticles from cell-encapsulating biodegradable nanobiomaterials.

    PubMed

    Xu, Feng; Inci, Fatih; Mullick, Omer; Gurkan, Umut Atakan; Sung, Yuree; Kavaz, Doga; Li, Baoqiang; Denkbas, Emir Baki; Demirci, Utkan

    2012-08-28

    The future of tissue engineering requires development of intelligent biomaterials using nanoparticles. Magnetic nanoparticles (MNPs) have several applications in biology and medicine; one example is Food and Drug Administration (FDA)-approved contrast agents in magnetic resonance imaging. Recently, MNPs have been encapsulated within cell-encapsulating hydrogels to create novel nanobiomaterials (i.e., M-gels), which can be manipulated and assembled in magnetic fields. The M-gels can be used as building blocks for bottom-up tissue engineering to create 3D tissue constructs. For tissue engineering applications of M-gels, it is essential to study the release of encapsulated MNPs from the hydrogel polymer network and the effect of MNPs on hydrogel properties, including mechanical characteristics, porosity, swelling behavior, and cellular response (e.g., viability, growth). Therefore, we evaluated the release of MNPs from photocrosslinkable gelatin methacrylate hydrogels as the polymer network undergoes biodegradation using inductively coupled plasma atomic emission spectroscopy. MNP release correlated linearly with hydrogel biodegradation rate with correlation factors (Pearson product moment correlation coefficient) of 0.96 ± 0.03 and 0.99 ± 0.01 for MNP concentrations of 1% and 5%, respectively. We also evaluated the effect of MNPs on hydrogel mechanical properties, porosity, and swelling behavior, as well as cell viability and growth in MNP-encapsulating hydrogels. Fibroblasts encapsulated with MNPs in hydrogels remained viable (>80% at t = 144 h) and formed microtissue constructs in culture (t = 144 h). These results indicated that MNP-encapsulating hydrogels show promise as intelligent nanobiomaterials, with great potential to impact broad areas of bioengineering, including tissue engineering, regenerative medicine, and pharmaceutical applications. PMID:22680777

  2. Translocation of Cell Penetrating Peptide Engrafted Nanoparticles Across Skin Layers

    PubMed Central

    Patlolla, Ram R; Desai, Pinaki; Belay, Kalayu; Singh, Mandip

    2010-01-01

    The objective of the current study was to evaluate the ability of cell penetrating peptides (CPP) to translocate the lipid payload into the skin layers. Fluorescent dye (DID-oil) encapsulated nano lipid crystal nanoparticles (FNLCN) were prepared using Compritol, Miglyol and DOGS-NTA-Ni lipids by hot melt homogenization technique. The FNLCN surface was coated with TAT peptide (FNLCNT) or control YKA peptide (FNLCNY) and in vitro rat skin permeation studies were performed using Franz diffusion cells. Observation of lateral skin sections obtained using cryotome with a confocal microscope demonstrated that skin permeation of FNLCNT was time dependent and after 24 h, fluorescence was observed upto a depth of 120 µm which was localized in the hair follicles and epidermis. In case of FNLCN and FNLCNY formulations fluorescence was mainly observed in the hair follicles. This observation was further supported by confocal Raman spectroscopy where higher fluorescence signal intensity was observed at 80 and 120 µm depth with FNLCNT treated skin and intensity of fluorescence peaks was in the ratio of 2:1:1 and 5:3:1 for FNLCNT, FNLCN, and FNLCNY treated skin sections, respectively. Furthermore, replacement of DID-oil with celecoxib (Cxb), a model lipophilic drug showed similar results and after 24 h, the CXBNT formulation increased the Cxb concentration in SC by 3 and 6 fold and in epidermis by 2 and 3 fold as compared to CXBN and CXBNY formulations respectively. Our results strongly suggest that CPP can translocate nanoparticles with their payloads into deeper skin layers. PMID:20413152

  3. Release of Magnetic Nanoparticles from Cell-Encapsulating Biodegradable Nanobiomaterials

    PubMed Central

    Xu, Feng; Inci, Fatih; Mullick, Omer; Gurkan, Umut Atakan; Sung, Yuree; Kavaz, Doga; Li, Baoqiang; Denkbas, Emir Baki; Demirci, Utkan

    2013-01-01

    The future of tissue engineering requires development of intelligent biomaterials using nanoparticles. Magnetic nanoparticles (MNPs) have several applications in biology and medicine; one example is Food and Drug Administration (FDA)-approved contrast agents in magnetic resonance imaging. Recently, MNPs have been encapsulated within cell-encapsulating hydrogels to create novel nanobiomaterials (i.e., M-gels), which can be manipulated and assembled in magnetic fields. The M-gels can be used as building blocks for bottom-up tissue engineering to create 3D tissue constructs. For tissue engineering applications of M-gels, it is essential to study the release of encapsulated MNPs from the hydrogel polymer network and the effect of MNPs on hydrogel properties, including mechanical characteristics, porosity, swelling behavior, and cellular response (e.g., viability, growth). Therefore, we evaluated the release of MNPs from photocrosslinkable gelatin methacrylate hydrogels as the polymer network undergoes biodegradation using inductively coupled plasma atomic emission spectroscopy. MNP release correlated linearly with hydrogel biodegradation rate with correlation factors (Pearson product moment correlation coefficient) of 0.96 ± 0.03 and 0.99 ± 0.01 for MNP concentrations of 1% and 5%, respectively. We also evaluated the effect of MNPs on hydrogel mechanical properties, porosity, and swelling behavior, as well as cell viability and growth in MNP-encapsulating hydrogels. Fibroblasts encapsulated with MNPs in hydrogels remained viable (>80% at t = 144 h) and formed microtissue constructs in culture (t = 144 h). These results indicated that MNP-encapsulating hydrogels show promise as intelligent nanobiomaterials, with great potential to impact broad areas of bioengineering, including tissue engineering, regenerative medicine, and pharmaceutical applications. PMID:22680777

  4. Cells derived from normal or cancer breast tissue exhibit different growth properties when deprived of arginine.

    PubMed

    Chiaviello, Angela; Paciello, Ida; Veneziani, Bianca Maria; Palumbo, Giuseppe; Aloj, Salvatore M

    2012-12-01

    Arginine deprivation impairs cell proliferation more strong in cancer than in normal cells; thus, it has been proposed that such an effect could be exploited for cancer therapy. We have compared the effect of arginine deprivation on normal and cancer cells, studying growth rate, morphology, and protein expression patterns in immortalized human MCF10a cells and in MCF7 cells. Arginine deprivation forces MCF10a cells into irreversible senescence while the vast majority of MCF7 cells become quiescent and resume normal growth following arginine re-addition. Arginine deprivation induced a significant burst of p21cip1 in both cell lines that were reversible in MCF7 and irreversible in MCF10 cells. In the latter cells, p21cip1 increase was accompanied by a time-dependent increase of p16INK4A. Similar effects could be obtained by treating both cell types with ?-difluoro-methyl-ornithine, but not with N?-hydroxy-L-arginine, drugs that interfere specifically but differently with the major pathways of arginine metabolism. Our data suggest that derangement in polyamine synthesis is the main consequence of arginine starvation. PMID:22183716

  5. Laser-induced cell detachment, patterning, and regrowth on gold nanoparticle functionalized surfaces.

    PubMed

    Kolesnikova, Tatiana A; Kohler, Dorothee; Skirtach, Andre G; Möhwald, Helmuth

    2012-11-27

    We report on the selective cell detachment from nanoengineered gold nanoparticle (AuNP) surfaces triggered by laser irradiation, which occurs in a nonthermal manner. The gold nanoparticle-based surfaces reveal good adhesion of NIH3T3 fibroblast cells. Patterning is achieved by lithographic microcontact printing, selective gold nanoparticle deposition, and by laser beam profiling. It is shown that the effectiveness of fibroblast cell detachment depends on the cell age, laser power, and AuNP patterning profile. Heat distribution and temperature rise around gold nanoparticle functionalized surfaces is modeled, revealing low heating of nanoparticles by laser illumination. The nonthermal photochemical mechanism of cell detachment due to production of reactive oxygen species under illumination of gold nanoparticles by green laser light is studied. We also demonstrate that cells migrate from unirradiated areas leading to their reattachment and surface recovery which is important for controlled spatial organization of cells in wound healing and tissue engineering. Research presented in this work is targeted at designing biointerfaces for cell cultures. PMID:23066742

  6. Effect of hydroxyapatite nanoparticles on osmotic responses of pig iliac endothelial cells.

    PubMed

    Yue, Cui; Zhao, Gang; Yi, Jingru; Gao, Cai; Shen, Lingxiao; Zhang, Yuntian; Wang, Zhen; Liu, Wei

    2014-10-01

    In order to fully explore the potential applications of nanoparticles in biopreservation, it is necessary to study the effect of nanoparticles on cell membrane permeabilities. The aim of this study is therefore to comparatively evaluate the osmotic responses of pig iliac endothelial cells in the absence and presence of commercially available hydroxyapatite nanoparticles. The results indicate that, after the introduction of 0.0 1 wt% hydroxyapatite nanoparticles, the dependence of cell membrane hydraulic conductivity (Lp) on temperature still obeys the Arrhenius relationship, while the reference value of the hydraulic conductivity of the cell membrane at 273.15K (Lpg) and the activation energy for water transport across cell membrane (ELp) change from 0.77 × 10(-14)m/Pa/s and 15.65 kJ/mol to 0.65 × 10(-14)m/Pa/s and 26.14 kJ/mol. That is to say, the reference value of the hydraulic conductivity of the cell membrane has been slightly decreased while the activation energy for water transport across cell membrane has been greatly enhanced, and thus it implies that the hydraulic conductivity of cell membrane are more sensitive to temperature in the presence of nanoparticles. These findings are of potential significance to the optimization of nanoparticles-aided cryopreservation. PMID:25111088

  7. Metallofullerene nanoparticles promote osteogenic differentiation of bone marrow stromal cells through BMP signaling pathway

    NASA Astrophysics Data System (ADS)

    Yang, Kangning; Cao, Weipeng; Hao, Xiaohong; Xue, Xue; Zhao, Jing; Liu, Juan; Zhao, Yuliang; Meng, Jie; Sun, Baoyun; Zhang, Jinchao; Liang, Xing-Jie

    2013-01-01

    Although endohedral metallofullerenol [Gd@C82(OH)22]n nanoparticles have anti-tumor efficiency and mostly deposit in the bones of mice, how these nanoparticles act in bone marrow stromal cells (MSCs) remains largely unknown. Herein, we observed that [Gd@C82(OH)22]n nanoparticles facilitated the differentiation of MSCs toward osteoblasts, as evidenced by the enhancement of alkaline phosphatase (ALP) activity and mineralized nodule formation upon [Gd@C82(OH)22]n nanoparticle treatment. Mechanistically, the effect of [Gd@C82(OH)22]n nanoparticles on ALP activity was inhibited by the addition of noggin as an inhibitor of the BMP signaling pathway. Moreover, the in vivo results of the ovariectomized rats further indicated that [Gd@C82(OH)22]n nanoparticles effectively improved bone density and prevented osteoporosis.Although endohedral metallofullerenol [Gd@C82(OH)22]n nanoparticles have anti-tumor efficiency and mostly deposit in the bones of mice, how these nanoparticles act in bone marrow stromal cells (MSCs) remains largely unknown. Herein, we observed that [Gd@C82(OH)22]n nanoparticles facilitated the differentiation of MSCs toward osteoblasts, as evidenced by the enhancement of alkaline phosphatase (ALP) activity and mineralized nodule formation upon [Gd@C82(OH)22]n nanoparticle treatment. Mechanistically, the effect of [Gd@C82(OH)22]n nanoparticles on ALP activity was inhibited by the addition of noggin as an inhibitor of the BMP signaling pathway. Moreover, the in vivo results of the ovariectomized rats further indicated that [Gd@C82(OH)22]n nanoparticles effectively improved bone density and prevented osteoporosis. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr33575a

  8. Effect of silver nanoparticles on human mesenchymal stem cell differentiation

    PubMed Central

    Diendorf, Jörg; Epple, Matthias; Schildhauer, Thomas A; Köller, Manfred

    2014-01-01

    Summary Background: Silver nanoparticles (Ag-NP) are one of the fastest growing products in nano-medicine due to their enhanced antibacterial activity at the nanoscale level. In biomedicine, hundreds of products have been coated with Ag-NP. For example, various medical devices include silver, such as surgical instruments, bone implants and wound dressings. After the degradation of these materials, or depending on the coating technique, silver in nanoparticle or ion form can be released and may come into close contact with tissues and cells. Despite incorporation of Ag-NP as an antibacterial agent in different products, the toxicological and biological effects of silver in the human body after long-term and low-concentration exposure are not well understood. In the current study, we investigated the effects of both ionic and nanoparticulate silver on the differentiation of human mesenchymal stem cells (hMSCs) into adipogenic, osteogenic and chondrogenic lineages and on the secretion of the respective differentiation markers adiponectin, osteocalcin and aggrecan. Results: As shown through laser scanning microscopy, Ag-NP with a size of 80 nm (hydrodynamic diameter) were taken up into hMSCs as nanoparticulate material. After 24 h of incubation, these Ag-NP were mainly found in the endo-lysosomal cell compartment as agglomerated material. Cytotoxicity was observed for differentiated or undifferentiated hMSCs treated with high silver concentrations (?20 µg·mL?1 Ag-NP; ?1.5 µg·mL?1 Ag+ ions) but not with low-concentration treatments (?10 µg·mL?1 Ag-NP; ?1.0 µg·mL?1 Ag+ ions). Subtoxic concentrations of Ag-NP and Ag+ ions impaired the adipogenic and osteogenic differentiation of hMSCs in a concentration-dependent manner, whereas chondrogenic differentiation was unaffected after 21 d of incubation. In contrast to aggrecan, the inhibitory effect of adipogenic and osteogenic differentiation was confirmed by a decrease in the secretion of specific biomarkers, including adiponectin (adipocytes) and osteocalcin (osteoblasts). Conclusion: Aside from the well-studied antibacterial effect of silver, little is known about the influence of nano-silver on cell differentiation processes. Our results demonstrate that ionic or nanoparticulate silver attenuates the adipogenic and osteogenic differentiation of hMSCs even at non-toxic concentrations. Therefore, more studies are needed to investigate the effects of silver species on cells at low concentrations during long-term treatment. PMID:25551033

  9. NANOPARTICLES AND THEIR APPLICATIONS IN CELL AND MOLECULAR BIOLOGY

    PubMed Central

    Wang, Edina C.; Wang, Andrew Z.

    2013-01-01

    Nanoparticles can be engineered with distinctive compositions, sizes, shapes, and surface chemistries to enable novel techniques in a wide range of biological applications. The unique properties of nanoparticles and their behavior in biological milieu also enable exciting and integrative approaches to studying fundamental biological questions. This review will provide an overview of various types of nanoparticles and concepts of targeting nanoparticles. We will also discuss the advantages and recent applications of using nanoparticles as tools for drug delivery, imaging, sensing, and for the understanding of basic biological processes. PMID:24104563

  10. Paracrine signalling events in embryonic stem cell renewal mediated by affinity targeted nanoparticles

    E-print Network

    Fahmy, Tarek

    Paracrine signalling events in embryonic stem cell renewal mediated by affinity targeted: Embryonic stem cell Biodegradable nanoparticles Paracrine delivery Renewal a b s t r a c t Stem cell growth and pluripotency of mouse embryonic stem cells. Leukaemia Inhibitory Factor (LIF) was encapsulated in biodegradable

  11. Mapping force of interaction between PLGA nanoparticle with cell membrane using optical tweezers

    NASA Astrophysics Data System (ADS)

    Chhajed, Suyash; Gu, Ling; Homayoni, Homa; Nguyen, Kytai; Mohanty, Samarendra

    2011-03-01

    Drug delivery using magnetic (Fe3O4) Poly Lactic-co-Glycolic Acid (PLGA) nanoparticles is finding increasing usage in therapeutic applications due to its biodegradability, biocompatibility and targeted localization. Since optical tweezers allow non-contact, highly sensitive force measurement, we utilized optical tweezers for studying interaction forces between the Fe3O4-PLGA nanoparticles with prostate cancer PC3 cells. Presence of Fe3O4 within the PLGA shell allowed efficient trapping of these nanoparticles in near-IR optical tweezers. The conglomerated PLGA nanoparticles could be dispersed by use of the optical tweezers. Calibration of trapping stiffness as a function of laser beam power was carried out using equipartition theorem method, where the mean square displacement was measured with high precision using time-lapse fluorescence imaging of the nanoparticles. After the trapped PLGA nanoparticle was brought in close vicinity of the PC3 cell membrane, displacement of the nanoparticle from trap center was measured as a function of time. In short time scale (< 30sec), while the force of interaction was within 0.2 pN, the force increased beyond 1pN at longer time scales (˜ 10 min). We will present the results of the time-varying force of interactions between PLGA nanoparticles with PC3 cells using optical tweezers.

  12. Lead sulfide nanoparticles increase cell wall chitin content and induce apoptosis in Saccharomyces cerevisiae.

    PubMed

    Sun, Meiqing; Yu, Qilin; Hu, Mengyuan; Hao, Zhenwei; Zhang, Chengdong; Li, Mingchun

    2014-05-30

    Although there have been numerous studies on bacterial toxicity, the cytotoxicity of nanoparticles toward fungi remains poorly understood. We investigated the toxicity of various sizes of lead sulfide particles against the important model fungus, Saccharomyces cerevisiae. The smallest particle exerted the highest toxicity, inhibiting cell growth and decreasing cell viability, likely reflecting reduced sedimentation and persistent cell wall attack. In response to cell wall stress, S. cerevisiae showed an increase in the cell wall chitin content and the overexpression of FKS2 and PRM5, two genes of the cell wall integrity signaling pathway. Cell wall stress increased the concentration of intracellular reactive oxygen species, leading to mitochondrial dysfunction and cell apoptosis. The contribution of dissolved lead ions to the overall toxicity was negligible. These findings provide the first demonstration of the physiological protective response of a fungus toward nanoparticles, thereby contributing useful information to the assessment of the environmental impact of metal nanoparticles. PMID:24704549

  13. Fixed endothelial cells exhibit physiologically relevant nanomechanics of the cortical actin web

    NASA Astrophysics Data System (ADS)

    Bodo Grimm, Kai; Oberleithner, Hans; Fels, Johannes

    2014-05-01

    It has been unknown whether cells retain their mechanical properties after fixation. Therefore, this study was designed to compare the stiffness properties of the cell cortex (the 50-100 nm thick zone below the plasma membrane) before and after fixation. Atomic force microscopy was used to acquire force indentation curves from which the nanomechanical cell properties were derived. Cells were pretreated with different concentrations of actin destabilizing agent cytochalasin D, which results in a gradual softening of the cell cortex. Then cells were studied ‘alive’ or ‘fixed’. We show that the cortical stiffness of fixed endothelial cells still reports functional properties of the actin web qualitatively comparable to those of living cells. Myosin motor protein activity, tested by blebbistatin inhibition, can only be detected, in terms of cortical mechanics, in living but not in fixed cells. We conclude that fixation interferes with motor proteins while maintaining a functional cortical actin web. Thus, fixation of cells opens up the prospect of differentially studying the actions of cellular myosin and actin.

  14. CD4+NKG2D+ T Cells Exhibit Enhanced Migratory and Encephalitogenic Properties in Neuroinflammation

    PubMed Central

    Ruck, Tobias; Bittner, Stefan; Gross, Catharina C.; Breuer, Johanna; Albrecht, Stefanie; Korr, Sabrina; Göbel, Kerstin; Pankratz, Susann; Henschel, Christian M.; Schwab, Nicholas; Staszewski, Ori; Prinz, Marco; Kuhlmann, Tanja

    2013-01-01

    Migration of encephalitogenic CD4+ T lymphocytes across the blood-brain barrier is an essential step in the pathogenesis of multiple sclerosis (MS). We here demonstrate that expression of the co-stimulatory receptor NKG2D defines a subpopulation of CD4+ T cells with elevated levels of markers for migration, activation, and cytolytic capacity especially when derived from MS patients. Furthermore, CD4+NKG2D+ cells produce high levels of proinflammatory IFN-? and IL-17 upon stimulation. NKG2D promotes the capacity of CD4+NKG2D+ cells to migrate across endothelial cells in an in vitro model of the blood-brain barrier. CD4+NKG2D+ T cells are enriched in the cerebrospinal fluid of MS patients, and a significant number of CD4+ T cells in MS lesions coexpress NKG2D. We further elucidated the role of CD4+NKG2D+ T cells in the mouse system. NKG2D blockade restricted central nervous system migration of T lymphocytes in vivo, leading to a significant decrease in the clinical and pathologic severity of experimental autoimmune encephalomyelitis, an animal model of MS. Blockade of NKG2D reduced killing of cultivated mouse oligodendrocytes by activated CD4+ T cells. Taken together, we identify CD4+NKG2D+ cells as a subpopulation of T helper cells with enhanced migratory, encephalitogenic and cytotoxic properties involved in inflammatory CNS lesion development. PMID:24282598

  15. Middle T antigen-transformed endothelial cells exhibit an increased activity of nitric oxide synthase

    PubMed Central

    1995-01-01

    Endothelioma cell lines transformed by polyoma virus middle T antigen (mTa) cause cavernous hemangiomas in syngeneic mice by recruitment of host cells. The production of nitric oxide (NO), as measured by nitrite and citrulline production, was significantly higher in mTa-transformed endothelial cells in comparison with nontransformed control cells. The maximal activity of NO synthase (NOS) was about 200-fold higher in cell lysates from the tEnd.1 endothelioma cell line than in lysates from nontransformed controls, whereas the affinity for arginine did not differ. The biochemical characterization of NOS and the study of mRNA transcripts indicate that tEnd.1 cells express both the inducible and the constitutive isoforms. NOS hyperactivity is not a simple consequence of cell transformation but needs a tissue-specific mTa expression. Since tEnd.1-conditioned medium induces NOS activity in normal endothelial cells, most likely NOS hyperactivity in endothelioma cells is attributable to the release of a soluble factor. This NOS- activating factor, which seems to be an anionic protein, could stimulate tEnd.1 cells to express NOS by an autocrine way. By the same mechanism, tEnd.1 cells could induce NOS in the neighboring endothelial cells, and NO release could play a role in the hemangioma development. Such hypothesis is confirmed by our in vivo experiments, showing that the administration of the NOS inhibitor L-canavanine to endothelioma- bearing mice significantly reduced both the volume and the relapse time of the tumor. PMID:7528781

  16. One-step bulk synthesis of stable, near unit-cell sized oxide nanoparticles and nanoparticle blends using KO2.

    PubMed

    Sutto, Thomas E

    2014-05-01

    Presented here is a novel one-step synthesis of oxide or hydroxide nanoparticles using, for the first time, potassium superoxide (KO2). This work demonstrates that the reaction of KO2 with different salt solutions produces grams of stable, near unit-cell sized nanoparticles. This new synthetic technique is applied to representative elements from across the periodic table to rapidly produce nanometer sized oxides or hydroxides of Mg, Al, Y, Ti, Mn, Fe, Co, Ni, Cu, Zn, Sn, Tl, Pb, and Ce. This technique is also used to produce blends of nanoparticles, demonstrating the ability to prepare complex materials such as nanoparticulate blends of a lithium cathode material (LiCoO2), the multiferroic compound (BiMnO(3+?)), and the superconducting YBa2Cu3O(7-?). PMID:24724979

