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ONTOLOGY SOURCE REFERENCE
Term Source NameMONPOUOChEBIPATONCIt
Term Source Filehttp://purl.bioontology.org/ontology/MOhttp://purl.bioontology.org/ontology/npohttp://purl.bioontology.org/ontology/UOhttp://purl.bioontology.org/ontology/CHEBIhttp://purl.bioontology.org/ontology/PATOhttp://ncit.nci.nih.gov/
Term Source Versionv. 1.3.1.1v. 2011-02-12v. 80v. 11.11d
Term Source DescriptionMGED OntologyNanoParticle OntologyUnit OntologyChemical Entities of Biological InterestPhenotype OntologyNCI Thesaurus
INVESTIGATION
Investigation IdentifierNCL200612A
Investigation TitleDendrimer-Based MRI Contrast Agents
Investigation DescriptionThe goal of this investigation is to characterize a PAMAM dendrimer with an associated gadolinium chelate MRI contrast agent.
Investigation Submission Date2002-11-30
Investigation Public Release Date2002-11-30
Investigation Disease
Investigation Disease Term Accession Number
Investigation Disease Term Source REF
Investigation Outcome
INVESTIGATION PUBLICATIONS
Investigation PubMed ID18095846
Investigation Publication DOI10.2217/17435889.2.6.789
Investigation Publication Author List10.2217/17435889.2.6.789
Investigation Publication TitleCharacterization of nanoparticles for therapeutics
Investigation Publication Statuspublished
Investigation Publication Status Term Accession Number
Investigation Publication Status Term Source REF
INVESTIGATION CONTACTS
Investigation Person Last NameDoe
Investigation Person First NameJohn
Investigation Person Mid InitialsE
Investigation Person Emaildoej@mail.nih.gov
Investigation Person Phone1231231234
Investigation Person Fax
Investigation Person AddressLaboratory Street, City, State 111111
Investigation Person AffiliationDoe Laboratories
Investigation Person Rolesinvestigator
Investigation Person Roles Term Accession Number
Investigation Person Roles Term Source REFMO
MATERIAL
Material File Namem_NCL-20.xlsm_NCL-21.xlsm_NCL-22.xlsm_NCL-23.xlsm_NCL-24.xlsm_NCL-25.xlsm_NCL-26.xls
Material Source NameNCL-20NCL-21NCL-22NCL-23NCL-24NCL-25NCL-26
STUDY
Study IdentifierNCL200612A-Size
Study TitleHydrodynamic Size/Size Distribution via Dynamic Light Scattering (DLS)
Study DescriptionDynamic light scattering(DLS) technique was used to measure the hydrodynamic size of dendritic nanomaterials. The effects of sample concentration, buffer and temperature on the hydrodynamic size (stability) also were measured.
Study Submission Date2002-11-30
Study Public Release Date2002-11-30
Study Disease
Study Disease Term Accession Number
Study Disease Term Source REF
Study OutcomeHydrodynamic size (diameter) of the dendrimer samples NCL22, NCL23 and NCL20 were measured in aqueous solutions using DLS at 25 °C and 37 °C. An instrument with a backscattering detector was used for these measurements in batch mode (no fractionation). This technique does not have the resolving power of differentiating monomers and dimers without fractionation. Samples were weighed, dissolved in deionized (DI) water, aliquoted, lyophilized and resuspended in desired buffer solutions to a final concentration of 1 mg/mL, and filtered through a 0.2-μm filter, unless otherwise indicated. The measurements were taken in saline (154 mM NaCl) and phosphate-buffered saline (PBS) at pH 7.4. Three measurements were taken for each sample. For NCL22, the size is slightly larger when dispersed in saline compared to PBS. In PBS, the size is independent of temperature. This is in contrast to NCL23, which is larger in PBS than in saline. NCL23 also shows temperature dependence, as its size decreases slightly with increased temperature in PBS. Finally, NCL20 is larger when dispersed in PBS compared to saline. For each sample, the intensity weighted mean diameter (Z-avg) derived from the cumulants analysis and the diameter after conversion to volume-weighted distribution are provided on the following pages.