  17. Aging Hematopoietic Stem Cells Decline in Function and Exhibit Epigenetic Dysregulation

    Microsoft Academic Search

    Stuart M Chambers; Chad A Shaw; Catherine Gatza; C. Joseph Fisk; Lawrence A Donehower; Margaret A Goodell

    2007-01-01

    Age-related defects in stem cells can limit proper tissue maintenance and hence contribute to a shortened lifespan. Using highly purified hematopoietic stem cells from mice aged 2 to 21 mo, we demonstrate a deficit in function yet an increase in stem cell number with advancing age. Expression analysis of more than 14,000 genes identified 1,500 that were age-induced and 1,600

  18. Prostate Cancer Cell Lines under Hypoxia Exhibit Greater Stem-Like Properties

    PubMed Central

    Ma, Yuanyuan; Liang, Dongming; Liu, Jian; Axcrona, Karol; Kvalheim, Gunnar; Stokke, Trond; Nesland, Jahn M.; Suo, Zhenhe

    2011-01-01

    Hypoxia is an important environmental change in many cancers. Hypoxic niches can be occupied by cancer stem/progenitor-like cells that are associated with tumor progression and resistance to radiotherapy and chemotherapy. However, it has not yet been fully elucidated how hypoxia influences the stem-like properties of prostate cancer cells. In this report, we investigated the effects of hypoxia on human prostate cancer cell lines, PC-3 and DU145. In comparison to normoxia (20% O2), 7% O2 induced higher expressions of HIF-1? and HIF-2?, which were associated with upregulation of Oct3/4 and Nanog; 1% O2 induced even greater levels of these factors. The upregulated NANOG mRNA expression in hypoxia was confirmed to be predominantly retrogene NANOGP8. Similar growth rates were observed for cells cultivated under hypoxic and normoxic conditions for 48 hours; however, the colony formation assay revealed that 48 hours of hypoxic pretreatment resulted in the formation of more colonies. Treatment with 1% O2 also extended the G0/G1 stage, resulting in more side population cells, and induced CD44 and ABCG2 expressions. Hypoxia also increased the number of cells positive for ABCG2 expression, which were predominantly found to be CD44bright cells. Correspondingly, the sorted CD44bright cells expressed higher levels of ABCG2, Oct3/4, and Nanog than CD44dim cells, and hypoxic pretreatment significantly increased the expressions of these factors. CD44bright cells under normoxia formed significantly more colonies and spheres compared with the CD44dim cells, and hypoxic pretreatment even increased this effect. Our data indicate that prostate cancer cells under hypoxia possess greater stem-like properties. PMID:22216200

  19. Antioxidant activity of Vaccinium stamineum: exhibition of anticancer capability in human lung and leukemia cells.

    PubMed

    Wang, Shiow Y; Feng, Rentian; Bowman, Linda; Lu, Yongju; Ballington, James R; Ding, Min

    2007-05-01

    Fruit of deerberry [Vaccinium stamineum L.] were evaluated for their antioxidant capacity and anticancer properties in JB6 P (+) mouse epidermal cells, human lung and leukemia cells. Deerberries contain potent free radical scavenging activities. Pretreatment of JB6 P (+) mouse epidermal cells with deerberry fruit extracts produced an inhibition on the activation of activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) induced by either 12- O-tetradecanoylphorbol 13-acetate (TPA) or ultraviolet-B (UVB). Deerberry fruit extracts also blocked TPA- or UVB-induced phosphorylation of ERKs and MEK 1/2, two upstream regulators of AP-1 and inhibited proliferation of human leukemia HL-60 cancer cells and human lung epithelial cancer A549 cells and induced apoptosis of HL-60 cells. These results suggest that the inhibition of TPA- or UVB-induced AP-1 and NF-kappaB activity, inhibition of HL-60 cells and cancer A549 cells proliferation and induction of apoptotic in human leukemia HL-60 cancer cells may be mediated through the ERKs and MEK 1/2 signal pathway. PMID:17394101

  20. Developmental factor IRF6 exhibits tumor suppressor activity in squamous cell carcinomas.

    PubMed

    Botti, Elisabetta; Spallone, Giulia; Moretti, Francesca; Marinari, Barbara; Pinetti, Valentina; Galanti, Sergio; De Meo, Paolo D'Onorio; De Nicola, Francesca; Ganci, Federica; Castrignanò, Tiziana; Pesole, Graziano; Chimenti, Sergio; Guerrini, Luisa; Fanciulli, Maurizio; Blandino, Giovanni; Karin, Michael; Costanzo, Antonio

    2011-08-16

    The transcription factor interferon regulatory factor 6 (IRF6) regulates craniofacial development and epidermal proliferation. We recently showed that IRF6 is a component of a regulatory feedback loop that controls the proliferative potential of epidermal cells. IRF6 is transcriptionally activated by p63 and induces its proteasome-mediated down-regulation, thereby limiting keratinocyte proliferative potential. We hypothesized that IRF6 may also be involved in skin carcinogenesis. Hence, we analyzed IRF6 expression in a large series of squamous cell carcinomas (SCCs) and found a strong down-regulation of IRF6 that correlated with tumor invasive and differentiation status. IRF6 down-regulation in SCC cell lines and primary tumors correlates with methylation on a CpG dinucleotide island located in its promoter region. To identify the molecular mechanisms regulating IRF6 potential tumor suppressive activity, we performed a genome-wide analysis by combining ChIP sequencing for IRF6 binding sites and gene expression profiling in primary human keratinocytes after siRNA-mediated IRF6 depletion. We observed dysregulation of cell cycle-related genes and genes involved in differentiation, cell adhesion, and cell-cell contact. Many of these genes were direct IRF6 targets. We also performed in vitro invasion assays showing that IRF6 down-regulation promotes invasive behavior and that reintroduction of IRF6 into SCC cells strongly inhibits cell growth. These results indicate a function for IRF6 in suppression of tumorigenesis in stratified epithelia. PMID:21807998

  1. Hematopoietic and mesenchymal stem cells: polymeric nanoparticle uptake and lineage differentiation

    PubMed Central

    Brüstle, Ivonne; Simmet, Thomas; Nienhaus, Gerd Ulrich; Landfester, Katharina

    2015-01-01

    Summary The combination of stem cell therapy and nanoparticles promises to enhance the effect of cellular therapies by using nanocarriers as drug delivery devices to guide the further differentiation or homing of stem cells. The impact of nanoparticles on primary cell types remains much more elusive as most groups study the nanoparticle–cell interaction in malignant cell lines. Here, we report on the influence of polymeric nanoparticles on human hematopoietic stem cells (hHSCs) and mesenchymal stem cells (hMSCs). In this study we systematically investigated the influence of polymeric nanoparticles on the cell functionality and differentiation capacity of hHSCs and hMSCs to obtain a deeper knowledge of the interaction of stem cells and nanoparticles. As model systems of nanoparticles, two sets of either bioinert (polystyrene without carboxylic groups on the surface) or biodegradable (PLLA without magnetite) particles were analyzed. Flow cytometry and microscopy analysis showed high uptake rates and no toxicity for all four tested particles in hMSCs and hHSCs. During the differentiation process, the payload of particles per cell decreased. The PLLA–Fe particle showed a significant increase in the IL-8 release in hMSCs but not in hHSCs. We assume that this is due to an increase of free intracellular iron ions but obviously also depends on the cell type. For hHSCs and hMSCs, lineage differentiation into erythrocytes, granulocytes, and megakaryocytes or adipocytes, osteocytes and chondrocytes, was not influenced by the particles when analyzed with lineage specific cluster of differentiation markers. On the other hand qPCR analysis showed significant changes in the expression of some (but not all) investigated lineage markers for both primary cell types. PMID:25821678

  2. Green synthesis of silver nanoparticles using cell extracts of Anabaena doliolum and screening of its antibacterial and antitumor activity.

    PubMed

    Singh, Garvita; Babele, Piyoosh K; Shahi, Shailesh K; Sinha, Rajeshwar P; Tyagi, Madhu B; Kumar, Ashok

    2014-10-01

    In the present work, we describe a simple, cheap, and unexplored method for "green" synthesis of silver nanoparticles using cell extracts of the cyanobacterium Anabaena doliolum. An attempt was also made to test the antimicrobial and antitumor activities of the synthesized nanoparticles. Analytical techniques, namely UV-vis spectroscopy, X-ray diffraction, Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), and TEMselected area electron diffraction, were used to elucidate the formation and characterization of silver-cyanobacterial nanoparticles (Ag-CNPs). Results showed that the original color of the cell extract changed from reddish blue to dark brown after addition of silver nitrate solution (1 mM) within 1 h, suggesting the synthesis of Ag-CNPs. That the formation Ag-CNPs indeed occurred was also evident from the spectroscopic analysis of the reaction mixture, wherein a prominent peak at 420 nm was noted. TEM images revealed well-dispersed, spherical Ag- CNPs with a particle size in the range of 10-50 nm. The X-ray diffraction spectrum suggested a crystalline nature of the Ag-CNPs. FTIR analysis indicated the utilization of a hydroxyl (-OH) group in the formation of Ag-CNPs. Ag-CNPs exhibited strong antibacterial activity against three multidrug-resistant bacteria. Additionally, Ag-CNPs strongly affected the survival of Dalton's lymphoma and human carcinoma colo205 cells at a very low concentration. The Ag-CNPs-induced loss of survival of both cell types may be due to the induction of reactive oxygen species generation and DNA fragmentation, resulting in apoptosis. Properties exhibited by the Ag-CNP suggest that it may be used as a potential antibacterial and antitumor agent. PMID:24986675

  3. Ectoenzyme switches the surface of magnetic nanoparticles for selective binding of cancer cells.

    PubMed

    Du, Xuewen; Zhou, Jie; Xu, Bing

    2015-06-01

    Enzymatic switch, such as phosphorylation and dephosphorylation of proteins, is the most important mechanism for cellular signal transductions. Inspired by Nature and encouraged by our recent unexpected observation of the dephosphorylation of d-tyrosine phosphate-contain small peptides, we modify the surface of magnetic nanoparticles (MNP) with d-tyrosine phosphate that is a substrate of alkaline phosphatase (ALP). Our studies find that ALP is able to remove the phosphate groups from the magnetic nanoparticles. Most importantly, placental alkaline phosphatase (ALPP), an ectoenzyme that locates on cell surface with catalytic domains outside the plasma membrane and is overexpressed on many cancer cells, dephosphorylate the d-tyrosine phosphates on the surface of the magnetic nanoparticle and enable the magnetic nanoparticles to adhere selectively to the cancer cells, such as HeLa cells. Unlikely commonly used antibodies, the selectivity of the magnetic nanoparticles to cancer cells originates from the enzymatic reaction catalyzed by ALPP. The use of enzymatic reaction to modulate the surface of various nanostructures may lead to a general method to broadly target cancer cells without relying on specific ligand-receptor interactions (e.g., antibodies). This work, thus, illustrates a fundamentally new concept to allow cells to actively engineer the surface of colloids materials, such as magnetic nanoparticles, for various applications. PMID:25586118

  4. Evaluating Cytotoxicity of Hyaluronate Targeted Solid Lipid Nanoparticles of Etoposide on SK-OV-3 Cells

    PubMed Central

    Varshosaz, Jaleh; Sadeghi Aliabadi, Hojatollah

    2014-01-01

    The epithelial ovarian carcinoma is one of the most fatal gynecological cancers. Etoposide is used in treating platinum-resistant ovarian cancer. Sodium hyaluronate is a substance that binds to the CD44 receptors overexpressed in SK-OV-3 cells of epithelial ovarian carcinoma. The aim of the present work was to study the cytotoxicity effect of hyaluronate targeted solid lipid nanoparticles (SLNs) of etoposide on SK-OV-3 cells. The cytotoxicity of the targeted and nontargeted SLNs of etoposide was compared to free drug on the SK-OV-3 cells by MTT assay method. The cellular uptake of the targeted and nontargeted nanoparticles containing sodium fluorescein was also studied. The difference of cell vitality between nontargeted nanoparticles and also targeted nanoparticles with free drug was significant. Targeted nanoparticles also caused more toxicity than nontargeted nanoparticles (P < 0.05). After 4 hours of incubating, the fluorescence was remarkably higher in the cells treated by targeted SLNs rather than nontargeted ones, and there was no observable fluorescence in cells incubated with pure sodium fluorescein. Hyaluronate targeted SLNs containing etoposide increased the cytotoxicity of etoposide on SK-OV-3 cells which may be a worthwhile potential method for reducing the prescribed dose and systemic side effects of this drug in epithelial ovarian carcinoma. PMID:24868467

  5. Live Cell Plasma Membranes Do Not Exhibit a Miscibility Phase Transition over a Wide Range of Temperatures.

    PubMed

    Lee, Il-Hyung; Saha, Suvrajit; Polley, Anirban; Huang, Hector; Mayor, Satyajit; Rao, Madan; Groves, Jay T

    2015-03-26

    Lipid/cholesterol mixtures derived from cell membranes as well as their synthetic reconstitutions exhibit well-defined miscibility phase transitions and critical phenomena near physiological temperatures. This suggests that lipid/cholesterol-mediated phase separation plays a role in the organization of live cell membranes. However, macroscopic lipid-phase separation is not generally observed in cell membranes, and the degree to which properties of isolated lipid mixtures are preserved in the cell membrane remain unknown. A fundamental property of phase transitions is that the variation of tagged particle diffusion with temperature exhibits an abrupt change as the system passes through the transition, even when the two phases are distributed in a nanometer-scale emulsion. We support this using a variety of Monte Carlo and atomistic simulations on model lipid membrane systems. However, temperature-dependent fluorescence correlation spectroscopy of labeled lipids and membrane-anchored proteins in live cell membranes shows a consistently smooth increase in the diffusion coefficient as a function of temperature. We find no evidence of a discrete miscibility phase transition throughout a wide range of temperatures: 14-37 °C. This contrasts the behavior of giant plasma membrane vesicles (GPMVs) blebbed from the same cells, which do exhibit phase transitions and macroscopic phase separation. Fluorescence lifetime analysis of a DiI probe in both cases reveals a significant environmental difference between the live cell and the GPMV. Taken together, these data suggest the live cell membrane may avoid the miscibility phase transition inherent to its lipid constituents by actively regulating physical parameters, such as tension, in the membrane. PMID:25747462

  6. Plasma-micropatterning of albumin nanoparticles: Substrates for enhanced cell-interactive display of ligands.

    PubMed

    Rossi, María Pía; Xu, Jing; Schwarzbauer, Jean; Moghe, Prabhas V

    2010-12-01

    The authors demonstrate a novel, efficient, and widely applicable approach to direct the patterning of ligand-functionalized organic nanoparticles derived from albumin on nonconductive, biodegradable polymeric substrates. In contrast to traditional deposition methods for inorganic nanoparticles, the approach involves oxygen plasma treatment of spatially restricted regions on a nonbiopermissive polymer. Albumin nanoparticles conjugated with a truncated fragment of fibronectin containing the Arg-Gly-Asp domain were successfully patterned and used as templates to elicit adhesion and spreading of human mesenchymal stem cells and fibroblasts. Attachment and spreading of both cell types into the plasma-exposed polymer areas was considerably more pronounced than with the ligand alone. The authors hypothesize that the underlying mechanism is oxygen plasma treatment-induced selective enhancement of ligand exposure from the deposited functionalized nanoparticles, which facilitates ligand receptor clustering at the cell membrane. The results highlight a promising nanoscale approach to modulate ligand presentation and spatially direct cell attachment and phenotypic behaviors. PMID:21219031

  7. Invasive breast carcinoma cells from patients exhibit MenaINV- and macrophage-dependent transendothelial migration

    PubMed Central

    Pignatelli, Jeanine; Goswami, Sumanta; Jones, Joan G.; Rohan, Thomas E.; Pieri, Evan; Chen, Xiaoming; Adler, Esther; Cox, Dianne; Maleki, Sara; Bresnick, Anne; Gertler, Frank B.; Condeelis, John S.; Oktay, Maja H.

    2014-01-01

    Metastasis is a complex, multistep process of cancer progression that has few treatment options. A critical event is the invasion of cancer cells into blood vessels (intravasation), through which cancer cells disseminate to distant organs. Breast cancer cells with increased abundance of Mena [an epidermal growth factor (EGF)–responsive cell migration protein] are present with macrophages at sites of intravasation, called TMEM sites (for tumor microenvironment of metastasis), in patient tumor samples. Furthermore, the density of these intravasation sites correlates with metastatic risk in patients. We found that intravasation of breast cancer cells may be prevented by blocking the signaling between cancer cells and macrophages. We obtained invasive breast ductal carcinoma cells of various subtypes by fine-needle aspiration (FNA) biopsies from patients and found that, in an in vitro transendothelial migration assay, cells that migrated through a layer of human endothelial cells were enriched for the transcript encoding MenaINV, an invasive isoform of Mena. This enhanced transendothelial migration required macrophages and occurred with all of the breast cancer subtypes. Using mouse macrophages and the human cancer cells from the FNAs, we identified paracrine and autocrine activation of colony-stimulating factor-1 receptor (CSF-1R). The paracrine or autocrine nature of the signal depended on the breast cancer cell subtype. Knocking down MenaINV or adding an antibody that blocks CSF-1R function prevented transendothelial migration. Our findings indicate that MenaINV and TMEM frequency are correlated prognostic markers and CSF-1 and MenaINV may be therapeutic targets to prevent metastasis of multiple breast cancer subtypes. PMID:25429076

  8. Invasive breast carcinoma cells from patients exhibit MenaINV- and macrophage-dependent transendothelial migration.

    PubMed

    Pignatelli, Jeanine; Goswami, Sumanta; Jones, Joan G; Rohan, Thomas E; Pieri, Evan; Chen, Xiaoming; Adler, Esther; Cox, Dianne; Maleki, Sara; Bresnick, Anne; Gertler, Frank B; Condeelis, John S; Oktay, Maja H

    2014-11-25

    Metastasis is a complex, multistep process of cancer progression that has few treatment options. A critical event is the invasion of cancer cells into blood vessels (intravasation), through which cancer cells disseminate to distant organs. Breast cancer cells with increased abundance of Mena [an epidermal growth factor (EGF)-responsive cell migration protein] are present with macrophages at sites of intravasation, called TMEM sites (for tumor microenvironment of metastasis), in patient tumor samples. Furthermore, the density of these intravasation sites correlates with metastatic risk in patients. We found that intravasation of breast cancer cells may be prevented by blocking the signaling between cancer cells and macrophages. We obtained invasive breast ductal carcinoma cells of various subtypes by fine-needle aspiration (FNA) biopsies from patients and found that, in an in vitro transendothelial migration assay, cells that migrated through a layer of human endothelial cells were enriched for the transcript encoding Mena(INV), an invasive isoform of Mena. This enhanced transendothelial migration required macrophages and occurred with all of the breast cancer subtypes. Using mouse macrophages and the human cancer cells from the FNAs, we identified paracrine and autocrine activation of colony-stimulating factor-1 receptor (CSF-1R). The paracrine or autocrine nature of the signal depended on the breast cancer cell subtype. Knocking down Mena(INV) or adding an antibody that blocks CSF-1R function prevented transendothelial migration. Our findings indicate that Mena(INV) and TMEM frequency are correlated prognostic markers and CSF-1 and Mena(INV) may be therapeutic targets to prevent metastasis of multiple breast cancer subtypes. PMID:25429076

  9. Colony Forming Unit Endothelial Cells Do not Exhibit Telomerase Alternative Splicing Variants and Activity

    PubMed Central

    Attar, Armin; Khosravi Maharlooi, Mohsen; Khoshkhou, Sara; Hosseini, Ahmad; Jaberipour, Mansoureh; Dehghan, Arman; Monabati, Ahmad

    2013-01-01

    Introduction: Endothelial progenitor colony forming unit-endothelial cells (CFU-EC) were first believed to be the progenitors of endothelial cells, named endothelial progenitor cells. Further studies revealed that they are monocytes regulating vasculogenesis. The main hindrance of these cells for therapeutic purposes is their low frequency and limited replicative potentials. This study was undertaken to determine telomerase activity and alternative splicing variants in CFU-EC as a potential cause of limited replicative capacity in these cells. Methods: CFU-EC were isolated from peripheral blood using a standard cell culture assay. Colonies were detached mechanically and alternative splicing variant mRNA were evaluated using real-time PCR. Telomerase enzyme activity was assessed using telomerase repeat amplification protocol. The same procedures were done on the cancer cell line Calu6 as the positive control. Results: The cultured peripheral blood mononuclear cells formed colonies with spindle-shaped monocytic cells sprouted from the clusters. These morphological characteristics fulfill the definition of CFU-EC. Telomere length amplification protocol assay revealed no telomerase activity and real-time PCR showed no expression of telomerase enzyme mRNA in CFU-EC. Both parameters were significantly higher in the cancer cell line Calu6 taken as the positive control. Conclusion: The absence of telomerase activity in the CFU-EC is a result of pre-transcriptional regulation of gene expression rather than other mechanisms for controlling telomerase activity such as post-transcriptional modifications. This finding can explain the limited proliferative activity of CFU-EC cells. We propose that absence of telomerase activity in CFU-EC can be attributable to their more mature monocytic nature that needs further investigations. PMID:23748893

  10. Mixing Effect of Gold and Silver Nanoparticles on Enhancement in Performance of Organic Thin-Film Solar Cells

    NASA Astrophysics Data System (ADS)

    Akiyama, Tsuyoshi; Yamamoto, Tomoki; Oku, Takeo; Yahiro, Masayuki; Kurihara, Takashi; Adachi, Chihaya; Yamada, Sunao

    2013-12-01

    Bulk-heterojunction organic thin-film solar cells incorporating gold and silver nanoparticles were fabricated and evaluated. These nanoparticles were embedded in the hole-transport layer of the solar cells. Plasmonic absorption peaks of isolated gold and silver nanoparticles were confirmed from extinction spectra even in the hole-transport material. The incorporation of gold and silver nanoparticles increased the photoelectric conversion efficiency of organic thin-film solar cells, whose enhancement ratio was further increased by mixing gold and silver nanoparticles.

  11. Pseudoislets in stirred-suspension culture exhibit enhanced cell survival, propagation and insulin secretion.

    PubMed

    Lock, Lye T; Laychock, Suzanne G; Tzanakakis, Emmanuel S

    2011-02-10

    Cells from primary islets and beta-cell lines form pseudoislets (PIs) in static cultures. Interestingly, MIN6 beta-cells with aberrant regulation of proliferation form PIs which cease to grow after a week in culture. This growth arrest is attributed to a pro-apoptotic and anti-proliferative PI environment. We hypothesized that cell necrosis due to poor nutrient transport in dishes rather than apoptosis effects the observed PI size restriction. Formation of beta-cell PIs was explored in stirred-suspension bioreactors with enhanced mass transfer. Cells in stirred-suspension proliferated continuously and the PI size increased for two weeks. Bioreactor PIs displayed regulated basal insulin secretion and enhanced responsivity to glucose and incretins. Compared to dishes, cell viability in the bioreactor was higher with lower released lactate dehydrogenase activity. Similar expression of p21 and p27 in monolayers and PIs did not suggest an anti-proliferative PI milieu. Caspase-2, -8 and -9 activities were comparable in dish and bioreactor PIs, and the latter continued to grow after one week of culture. Thus, apoptosis is not sufficient to explain the differences in PI size between dishes and bioreactor. Moreover, the bioreactor method described here may be used to generate PIs with increased cell viability and function for research and clinical applications. PMID:21185337

  12. Targeting Mantle Cell Lymphoma with Anti-SYK Nanoparticles

    PubMed Central

    Cely, Ingrid; Yiv, Seang; Yin, Qian; Shahidzadeh, Anoush; Tang, Li; Cheng, Jianjun; Uckun, Fatih M.

    2013-01-01

    The pentapeptide mimic 1,4-bis(9-O-dihydroquinidinyl)phthalazine / hydroquinidine 1,4-phathalazinediyl diether (“compound 61”) (C-61) is the first reported inhibitor targeting the P-site of SYK. Here we report a nanotechnology platform to target C-61 to mantle cell lymphoma (MCL) cells. Liposomal nanoparticles (NP) loaded with C-61 were prepared using the standard thin film evaporation method. The entrapment of C-61 was obtained using the pH gradient procedure with lactobionic acid (LBA) being used as a low pH buffer inside the NP. Formulation F6A was selected as a lead candidate for further biological testing. The average diameter, zeta potential and C-61 content of the F6A NP was 40 nm, 0.1 mV, and 12.6 mg/ml, respectively. F6A induces apoptosis in SYK+ but not SYK? leukemia/lymphoma cells. We also evaluated the cytotoxic activity of F6A in the context of an in vitro artificial bone marrow assay platform based on a 3D scaffold with inverted colloidal crystal geometry mimicking the structural topology of actual bone marrow matrix. The ability of C-61 to induce apoptosis in ALL-1 cells was not adversely affected by the scaffolds. F6A, but not the drug-free NP formulation F6B, caused apoptosis of MCL cell lines MAVER-1 and MINO within 24h. Further development of rationally designed SYK inhibitors and their nanoscale formulations may provide the foundation for therapeutic innovation against a broad spectrum of lymphoid malignancies, including MCL. PMID:23730399

  13. Targeting Mantle Cell Lymphoma with Anti-SYK Nanoparticles.

    PubMed

    Cely, Ingrid; Yiv, Seang; Yin, Qian; Shahidzadeh, Anoush; Tang, Li; Cheng, Jianjun; Uckun, Fatih M

    2012-06-25

    The pentapeptide mimic 1,4-bis(9-O-dihydroquinidinyl)phthalazine / hydroquinidine 1,4-phathalazinediyl diether ("compound 61") (C-61) is the first reported inhibitor targeting the P-site of SYK. Here we report a nanotechnology platform to target C-61 to mantle cell lymphoma (MCL) cells. Liposomal nanoparticles (NP) loaded with C-61 were prepared using the standard thin film evaporation method. The entrapment of C-61 was obtained using the pH gradient procedure with lactobionic acid (LBA) being used as a low pH buffer inside the NP. Formulation F6A was selected as a lead candidate for further biological testing. The average diameter, zeta potential and C-61 content of the F6A NP was 40 nm, 0.1 mV, and 12.6 mg/ml, respectively. F6A induces apoptosis in SYK(+) but not SYK(-) leukemia/lymphoma cells. We also evaluated the cytotoxic activity of F6A in the context of an in vitro artificial bone marrow assay platform based on a 3D scaffold with inverted colloidal crystal geometry mimicking the structural topology of actual bone marrow matrix. The ability of C-61 to induce apoptosis in ALL-1 cells was not adversely affected by the scaffolds. F6A, but not the drug-free NP formulation F6B, caused apoptosis of MCL cell lines MAVER-1 and MINO within 24h. Further development of rationally designed SYK inhibitors and their nanoscale formulations may provide the foundation for therapeutic innovation against a broad spectrum of lymphoid malignancies, including MCL. PMID:23730399

  14. PEG-detachable lipid-polymer hybrid nanoparticle for delivery of chemotherapy drugs to cancer cells.

    PubMed

    Du, Jiang-bo; Song, Yan-feng; Ye, Wei-liang; Cheng, Ying; Cui, Han; Liu, Dao-zhou; Liu, Miao; Zhang, Bang-le; Zhou, Si-yuan

    2014-08-01

    The experiment aimed to increase the drug-delivery efficiency of poly-lactic-co-glycolic acid (PLGA) nanoparticles. Lipid-polymer hybrid nanoparticles (LPNs-1) were prepared using PLGA as a hydrophobic core and FA-PEG-hyd-DSPE as an amphiphilic shell. Uniform and spherical nanoparticles with an average size of 185 nm were obtained using the emulsification solvent evaporation method. The results indicated that LPNs-1 showed higher drug loading compared with naked PLGA nanoparticles (NNPs). Drug release from LPNs-1 was faster in an acidic environment than in a neutral environment. LPNs-1 showed higher cytotoxicity on KB cells, A549 cells, MDA-MB-231 cells, and MDA-MB-231/ADR cells compared with free doxorubicin (DOX) and NNPs. The results also showed that, compared with free DOX and NNPs, LPNs-1 delivered more DOX to the nuclear of KB cells and MDA-MB-231/ADR cells. LPNs-1 induced apoptosis in KB cells and MDA-MB-231/ADR cells in a dose-dependent manner. The above data indicated that DOX-loaded LPNs-1 could kill not only normal tumor cells but also drug-resistant tumor cells. These results indicated that modification of PLGA nanoparticles with FA-PEG-hyd-DSPE could considerably increase the drug-delivery efficiency and LPNs-1 had potential in the delivery of chemotherapeutic agents in the treatment of cancer. PMID:24590167

  15. In vitro developmental toxicity test detects inhibition of stem cell differentiation by silica nanoparticles

    SciTech Connect

    Park, Margriet V.D.Z. [Laboratory for Health Protection Research, National Institute for Public Health and the Environment, 3720 BA, Bilthoven (Netherlands); Department of Health Risk Analysis and Toxicology, Maastricht University, 6200 MD Maastricht (Netherlands)], E-mail: Margriet.Park@rivm.nl; Annema, Wijtske [Laboratory for Health Protection Research, National Institute for Public Health and the Environment, 3720 BA, Bilthoven (Netherlands); Salvati, Anna; Lesniak, Anna [Centre for BioNano Interactions, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4 (Ireland); Elsaesser, Andreas; Barnes, Clifford; McKerr, George; Howard, C. Vyvyan [Centre for Molecular Bioscience, University of Ulster, Coleraine, BT52 1SA (United Kingdom); Lynch, Iseult; Dawson, Kenneth A. [Centre for BioNano Interactions, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4 (Ireland); Piersma, Aldert H. [Laboratory for Health Protection Research, National Institute for Public Health and the Environment, 3720 BA, Bilthoven (Netherlands); Institute for Risk Assessment Sciences, University of Utrecht, 3508 TD, Utrecht (Netherlands); Jong, Wim H. de [Laboratory for Health Protection Research, National Institute for Public Health and the Environment, 3720 BA, Bilthoven (Netherlands)

    2009-10-01

    While research into the potential toxic properties of nanomaterials is now increasing, the area of developmental toxicity has remained relatively uninvestigated. The embryonic stem cell test is an in vitro screening assay used to investigate the embryotoxic potential of chemicals by determining their ability to inhibit differentiation of embryonic stem cells into spontaneously contracting cardiomyocytes. Four well characterized silica nanoparticles of various sizes were used to investigate whether nanomaterials are capable of inhibition of differentiation in the embryonic stem cell test. Nanoparticle size distributions and dispersion characteristics were determined before and during incubation in the stem cell culture medium by means of transmission electron microscopy (TEM) and dynamic light scattering. Mouse embryonic stem cells were exposed to silica nanoparticles at concentrations ranging from 1 to 100 {mu}g/ml. The embryonic stem cell test detected a concentration dependent inhibition of differentiation of stem cells into contracting cardiomyocytes by two silica nanoparticles of primary size 10 (TEM 11) and 30 (TEM 34) nm while two other particles of primary size 80 (TEM 34) and 400 (TEM 248) nm had no effect up to the highest concentration tested. Inhibition of differentiation of stem cells occurred below cytotoxic concentrations, indicating a specific effect of the particles on the differentiation of the embryonic stem cells. The impaired differentiation of stem cells by such widely used particles warrants further investigation into the potential of these nanoparticles to migrate into the uterus, placenta and embryo and their possible effects on embryogenesis.

  16. Managing magnetic nanoparticle aggregation and cellular uptake: a precondition for efficient stem-cell differentiation and MRI tracking.

    PubMed

    Fayol, Delphine; Luciani, Nathalie; Lartigue, Lenaic; Gazeau, Florence; Wilhelm, Claire

    2013-02-01

    The labeling of stem cells with iron oxide nanoparticles is increasingly used to enable MRI cell tracking and magnetic cell manipulation, stimulating the fields of tissue engineering and cell therapy. However, the impact of magnetic labeling on stem-cell differentiation is still controversial. One compromising factor for successful differentiation may arise from early interactions of nanoparticles with cells during the labeling procedure. It is hypothesized that the lack of control over nanoparticle colloidal stability in biological media may lead to undesirable nanoparticle localization, overestimation of cellular uptake, misleading MRI cell tracking, and further impairment of differentiation. Herein a method is described for labeling mesenchymal stem cells (MSC), in which the physical state of citrate-coated nanoparticles (dispersed versus aggregated) can be kinetically tuned through electrostatic and magnetic triggers, as monitored by diffusion light scattering in the extracellular medium and by optical and electronic microscopy in cells. A set of statistical cell-by-cell measurements (flow cytometry, single-cell magnetophoresis, and high-resolution MRI cellular detection) is used to independently quantify the nanoparticle cell uptake and the effects of nanoparticle aggregation. Such aggregation confounds MRI cell detection as well as global iron quantification and has adverse effects on chondrogenetic differentiation. Magnetic labeling conditions with perfectly stable nanoparticles-suitable for obtaining differentiation-capable magnetic stem cells for use in cell therapy-are subsequently identified. PMID:23184893

  17. In vitro interaction of colloidal nanoparticles with mammalian cells: What have we learned thus far?

    PubMed Central

    Nazarenus, Moritz; Zhang, Qian; Soliman, Mahmoud G; del Pino, Pablo; Pelaz, Beatriz; Carregal-Romero, Susana; Rejman, Joanna; Rothen-Rutishauser, Barbara; Clift, Martin J D; Zellner, Reinhard; Nienhaus, G Ulrich; Delehanty, James B; Medintz, Igor L

    2014-01-01

    Summary The interfacing of colloidal nanoparticles with mammalian cells is now well into its second decade. In this review our goal is to highlight the more generally accepted concepts that we have gleaned from nearly twenty years of research. While details of these complex interactions strongly depend, amongst others, upon the specific properties of the nanoparticles used, the cell type, and their environmental conditions, a number of fundamental principles exist, which are outlined in this review. PMID:25247131

  18. A Magnetic Nanoparticle-Based Multiple-Gene Delivery System for Transfection of Porcine Kidney Cells

    PubMed Central

    Wang, Yan; Cui, Haixin; Li, Kui; Sun, Changjiao; Du, Wei; Cui, Jinhui; Zhao, Xiang; Chen, Wenjie

    2014-01-01

    Superparamagnetic nanoparticles are promising candidates for gene delivery into mammalian somatic cells and may be useful for reproductive cloning using the somatic cell nuclear transfer technique. However, limited investigations of their potential applications in animal genetics and breeding, particularly multiple-gene delivery by magnetofection, have been performed. Here, we developed a stable, targetable and convenient system for delivering multiple genes into the nuclei of porcine somatic cells using magnetic Fe3O4 nanoparticles as gene carriers. After surface modification by polyethylenimine, the spherical magnetic Fe3O4 nanoparticles showed strong binding affinity for DNA plasmids expressing the genes encoding a green (DNAGFP) or red (DNADsRed) fluorescent protein. At weight ratios of DNAGFP or DNADsRed to magnetic nanoparticles lower than or equal to 10?1 or 5?1, respectively, the DNA molecules were completely bound by the magnetic nanoparticles. Atomic force microscopy analyses confirmed binding of the spherical magnetic nanoparticles to stretched DNA strands up to several hundred nanometers in length. As a result, stable and efficient co-expression of GFP and DsRed in porcine kidney PK-15 cells was achieved by magnetofection. The results presented here demonstrate the potential application of magnetic nanoparticles as an attractive delivery system for animal genetics and breeding studies. PMID:25048709

  19. Defining the Subcellular Interface of Nanoparticles by Live-Cell Imaging

    PubMed Central

    Hemmerich, Peter H.; von Mikecz, Anna H.

    2013-01-01

    Understanding of nanoparticle-bio-interactions within living cells requires knowledge about the dynamic behavior of nanomaterials during their cellular uptake, intracellular traffic and mutual reactions with cell organelles. Here, we introduce a protocol of combined kinetic imaging techniques that enables investigation of exemplary fluorochrome-labelled nanoparticles concerning their intracellular fate. By time-lapse confocal microscopy we observe fast, dynamin-dependent uptake of polystyrene and silica nanoparticles via the cell membrane within seconds. Fluorescence recovery after photobleaching (FRAP) experiments reveal fast and complete exchange of the investigated nanoparticles at mitochondria, cytoplasmic vesicles or the nuclear envelope. Nuclear translocation is observed within minutes by free diffusion and active transport. Fluorescence correlation spectroscopy (FCS) and raster image correlation spectroscopy (RICS) indicate diffusion coefficients of polystyrene and silica nanoparticles in the nucleus and the cytoplasm that are consistent with particle motion in living cells based on diffusion. Determination of the apparent hydrodynamic radii by FCS and RICS shows that nanoparticles exert their cytoplasmic and nuclear effects mainly as mobile, monodisperse entities. Thus, a complete toolkit of fluorescence fluctuation microscopy is presented for the investigation of nanomaterial biophysics in subcellular microenvironments that contributes to develop a framework of intracellular nanoparticle delivery routes. PMID:23637951

  20. Magnetic nanoparticle-mediated massively-parallel mechanical modulation of single-cell behavior

    PubMed Central

    Tseng, Peter; Judy, Jack W.; Di Carlo, Dino

    2012-01-01

    We report a technique for generating controllable, time-varying and localizable forces on arrays of cells in a massively parallel fashion. To achieve this, we grow magnetic nanoparticle-dosed cells in defined patterns on micro-magnetic substrates. By manipulating and coalescing nanoparticles within cells, we apply localized nanoparticle-mediated forces approaching cellular yield tensions on the cortex of HeLa cells. We observed highly coordinated responses in cellular behavior, including the p21-activated kinase (PAK)-dependent generation of active, leading-edge type filopodia, and biasing of the metaphase plate during mitosis. The large sample size and rapid sample generation inherent to this approach allow the analysis of cells at an unprecedented rate; a single experiment can potentially stimulate tens of thousands of cells for high statistical accuracy in measurements. This technique shows promise as a tool for both cell analysis and control. PMID:23064517

  1. Self-patterned nanoparticle layers for vertical interconnects: application in tandem solar cells.

    PubMed

    Niesen, Bjoern; Blondiaux, Nicolas; Boccard, Mathieu; Stuckelberger, Michael; Pugin, Raphaël; Scolan, Emmanuel; Meillaud, Fanny; Haug, Franz-Josef; Hessler-Wyser, Aïcha; Ballif, Christophe

    2014-09-10

    We demonstrate self-patterned insulating nanoparticle layers to define local electrical interconnects in thin-film electronic devices. We show this with thin-film silicon tandem solar cells, where we introduce between the two component cells a solution-processed SiO2 nanoparticle layer with local openings to allow for charge transport. Because of its low refractive index, high transparency, and smooth surface, the SiO2 nanoparticle layer acts as an excellent intermediate reflector allowing for efficient light management. PMID:25102168

  2. Cells from long-lived mutant mice exhibit enhanced repair of UV lesions

    PubMed Central

    Salmon, Adam B.; Ljungman, Mats; Miller, Richard A.

    2009-01-01

    Fibroblasts isolated from long-lived hypopituitary dwarf mice are resistant to many cell stresses, including UV light and MMS, which induce cell death by producing DNA damage. Here we report that cells from Snell dwarf mice recover more rapidly than controls from the inhibition of RNA synthesis induced by UV damage. Recovery of mRNA synthesis in particular is more rapid in dwarf cells, suggesting enhanced repair of the actively transcribing genes in dwarf-derived cells. At early timepoints, there was no difference in the repair of CPD or 6-4PP in the whole genome, nor was there any significant difference in the repair of UV lesions in specific genes. However, at later time points we found that more lesions had been removed from the genome of dwarf-derived cells. We have also found that cells from dwarf mice express higher levels of the nucleotide excision repair proteins XPC and CSA, suggesting a causal link to enhanced DNA repair. Overall, these data suggest a mechanism for the UV resistance of Snell dwarf-derived fibroblasts that could contribute to the delay of aging and neoplasia in these mice. PMID:18375871

  3. Antiandrogen gold nanoparticles dual-target and overcome treatment resistance in hormone-insensitive prostate cancer cells.

    PubMed

    Dreaden, Erik C; Gryder, Berkley E; Austin, Lauren A; Tene Defo, Brice A; Hayden, Steven C; Pi, Min; Quarles, L Darryl; Oyelere, Adegboyega K; El-Sayed, Mostafa A

    2012-08-15

    Prostate cancer is the most commonly diagnosed cancer among men in the developed countries.(1) One in six males in the U.S.(2) and one in nine males in the U.K.(3) will develop the disease at some point during their lifetime. Despite advances in prostate cancer screening, more than a quarter of a million men die from the disease every year(1) due primarily to treatment-resistance and metastasis. Colloidal nanotechnologies can provide tremendous enhancements to existing targeting/treatment strategies for prostate cancer to which malignant cells are less sensitive. Here, we show that antiandrogen gold nanoparticles--multivalent analogues of antiandrogens currently used in clinical therapy for prostate cancer--selectively engage two distinct receptors, androgen receptor (AR), a target for the treatment of prostate cancer, as well as a novel G-protein coupled receptor, GPRC6A, that is also upregulated in prostate cancer. These nanoparticles selectively accumulated in hormone-insensitive and chemotherapy-resistant prostate cancer cells, bound androgen receptor with multivalent affinity, and exhibited greatly enhanced drug potency versus monovalent antiandrogens currently in clinical use. Further, antiandrogen gold nanoparticles selectively stimulated GPRC6A with multivalent affinity, demonstrating that the delivery of nanoscale antiandrogens can also be facilitated by the transmembrane receptor in order to realize increasingly selective, increasingly potent therapy for treatment-resistant prostate cancers. PMID:22768914

  4. Biocompatibility of Fe3O4 nanoparticles evaluated by in vitro cytotoxicity assays using normal, glia and breast cancer cells

    NASA Astrophysics Data System (ADS)

    Ankamwar, B.; Lai, T. C.; Huang, J. H.; Liu, R. S.; Hsiao, M.; Chen, C. H.; Hwu, Y. K.

    2010-02-01

    In order to reveal the biocompatibility of Fe3O4 nanoparticles and bipolar surfactant tetramethylammonium 11-aminoundecanoate cytotoxicity tests were performed as a function of concentration from low (0.1 µg ml-1) to higher concentration (100 µg ml-1) using various human glia, human breast cancer and normal cell lines. Cytotoxicity tests for human glia (D54MG, G9T, SF126, U87, U251, U373), human breast cancer (MB157, SKBR3, T47D) and normal (H184B5F5/M10, WI-38, SVGp12) cell lines exhibited almost nontoxicity and reveal biocompatibility of Fe3O4 nanoparticles in the concentration range of 0.1-10 µg ml-1, while accountable cytotoxicity can be seen at 100 µg ml-1. The results of our studies suggest that Fe3O4 nanoparticles coated with bipolar surfactant tetramethylammonium 11-aminoundecanoate are biocompatible and promising for bio-applications such as drug delivery, magnetic resonance imaging and magnetic hyperthermia.

  5. Biocompatibility of Fe(3)O(4) nanoparticles evaluated by in vitro cytotoxicity assays using normal, glia and breast cancer cells.

    PubMed

    Ankamwar, B; Lai, T C; Huang, J H; Liu, R S; Hsiao, M; Chen, C H; Hwu, Y K

    2010-02-19

    In order to reveal the biocompatibility of Fe(3)O(4) nanoparticles and bipolar surfactant tetramethylammonium 11-aminoundecanoate cytotoxicity tests were performed as a function of concentration from low (0.1 microg ml(-1)) to higher concentration (100 microg ml(-1)) using various human glia, human breast cancer and normal cell lines. Cytotoxicity tests for human glia (D54MG, G9T, SF126, U87, U251, U373), human breast cancer (MB157, SKBR3, T47D) and normal (H184B5F5/M10, WI-38, SVGp12) cell lines exhibited almost nontoxicity and reveal biocompatibility of Fe(3)O(4) nanoparticles in the concentration range of 0.1-10 microg ml(-1), while accountable cytotoxicity can be seen at 100 microg ml(-1). The results of our studies suggest that Fe(3)O(4) nanoparticles coated with bipolar surfactant tetramethylammonium 11-aminoundecanoate are biocompatible and promising for bio-applications such as drug delivery, magnetic resonance imaging and magnetic hyperthermia. PMID:20090199

  6. Immuno Nanoparticles Integrated Electrical Control of Targeted Cancer Cell Development Using Whole Cell Bioelectronic Device

    PubMed Central

    Hondroulis, Evangelia; Zhang, Rui; Zhang, Chengxiao; Chen, Chunying; Ino, Kosuke; Matsue, Tomokazu; Li, Chen-Zhong

    2014-01-01

    Electrical properties of cells determine most of the cellular functions, particularly ones which occur in the cell's membrane. Manipulation of these electrical properties may provide a powerful electrotherapy option for the treatment of cancer as cancerous cells have been shown to be more electronegative than normal proliferating cells. Previously, we used an electrical impedance sensing system (EIS) to explore the responses of cancerous SKOV3 cells and normal HUVEC cells to low intensity (<2 V/cm) AC electric fields, determining that the optimal frequency for SKOV3 proliferation arrest was 200 kHz, without harming the non-cancerous HUVECs. In this study, to determine if these effects are cell type dependant, human breast adenocarcinoma cells (MCF7) were subjected to a range of frequencies (50 kHz-2 MHz) similar to the previously tested SKOV3. For the MCF7, an optimal frequency of 100 kHz was determined using the EIS, indicating a higher sensitivity towards the applied field. Further experiments specifically targeting the two types of cancer cells using HER2 antibody functionalized gold nanoparticles (HER2-AuNPs) were performed to determine if enhanced electric field strength can be induced via the application of nanoparticles, consequently leading to the killing of the cancerous cells without affecting non cancerous HUVECs and MCF10a providing a platform for the development of a non-invasive cancer treatment without any harmful side effects. The EIS was used to monitor the real-time consequences on cellular viability and a noticeable decrease in the growth profile of the MCF7 was observed with the application of the HER2-AuNPs and the electric fields indicating specific inhibitory effects on dividing cells in culture. To further understand the effects of the externally applied field to the cells, an Annexin V/EthD-III assay was performed to determine the cell death mechanism indicating apoptosis. The zeta potential of the SKOV3 and the MCF7 before and after incorporation of the HER2-AuNPs was also obtained indicating a decrease in zeta potential with the incorporation of the nanoparticles. The outcome of this research will improve our fundamental understanding of the behavior of cancer cells and define optimal parameters of electrotherapy for clinical and drug delivery applications. PMID:25057316

  7. Microbubbles-overlapping mode for laser killing of cancer cells with absorbing nanoparticle clusters

    NASA Astrophysics Data System (ADS)

    Zharov, V. P.; Letfullin, R. R.; Galitovskaya, E. N.