Study File Names_size-DLS.xls
STUDY DESIGN DESCRIPTORS
Study Design Typecomparison
Study Design Type Term Accession Number
Study Design Type Term Source REF
STUDY PUBLICATIONS
Study PubMed ID18095846
Study Publication DOI10.2217/17435889.2.6.789
Study Publication Author ListHall JB; Dobrovolskaia MA; Patri AK; McNeil SE
Study Publication TitleCharacterization of nanoparticles for therapeutics
Study Publication Statuspublished
Study Publication Status Term Accession Number
Study Publication Status Term Source REF
STUDY FACTORS
Study Factor Nametemperaturesolvent medium
Study Factor Typetemperaturesolvent medium
Study Factor Type Term Accession NumberPATO_0000146NPO_1855
Study Factor Type Term Source REFPATONPO
STUDY ASSAYS
Study Assay Measurement Typehydrodynamic size
Study Assay Measurement Type Term Accession Number
Study Assay Measurement Type Term Source REF
Study Assay Technology Typedynamic light scattering(DLS)
Study Assay Technology Type Term Accession NumberNPO_1469
Study Assay Technology Type Term Source REFNPO
Study Assay Technology Platform
Study Assay Measurement Namehydrodynamic diameter; peak size; PDI
Study Assay Measurement Name Term Accession Number; ; NPO_1155
Study Assay Measurement Name Term Source REF; ; NPO
Study Assay File Namea_size-DLS.xls
STUDY PROTOCOLS
Study Protocol NameMeasuring the size of nanoparticles in aqueous media using batch-mode dynamic light scattering
Study Protocol Typesample preparation procedure; particle size measurement procedure
Study Protocol Type Term Accession Number
Study Protocol Type Term Source REF
Study Protocol DescriptionThis assay protocol outlines procedures for sample preparation and the determination of mean nanoparticle size (hydrodynamic diameter) using batch-mode dynamic light scattering (DLS) in dilute aqueous suspensions. Although particle size is the primary determinant of the measured diffusion coefficient, other parameters can impact these measurements and influence the measured size. Therefore, guidelines for making successfully size measurements in the nanosize range are provided, as well as a discussion of relevant standards and data analysis. This protocol can be applied to any suitable DLS instrument with batch measurement capability.
Study Protocol URINCL_Method_PCC-1.pdf
Study Protocol Version1.1
Study Protocol Parameters NamepH; NaCl concentration; particle concentration
Study Protocol Parameters Name Term Accession NumberUO_0000196; ; NPO_1830
Study Protocol Parameters Name Term Source REFUO; ;NPO
Study Protocol Components NameMastersizer 2000 (Malvern DLS)
Study Protocol Components TypeDLS instrument
Study Protocol Components Type Term Accession NumberNPO_1766
Study Protocol Components Type Term Source REFNPO
STUDY CONTACTS
Study Person Last NameSmith
Study Person First NameJane
Study Person Mid InitialsK
Study Person Emailsmithj@mail.nih.gov
Study Person Phone1231231235
Study Person Fax
Study Person AddressLaboratory Street, City, State 111111
Study Person AffiliationDoe Laboratories
Study Person Rolesinvestigator
Study Person Roles Term Accession Number
Study Person Roles Term Source REFMO
STUDY
Study IdentifierNCL200612A-CytoxicityLLC-PK1
Study TitleCytotoxicity characterization in LLC-PK1 cells
Study DescriptionNanoparticle biocompatibility was evaluated in the porcine renal proximal tubule cell line, LLC-PK1. Cytotoxicity was determined as described in the NCL protocol for LLC-PK1 Kidney Cytotoxicity Assay(GTA-1). Briefly, test materials were diluted to the desired assay concentrations in cell culture media. Cells were preincubated for 24 h prior to adding test material, reaching an approximate confluence of 80%. Cells were exposed to test material for 6, 24 and 48 h, and cytotoxicity was determined using the MTT cell viability and LDH membrane integrity assays.