    2005-08-01

    Laser-induced bubble formation around nanoparticles may play a crucial role in selective laser nanophotothermolysis of cancer cells targeted with nanoparticles. In this paper, we propose theoretically, and confirm experimentally, a new dynamic mode for selective cancer treatment that involves the overlapping of bubbles inside the cell volume. This bubbles-overlapping mode (BOM) can dramatically increase the efficiency of cancer treatment by laser-heated nanoparticles as a result of the large damage range. On the basis of nanoparticle optics below the diffraction limit and the kinetic model of bubble dynamics, we found the criteria and conditions (interparticle distance and particle size and concentration) for BOM initiation in cancer cells by laser radiation. Using MDA-MB-231 breast cancer cells, we showed that the optimal size range of the gold nanoparticles for effective laser initiation of BOM is 30-40 nm and the lower concentration limit is n ap 2.44 × 1011 cm-3 (i.e. the absolute number of particles homogeneously distributed inside a tumour cell is n ap 430). It was demonstrated that the formation of nanoclusters on the cell surface with sizes larger than the sizes of individual nanoparticles, may further increase the efficiency of the laser treatment of cancer.

  8. Surface engineering of silica nanoparticles for oral insulin delivery: characterization and cell toxicity studies.

    PubMed

    Andreani, Tatiana; Kiill, Charlene P; de Souza, Ana Luiza R; Fangueiro, Joana F; Fernandes, Lisete; Doktorovová, Slavomira; Santos, Dario L; Garcia, Maria L; Gremião, Maria Palmira D; Souto, Eliana B; Silva, Amélia M

    2014-11-01

    The present work aimed at studying the interaction between insulin and SiNP surfaced with mucoadhesive polymers (chitosan, sodium alginate or polyethylene glycol) and the evaluation of their biocompatibility with HepG2 and Caco-2 cell lines, which mimic in vivo the target of insulin-loaded nanoparticles upon oral administration. Thus, a systematic physicochemical study of the surface-modified insulin-silica nanoparticles (Ins-SiNP) using mucoadhesive polymers has been described. The surfacing of nanoparticle involved the coating of silica nanoparticles (SiNP) with different mucoadhesive polymers, to achieve high contact between the systems and the gut mucosa to enhance the oral insulin bioavailability. SiNP were prepared by a modified Stöber method at room temperature via hydrolysis and condensation of tetraethyl orthosilicate (TEOS). Interaction between insulin and nanoparticles was assessed by differential scanning calorimetry (DSC), X-ray and Fourier-transform infrared (FTIR) studies. The high efficiency of nanoparticles' coating resulted in more stable system. FTIR spectra of insulin-loaded nanoparticles showed amide absorption bands which are characteristic of ?-helix content. In general, all developed nanoparticles demonstrated high biocompatible, at the tested concentrations (50-500 ?g/mL), revealing no or low toxicity in the two human cancer cell lines (HepG2 and Caco-2). In conclusion, the developed insulin-loaded SiNP surfaced with mucoadhesive polymers demonstrated its added value for oral administration of proteins. PMID:25466464

  9. Biocompatibility of chitosan-coated iron oxide nanoparticles with osteoblast cells

    PubMed Central

    Shi, Si-Feng; Jia, Jing-Fu; Guo, Xiao-Kui; Zhao, Ya-Ping; Chen, De-Sheng; Guo, Yong-Yuan; Cheng, Tao; Zhang, Xian-Long

    2012-01-01

    Background: Bone disorders (including osteoporosis, loosening of a prosthesis, and bone infections) are of great concern to the medical community and are difficult to cure. Therapies are available to treat such diseases, but all have drawbacks and are not specifically targeted to the site of disease. Chitosan is widely used in the biomedical community, including for orthopedic applications. The aim of the present study was to coat chitosan onto iron oxide nanoparticles and to determine its effect on the proliferation and differentiation of osteoblasts. Methods: Nanoparticles were characterized using transmission electron microscopy, dynamic light scattering, x-ray diffraction, zeta potential, and vibrating sample magnetometry. Uptake of nanoparticles by osteoblasts was studied by transmission electron microscopy and Prussian blue staining. Viability and proliferation of osteoblasts were measured in the presence of uncoated iron oxide magnetic nanoparticles or those coated with chitosan. Lactate dehydrogenase, alkaline phosphatase, total protein synthesis, and extracellular calcium deposition was studied in the presence of the nanoparticles. Results: Chitosan-coated iron oxide nanoparticles enhanced osteoblast proliferation, decreased cell membrane damage, and promoted cell differentiation, as indicated by an increase in alkaline phosphatase and extracellular calcium deposition. Chitosan-coated iron oxide nanoparticles showed good compatibility with osteoblasts. Conclusion: Further research is necessary to optimize magnetic nanoparticles for the treatment of bone disease. PMID:23118539

  10. Surface plasma resonant effect of gold nanoparticles on the photoelectrodes of dye-sensitized solar cells

    PubMed Central

    2013-01-01

    In this study, we prepared different shapes of gold nanoparticles by seed-mediated growth method and applied them on the photoelectrodes of dye-sensitized solar cells (DSSCs) to study the surface plasma resonant (SPR) effect of gold nanoparticles on the photoelectrodes of dye-sensitized solar cells. The analyses of field emission scanning electron microscopy show that the average diameter of the spherical gold nanoparticles is 45 nm, the average length and width of the short gold nanorods were 55 and 22 nm, respectively, and the average length and width of the long gold nanorods were 55 and 14 nm, respectively. The aspect ratio of the short and long gold nanorods was about 2.5 and 4, respectively. The results of ultraviolet–visible absorption spectra show that the absorption wavelength is about 540 nm for spherical gold nanoparticles, and the absorption of the gold nanorods reveals two peaks. One is about 510 to 520 nm, and the other is about 670 and 710 nm for the short and long gold nanorods, respectively. The best conversion efficiency of the dye-sensitized solar cells with spherical gold nanoparticles and short and long gold nanorods added in is 6.77%, 7.08%, and 7.29%, respectively, and is higher than that of the cells without gold nanoparticles, which is 6.21%. This result indicates that the effect of gold nanoparticles on the photoelectrodes can increase the conductivity and reduce the recombination of charges in the photoelectrodes, resulting in the increase of conversion efficiency for DSSCs. In addition, the long gold nanorods have stronger SPR effect than the spherical gold nanoparticles and short gold nanorods at long wavelength. This may be the reason for the higher conversion efficiency of DSSCs with long gold nanorods than those of the cells with spherical gold nanoparticles and short gold nanorods. PMID:24172147

  11. High resolution SEM imaging of gold nanoparticles in cells and tissues.

    PubMed

    Goldstein, A; Soroka, Y; Fruši?-Zlotkin, M; Popov, I; Kohen, R

    2014-12-01

    The growing demand of gold nanoparticles in medical applications increases the need for simple and efficient characterization methods of the interaction between the nanoparticles and biological systems. Due to its nanometre resolution, modern scanning electron microscopy (SEM) offers straightforward visualization of metallic nanoparticles down to a few nanometre size, almost without any special preparation step. However, visualization of biological materials in SEM requires complicated preparation procedure, which is typically finished by metal coating needed to decrease charging artefacts and quick radiation damage of biomaterials in the course of SEM imaging. The finest conductive metal coating available is usually composed of a few nanometre size clusters, which are almost identical to the metal nanoparticles employed in medical applications. Therefore, SEM monitoring of metal nanoparticles within cells and tissues is incompatible with the conventional preparation methods. In this work, we show that charging artefacts related to non-conductive biological specimen can be successfully eliminated by placing the uncoated biological sample on a conductive substrate. By growing the cells on glass pre-coated with a chromium layer, we were able to observe the uptake of 10 nm gold nanoparticles inside uncoated and unstained macrophages and keratinocytes cells. Imaging in back scattered electrons allowed observation of gold nanoparticles located inside the cells, while imaging in secondary electron gave information on gold nanoparticles located on the surface of the cells. By mounting a skin cross-section on an improved conductive holder, consisting of a silicon substrate coated with copper, we were able to observe penetration of gold nanoparticles of only 5 nm size through the skin barrier in an uncoated skin tissue. The described method offers a convenient modification in preparation procedure for biological samples to be analyzed in SEM. The method provides high conductivity without application of surface coating and requires less time and a reduced use of toxic chemicals. PMID:25228335

  12. Surface plasma resonant effect of gold nanoparticles on the photoelectrodes of dye-sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Meen, Teen-Hang; Tsai, Jenn-Kai; Chao, Shi-Mian; Lin, Yu-Chien; Wu, Tien-Chuan; Chang, Tang-Yun; Ji, Liang-Wen; Water, Walter; Chen, Wen-Ray; Tang, I.-Tseng; Huang, Chien-Jung

    2013-10-01

    In this study, we prepared different shapes of gold nanoparticles by seed-mediated growth method and applied them on the photoelectrodes of dye-sensitized solar cells (DSSCs) to study the surface plasma resonant (SPR) effect of gold nanoparticles on the photoelectrodes of dye-sensitized solar cells. The analyses of field emission scanning electron microscopy show that the average diameter of the spherical gold nanoparticles is 45 nm, the average length and width of the short gold nanorods were 55 and 22 nm, respectively, and the average length and width of the long gold nanorods were 55 and 14 nm, respectively. The aspect ratio of the short and long gold nanorods was about 2.5 and 4, respectively. The results of ultraviolet-visible absorption spectra show that the absorption wavelength is about 540 nm for spherical gold nanoparticles, and the absorption of the gold nanorods reveals two peaks. One is about 510 to 520 nm, and the other is about 670 and 710 nm for the short and long gold nanorods, respectively. The best conversion efficiency of the dye-sensitized solar cells with spherical gold nanoparticles and short and long gold nanorods added in is 6.77%, 7.08%, and 7.29%, respectively, and is higher than that of the cells without gold nanoparticles, which is 6.21%. This result indicates that the effect of gold nanoparticles on the photoelectrodes can increase the conductivity and reduce the recombination of charges in the photoelectrodes, resulting in the increase of conversion efficiency for DSSCs. In addition, the long gold nanorods have stronger SPR effect than the spherical gold nanoparticles and short gold nanorods at long wavelength. This may be the reason for the higher conversion efficiency of DSSCs with long gold nanorods than those of the cells with spherical gold nanoparticles and short gold nanorods.

  13. Aneuploid yeast strains exhibit defects in cell growth and passage through START

    E-print Network

    Thorburn, Rebecca Ruth

    Aneuploidy, a chromosome content that is not a multiple of the haploid karyotype, is associated with reduced fitness in all organisms analyzed to date. In budding yeast aneuploidy causes cell proliferation defects, with ...

  14. Nitric oxide releasing iron oxide magnetic nanoparticles for biomedical applications: cell viability, apoptosis and cell death evaluations

    NASA Astrophysics Data System (ADS)

    de Lima, R.; de Oliveira, J. L.; Ludescher, A.; Molina, M. M.; Itri, R.; Seabra, A. B.; Haddad, P. S.

    2013-04-01

    Nitric oxide (NO) is involved in several physiological and pathophysiological processes, such as control of vascular tone and immune responses against microbes. Thus, there is great interest in the development of NO-releasing materials to carry and deliver NO for biomedical applications. Magnetic iron oxide nanoparticles have been used in important pharmacological applications, including drug-delivery. In this work, magnetic iron oxide nanoparticles were coated with thiol-containing hydrophilic ligands: mercaptosuccinic acid (MSA) and dimercaptosuccinic acid (DMSA). Free thiol groups on the surface of MSA- or DMSA- coated nanoparticles were nitrosated, leading to the formation of NO-releasing iron oxide nanoparticles. The cytotoxicity of MSA- or DMSA-coated magnetic nanoparticles (MNP) (thiolated nanoparticles) and nitrosated MSA- or nitrosated DMSA- coated MNPs (NO-releasing nanoparticles) were evaluated towards human lymphocytes. The results showed that MNP-MSA and MNP-DMSA have low cytotoxicity effects. On the other hand, NO-releasing MNPs were found to increase apoptosis and cell death compared to free NO-nanoparticles. Therefore, the cytotoxicity effects observed for NO-releasing MNPs may result in important biomedical applications, such as the treatment of tumors cells.

  15. Labeling of immune cells for in vivo imaging using magnetofluorescent nanoparticles.

    PubMed

    Pittet, Mikael J; Swirski, Filip K; Reynolds, Frederick; Josephson, Lee; Weissleder, Ralph

    2006-01-01

    Observation of immune and stem cells in their native microenvironments requires the development of imaging agents to allow their in vivo tracking. We describe here the synthesis of magnetofluorescent nanoparticles for cell labeling in vitro and for multimodality imaging of administered cells in vivo. MION-47, a prototype monocrystalline iron oxide nanoparticle, was first converted to an intermediate bearing a fluorochrome and amine groups, then reacted with either HIV-Tat peptide or protamine to yield a nanoparticle with membrane-translocating properties. We describe how to assess optimal cell labeling with tests of cell phenotype and function. Synthesis of magnetofluorescent nanoparticles and cell-labeling optimization can be realized in 48 h, whereas nanoparticle uptakes and retention studies may generally take up to 120 h. Labeled cells can be detected by magnetic resonance imaging, fluorescence reflectance imaging, fluorescence-mediated tomography, confocal microscopy and flow cytometry, and can be purified based on their fluorescent or magnetic properties. The present protocol focuses on T-cell labeling but can be used for labeling a variety of circulating cells. PMID:17406214

  16. Analysis of the Influence of Cell Heterogeneity on Nanoparticle Dose Response

    PubMed Central

    2014-01-01

    Understanding the effect of variability in the interaction of individual cells with nanoparticles on the overall response of the cell population to a nanoagent is a fundamental challenge in bionanotechnology. Here, we show that the technique of time-resolved, high-throughput microscopy can be used in this endeavor. Mass measurement with single-cell resolution provides statistically robust assessments of cell heterogeneity, while the addition of a temporal element allows assessment of separate processes leading to deconvolution of the effects of particle supply and biological response. We provide a specific demonstration of the approach, in vitro, through time-resolved measurement of fibroblast cell (HFF-1) death caused by exposure to cationic nanoparticles. The results show that heterogeneity in cell area is the major source of variability with area-dependent nanoparticle capture rates determining the time of cell death and hence the form of the exposure–response characteristic. Moreover, due to the particulate nature of the nanoparticle suspension, there is a reduction in the particle concentration over the course of the experiment, eventually causing saturation in the level of measured biological outcome. A generalized mathematical description of the system is proposed, based on a simple model of particle depletion from a finite supply reservoir. This captures the essential aspects of the nanoparticle–cell interaction dynamics and accurately predicts the population exposure–response curves from individual cell heterogeneity distributions. PMID:24923782

  17. Nanoparticles containing allotropes of carbon have genotoxic effects on glioblastoma multiforme cells

    PubMed Central

    Hinzmann, Mateusz; Jaworski, S?awomir; Kutwin, Marta; Jagie??o, Joanna; Kozi?ski, Rafa?; Wierzbicki, Mateusz; Grodzik, Marta; Lipi?ska, Ludwika; Sawosz, Ewa; Chwalibog, Andrè

    2014-01-01

    The carbon-based nanomaterial family consists of nanoparticles containing allotropes of carbon, which may have a number of interactions with biological systems. The objective of this study was to evaluate the toxicity of nanoparticles comprised of pristine graphene, reduced graphene oxide, graphene oxide, graphite, and ultradispersed detonation diamond in a U87 cell line. The scope of the work consisted of structural analysis of the nanoparticles using transmission electron microscopy, evaluation of cell morphology, and assessment of cell viability by Trypan blue assay and level of DNA fragmentation of U87 cells after 24 hours of incubation with 50 ?g/mL carbon nanoparticles. DNA fragmentation was studied using single-cell gel electrophoresis. Incubation with nanoparticles containing the allotropes of carbon did not alter the morphology of the U87 cancer cells. However, incubation with pristine graphene and reduced graphene oxide led to a significant decrease in cell viability, whereas incubation with graphene oxide, graphite, and ultradispersed detonation diamond led to a smaller decrease in cell viability. The results of a comet assay demonstrated that pristine graphene, reduced graphene oxide, graphite, and ultradispersed detonation diamond caused DNA damage and were therefore genotoxic in U87 cells, whereas graphene oxide was not. PMID:24876774

  18. Chimeric antigen receptor T Cells with dissociated signaling domains exhibit focused antitumor activity with reduced potential for toxicity in vivo.

    PubMed

    Lanitis, Evripidis; Poussin, Mathilde; Klattenhoff, Alex W; Song, Degang; Sandaltzopoulos, Raphael; June, Carl H; Powell, Daniel J

    2013-07-01

    Adoptive immunotherapy using T lymphocytes genetically modified to express a chimeric antigen receptor (CAR-T) holds considerable promise for the treatment of cancer. However, CAR-based therapies may involve on-target toxicity against normal tissues expressing low amounts of the targeted tumor-associated antigen (TAA). To specify T cells for robust effector function that is selective for tumor but not normal tissue, we developed a trans-signaling CAR strategy, whereby T-cell activation signal 1 (CD3z) is physically dissociated from costimulatory signal 2 (CD28) in two CARs of differing antigen specificity: mesothelin and a-folate receptor (FRa). Human T cells were genetically modified to coexpress signal 1 (anti-Meso scFv-CD3z) and signal 2 (anti-FRa scFv-CD28) CARs in trans. Trans-signaling CAR-T cells showed weak cytokine secretion against target cells expressing only one TAA in vitro, similar to first-generation CAR-T cells bearing CD3z only, but showed enhanced cytokine secretion upon encountering natural or engineered tumor cells coexpressing both antigens, equivalent to that of second-generation CAR-T cells with dual signaling in cis. CAR-T cells with dual specificity also showed potent anticancer activity and persistence in vivo, which was superior to first-generation CAR-T cells and equivalent to second-generation CARs. Importantly, second-generation CAR-T cells exhibited potent activity against cells expressing mesothelin alone, recapitulating normal tissue, whereas trans-signaling CAR-T cells did not. Thus, a dual specificity, trans-signaling CAR approach can potentiate the therapeutic efficacy of CAR-T cells against cancer while minimizing parallel reactivity against normal tissues bearing single antigen. PMID:24409448

  19. Targeting prostate cancer cells with PSMA inhibitor-guided gold nanoparticles

    PubMed Central

    Kasten, Benjamin B.; Liu, Tiancheng; Nedrow-Byers, Jessie R.; Benny, Paul D.; Berkman, Clifford E.

    2012-01-01

    Prostate-specific membrane antigen (PSMA) is a notable biomarker for diagnostic and therapeutic applications in prostate cancer. Gold nanoparticles (AuNPs) provide an attractive nanomaterial platform for combining a variety of targeting, imaging, and cytotoxic agents into a unified device for biomedical research. In this study, we present the generation and evaluation of the first AuNP system functionalized with a small molecule phosphoramidate peptidomimetic inhibitor for the targeted delivery to PSMA-expressing prostate cancer cells. The general approach involved the conjugation of streptavidin-coated AuNPs with a biotin-linked PSMA inhibitor (CTT54) to generate PSMA-targeted AuNPs. In vitro evaluations of these targeted AuNPs were conducted to determine PSMA-mediated and time-dependent binding to PSMA-positive LNCaP cells. The PSMA-targeted AuNPs exhibited significantly higher and selective binding to LNCaP cells compared to control non-targeted AuNPs, thus demonstrating the feasibility of this approach. PMID:23232055

  20. Targeting prostate cancer cells with PSMA inhibitor-guided gold nanoparticles.

    PubMed

    Kasten, Benjamin B; Liu, Tiancheng; Nedrow-Byers, Jessie R; Benny, Paul D; Berkman, Clifford E

    2013-01-15

    Prostate-specific membrane antigen (PSMA) is a notable biomarker for diagnostic and therapeutic applications in prostate cancer. Gold nanoparticles (AuNPs) provide an attractive nanomaterial platform for combining a variety of targeting, imaging, and cytotoxic agents into a unified device for biomedical research. In this study, we present the generation and evaluation of the first AuNP system functionalized with a small molecule phosphoramidate peptidomimetic inhibitor for the targeted delivery to PSMA-expressing prostate cancer cells. The general approach involved the conjugation of streptavidin-coated AuNPs with a biotin-linked PSMA inhibitor (CTT54) to generate PSMA-targeted AuNPs. In vitro evaluations of these targeted AuNPs were conducted to determine PSMA-mediated and time-dependent binding to PSMA-positive LNCaP cells. The PSMA-targeted AuNPs exhibited significantly higher and selective binding to LNCaP cells compared to control non-targeted AuNPs, thus demonstrating the feasibility of this approach. PMID:23232055

  1. Toxicity and antibacterial assessment of chitosancoated silver nanoparticles on human pathogens and macrophage cells

    PubMed Central

    Jena, Prajna; Mohanty, Soumitra; Mallick, Rojee; Jacob, Biju; Sonawane, Avinash

    2012-01-01

    Background Pathogenic bacteria are able to develop various strategies to counteract the bactericidal action of antibiotics. Silver nanoparticles (AgNPs) have emerged as a potential alternative to conventional antibiotics because of their potent antimicrobial properties. The purpose of this study was to synthesize chitosan-stabilized AgNPs (CS-AgNPs) and test for their cytotoxic, genotoxic, macrophage cell uptake, antibacterial, and antibiofilm activities. Methods AgNPs were synthesized using chitosan as both a stabilizing and a reducing agent. Antibacterial activity was determined by colony-forming unit assay and scanning electron microscopy. Genotoxic and cytotoxic activity were determined by DNA fragmentation, comet, and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays. Cellular uptake and intracellular antibacterial activity were tested on macrophages. Results CS-AgNPs exhibited potent antibacterial activity against different human pathogens and also impeded bacterial biofilm formation. Scanning electron microscopy analysis indicated that CS-AgNPs kill bacteria by disrupting the cell membrane. CS-AgNPs showed no significant cytotoxic or DNA damage effect on macrophages at the bactericidal dose. Propidium iodide staining indicated active endocytosis of CS-AgNPs resulting in reduced intracellular bacterial survival in macrophages. Conclusion The present study concludes that at a specific dose, chitosan-based AgNPs kill bacteria without harming the host cells, thus representing a potential template for the design of antibacterial agents to decrease bacterial colonization and to overcome the problem of drug resistance. PMID:22619529

  2. Enhanced performance of dye-sensitized solar cells using gold nanoparticles modified fluorine tin oxide electrodes

    NASA Astrophysics Data System (ADS)

    Zhang, Dingwen; Wang, Milton; Brolo, Alexandre G.; Shen, Jie; Li, Xiaodong; Huang, Sumei

    2013-01-01

    We have investigated plasmon-assisted energy conversion in dye-sensitized solar cells (DSCs) applying gold nanoparticles (NPs) modified fluorine tin oxide (FTO) electrodes. A series of Au NPs with different sizes (15-80 nm) were synthesized and immobilized onto FTO glass slides. Photoanodes were prepared on these Au modified FTO substrates using P25 TiO2 powders and by the screen-printing method. The size effects of Au NPs on the photovoltaic performance of the formed DSCs were investigated systematically. Structural and photoelectrochemical properties of the formed photoanodes were examined by field emission scanning electron microscopy and electrochemical impedance spectroscopy. It was found that the energy conversion efficiency of the DSC was highly dependent on the Au particle size. When the particle size was not greater than 60 nm, the DSC based on the Au NP-FTO composite electrode showed a higher short-circuit current density and better photovoltaic (PV) performance than the cell based on the bare FTO. The best cell was achieved using 25 nm sized Au NPs modified FTO. It exhibited a conversion efficiency of 6.69%, which was 15% higher than that of DSCs without Au NPs. The related PV performance enhancement mechanisms, photoelectrochemical processes and surface-plasmon resonances in DSCs with Au nanostructures are analysed and discussed.

  3. Iron nanoparticles increase 7-ketocholesterol-induced cell death, inflammation, and oxidation on murine cardiac HL1-NB cells

    PubMed Central

    Kahn, Edmond; Baarine, Mauhamad; Pelloux, Sophie; Riedinger, Jean-Marc; Frouin, Frédérique; Tourneur, Yves; Lizard, Gérard

    2010-01-01

    Objective: To evaluate the cytotoxicity of iron nanoparticles on cardiac cells and to determine whether they can modulate the biological activity of 7-ketocholesterol (7KC) involved in the development of cardiovascular diseases. Nanoparticles of iron labeled with Texas Red are introduced in cultures of nonbeating mouse cardiac cells (HL1-NB) with or without 7-ketocholesterol 7KC, and their ability to induce cell death, pro-inflammatory and oxidative effects are analyzed simultaneously. Study design: Flow cytometry (FCM), confocal laser scanning microscopy (CLSM), and subsequent factor analysis image processing (FAMIS) are used to characterize the action of iron nanoparticles and to define their cytotoxicity which is evaluated by enhanced permeability to SYTOX Green, and release of lactate deshydrogenase (LDH). Pro-inflammatory effects are estimated by ELISA in order to quantify IL-8 and MCP-1 secretions. Pro-oxidative effects are measured with hydroethydine (HE). Results: Iron Texas Red nanoparticles accumulate at the cytoplasmic membrane level. They induce a slight LDH release, and have no inflammatory or oxidative effects. However, they enhance the cytotoxic, pro-inflammatory and oxidative effects of 7KC. The accumulation dynamics of SYTOX Green in cells is measured by CLSM to characterize the toxicity of nanoparticles. The emission spectra of SYTOX Green and nanoparticles are differentiated, and corresponding factor images specify the possible capture and cellular localization of nanoparticles in cells. Conclusion: The designed protocol makes it possible to show how Iron Texas Red nanoparticles are captured by cardiomyocytes. Interestingly, whereas these fluorescent iron nanoparticles have no cytotoxic, pro-inflammatory or oxidative activities, they enhance the side effects of 7KC. PMID:20463934

  4. Oritavancin exhibits dual mode of action to inhibit cell-wall biosynthesis in Staphylococcus aureus

    PubMed Central

    Kim, Sung Joon; Cegelski, Lynette; Stueber, Dirk; Singh, Manmilan; Dietrich, Evelyne; Tanaka, Kelly S. E.; Parr, Thomas R.; Far, Adel Rafai; Schaefer, Jacob

    2008-01-01

    Solid-state NMR measurements performed on intact whole cells of S. aureus labeled selectively in vivo have established that des-N-methylleucyl oritavancin (which has antimicrobial activity) binds to the cell-wall peptidoglycan, even though removal of the terminal N-methylleucyl residue destroys the D-Ala-D-Ala binding pocket. By contrast, the des-N-methylleucyl form of vancomycin (which has no antimicrobial activity) does not bind to the cell wall. Solid-state NMR has also determined that oritavancin and vancomycin are comparable inhibitors of transglycosylation, but that oritavancin is a more potent inhibitor of transpeptidation. This combination of effects on cell-wall binding and biosynthesis is interpreted in terms of a recent proposal that oritavancin-like glycopeptides have two cell-wall binding sites: the well-known peptidoglycan D-Ala-D-Ala pentapeptide stem terminus, and the pentaglycyl bridging segment. The resulting dual mode of action provides a structural framework for coordinated cell-wall assembly that accounts for the enhanced potency of oritavancin and oritavancin-like analogues against vancomycin-resistant organisms. PMID:18258256

  5. The natural products parthenolide and andrographolide exhibit anti-cancer stem cell activity in multiple myeloma.

    PubMed

    Gunn, Ellen J; Williams, John T; Huynh, Daniel T; Iannotti, Michael J; Han, Changho; Barrios, Francis J; Kendall, Stephen; Glackin, Carlotta A; Colby, David A; Kirshner, Julia

    2011-06-01

    Multiple myeloma (MM) is an incurable plasma cell malignancy where nearly all patients succumb to a relapse. The current preclinical models of MM target the plasma cells, constituting the bulk of the tumor, leaving the cancer stem cells to trigger a relapse. Utilizing a three-dimensional tissue culture system where cells were grown in extracellular matrix designed to reconstruct human bone marrow, we tested the anti-multiple myeloma cancer stem cell (MM-CSC) potential of two natural product inhibitors of nuclear factor ?B (NF?B). Here we show that parthenolide and andrographolide are potent anti-MM-CSC agents. Both natural products demonstrated preferential toxicity toward MM-CSCs over non-tumorigenic MM cells. Addition of the bone marrow stromal compartment abrogated andrographolide activity while having no effect on parthenolide cytoxicity. This is the first report of a natural product with anti-CSC activity in myeloma, suggesting that it has the potential to improve the survival of patients with MM by eliminating the relapse-causing MM-CSCs. PMID:21417826

  6. Gold nanoparticles as physiological markers of urine internalization into urothelial cells in vivo

    PubMed Central

    Hudoklin, Samo; Zupan?i?, Daša; Makovec, Darko; Kreft, Mateja Erdani; Romih, Rok

    2013-01-01

    Background Urothelial bladder is the reservoir of urine and the urothelium minimizes the exchange of urine constituents with this tissue. Our aim was to test 1.9 nm biocompatible gold nanoparticles as a novel marker of internalization into the urothelial cells under physiological conditions in vivo. Methods We compared normal and neoplastic mice urothelium. Neoplastic lesions were induced by 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in drinking water for 10 weeks. Nanoparticles, intravenously injected into normal and BBN-treated mice, were filtered through the kidneys and became constituents of the urine within 90 minutes after injection. Results Gold nanoparticles were densely accumulated in the urine, while their internalization into urothelial cells depended on the cell differentiation stage. In the terminally differentiated superficial urothelial cells of normal animals, nanoparticles were occasionally found in the endosomes, but not in the fusiform vesicles. Regions of exfoliated cells were occasionally found in the normal urothelium. Superficial urothelial cells located next to exfoliated regions contained gold nanoparticles in the endosomes and in the cytosol beneath the apical plasma membrane. The urothelium of BBN-treated animals developed fat hyperplasia with moderate dysplasia. The superficial cells of BBN-treated animals were partially differentiated as demonstrated by the lack of fusiform vesicles. These cells contained the gold nanoparticles distributed in the endosomes and throughout their cytosol. Conclusion Gold nanoparticles are a valuable marker to study urine internalization into urothelial cells in vivo. Moreover, they can be used as a sensitive marker of differentiation and functionality of urothelial cells. PMID:24143099

  7. Nanoparticles, [Gd@C82(OH)22]n, induces dendritic cell maturation and activates Th1 immune responses

    PubMed Central

    Yang, De; Zhao, Yuliang; Guo, Hua; Li, Yana; Tewary, Poonam; Xing, Gengmei; Hou, Wei; Oppenheim, Joost J.; Zhang, Ning

    2010-01-01

    Dendritic cells play a pivotal role in host immune defense, such as elimination of foreign pathogen and inhibition of tumorigenesis. In this paper, we report that [Gd@C82(OH)22]n could induce phenotypic maturation of dendritic cells by stimulating DC production of cytokines including IL-12p70, upregulating DC costimulatory (CD80, CD83, and CD86) and MHC (HLA-A,B,C and HLA-DR) molecules, and switching DCs from a CCL5-responsive to a CCL19-responsive phenotype. We found that [Gd@C82(OH)22]n can induce dendritic cells to become functionally mature as illustrated by their capacity to activate allogeneic T cells. Mice immunized with ovalbumin in the presence of [Gd@C82(OH)22]n exhibit enhanced ovalbumin-specific Th1-polarized immune response as evidenced by the predominantly increased production of IFN?, IL-1?, and IL-2. The [Gd@C82(OH)22]n nanoparticle is a potent activator of dendritic cells and Th1 immune responses. These new findings also provide a rational understanding of the potent anticancer activities of [Gd@C82(OH)22]n nanoparticles reported previously. PMID:20121217

  8. A Micro-Raman Study of Live, Single Red Blood Cells (RBCs) Treated with AgNO3 Nanoparticles

    PubMed Central

    Bankapur, Aseefhali; Barkur, Surekha; Chidangil, Santhosh; Mathur, Deepak

    2014-01-01

    Silver nanoparticles (Ag NPs) are known to exhibit broad antimicrobial activity. However, such activity continues to raise concerns in the context of the interaction of such NPs with biomolecules. In a physiological environment NPs interact with individual biological cells either by penetrating through the cell membrane or by adhering to the membrane. We have explored the interaction of Ag NPs with single optically-trapped, live erythrocytes (red blood cells, RBCs) using Raman Tweezers spectroscopy. Our experiments reveal that Ag NPs induce modifications within an RBC that appear to be irreversible. In particular we are able to identify that the heme conformation in an RBC transforms from the usual R-state (oxy-state) to the T-state (deoxy-state). We rationalize our observations by proposing a model for the nanoparticle cytotoxicity pathway when the NP size is larger than the membrane pore size. We propose that the interaction of Ag NPs with the cell surface induces damage brought about by alteration of intracellular pH caused by the blockage of the cell membrane transport. PMID:25057913

  9. Multicomponent polymeric nanoparticles enhancing intracellular drug release in cancer cells.

    PubMed

    Ahmed, Arsalan; Liu, Sen; Pan, Yutong; Yuan, Shanmei; He, Jian; Hu, Yong

    2014-12-10

    Three kinds of amphiphilic copolymer, that is, poly(?-caprolactone)-SS-poly(ethylene glycol) (PCL-SS-PEG), poly(?-caprolactone)-polyethylenimine (PCL-PEI), and poly(?-caprolactone)-polyethylenimine-folate (PCL-PEI-Fol) were synthesized and self-assembled into surface engineered hybrid nanoparticles (NPs). Morphological studies elucidated the stable, spherical, and uniform sandwich structure of the NPs. PCL-PEI and PCL-SS-PEG segments have introduced pH and reduction responsive characteristics in these NPs, while PCL-PEI-FA copolymers could provide specific targeting capability to cancer cells. The stimuli responsive capabilities of these NPs were carried out. Negative-to-positive charge reversible property, in response to the pH change, was investigated by zeta potential and nuclear magnetic resonance (NMR) measurements. The structure cleavage, due to redox gradient, was studied by dynamic light scattering (DLS) and transmission electron microscopy (TEM). These NPs showed controlled degradation, better drug release, less toxicity, and effective uptake in MCF-7 breast cancer cells. These multifunctional NPs showed promising potential in the treatment of cancer. PMID:25333325

  10. Evaluation of cytotoxic, genotoxic and inflammatory responses of nanoparticles from photocopiers in three human cell lines

    PubMed Central

    2013-01-01

    Background Photocopiers emit nanoparticles with complex chemical composition. Short-term exposures to modest nanoparticle concentrations triggered upper airway inflammation and oxidative stress in healthy human volunteers in a recent study. To further understand the toxicological properties of copier-emitted nanoparticles, we studied in-vitro their ability to induce cytotoxicity, pro-inflammatory cytokine release, DNA damage, and apoptosis in relevant human cell lines. Methods Three cell types were used: THP-1, primary human nasal- and small airway epithelial cells. Following collection in a large volume photocopy center, nanoparticles were extracted, dispersed and characterized in the cell culture medium. Cells were doped at 30, 100 and 300 ?g/mL administered doses for up to 24 hrs. Estimated dose delivered to cells, was ~10% and 22% of the administered dose at 6 and 24 hrs, respectively. Gene expression analysis of key biomarkers was performed using real time quantitative PCR (RT-qPCR) in THP-1 cells at 5 ?g nanoparticles/mL for 6-hr exposure for confirmation purposes. Results Multiple cytokines, GM-CSF, IL-1?, IL-6, IL-8, IFN?, MCP-1, TNF-? and VEGF, were significantly elevated in THP-1 cells in a dose-dependent manner. Gene expression analysis confirmed up-regulation of the TNF-? gene in THP-1 cells, consistent with cytokine findings. In both primary epithelial cells, cytokines IL-8, VEGF, EGF, IL-1?, TNF-?, IL-6 and GM-CSF were significantly elevated. Apoptosis was induced in all cell lines in a dose-dependent manner, consistent with the significant up-regulation of key apoptosis-regulating genes P53 and Casp8 in THP-1 cells. No significant DNA damage was found at any concentration with the comet assay. Up-regulation of key DNA damage and repair genes, Ku70 and Rad51, were also observed in THP-1 cells, albeit not statistically significant. Significant up-regulation of the key gene HO1 for oxidative stress, implicates oxidative stress induced by nanoparticles. Conclusions Copier-emitted nanoparticles induced the release of pro-inflammatory cytokines, apoptosis and modest cytotoxicity but no DNA damage in all three-human cell lines. Taken together with gene expression data in THP-1 cells, we conclude that these nanoparticles are directly responsible for inflammation observed in human volunteers. Further toxicological evaluations of these nanoparticles, including across different toner formulations, are warranted. PMID:23968360

  11. Copper Oxide Nanoparticles Induce Autophagic Cell Death in A549 Cells

    PubMed Central

    Zhao, Yan; Guo, Feng; Jiang, Chengyu

    2012-01-01

    Metal oxide nanoparticles (NPs) are among the most highly produced nanomaterials, and have many diverse functions in catalysis, environmental remediation, as sensors, and in the production of personal care products. In this study, the toxicity of several widely used metal oxide NPs such as copper oxide, silica, titanium oxide and ferric oxide NPs, were evaluated In vitro. We exposed A549, H1650 and CNE-2Z cell lines to metal oxide NPs, and found CuO NPs to be the most toxic, SiO2 mild toxic, while the other metal oxide NPs had little effect on cell viability. Furthermore, the autophagic biomarker LC3-II significantly increased in A549 cells treated with CuO NPs, and the use of the autophagy inhibitors wortmannin and 3-methyladenin significantly improved cell survival. These results indicate that the cytoxicity of CuO NPs may involve the autophagic pathway in A549 cells. PMID:22916263

  12. Design of a TiO2 nanosheet/nanoparticle gradient film photoanode and its improved performance for dye-sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Wang, Wenguang; Zhang, Haiyan; Wang, Rong; Feng, Ming; Chen, Yiming

    2014-01-01

    A TiO2 film photoanode with gradient structure in nanosheet/nanoparticle concentration on the fluorine-doped tin oxide glass from substrate to surface was prepared by a screen printing method. The as-prepared dye-sensitized solar cell (DSSC) based on the gradient film electrode exhibited an enhanced photoelectric conversion efficiency of 6.48%, exceeding that of a pure nanoparticle-based DSSC with the same film thickness by a factor of 2.6. The enhanced photovoltaic performance of the gradient film-based DSSC was attributed to the superior light scattering ability of TiO2 nanosheets within the gradient structure, which was beneficial to light harvesting. Furthermore, the TiO2 nanosheets with exposed {001} facets facilitated the electron transport from dye molecules to the conduction band of TiO2 and further to the conductive glass. Meanwhile, the high specific surface area of TiO2 nanosheets helped the adsorption of dye molecules, and the TiO2 nanoparticle underlayer ensured good electronic contact between the TiO2 film and the fluorine-doped tin oxide glass substrate. The electrochemical impedance spectroscopy measurements further confirmed the electron transport differences between DSSCs based on nanosheet/nanoparticle gradient film electrodes and DSSCs based on nanosheet/nanoparticle homogeneous mixtures, pure TiO2 nanoparticles and pure TiO2 nanosheets with the same film thickness.

  13. Design of a TiO2 nanosheet/nanoparticle gradient film photoanode and its improved performance for dye-sensitized solar cells.

    PubMed

    Wang, Wenguang; Zhang, Haiyan; Wang, Rong; Feng, Ming; Chen, Yiming

    2014-02-21

    A TiO2 film photoanode with gradient structure in nanosheet/nanoparticle concentration on the fluorine-doped tin oxide glass from substrate to surface was prepared by a screen printing method. The as-prepared dye-sensitized solar cell (DSSC) based on the gradient film electrode exhibited an enhanced photoelectric conversion efficiency of 6.48%, exceeding that of a pure nanoparticle-based DSSC with the same film thickness by a factor of 2.6. The enhanced photovoltaic performance of the gradient film-based DSSC was attributed to the superior light scattering ability of TiO2 nanosheets within the gradient structure, which was beneficial to light harvesting. Furthermore, the TiO2 nanosheets with exposed {001} facets facilitated the electron transport from dye molecules to the conduction band of TiO2 and further to the conductive glass. Meanwhile, the high specific surface area of TiO2 nanosheets helped the adsorption of dye molecules, and the TiO2 nanoparticle underlayer ensured good electronic contact between the TiO2 film and the fluorine-doped tin oxide glass substrate. The electrochemical impedance spectroscopy measurements further confirmed the electron transport differences between DSSCs based on nanosheet/nanoparticle gradient film electrodes and DSSCs based on nanosheet/nanoparticle homogeneous mixtures, pure TiO2 nanoparticles and pure TiO2 nanosheets with the same film thickness. PMID:24435106

  14. Laser-ablated titania nanoparticles for aqueous processed hybrid solar cells

    NASA Astrophysics Data System (ADS)

    Körstgens, V.; Pröller, S.; Buchmann, T.; Moseguí González, D.; Song, L.; Yao, Y.; Wang, W.; Werhahn, J.; Santoro, G.; Roth, S. V.; Iglev, H.; Kienberger, R.; Müller-Buschbaum, P.

    2015-02-01

    Titania nanoparticles are produced by laser ablation in liquid in order to initiate functionalization of titania with the polymer for the active layer. By combining these titania nanoparticles and water-soluble poly[3-(potassium-6-hexanoate)thiophene-2,5-diyl] (P3P6T) hybrid solar cells are realized.Titania nanoparticles are produced by laser ablation in liquid in order to initiate functionalization of titania with the polymer for the active layer. By combining these titania nanoparticles and water-soluble poly[3-(potassium-6-hexanoate)thiophene-2,5-diyl] (P3P6T) hybrid solar cells are realized. Electronic supplementary information (ESI) available: Full scheme of the production of solar cells, additional spectra and details of the measurement techniques. See DOI: 10.1039/c4nr06782g

  15. In vitro nanoparticle toxicity to rat alveolar cells and coelomocytes from the earthworm Lumbricus rubellus.

    PubMed

    van der Ploeg, Merel J C; van den Berg, Johannes H J; Bhattacharjee, Sourav; de Haan, Laura H J; Ershov, Dmitry S; Fokkink, Remco G; Zuilhof, Han; Rietjens, Ivonne M C M; van den Brink, Nico W

    2014-02-01

    Sensitivity of immune cells (coelomocytes) of Lumbricus rubellus earthworms was investigated for exposure to selected nanoparticles, in order to obtain further insight in mechanisms of effects observed after in vivo C60 exposure. In the in vivo study, tissue damage appeared to occur without accompanying increased immune responses. Coelomocytes exposed in vitro to C60 showed no decrease of their cellular viability, but demonstrated a decrease in gene expression of the cytokine-like protein CCF-1, indicating immunosuppression. Experiments with NR8383 rat macrophage cells and tri-block copolymer nanoparticles were used to compare sensitivity and to demonstrate the usefulness of coelomocytes as a test system for nano-immunotoxicity, respectively. Overall, the results imply that sensitivity towards nanoparticles differs between cell types and nanoparticles. Moreover, this study indicates that injuries in absence of an immune response, observed after in vivo C60 exposure in our earlier work, are caused by immunosuppression rather than coelomocyte mortality. PMID:23102209

  16. Uropathogenic Escherichia coli isolates with different virulence genes content exhibit similar pathologic influence on Vero cells.

    PubMed

    Obaid, Jamil M A S; Mansour, Samira R; Elshahedy, Mohammed S; Rabie, Tarik E; Azab, Adel M H

    2014-01-01

    Uropathogenic Escherichia coli are the major causative agent of urinary tract infection--they may simultaneously express a number of virulence factors to cause disease. The aim of this study was to investigate the relation between virulence factors content of fifteen UPEC isolates and their pathogenic potential. The isolates belonged to the five serotypes O78:K80, O114:K90, O142:K86, O164 and O157. Nine of the virulence factors have been explored, ibeA, pap, sfa/foc, cnfl, hly, fyuA, pil, ompT and traT. Virulence factors profiling of the isolates revealed a different content ranging from 22% to 100% of the virulence genes explored. The pathogenic capacity of all fifteen isolates when tested on Vero cells showed that the cytotoxicity for all tested strains on Vero cells was approximately equal and enhanced after growth in syncase broth, leading mainly to cell lysis. The toxic effects reduced slightly after heat treatment of the toxin, and greatly after formalin detoxification, but not all the deleterious effect was abolished. Endotoxin also has cytotoxic effects on Vero cells, but longer time is needed for cytolysis which is greatly diminished with formalin treatment. In conclusion, our study revealed that pathogenic strains of UPEC can exert their pathogenic effect on live cells or system with limited virulence factors gene content. PMID:25033661

  17. The ALDH1+ subpopulation of the human NMFH?1 cell line exhibits cancer stem-like characteristics.

    PubMed

    Li, Dejian; Zhang, Tao; Gu, Wenguang; Li, Peng; Cheng, Xiangyang; Tong, Tiejun; Wang, Wenbo

    2015-05-01

    Cancer stem cells (CSCs) have been reported in many tissues. However, CSCs have yet to be identified in a human malignant fibrous histiocytoma (MFH) cell line. Elevated aldehyde dehydrogenase 1 (ALDH1) has been proposed as a stem cell marker for isolating CSCs from cancer. The aim of the present study was to identify a population with elevated ALDH in the human NMFH-1 cell line. ALDH+ and ALDH- cell populations were isolated and compared for CSC characteristics. ALDH enzymatic activity was used as a marker to identify the cells in the NMFH-1 line. Self-renewal, differentiation capacity, and tumorigenicity of the NMFH-1 ALDH+ cell population were then examined using a spheroid formation assay and xenograft model in nude mice. Chemoresistance levels, ABCG2 drug transport gene expression, and stem cell?associated gene expression were compared in these NMFH-1 populations. The ALDH+ population was better able to form spheres in anchorage-independent serum?starved conditions. Furthermore, the mRNA expression of key stem cell-related genes was enhanced in these cells. Increased expression of the drug transporter gene, ABCG2, was detected. Compared with ALDH-, the ALDH+ subpopulation had higher levels of chemoresistance to doxorubicin (DXR) and cisplatin (CDDP). Additionally, the ALDH+ cells more efficiently formed tumors when implanted into BALB/c nude mice. ALDH1 may therefore be used as a marker for the isolation of cells that exhibit several characteristics of CSCs from the NMFH-1 cell line. This finding may lead to the development of novel therapies to specifically kill ALDH1+ subpopulations (CSCs). PMID:25760144

  18. Enhanced cellular uptake of aminosilane-coated superparamagnetic iron oxide nanoparticles in mammalian cell lines

    PubMed Central

    Zhu, Xiao-Ming; Wang, Yi-Xiang J; Leung, Ken Cham-Fai; Lee, Siu-Fung; Zhao, Feng; Wang, Da-Wei; Lai, Josie MY; Wan, Chao; Cheng, Christopher HK; Ahuja, Anil T

    2012-01-01

    Purpose To compare the cellular uptake efficiency and cytotoxicity of aminosilane (SiO2-NH2)-coated superparamagnetic iron oxide (SPIO@SiO2-NH2) nanoparticles with three other types of SPIO nanoparticles coated with SiO2 (SPIO@SiO2), dextran (SPIO@dextran), or bare SPIO in mammalian cell lines. Materials and methods Four types of monodispersed SPIO nanoparticles with a SPIO core size of 7 nm and an overall size in a range of 7–15 nm were synthesized. The mammalian cell lines of MCF-7, MDA-MB-231, HT-29, RAW264.7, L929, HepG2, PC-3, U-87 MG, and mouse mesenchymal stem cells (MSCs) were incubated with four types of SPIO nanoparticles for 24 hours in the serum-free culture medium Dulbecco’s modified Eagle’s medium (DMEM) with 4.5 ?g/mL iron concentration. The cellular uptake efficiencies of SPIO nanoparticles were compared by Prussian blue staining and intracellular iron quantification. In vitro magnetic resonance imaging of MSC pellets after SPIO labeling was performed at 3 T. The effect of each SPIO nanoparticle on the cell viability of RAW 264.7 (mouse monocyte/macrophage) cells was also evaluated. Results Transmission electron microscopy demonstrated surface coating with SiO2-NH2, SiO2, and dextran prevented SPIO nanoparticle aggregation in DMEM culture medium. MCF-7, MDA-MB-231, and HT-29 cells failed to show notable iron uptake. For all the remaining six cell lines, Prussian blue staining and intracellular iron quantification demonstrated that SPIO@ SiO2-NH2 nanoparticles had the highest cellular uptake efficiency. SPIO@SiO2-NH2, bare SPIO, and SPIO@dextran nanoparticles did not affect RAW 264.7 cell viability up to 200 ?g Fe/mL, while SPIO@SiO2 reduced RAW 264.7 cell viability from 10 to 200 ?g Fe/mL in a dose-dependent manner. Conclusion Cellular uptake efficiency of SPIO nanoparticles depends on both the cell type and SPIO surface characteristics. Aminosilane surface coating enhanced the cellular uptake efficiency without inducing cytotoxicity in a number of cell lines. PMID:22393292

  19. CSMD1 exhibits antitumor activity in A375 melanoma cells through activation of the Smad pathway.

    PubMed

    Tang, Ming-Rui; Wang, Yu-Xin; Guo, Shu; Han, Si-Yuan; Wang, Di

    2012-09-01

    In this work, we studied the effects of CUB and Sushi multiple domains 1 gene (CSMD1) expression in A375 melanoma cells in vivo and in vitro. CSDM1 expression decreased proliferation and migration, and increased apoptosis and G(1) arrest in A375 cells in vitro. Expression of CSDM1 in established xenografted tumors decreased tumor size and weight, and decreased the density of intratumor microvessels. The survival rate of mice with tumors expressing CSMD1 was significantly higher than mice with tumors that did not express CSDM1. These results confirm the role of CSDM1 as a tumor suppressor gene in melanoma cells. Furthermore, we found that CSMD1 can interact with Smad3, activate Smad1, Smad2, and Smad3, and increase the expression of Smad4. These results might prove helpful for the development of novel therapies for melanoma treatment. PMID:22538441

  20. Metre-long cell-laden microfibres exhibit tissue morphologies and functions

    NASA Astrophysics Data System (ADS)

    Onoe, Hiroaki; Okitsu, Teru; Itou, Akane; Kato-Negishi, Midori; Gojo, Riho; Kiriya, Daisuke; Sato, Koji; Miura, Shigenori; Iwanaga, Shintaroh; Kuribayashi-Shigetomi, Kaori; Matsunaga, Yukiko T.; Shimoyama, Yuto; Takeuchi, Shoji

    2013-06-01

    Artificial reconstruction of fibre-shaped cellular constructs could greatly contribute to tissue assembly in vitro. Here we show that, by using a microfluidic device with double-coaxial laminar flow, metre-long core-shell hydrogel microfibres encapsulating ECM proteins and differentiated cells or somatic stem cells can be fabricated, and that the microfibres reconstitute intrinsic morphologies and functions of living tissues. We also show that these functional fibres can be assembled, by weaving and reeling, into macroscopic cellular structures with various spatial patterns. Moreover, fibres encapsulating primary pancreatic islet cells and transplanted through a microcatheter into the subrenal capsular space of diabetic mice normalized blood glucose concentrations for about two weeks. These microfibres may find use as templates for the reconstruction of fibre-shaped functional tissues that mimic muscle fibres, blood vessels or nerve networks in vivo.

  1. Growth of Pt nanoparticle for proton-exchange-membrane fuel cells by

    E-print Network

    PEMFC Growth of Pt nanoparticle for proton-exchange-membrane fuel cells at anode side of a polymer electrolyte membrane (PEM) fuel cell. With a Pt loading of 25 g-Pt/cm2 , current of the electrochemical test result and fuel cell performance agree with each other. Key word : Pulsed laser deposition

  2. Targeting of peptide conjugated magnetic nanoparticles to urokinase plasminogen activator receptor (uPAR) expressing cells

    NASA Astrophysics Data System (ADS)

    Hansen, Line; Unmack Larsen, Esben Kjær; Nielsen, Erik Holm; Iversen, Frank; Liu, Zhuo; Thomsen, Karen; Pedersen, Michael; Skrydstrup, Troels; Nielsen, Niels Chr.; Ploug, Michael; Kjems, Jørgen

    2013-08-01

    Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles are currently being used as a magnetic resonance imaging (MRI) contrast agent in vivo, mainly by their passive accumulation in tissues of interest. However, a higher specificity can ideally be achieved when the nanoparticles are targeted towards cell specific receptors and this may also facilitate specific drug delivery by an enhanced target-mediated endocytosis. We report efficient peptide-mediated targeting of magnetic nanoparticles to cells expressing the urokinase plasminogen activator receptor (uPAR), a surface biomarker for poor patient prognosis shared by several cancers including breast, colorectal, and gastric cancers. Conjugation of a uPAR specific targeting peptide onto polyethylene glycol (PEG) coated USPIO nanoparticles by click chemistry resulted in a five times higher uptake in vitro in a uPAR positive cell line compared to nanoparticles carrying a non-binding control peptide. In accordance with specific receptor-mediated recognition, a low uptake was observed in the presence of an excess of ATF, a natural ligand for uPAR. The uPAR specific magnetic nanoparticles can potentially provide a useful supplement for tumor patient management when combined with MRI and drug delivery.Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles are currently being used as a magnetic resonance imaging (MRI) contrast agent in vivo, mainly by their passive accumulation in tissues of interest. However, a higher specificity can ideally be achieved when the nanoparticles are targeted towards cell specific receptors and this may also facilitate specific drug delivery by an enhanced target-mediated endocytosis. We report efficient peptide-mediated targeting of magnetic nanoparticles to cells expressing the urokinase plasminogen activator receptor (uPAR), a surface biomarker for poor patient prognosis shared by several cancers including breast, colorectal, and gastric cancers. Conjugation of a uPAR specific targeting peptide onto polyethylene glycol (PEG) coated USPIO nanoparticles by click chemistry resulted in a five times higher uptake in vitro in a uPAR positive cell line compared to nanoparticles carrying a non-binding control peptide. In accordance with specific receptor-mediated recognition, a low uptake was observed in the presence of an excess of ATF, a natural ligand for uPAR. The uPAR specific magnetic nanoparticles can potentially provide a useful supplement for tumor patient management when combined with MRI and drug delivery. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr32922d

  3. The antibacterial substance taurolidine exhibits anti-neoplastic action based on a mixed type of programmed cell death.

    PubMed

    Stendel, Ruediger; Biefer, Hector Rodriguez Cetina; Dékány, Gabriela Marta; Kubota, Hisashi; Münz, Christian; Wang, Sheng; Mohler, Hanns; Yonekawa, Yasuhiro; Frei, Karl

    2009-02-01

    The antibacterial amino-acid derivative taurolidine (TAU) has been recently shown to exhibit anti-neoplastic activity based on a mechanism, which is still unknown in detail. Cytotoxicity and clonogenic assays were performed and the impact of apoptosis modulators, a radical scavenger, autophagy inhibitors, silencing of apoptosis inducing actor (AIF) and cytochrome-c (Cyt-C) by siRNA, and knockdown of autophagy related genes were evaluated in vitro. The intracellular ATP-content, release of AIF and Cyt-C, and DNA-laddering were investigated. This study could demonstrate cell killing, inhibition of proliferation, and inhibition or prevention of colony formation in human glioma cell lines and ex vivo glioblastoma cells after incubation with TAU. This effect is based on the induction of a mixed type of programmed cell death with the main preference of autophagy, and involvement of senescence, necroptosis and necrosis. This mechanism of action may open a new approach for therapeutic intervention. PMID:19066471

  4. Detection of single photoluminescent diamond nanoparticles in cells and study of the internalization pathway.

    PubMed

    Faklaris, Orestis; Garrot, Damien; Joshi, Vandana; Druon, Frédéric; Boudou, Jean-Paul; Sauvage, Thierry; Georges, Patrick; Curmi, Patrick A; Treussart, François

    2008-12-01

    Diamond nanoparticles are promising photoluminescent probes for tracking intracellular processes, due to embedded, perfectly photostable color centers. In this work, the spontaneous internalization of such nanoparticles (diameter 25 nm) in HeLa cancer cells is investigated by confocal microscopy and time-resolved techniques. Nanoparticles are observed inside the cell cytoplasm at the single-particle and single-color-center level, assessed by time-correlation intensity measurements. Improvement of the nanoparticle signal-to-noise ratio inside the cell is achieved using a pulsed-excitation laser and time-resolved detection taking advantage of the long radiative lifetime of the color-center excited state as compared to cell autofluorescence. The internalization pathways are also investigated, with endosomal marking and colocalization analyses. The low colocalization ratio observed proves that nanodiamonds are not trapped in endosomes, a promising result in prospect of drug delivery by these nanoparticles. Low cytotoxicity of these nanoparticles in this cell line is also shown. PMID:18989862

  5. The influence of surface functionalization on the enhanced internalization of magnetic nanoparticles in cancer cells

    NASA Astrophysics Data System (ADS)

    Villanueva, Angeles; Cañete, Magdalena; Roca, Alejandro G; Calero, Macarena; Veintemillas-Verdaguer, Sabino; Serna, Carlos J; del Puerto Morales, María; Miranda, Rodolfo

    2009-03-01

    The internalization and biocompatibility of iron oxide nanoparticles surface functionalized with four differently charged carbohydrates have been tested in the human cervical carcinoma cell line (HeLa). Neutral, positive, and negative iron oxide nanoparticles were obtained by coating with dextran, aminodextran, heparin, and dimercaptosuccinic acid, resulting in colloidal suspensions stable at pH 7 with similar aggregate size. No intracellular uptake was detected in cells incubated with neutral charged nanoparticles, while negative particles showed different behaviour depending on the nature of the coating. Thus, dimercaptosuccinic-coated nanoparticles showed low cellular uptake with non-toxic effects, while heparin-coated particles showed cellular uptake only at high nanoparticle concentrations and induced abnormal mitotic spindle configurations. Finally, cationic magnetic nanoparticles show excellent properties for possible in vivo biomedical applications such as cell tracking by magnetic resonance imaging (MRI) and cancer treatment by hyperthermia: (i) they enter into cells with high effectiveness, and are localized in endosomes; (ii) they can be easily detected inside cells by optical microscopy, (iii) they are retained for relatively long periods of time, and (iv) they do not induce any cytotoxicity.

  6. Theranostic Mesoporous Silica Nanoparticles Biodegrade after Pro-Survival Drug Delivery and Ultrasound/Magnetic Resonance Imaging of Stem Cells

    PubMed Central

    Kempen, Paul J.; Greasley, Sarah; Parker, Kelly A.; Campbell, Jos L.; Chang, Huan-Yu; Jones, Julian R.; Sinclair, Robert; Gambhir, Sanjiv S.; Jokerst, Jesse V.

    2015-01-01

    Increasing cell survival in stem cell therapy is an important challenge for the field of regenerative medicine. Here, we report theranostic mesoporous silica nanoparticles that can increase cell survival through both diagnostic and therapeutic approaches. First, the nanoparticle offers ultrasound and MRI signal to guide implantation into the peri-infarct zone and away from the most necrotic tissue. Second, the nanoparticle serves as a slow release reservoir of insulin-like growth factor (IGF)—a protein shown to increase cell survival. Mesenchymal stem cells labeled with these nanoparticles had detection limits near 9000 cells with no cytotoxicity at the 250 µg/mL concentration required for labeling. We also studied the degradation of the nanoparticles and showed that they clear from cells in approximately 3 weeks. The presence of IGF increased cell survival up to 40% (p<0.05) versus unlabeled cells under in vitro serum-free culture conditions.

  7. Enhancing the efficiency of gold nanoparticles treatment of cancer by increasing their rate of endocytosis and cell accumulation using rifampicin.

    PubMed

    Ali, Moustafa R K; Panikkanvalappil, Sajanlal R; El-Sayed, Mostafa A

    2014-03-26

    To minimize the toxicity of gold nanoparticles (AuNPs) in cancer treatment, we have developed a technique, which utilizes lesser amount of AuNPs while exhibiting increased treatment efficiency. Rifampicin (RF) is known for its ability to enhance the accumulation of anticancer drugs in multidrug resistant (MDR) cancer cells. In this work we have shown that RF-conjugated AuNPs can greatly enhance the rate as well as efficiency of endocytosis of NPs and hence their concentration inside the cancer cell. Cell viability results showed a remarkable enhancement in the photothermal therapeutic effect of Au nanorods in presence of RF. This is expected to decrease the demand on the overall amount of AuNPs needed for treating cancer and thus decreasing its toxicity. PMID:24467386

  8. Effect of magnetite nanoparticles on living rate of MCF-7 human breast cancer cells.

    PubMed

    Baba, Daisuke; Seiko, Yasuhiro; Nakanishi, Takuya; Zhang, Hong; Arakaki, Atsushi; Matsunaga, Tadashi; Osaka, Tetsuya

    2012-06-15

    Superparamagnetic and ferromagnetic magnetite nanoparticles, with diameters of approximately 13 and 44 nm, respectively, were synthesized and their uptake amount and heating efficiency were evaluated for application to magnetic hyperthermia. Both nanoparticles had almost the same zeta-potential (+10.2 mV) and hydrodynamic size (?1 ?m) and there was no significant difference in their uptake amount 18 h after they were added to the medium. After internalization, the ferromagnetic nanoparticles incorporated in human breast cancer cells (MCF-7) showed a higher heating efficiency than the superparamagnetic nanoparticles when an external magnetic field (4 kW, 250 kHz) high enough to produce heat by hysteresis loss was applied, followed by cellular death of MCF-7 with high ferromagnetic nanoparticle content. PMID:22486944

  9. Novel curcumin analogue IHCH exhibits potent anti?proliferative effects by inducing autophagy in A549 lung cancer cells.

    PubMed

    Zhou, Guang-Zhou; Xu, Su-Li; Sun, Gang-Chun; Chen, Xiao-Bing

    2014-07-01

    Curcumin is a natural polyphenolic compound that exhibits strong antioxidant and anticancer activities; however, low bioavailability has restricted its application in chemotherapeutic trials. The present study aimed to investigate the anticancer effect of the novel curcumin derivative 2E,6E?2?(1H?indol?3?yl) methylene)?6?(4?hydroxy?3?methoxy benzylidene)?cyclohexanone (IHCH) on A549 lung cancer cells. Cells were treated with IHCH at different concentrations (1?40 µM) for different time periods (1?36 h). Microscopic analysis revealed that IHCH inhibited A549 cell growth and induced the formation of characteristic autophagolysosomes in a dose? and time?dependent manner. Furthermore, the inhibitory rate of IHCH (40 µM) on A549 cell viability was 77.34% after 36 h of treatment. Acridine orange staining revealed an increase in autophagic vacuoles in the IHCH?treated A549 cells. Monodansylcadaverine staining was used to analyze autophagy rate. Immunocytochemistry revealed an increase in light chain (LC) 3 protein expression in the IHCH?treated cells and western blot analysis detected the conversion of LC3?I to LC3?II, as well as the recruitment of LC3 to autophagosomes in the cytoplasmatic compartment, suggesting the occurrence of autophagy. These findings show that IHCH induced autophagy in A549 cells, which is a novel cell death mechanism induced by curcumin derivatives. PMID:24788478

  10. Sodium metavanadate exhibits carcinogenic tendencies in vitro in immortalized human bronchial epithelial cells

    PubMed Central

    Passantino, Lisa; Muñoz, Alexandra B.

    2014-01-01

    Pentavalent vanadium compounds induce intracellular changes in vitro that are consistent with those of other carcinogenic substances. While there is no clear evidence that vanadium compounds cause cancer in humans, vanadium pentoxide causes lung cancer in rodents after long-term inhalation exposures and in turn IARC has categorized it as a group 2B possible human carcinogen. The goal of this study was to investigate the carcinogenicity of NaVO3 in the human immortalized bronchial epithelial cell line, Beas-2B. Cells were treated with 10 ?M NaVO3 for 5 weeks, with or without recovery time, followed by gene expression microarray analysis. In a separate experiment, cells were exposed to 1–10 ?M NaVO3 for 4 weeks and then grown in soft agar to test for anchorage-independent growth. A dose-dependent increase in the number of colonies was observed. In scratch tests, NaVO3-transformed clones could repair a wound faster than controls. In a gene expression microarray analysis of soft agar clones there were 2010 differentially expressed genes (DEG) (adjusted p-value ? 0.05) in NaVO3-transformed clones relative to control clones. DEG from this experiment were compared with the DEG of 5 week NaVO3 exposure with or without recovery, all with adjusted p-values < 0.05, and 469 genes were altered in the same direction for transformed clones, 5 week NaVO3-treated cells, and the recovered cells. The data from this study imply that chronic exposure to NaVO3 causes changes that are consistent with cellular transformation including anchorage-independent growth, enhanced migration ability, and gene expression changes that were likely epigenetically inherited. PMID:23963610

  11. Statistical prediction of nanoparticle delivery: from culture media to cell

    NASA Astrophysics Data System (ADS)

    Rowan Brown, M.; Hondow, Nicole; Brydson, Rik; Rees, Paul; Brown, Andrew P.; Summers, Huw D.

    2015-04-01

    The application of nanoparticles (NPs) within medicine is of great interest; their innate physicochemical characteristics provide the potential to enhance current technology, diagnostics and therapeutics. Recently a number of NP-based diagnostic and therapeutic agents have been developed for treatment of various diseases, where judicious surface functionalization is exploited to increase efficacy of administered therapeutic dose. However, quantification of heterogeneity associated with absolute dose of a nanotherapeutic (NP number), how this is trafficked across biological barriers has proven difficult to achieve. The main issue being the quantitative assessment of NP number at the spatial scale of the individual NP, data which is essential for the continued growth and development of the next generation of nanotherapeutics. Recent advances in sample preparation and the imaging fidelity of transmission electron microscopy (TEM) platforms provide information at the required spatial scale, where individual NPs can be individually identified. High spatial resolution however reduces the sample frequency and as a result dynamic biological features or processes become opaque. However, the combination of TEM data with appropriate probabilistic models provide a means to extract biophysical information that imaging alone cannot. Previously, we demonstrated that limited cell sampling via TEM can be statistically coupled to large population flow cytometry measurements to quantify exact NP dose. Here we extended this concept to link TEM measurements of NP agglomerates in cell culture media to that encapsulated within vesicles in human osteosarcoma cells. By construction and validation of a data-driven transfer function, we are able to investigate the dynamic properties of NP agglomeration through endocytosis. In particular, we statistically predict how NP agglomerates may traverse a biological barrier, detailing inter-agglomerate merging events providing the basis for predictive modelling of nanopharmacology.

  12. Alveolar Epithelial Cell Injury Due to Zinc Oxide Nanoparticle Exposure

    PubMed Central

    Kim, Yong Ho; Fazlollahi, Farnoosh; Kennedy, Ian M.; Yacobi, Nazanin R.; Hamm-Alvarez, Sarah F.; Borok, Zea; Kim, Kwang-Jin; Crandall, Edward D.

    2010-01-01

    Rationale: Although inhalation of zinc oxide (ZnO) nanoparticles (NPs) is known to cause systemic disease (i.e., metal fume fever), little is known about mechanisms underlying injury to alveolar epithelium. Objectives: Investigate ZnO NP–induced injury to alveolar epithelium by exposing primary cultured rat alveolar epithelial cell monolayers (RAECMs) to ZnO NPs. Methods: RAECMs were exposed apically to ZnO NPs or, in some experiments, to culture fluid containing ZnCl2 or free Zn released from ZnO NPs. Transepithelial electrical resistance (RT) and equivalent short-circuit current (IEQ) were assessed as functions of concentration and time. Morphologic changes, lactate dehydrogenase release, cell membrane integrity, intracellular reactive oxygen species (ROS), and mitochondrial activity were measured. Measurements and Main Results: Apical exposure to 176 ?g/ml ZnO NPs decreased RT and IEQ of RAECMs by 100% over 24 hours, whereas exposure to 11 ?g/ml ZnO NPs had little effect. Changes in RT and IEQ caused by 176 ?g/ml ZnO NPs were irreversible. ZnO NP effects on RT yielded half-maximal concentrations of approximately 20 ?g/ml. Apical exposure for 24 hours to 176 ?g/ml ZnO NPs induced decreases in mitochondrial activity and increases in lactate dehydrogenase release, permeability to fluorescein sulfonic acid, increased intracellular ROS, and translocation of ZnO NPs from apical to basolateral fluid (most likely across injured cells and/or damaged paracellular pathways). Conclusions: ZnO NPs cause severe injury to RAECMs in a dose- and time-dependent manner, mediated, at least in part, by free Zn released from ZnO NPs, mitochondrial dysfunction, and increased intracellular ROS. PMID:20639441

  13. Statistical prediction of nanoparticle delivery: from culture media to cell.

    PubMed

    Brown, M Rowan; Hondow, Nicole; Brydson, Rik; Rees, Paul; Brown, Andrew P; Summers, Huw D

    2015-04-17

    The application of nanoparticles (NPs) within medicine is of great interest; their innate physicochemical characteristics provide the potential to enhance current technology, diagnostics and therapeutics. Recently a number of NP-based diagnostic and therapeutic agents have been developed for treatment of various diseases, where judicious surface functionalization is exploited to increase efficacy of administered therapeutic dose. However, quantification of heterogeneity associated with absolute dose of a nanotherapeutic (NP number), how this is trafficked across biological barriers has proven difficult to achieve. The main issue being the quantitative assessment of NP number at the spatial scale of the individual NP, data which is essential for the continued growth and development of the next generation of nanotherapeutics. Recent advances in sample preparation and the imaging fidelity of transmission electron microscopy (TEM) platforms provide information at the required spatial scale, where individual NPs can be individually identified. High spatial resolution however reduces the sample frequency and as a result dynamic biological features or processes become opaque. However, the combination of TEM data with appropriate probabilistic models provide a means to extract biophysical information that imaging alone cannot. Previously, we demonstrated that limited cell sampling via TEM can be statistically coupled to large population flow cytometry measurements to quantify exact NP dose. Here we extended this concept to link TEM measurements of NP agglomerates in cell culture media to that encapsulated within vesicles in human osteosarcoma cells. By construction and validation of a data-driven transfer function, we are able to investigate the dynamic properties of NP agglomeration through endocytosis. In particular, we statistically predict how NP agglomerates may traverse a biological barrier, detailing inter-agglomerate merging events providing the basis for predictive modelling of nanopharmacology. PMID:25797791

  14. Fluorescence Modified Chitosan-Coated Magnetic Nanoparticles for High-Efficient Cellular Imaging

    NASA Astrophysics Data System (ADS)

    Ge, Yuqing; Zhang, Yu; He, Shiying; Nie, Fang; Teng, Gaojun; Gu, Ning

    2009-04-01

    Labeling of cells with nanoparticles for living detection is of interest to various biomedical applications. In this study, novel fluorescent/magnetic nanoparticles were prepared and used in high-efficient cellular imaging. The nanoparticles coated with the modified chitosan possessed a magnetic oxide core and a covalently attached fluorescent dye. We evaluated the feasibility and efficiency in labeling cancer cells (SMMC-7721) with the nanoparticles. The nanoparticles exhibited a high affinity to cells, which was demonstrated by flow cytometry and magnetic resonance imaging. The results showed that cell-labeling efficiency of the nanoparticles was dependent on the incubation time and nanoparticles’ concentration. The minimum detected number of labeled cells was around 104 by using a clinical 1.5-T MRI imager. Fluorescence and transmission electron microscopy instruments were used to monitor the localization patterns of the magnetic nanoparticles in cells. These new magneto-fluorescent nanoagents have demonstrated the potential for future medical use.

  15. Comparative in vitro cytotoxicity study on uncoated magnetic nanoparticles: effects on cell viability, cell morphology, and cellular uptake.

    PubMed

    Li, L; Mak, K Y; Shi, J; Koon, H K; Leung, C H; Wong, C M; Leung, C W; Mak, C S K; Chan, N M M; Zhong, W; Lin, K W; Wu, E X; Pong, P W T

    2012-12-01

    Magnetic iron oxide nanoparticles (MIONPs) must be biocompatible, and a thorough knowledge on their potential cytotoxicity is crucial for their biomedical applications. However, the detailed study about the effects of iron oxide nanoparticles on cell viability, cell morphology, and cellular uptake of different mammalian cells is still insufficient. In this paper, comparative cytotoxicity study of uncoated magnetite nanoparticles at different concentrations was performed on human cervical cancer cell line (HeLa) and immortalized normal human retinal pigment epithelial cell line (RPE). The size, structure, and magnetic behavior of the MIONPs were characterized using transmission electron microscopy (TEM), X-ray diffractometry (XRD), and vibrating sample magnetometry (VSM) respectively. After 24-hour incubation with the MIONPs, the cell viability was determined by live/dead assay, the cell morphology at high magnification was observed under scanning electron microscopy (SEM), and the cellular uptake of MIONPs was measured under TEM and verified by energy-dispersive X-ray spectroscopy (EDX) analysis. Our results indicate that the uncoated MIONPs at a high concentration (0.40 mg/ml) were toxic to both HeLa and RPE cells. However, the cytotoxicity of uncoated MIONPs at low concentrations was cell-type specific, and RPE cells were more susceptible to these MIONPs than HeLa cells. The effects of the MIONPs on cell morphology and the nanoparticles uptake also showed different features between these two cell lines. Hence cell type should be taken into consideration in the in vitro cytotoxicity study of uncoated MIONPs. Additionally, it should be noticed that the cell morphological changes and the uptake of nanoparticles can take place even though no toxic effect of these MIONPs at low concentrations was reflected in the traditional cell viability assay. PMID:23447952

  16. Satellite cells isolated from aged or dystrophic muscle exhibit a reduced capacity to promote angiogenesis in vitro.

    PubMed

    Rhoads, R P; Flann, K L; Cardinal, T R; Rathbone, C R; Liu, X; Allen, R E

    2013-10-25

    Deficits in skeletal muscle function exist during aging and muscular dystrophy, and suboptimal function has been related to factors such as atrophy, excessive inflammation and fibrosis. Ineffective muscle regeneration underlies each condition and has been attributed to a deficit in myogenic potential of resident stem cells or satellite cells. In addition to reduced myogenic activity, satellite cells may also lose the ability to communicate with vascular cells for coordination of myogenesis and angiogenesis and restoration of proper muscle function. Objectives of the current study were to determine the angiogenic-promoting capacity of satellite cells from two states characterized by dysfunctional skeletal muscle repair, aging and Duchenne muscular dystrophy. An in vitro culture model composed of satellite cells or their conditioned media and rat adipose tissue microvascular fragments (MVF) was used to examine this relationship. Microvascular fragments cultured in the presence of rat satellite cells from adult muscle donors (9-12 month of age) exhibited greater indices of angiogenesis (endothelial cell sprouting, tubule formation and extensive branching) than MVF co-cultured with satellite cells from aged muscle donors (24 month of age). We sought to determine if the differential degree of angiogenesis we observed in the co-culture setting was due to soluble factors produced by each satellite cell age group. Similar to the co-culture experiment, conditioned media produced by adult satellite cells promoted greater angiogenesis than that of aged satellite cells. Next, we examined differences in angiogenesis-stimulating ability of satellite cells from 12 mo old MDX mice or age-matched wild-type mice. A reduction in angiogenesis activity of media conditioned by satellite cells from dystrophic muscle was observed as compared to healthy muscle. Finally, we found reduced gene expression of hypoxia-inducible factor 1? (HIF-1?) and vascular endothelial growth factor (VEGF) in both aged and dystrophic satellite cells compared to their adult and normal counterparts, respectively. These results indicate that functional deficits in satellite cell activities during aging and diseased muscle may extend to their ability to communicate with other cells in their environment, in this case cells involved in angiogenesis. PMID:24070607

  17. Detection of circulating tumor cells using targeted surface-enhanced Raman scattering nanoparticles and magnetic enrichment.

    PubMed

    Shi, Wei; Paproski, Robert J; Moore, Ronald; Zemp, Roger

    2014-05-01

    While more than 90% of cancer deaths are due to metastases, our ability to detect circulating tumor cells (CTCs) is limited by low numbers of these cells in the blood and factors confounding specificity of detection. We propose a magnetic enrichment and detection technique for detecting CTCs with high specificity. We targeted both magnetic and surface-enhanced Raman scattering (SERS) nanoparticles to cancer cells. Only cells that are dual-labeled with both kinds of nanoparticles demonstrate an increasing SERS signal over time due to magnetic trapping. PMID:24858132

  18. Identification of live liver cancer cells in a mixed cell system using galactose-conjugated fluorescent nanoparticles.

    PubMed

    Peng, Jiaofeng; Wang, Kemin; Tan, Weihong; He, Xiaoxiao; He, Chunmei; Wu, Ping; Liu, Fang

    2007-02-15

    Bio-functioned fluorescent silica nanoparticles have been synthesized for cell labeling and cell differentiation and have shown great promise as novel fluorescent probes. The galactose-conjugated fluorescent nanoparticles (GCFNPs) have been obtained by the conjugation of amino-modified fluorescent silica nanoparticles with lactobionic acid (LA) through EDAC linkage. The GCFNPs retain excellent biological activity and can be used in bioanalysis as an immunofluorescence assay. The specific identification of target cells from the background cells have been directly demonstrated in a simple model system by a laser confocal scanning microscope, because the specific and non-specific labeling can simultaneously visualized in a given microscopic field of view. The flow cytometric analysis has proved that GCFNPs can effectively recognize target cells in the mixed cell system. The demonstration of precise identification of few liver cancer cells in the blood confirmed the excellent capability of GCFNPs in identifying specific cells in a large host cell background. The nanoparticle's excellent photostability, good biocompatibility and significant signal amplification make them well-suited for the identification of individual cells sensitively for a variety of biomedical studies such as cancer metastasis and stem cell progeny in vivo. PMID:19071382

  19. Analytically monitored digestion of silver nanoparticles and their toxicity on human intestinal cells.

    PubMed

    Böhmert, Linda; Girod, Matthias; Hansen, Ulf; Maul, Ronald; Knappe, Patrick; Niemann, Birgit; Weidner, Steffen M; Thünemann, Andreas F; Lampen, Alfonso

    2014-09-01

    Orally ingested nanoparticles may overcome the gastrointestinal barrier, reach the circulatory system, be distributed in the organism and cause adverse health effects. However, ingested nanoparticles have to pass through different physicochemical environments, which may alter their properties before they reach the intestinal cells. In this study, silver nanoparticles are characterised physicochemically during the course of artificial digestion to simulate the biochemical processes occurring during digestion. Their cytotoxicity on intestinal cells was investigated using the Caco-2 cell model. Using field-flow fractionation combined with dynamic light scattering and small-angle X-ray scattering, the authors found that particles only partially aggregate as a result of the digestive process. Cell viabilities were determined by means of CellTiter-Blue® assay, 4',6-diamidino-2-phenylindole-staining and real-time impedance. These measurements reveal small differences between digested and undigested particles (1-100 µg/ml or 1-69 particles/cell). The findings suggest that silver nanoparticles may indeed overcome the gastrointestinal juices in their particulate form without forming large quantities of aggregates. Consequently, the authors presume that the particles can reach the intestinal epithelial cells after ingestion with only a slight reduction in their cytotoxic potential. The study indicates that it is important to determine the impact of body fluids on the nanoparticles of interest to provide a reliable interpretation of their nano-specific cytotoxicity testing in vivo and in vitro. PMID:23763544

  20. 3D Graphene Oxide-encapsulated Gold Nanoparticles to Detect Neural Stem Cell Differentiation

    PubMed Central

    Kim, Tae-Hyung; Lee, Ki-Bum; Choi, Jeong-Woo

    2013-01-01

    Monitoring of stem cell differentiation and pluripotency is an important step for the practical use of stem cells in the field of regenerative medicine. Hence, a new non-destructive detection tool capable of in situ monitoring of stem cell differentiation is highly needed. In this study, we report a 3D graphene oxide-encapsulated gold nanoparticle that is very effective for the detection of the differentiation potential of neural stem cells (NSCs) based on surface-enhanced Raman spectroscopy (SERS). A new material, 3D GO-encapsulated gold nanoparticle, is developed to induce the double enhancement effect of graphene oxide and gold nanoparticle on SERS signals which is only effective for undifferentiated NSCs. The Raman peaks achieved from undifferentiated NSCs on the graphene oxide (GO)-encapsulated gold nanoparticles were 3.5 times higher than peaks obtained from normal metal structures and were clearly distinguishable from those of differentiated cells. The number of C=C bonds and the raman instensity at 1656cm?1 was found to show a positive correlation, which matches the differentiation state of the NSCs. Moreover, the substrate composed of 3D GO-encapsulated gold nanoparticles was also effective at distinguishing the differentiation state of single NSC by using electrochemical and electrical techniques. Hence, the proposed technique can be used as a powerful non-destructive in situ monitoring tool for the identification of the differentiation potential of various kinds of stem cells (mesenchymal, hematopoietic, and neural stem cells). PMID:23937915

  1. VMP1-deficient Chlamydomonas exhibits severely aberrant cell morphology and disrupted cytokinesis

    PubMed Central

    2014-01-01

    Background The versatile Vacuole Membrane Protein 1 (VMP1) has been previously investigated in six species. It has been shown to be essential in macroautophagy, where it takes part in autophagy initiation. In addition, VMP1 has been implicated in organellar biogenesis; endo-, exo- and phagocytosis, and protein secretion; apoptosis; and cell adhesion. These roles underly its proven involvement in pancreatitis, diabetes and cancer in humans. Results In this study we analyzed a VMP1 homologue from the green alga Chlamydomonas reinhardtii. CrVMP1 knockdown lines showed severe phenotypes, mainly affecting cell division as well as the morphology of cells and organelles. We also provide several pieces of evidence for its involvement in macroautophagy. Conclusion Our study adds a novel role to VMP1's repertoire, namely the regulation of cytokinesis. Though the directness of the observed effects and the mechanisms underlying them remain to be defined, the protein's involvement in macroautophagy in Chlamydomonas, as found by us, suggests that CrVMP1 shares molecular characteristics with its animal and protist counterparts. PMID:24885763

  2. Preparation of epidermal growth factor (EGF) conjugated iron oxide nanoparticles and their internalization into colon cancer cells

    NASA Astrophysics Data System (ADS)

    Creixell, Mar; Herrera, Adriana P.; Ayala, Vanessa; Latorre-Esteves, Magda; Pérez-Torres, Marianela; Torres-Lugo, Madeline; Rinaldi, Carlos

    2010-08-01

    Epidermal growth factor (EGF) was conjugated with carboxymethyldextran (CMDx) coated iron oxide magnetic nanoparticles using carbodiimide chemistry to obtain magnetic nanoparticles that target the epidermal growth factor receptor (EGFR). Epidermal growth factor modified magnetic nanoparticles were colloidally stable when suspended in biological buffers such as PBS and cell culture media. Both targeted and non-targeted nanoparticles were incubated with CaCo-2 cancer cells, known to overexpress EGFR. Nanoparticle localization within the cell was visualized by confocal laser scanning microscopy and light microscopy using Prussian blue stain. Results showed that targeted magnetic nanoparticles were rapidly accumulated in both flask-shaped small vesicles and large circular endocytic structures. Internalization patterns suggest that both clathrin-dependent and clathrin-independent receptors mediated endocytosis mechanisms are responsible for nanoparticle internalization.

  3. Multi-ligand poly(L-lactic-co-glycolic acid) nanoparticles inhibit activation of endothelial cells.

    PubMed

    Xu, Hao; Kona, Soujanya; Su, Lee-Chun; Tsai, Yi-Ting; Dong, Jing-Fei; Brilakis, Emmanouil S; Tang, Liping; Banerjee, Subhash; Nguyen, Kytai T

    2013-08-01

    Endothelial cell (EC) activation and inflammation is a key step in the initiation and progression of many cardiovascular diseases. Targeted delivery of therapeutic reagents to inflamed EC using nanoparticles is challenging as nanoparticles do not arrest on EC efficiently under high shear stress. In this study, we developed a novel polymeric platelet-mimicking nanoparticle for strong particle adhesion onto ECs and enhanced particle internalization by ECs. This nanoparticle was encapsulated with dexamethasone as the anti-inflammatory drug, and conjugated with polyethylene glycol, glycoprotein 1b, and trans-activating transcriptional peptide. The multi-ligand nanoparticle showed significantly greater adhesion on P-selectin, von Willebrand Factor, than the unmodified particles, and activated EC in vitro under both static and flow conditions. Treatment of injured rat carotid arteries with these multi-ligand nanoparticles suppressed neointimal stenosis more than unconjugated nanoparticles did. These results indicate that this novel multi-ligand nanoparticle is efficient to target inflamed EC and inhibit inflammation and subsequent stenosis. PMID:23640308

  4. Proteomic study of human bronchial epithelial cells exposed to SiC nanoparticles

    NASA Astrophysics Data System (ADS)

    Tokarski, Caroline; Hirano, Seishiro; Rolando, Christian

    2011-07-01

    The presented work proposes an optimized methodology for the study of cell exposure to nanomaterials at protein level. The study was investigated on proteins extracted from human bronchial epithelial cells exposed and non-exposed to silicon carbide nanoparticles (SiC). The analytical strategy was based on high resolution measurement using Fourier transform mass spectrometer 9.4 T. The methodology proposed succeeds in identifying over 300 proteins; most of the identified proteins are present in both exposed and non exposed cells to SiC nanoparticles. More interestingly, cytokines as Macrophage migration inhibitory factor protein could be identified only in the cells exposed to SiC nanoparticles indicating cell inflammatory response.

  5. Dendrimer-like alpha-d-glucan nanoparticles activate dendritic cells and are effective vaccine adjuvants.

    PubMed

    Lu, Fangjia; Mencia, Alejandra; Bi, Lin; Taylor, Aaron; Yao, Yuan; HogenEsch, Harm

    2015-04-28

    The use of nanoparticles for delivery of vaccine antigens and as vaccine adjuvants is appealing because their size allows efficient uptake by dendritic cells and their biological properties can be tailored to the desired function. Here, we report the effect of chemically modified phytoglycogen, a dendrimer-like ?-d-glucan nanoparticle, on dendritic cells in vitro, and the utility of this type of nanoparticle as a vaccine adjuvant in vivo. The modified phytoglycogen nanoparticle, termed Nano-11, has a positive surface charge which enabled electrostatic adsorption of negatively charged protein antigens. The Nano-11-antigen complexes were efficiently phagocytized by dendritic cells. Nano-11 induced increased expression of costimulatory molecules and the secretion of IL-1? and IL-12p40 by dendritic cells. Intramuscular injection of Nano-11-antigen formulations induced a significantly enhanced immune response to two different protein antigens. Examination of the injection site revealed numerous monocytes and relatively few neutrophils at one day after injection. The inflammation had nearly completely disappeared by 2weeks after injection. These studies indicate that Nano-11 is an effective vaccine delivery vehicle that significantly enhances the immune response. This type of plant based nanoparticle is considered highly cost-effective compared with fully synthetic nanoparticles and appears to have an excellent safety profile making them an attractive adjuvant candidate for prophylactic vaccines. PMID:25747143

  6. TiO2 nanoparticles and bulk material stimulate human peripheral blood mononuclear cells?

    PubMed Central

    Becker, Kathrin; Schroecksnadel, Sebastian; Geisler, Simon; Carriere, Marie; Gostner, Johanna M.; Schennach, Harald; Herlin, Nathalie; Fuchs, Dietmar

    2014-01-01

    Nanomaterials are increasingly produced and used throughout recent years. Consequently the probability of exposure to nanoparticles has risen. Because of their small 1–100 nm size, the physicochemical properties of nanomaterials may differ from standard bulk materials and may pose a threat to human health. Only little is known about the effects of nanoparticles on the human immune system. In this study, we investigated the effects of TiO2 nanoparticles and bulk material in the in vitro model of human peripheral blood mononuclear cells (PBMC) and cytokine-induced neopterin formation and tryptophan breakdown was monitored. Both biochemical processes are closely related to the course of diseases like infections, atherogenesis and neurodegeneration. OCTi60 (25 nm diameter) TiO2 nanoparticles and bulk material increased neopterin production in unstimulated PBMC and stimulated cells significantly, the effects were stronger for OCTi60 compared to bulk material, while P25 TiO2 (25 nm diameter) nanoparticles had only little influence. No effect of TiO2 nanoparticles on tryptophan breakdown was detected in unstimulated cells, whereas in stimulated cells, IDO activity and IFN-? production were suppressed but only at the highest concentrations tested. Because neopterin was stimulated and tryptophan breakdown was suppressed in parallel, data suggests that the total effect of particles would be strongly pro-inflammatory. PMID:24361406

  7. Application of magnetic field hyperthermia and superparamagnetic iron oxide nanoparticles to HIV-1-specific T-cell cytotoxicity

    PubMed Central

    Williams, James P; Southern, Paul; Lissina, Anya; Christian, Helen C; Sewell, Andrew K; Phillips, Rodney; Pankhurst, Quentin; Frater, John

    2013-01-01

    The latent HIV-1 reservoir remains the major barrier to HIV-1 eradication. Although successful at limiting HIV replication, highly active antiretroviral therapy is unable to cure HIV infection, thus novel therapeutic strategies are needed to eliminate the virus. Magnetic field hyperthermia (MFH) generates thermoablative cytotoxic temperatures in target-cell populations, and has delivered promising outcomes in animal models, as well as in several cancer clinical trials. MFH has been proposed as a strategy to improve the killing of HIV-infected cells and for targeting the HIV latent reservoirs. We wished to determine whether MFH could be used to enhance cytotoxic T-lymphocyte (CTL) targeting of HIV-infected cells in a proof-of-concept study. Here, for the first time, we apply MFH to an infectious disease (HIV-1) using the superparamagnetic iron oxide nanoparticle FeraSpin R. We attempt to improve the cytotoxic potential of T-cell receptor-transfected HIV-specific CTLs using thermotherapy, and assess superparamagnetic iron oxide nanoparticle toxicity, uptake, and effect on cell function using more sensitive methods than previously described. FeraSpin R exhibited only limited toxicity, demonstrated efficient uptake and cell-surface attachment, and only modestly impacted T-cell function. In contrast to the cancer models, insufficient MFH was generated to enhance CTL killing of HIV-infected cells. MFH remains an exciting new technology in the field of cancer therapeutics, which, as technology improves, may have significant potential to enhance CTL function and act as an adjunctive therapy in the eradication of latently infected HIV-positive cells. PMID:23901272

  8. Secondary structure of corona proteins determines the cell surface receptors used by nanoparticles.

    PubMed

    Fleischer, Candace C; Payne, Christine K

    2014-12-11

    Nanoparticles used for biological and biomedical applications encounter a host of extracellular proteins. These proteins rapidly adsorb onto the nanoparticle surface, creating a protein corona. Poly(ethylene glycol) can reduce, but not eliminate, the nonspecific adsorption of proteins. As a result, the adsorbed proteins, rather than the nanoparticle itself, determine the cellular receptors used for binding, the internalization mechanism, the intracellular transport pathway, and the subsequent immune response. Using fluorescence microscopy and flow cytometry, we first characterize a set of polystyrene nanoparticles in which the same adsorbed protein, bovine serum albumin, leads to binding to two different cell surface receptors: native albumin receptors and scavenger receptors. Using a combination of circular dichroism spectroscopy, isothermal titration calorimetry, and fluorescence spectroscopy, we demonstrate that the secondary structure of the adsorbed bovine serum albumin protein controls the cellular receptors used by the protein-nanoparticle complexes. These results show that protein secondary structure is a key parameter in determining the cell surface receptor used by a protein-nanoparticle complex. We expect this link between protein structure and cellular outcomes will provide a molecular basis for the design of nanoparticles for use in biological and biomedical applications. PMID:24779411

  9. nanoparticles

    NASA Astrophysics Data System (ADS)

    Andreu-Cabedo, Patricia; Mondragon, Rosa; Hernandez, Leonor; Martinez-Cuenca, Raul; Cabedo, Luis; Julia, J. Enrique

    2014-10-01

    Thermal energy storage (TES) is extremely important in concentrated solar power (CSP) plants since it represents the main difference and advantage of CSP plants with respect to other renewable energy sources such as wind, photovoltaic, etc. CSP represents a low-carbon emission renewable source of energy, and TES allows CSP plants to have energy availability and dispatchability using available industrial technologies. Molten salts are used in CSP plants as a TES material because of their high operational temperature and stability of up to 500°C. Their main drawbacks are their relative poor thermal properties and energy storage density. A simple cost-effective way to improve thermal properties of fluids is to dope them with nanoparticles, thus obtaining the so-called salt-based nanofluids. In this work, solar salt used in CSP plants (60% NaNO3 + 40% KNO3) was doped with silica nanoparticles at different solid mass concentrations (from 0.5% to 2%). Specific heat was measured by means of differential scanning calorimetry (DSC). A maximum increase of 25.03% was found at an optimal concentration of 1 wt.% of nanoparticles. The size distribution of nanoparticle clusters present in the salt at each concentration was evaluated by means of scanning electron microscopy (SEM) and image processing, as well as by means of dynamic light scattering (DLS). The cluster size and the specific surface available depended on the solid content, and a relationship between the specific heat increment and the available particle surface area was obtained. It was proved that the mechanism involved in the specific heat increment is based on a surface phenomenon. Stability of samples was tested for several thermal cycles and thermogravimetric analysis at high temperature was carried out, the samples being stable.

  10. Mesenchymal stromal cells derived from acute myeloid leukemia bone marrow exhibit aberrant cytogenetics and cytokine elaboration.

    PubMed

    Huang, J C; Basu, S K; Zhao, X; Chien, S; Fang, M; Oehler, V G; Appelbaum, F R; Becker, P S

    2015-01-01

    Bone marrow-derived mesenchymal stromal cells (BM-MSCs) play a fundamental role in the BM microenvironment (BME) and abnormalities of these cells may contribute to acute myeloid leukemia (AML) pathogenesis. The aim of the study was to characterize the cytokine and gene expression profile, immunophenotype and cytogenetics of BM-MSCs from AML patients compared to normal BM-MSCs from healthy donors. AML BM-MSCs showed decreased monocyte chemoattractant protein-1 levels compared to normal BM-MSCs. AML BM-MSCs expressed similar ?1 integrin, CD44, CD73, CD90 and E-cadherin compared to normal BM-MSCs. Cytogenetic analysis revealed chromosomal aberrations in AML BM-MSCs, some overlapping with and others distinct from their corresponding AML blasts. No significant difference in gene expression was detected between AML BM-MSCs compared to normal BM-MSCs; however, comparing the differences between AML and MSCs from AML patients with the differences between normal hematopoietic cells and normal MSCs by Ingenuity pathway analysis showed key distinctions of the AML setting: (1) upstream gene regulation by transforming growth factor beta 1, tumor necrosis factor, tissue transglutaminase 2, CCAAT/enhancer binding protein alpha and SWItch/Sucrose NonFermentable related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4; (2) integrin and interleukin 8 signaling as overrepresented canonical pathways; and (3) upregulation of transcription factors FBJ murine osteosarcoma viral oncogene homolog and v-myb avian myeloblastosis viral oncogene homolog. Thus, phenotypic abnormalities of AML BM-MSCs highlight a dysfunctional BME that may impact AML survival and proliferation. PMID:25860293

  11. Cellular uptake and mutagenic potential of metal oxide nanoparticles in bacterial cells.

    PubMed

    Kumar, Ashutosh; Pandey, Alok K; Singh, Shashi S; Shanker, Rishi; Dhawan, Alok

    2011-05-01

    Extensive production and consumption of nanomaterials such as ZnO and TiO(2) has increased their release and disposal into the environment. The accumulation of nanoparticles (NPs) in ecosystem is likely to pose threat to non-specific targets such as bacteria. The present study explored the effect of ZnO and TiO(2) NPs in a model bacterium, Salmonella typhimurium. The uptake of ZnO and TiO(2) bare NPs in nano range without agglomeration was observed in S. typhimurium. TEM analysis demonstrated the internalization and uniform distribution of NPs inside the cells. Flow cytometry data also demonstrates that both ZnO and TiO(2) NPs were significantly internalized in the S. typhimurium cells in a concentration dependent manner. A significant increase in uptake was observed in the S. typhimurium treated even with 8 and 80 ng mL(-1) of ZnO and TiO(2) NPs with S9 after 60 min, possibly the formation of micelles or protein coat facilitated entry of NPs. These NPs exhibited weak mutagenic potential in S. typhimurium strains TA98, TA1537 and Escherichia coli (WP2uvrA) of Ames test underscoring the possible carcinogenic potential similar to certain mutagenic chemicals. Our study reiterates the need for re-evaluating environmental toxicity of ZnO and TiO(2) NPs presumably considered safe in environment. PMID:21310462

  12. Human Trabecular Meshwork Cells Exhibit Several Characteristics of, but Are Distinct from, Adipose-Derived Mesenchymal Stem Cells

    PubMed Central

    Morgan, Joshua T.; Wood, Joshua A.; Walker, Naomi J.; Raghunathan, Vijay Krishna; Borjesson, Dori L.; Murphy, Christopher J.

    2014-01-01

    Abstract Purpose: To support the growing promise of regenerative medicine in glaucoma, we characterized the similarities and differences between human trabecular meshwork (HTM) cells and human mesenchymal stem cells (hMSCs). Methods: HTM cells and hMSCs were phenotypically characterized by flow cytometry. Using quantitative polymerase chain reaction, the expression of myoc, angptl7, sox2, pou5f1, and notch1 was determined in both cell types with and without dexamethasone (Dex). Immunosuppressive behavior of HTM cells and hMSCs was determined using T cells activated with phytohemagglutinin. T-cell proliferation was determined using BrdU incorporation and flow cytometry. Multipotency of HTM cells and hMSCs was determined using adipogenic and osteogenic differentiation media as well as aqueous humor (AH). Alpha-smooth muscle actin (?SMA) expression was determined in HTM cells, hMSCs, and HTM tissue. Results: Phenotypically, HTM and hMSCs expressed CD73, CD90, CD105, and CD146 but not CD31, CD34, and CD45 and similar sox2, pou5f1, and notch1 expression. Both cell types suppressed T-cell proliferation. However, HTM cells, but not hMSCs, upregulated myoc and angptl7 in response to Dex. Additionally, HTM cells did not differentiate into adipocytes or osteocytes. Culture of hMSCs in 20%, but not 100%, AH potently induced alkaline phosphatase activity. HTM cells in culture possessed uniformly strong expression of ?SMA, which contrasted with the limited expression in hMSCs and spatially discrete expression in HTM tissue. Conclusions: HTM cells possess a number of important similarities with hMSCs but lack multipotency, one of the defining characteristics of stem cells. Further work is needed to explore the molecular mechanisms and functional implications underlying the phenotypic similarities. PMID:24456002

  13. The significance of a Cripto-1-positive subpopulation of human melanoma cells exhibiting stem cell-like characteristics

    PubMed Central

    Strizzi, Luigi; Margaryan, Naira V.; Gilgur, Alina; Hardy, Katharine M.; Normanno, Nicola; Salomon, David S.; Hendrix, Mary J.C.

    2013-01-01

    Cripto-1 (CR-1) protein function differs according to cellular or extracellular expression. In this study, we explore the significance of cell surface CR-1 expression in human melanoma cells. Cell surface CR-1-expressing human melanoma cells (CR1-CS+) were selected by fluorescence-activated cell sorting (FACS) and grown in vitro and in vivo in nude mice to study their growth characteristics. The CR1-CS+ melanoma cells were found to express increased levels of Oct4, MDR-1 and activated c-Src compared with cells lacking this subpopulation (CR1-CS?) or unsorted cells, used as control. CR1-CS+ show reduced proliferation rates and diminished spherical colony formation compared with control cells when cultured in vitro. Orthotopic injections of CR1-CS+ in nude mice formed slow growing tumors with histologic variability across different areas of the CR1-CS+ xenografts. CR-1-expressing cells from first generation CR1-CS+ tumors showed significantly increased tumor-forming rate and aggressiveness following subsequent transplants in nude mice. These data demonstrate that within a heterogeneous melanoma cell population there resides a slow proliferating, cell surface CR-1-expressing subpopulation capable of giving rise to a fast growing, aggressive progeny that may contribute to disease recurrence and progression. PMID:23574716

  14. Mature Dendritic Cells Infected with Herpes Simplex Virus Type 1 Exhibit Inhibited T-Cell Stimulatory Capacity

    Microsoft Academic Search

    MONIKA KRUSE; OLAF ROSORIUS; FRIEDRICH KRATZER; GERHARD STELZ; CHRISTINE KUHNT; GEROLD SCHULER; JOACHIM HAUBER; ALEXANDER STEINKASSERER

    2000-01-01

    Mature dendritic cells (DC) are the most potent antigen-presenting cells within the entire immune system. Interference with the function of these cells therefore constitutes a very powerful mechanism for viruses to escape immune responses. Several members of the Herpesviridae family have provided examples of such escape strategies, including interference with antigen presentation and production of homologous cytokines. In this study

  15. A comprehensive review of the application of chalcogenide nanoparticles in polymer solar cells

    NASA Astrophysics Data System (ADS)

    Freitas, Jilian N.; Gonçalves, Agnaldo S.; Nogueira, Ana F.

    2014-05-01

    In this review the use of solution-processed chalcogenide quantum dots (CdS, CdSe, PbS, etc.) in hybrid organic-inorganic solar cells is explored. Such devices are known as potential candidates for low-cost and efficient solar energy conversion, and compose the so-called third generation solar cells. The incorporation of oxides and metal nanoparticles has also been successfully achieved in this new class of photovoltaic devices; however, we choose to explore here chalcogenide quantum dots in light of their particularly attractive optical and electronic properties. We address herein a comprehensive review of the historical background and state-of-the-art comprising the incorporation of such nanoparticles in polymer matrices. Later strategies for surface chemistry manipulation, in situ synthesis of nanoparticles, use of continuous 3D nanoparticles network (aerogels) and ternary systems are also reviewed.

  16. A comprehensive review of the application of chalcogenide nanoparticles in polymer solar cells.

    PubMed

    Freitas, Jilian N; Gonçalves, Agnaldo S; Nogueira, Ana F

    2014-06-21

    In this review the use of solution-processed chalcogenide quantum dots (CdS, CdSe, PbS, etc.) in hybrid organic-inorganic solar cells is explored. Such devices are known as potential candidates for low-cost and efficient solar energy conversion, and compose the so-called third generation solar cells. The incorporation of oxides and metal nanoparticles has also been successfully achieved in this new class of photovoltaic devices; however, we choose to explore here chalcogenide quantum dots in light of their particularly attractive optical and electronic properties. We address herein a comprehensive review of the historical background and state-of-the-art comprising the incorporation of such nanoparticles in polymer matrices. Later strategies for surface chemistry manipulation, in situ synthesis of nanoparticles, use of continuous 3D nanoparticles network (aerogels) and ternary systems are also reviewed. PMID:24839190

  17. Effect of cerium oxide nanoparticles on inflammation in vascular endothelial cells

    PubMed Central

    Gojova, Andrea; Lee, Jun-Tae; Jung, Heejung S.; Guo, Bing; Barakat, Abdul I.; Kennedy, Ian M.

    2010-01-01

    Because vascular endothelial cell inflammation is critical in the development of cardiovascular pathology, we hypothesized that direct exposure of human aortic endothelial cells (HAECs) to ultrafine particles induces an inflammatory response. To test the hypothesis, we incubated HAECs for 4 h with different concentrations (0.001–50 ?g/ml) of CeO2 nanoparticles and subsequently measured mRNA levels of the three inflammatory markers intercellular adhesion molecule 1 (ICAM-1), interleukin (IL)-8, and monocyte chemotactic protein (MCP-1) using real-time polymerase chain reaction (PCR). Ceria nanoparticles caused very little inflammatory response in HAECs, even at the highest dose. This material is apparently rather benign in comparison with Y2O3 and ZnO nanoparticles that we have studied previously. These results suggest that inflammation in HAECs following acute exposure to metal oxide nanoparticles depends strongly on particle composition. PMID:19558244

  18. Three-dimensional nanoparticle tracking and simultaneously membrane profiling during endocytosis of living cells

    NASA Astrophysics Data System (ADS)

    Wang, Chun-Chieh; Liang, Chia-Pin; Lee, Chau-Hwang

    2009-11-01

    By utilizing a wide-field optical profiling technique and a bright-field particle-tracking algorithm, we record the tracks of gold nanoparticles as well as the topography of cell membranes during the endocytosis processes. The motion of 60 nm gold particles relative to the membrane is directly visualized without fluorescence labeling. The positioning accuracy of gold nanoparticles in three dimensions is nearly 20 nm. On the basis of the simultaneous particle tracks and membrane topography, we estimate the internalization efficiency of transferrin-coated gold nanoparticles on fibroblasts.

  19. Edge-truncated cubic platinum nanoparticles as anode catalysts for direct methanol fuel cells

    NASA Astrophysics Data System (ADS)

    Liu, Y. G.; Shi, S. L.; Xue, X. Y.; Zhang, J. Y.; Wang, Y. G.; Wang, T. H.

    2008-05-01

    The edge-truncated cubic platinum nanoparticles (ECPs) are synthesized with the addition of silver ions. The nanoparticle is closed by 6 {100} facets and 12 {110} facets, confirmed from the transmission electron microscopy and cyclic voltammogram (CV) results. The current density increases up to a maximum of 1.05mAcm-2 (Jf) during the forward sweep. A current density ratio value of 1.12 and the sharp initial current drop in the CV reveal that the tailored facets of the ECPs dominate the methanol oxidation behaviors. Our results show promising anode catalysts of truncated-edge cubic platinum nanoparticles for direct methanol fuel cells.

  20. Intracellular heating of living cells through Néel relaxation of magnetic nanoparticles.

    PubMed

    Fortin, Jean-Paul; Gazeau, Florence; Wilhelm, Claire

    2008-02-01

    Maghemite and cobalt ferrite anionic magnetic nanoparticles enter tumor cells and can be used as heat sources when exposed to a high-frequency magnetic field. Comparative studies of the two particles enable to unravel the magnetic heating mechanisms (Néel relaxation vs. Brown relaxation) responsible for the cellular temperature rise, and also to establish a simple model, adjusted to the experimental results, allowing to predict the intracellular heating efficiency of iron oxide nanoparticles. Hence, we are able to derive the best nanoparticle design for a given material with a view to intracellular hyperthermia-based applications. PMID:17641885

  1. Cell Penetrating Synthetic Antimicrobial Peptides (SAMPs) Exhibiting Potent and Selective Killing of Mycobacterium by Targeting Its DNA.

    PubMed

    Sharma, Aashish; Pohane, Amol Arunrao; Bansal, Sandhya; Bajaj, Avinash; Jain, Vikas; Srivastava, Aasheesh

    2015-02-23

    Naturally occurring antimicrobial peptides (AMPs) are powerful defence tools to tackle pathogenic microbes. However, limited natural production and high synthetic costs in addition to poor selectivity limit large-scale use of AMPs in clinical settings. Here, we present a series of synthetic AMPs (SAMPs) that exhibit highly selective and potent killing of Mycobacterium (minimum inhibitory concentration <20??g?mL(-1) ) over E.?coli or mammalian cells. These SAMPs are active against rapidly multiplying as well as growth saturated Mycobacterium cultures. These SAMPs are not membrane-lytic in nature, and are readily internalized by Mycobacterium and mammalian cells; whereas in E.?coli, the lipopolysaccharide layer inhibits their cellular uptake, and hence, their antibacterial action. Upon internalization, these SAMPs interact with the unprotected genomic DNA of mycobacteria, and impede DNA-dependent processes, leading to bacterial cell death. PMID:25608020

  2. DNAM-1-based chimeric antigen receptors enhance T cell effector function and exhibit in vivo efficacy against melanoma.

    PubMed

    Wu, Ming-Ru; Zhang, Tong; Alcon, Andre; Sentman, Charles L

    2015-04-01

    Chimeric antigen receptor (CAR) T cell therapies hold great potential for treating cancers, and new CARs that can target multiple tumor types and have the potential to target non-hematological malignancies are needed. In this study, the tumor recognition ability of a natural killer cell-activating receptor, DNAM-1 was harnessed to design CARs that target multiple tumor types. DNAM-1 ligands, PVR and nectin-2, are expressed on primary human leukemia, myeloma, ovarian cancer, melanoma, neuroblastoma, and Ewing sarcoma. DNAM-1 CARs exhibit high tumor cell cytotoxicity but low IFN-? secretion in vitro. In contrast to other CAR designs, co-stimulatory domains did not improve the expression and function of DNAM-1 CARs. A DNAM-1/CD3zeta CAR reduced tumor burden in a murine melanoma model in vivo. In conclusion, DNAM-1-based CARs may have the potential to treat PVR and nectin-2 expressing hematological and solid tumors. PMID:25549845

  3. Quantitative photoacoustics to measure single cell melanin production and nanoparticle attachment

    NASA Astrophysics Data System (ADS)

    Bhattacharyya, Kiran; Eshein, Adam; Chandrasekhar, Anand; Viator, John A.

    2015-04-01

    Photoacoustics can be used as a label-free spectroscopic method of identifying pigmented proteins and characterizing their intracellular concentration over time in a single living cell. The authors use a microscopic laser irradiation system with a 5 ns, Q-switched laser focused onto single cells in order to collect photoacoustic responses of melanoma cells from the HS936 cell line and gold nanoparticle labeled breast cancer cells from the T47D cell line. The volume averaged intracellular concentration of melanin is found to range from 29–270 mM for single melanoma cells and the number of gold nanoparticles (AuNP) is shown to range from 850–5900 AuNPs/cell. Additionally, the melanin production response to UV-A light stimulus is measured in four melanoma cells to find a mass production rate of 5.7 pg of melanin every 15 min.

  4. Quantitative photoacoustics to measure single cell melanin production and nanoparticle attachment.

    PubMed

    Bhattacharyya, Kiran; Eshein, Adam; Chandrasekhar, Anand; Viator, John A

    2015-04-21

    Photoacoustics can be used as a label-free spectroscopic method of identifying pigmented proteins and characterizing their intracellular concentration over time in a single living cell. The authors use a microscopic laser irradiation system with a 5 ns, Q-switched laser focused onto single cells in order to collect photoacoustic responses of melanoma cells from the HS936 cell line and gold nanoparticle labeled breast cancer cells from the T47D cell line. The volume averaged intracellular concentration of melanin is found to range from 29-270 mM for single melanoma cells and the number of gold nanoparticles (AuNP) is shown to range from 850-5900 AuNPs/cell. Additionally, the melanin production response to UV-A light stimulus is measured in four melanoma cells to find a mass production rate of 5.7 pg of melanin every 15 min. PMID:25803095

  5. A spiking network model of cerebellar Purkinje cells and molecular layer interneurons exhibiting irregular firing

    PubMed Central

    Lennon, William; Hecht-Nielsen, Robert; Yamazaki, Tadashi

    2014-01-01

    While the anatomy of the cerebellar microcircuit is well-studied, how it implements cerebellar function is not understood. A number of models have been proposed to describe this mechanism but few emphasize the role of the vast network Purkinje cells (PKJs) form with the molecular layer interneurons (MLIs)—the stellate and basket cells. We propose a model of the MLI-PKJ network composed of simple spiking neurons incorporating the major anatomical and physiological features. In computer simulations, the model reproduces the irregular firing patterns observed in PKJs and MLIs in vitro and a shift toward faster, more regular firing patterns when inhibitory synaptic currents are blocked. In the model, the time between PKJ spikes is shown to be proportional to the amount of feedforward inhibition from an MLI on average. The two key elements of the model are: (1) spontaneously active PKJs and MLIs due to an endogenous depolarizing current, and (2) adherence to known anatomical connectivity along a parasagittal strip of cerebellar cortex. We propose this model to extend previous spiking network models of the cerebellum and for further computational investigation into the role of irregular firing and MLIs in cerebellar learning and function. PMID:25520646

  6. In vitro labeling of endothelial progenitor cells isolated from peripheral blood with superparamagnetic iron oxide nanoparticles.

    PubMed

    Sun, Jun-Hui; Zhang, Yue-Lin; Nie, Chun-Hui; Qian, Su-Ping; Yu, Xiao-Bo; Xie, Hai-Yang; Zhou, Lin; Zheng, Shu-Sen

    2012-08-01

    The transplantation of endothelial progenitor cells (EPCs) provides a novel method for the treatment of human tumors or vascular diseases. Magnetic resonance imaging (MRI) has proven to be effective in tracking transplanted stem cells by labeling the cells with superparamagnetic iron oxide (SPIO) nanoparticles. The SPIO has been used to label and track the EPCs; however, the effect of SPIO upon EPCs remains unclear on a cellular level. In the present study, EPCs were labeled with home-synthesized SPIO nanoparticles in vitro and the biological characteristics of the labeled EPCs were evaluated. The EPCs were isolated from the peripheral blood of New Zealand rabbits and cultured in fibronectin-coated culture flasks. The EPCs were labeled with home-synthesized SPIO nanoparticles at a final iron concentration of 20 µg/ml. Labeled EPCs were confirmed with transmission electron microscopy and Prussian blue staining. The quantity of iron/cell was detected by atomic absorption spectrometry. The membranous antigens of EPCs were detected by cytofluorimetric analysis. Cell viability and proliferative capability between the labeled and unlabeled EPCs were compared. The rabbit EPCs were effectively labeled and the labeling efficiency was approximately 95%. The SPIO nanoparticles were localized in the endosomal vesicles of the EPCs, which were confirmed by transmission electron microscopy. No significant differences were found in cell viability and proliferative capability between labeled and unlabeled EPCs (P>0.05). In conclusion, rabbit peripheral blood EPCs were effectively labeled by home-synthesized SPIO nanoparticles, without influencing their main biological characteristics. PMID:22580964

  7. Study the cytotoxicity of different kinds of water-soluble nanoparticles in human osteoblast-like MG-63 cells

    SciTech Connect

    Niu, Lu [Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012 (China)] [Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012 (China); Li, Yang; Li, Xiaojie [Department of Pathophysiology, Prostate Diseases Prevention and Treatment Research Centre, Norman Bethune Medical School, Jilin University, Changchun 130012 (China)] [Department of Pathophysiology, Prostate Diseases Prevention and Treatment Research Centre, Norman Bethune Medical School, Jilin University, Changchun 130012 (China); Gao, Xue [Department of Analytical Chemistry, College of Chemistry, Jilin University, Changch