Study Submission Date2002-11-30
Study Public Release Date2002-11-30
Study Disease
Study Disease Term Accession Number
Study Disease Term Source REF
Study OutcomeNCL22, NCL23 and NCL24 were found to be minimally cytotoxic, under the testing conditions utilized.
Study File Names_cytotoxicity-LLC-PK1.xls
STUDY DESIGN DESCRIPTORS
Study Design Typecomparison
Study Design Type Term Accession Number
Study Design Type Term Source REF
STUDY PUBLICATIONS
Study PubMed ID18095846
Study Publication DOI10.2217/17435889.2.6.789
Study Publication Author ListHall JB; Dobrovolskaia MA; Patri AK; McNeil SE
Study Publication TitleCharacterization of nanoparticles for therapeutics
Study Publication Statuspublished
Study Publication Status Term Accession Number
Study Publication Status Term Source REF
STUDY FACTORS
Study Factor Namenanoparticle sampleparticle concentrationtime of exposure
Study Factor Typenanoparticle sampleparticle concentrationtime of exposure
Study Factor Type Term Accession NumberNPO_1830
Study Factor Type Term Source REFNPO
STUDY ASSAYS
Study Assay Measurement TypeMTT AssayLDH Release Assay
Study Assay Measurement Type Term Accession NumberNPO_1709
Study Assay Measurement Type Term Source REFNPO
Study Assay Technology Type
Study Assay Technology Type Term Accession Number
Study Assay Technology Type Term Source REF
Study Assay Technology Platform
Study Assay Measurement Namecell viabilityLDH release
Study Assay Measurement Name Term Accession NumberNPO_1343
Study Assay Measurement Name Term Source REFNPO
Study Assay File Namea_MTT-LLCPK1.xlsa_LDH-LLCPK1.xls
STUDY PROTOCOLS
Study Protocol NameTime-6-24-48 plate MTT assayTest plate LDH assayTime zero plate MTT assaycell preparation in four 96-well platesAPAP positive control preparationTriton-X-100 positive control preparationMTT assay reagent preparationTest sample and positive control additionLDH assay reagent preparation
Study Protocol Typecell preparationcontrol preparationcontrol preparationreagent preparationreagent preparation
Study Protocol Type Term Accession Number
Study Protocol Type Term Source REF
Study Protocol DescriptionTest Plates: 6, 24 and 48 hour exposures (MTT Assay) 5.4.1 Remove appropriate test plate from incubator and replace media from Triton-X positive control wells (see plate format in Appendix) with 200 mL 1% Triton-X (made in Step 4.1.2). Let the plate set for 10 minutes at room temperature. Spin plate at 700 x g for 3 minutes. 5.4.2 Remove 100 mL of media from each well and transfer it to another plate, maintaining plate format. Use this plate immediately for the LDH assay (see Section 5.5). 5.4.3 Remove remaining media from original plate and discard. 5.4.4 Add 200 mL of fresh media to all wells. 5.4.5 Add 50 mL of MTT to all wells. 5.4.6 Cover in aluminum foil and incubate for 37°C for 4 hours. 5.4.7 Remove plate from incubator and spin at 700 x g for 3 minutes. 5.4.8 Remove media and MTT. 5.4.9 Add 200 mL of DMSO to each well. 5.4.10 Add 25 mL of glycine buffer to each well. Place on shaker to mix. 5.4.11 Read absorbance at 570 nm on plate reader using a reference wavelength of 680 nm.Test Plates: 0, 6, 24 and 48 hour exposures (LDH Assay) (Adapted from Biovision LDH Cytotoxicity Assay Kit, K311-400) 5.5.1 Add 100 mL of the Reaction Mixture (step 4.3.2) to each well of transfer plate. Shake plate on an orbital shaker briefly to mix samples. 5.5.2 Incubate at room temperature for up to 20 minutes in the dark. 5.5.3 Read the plate on plate reader at 490 nm using a reference wavelength of 680 nm.5.2.1 Remove time zero plate from incubator and replace media from Triton-X positive control wells (see plate format in Appendix) with 200 mL 1% Triton-X (made in Step 4.1.2). Add 100 mL of media to the remaining wells. Let the plate set for 10 minutes at room temperature. Spin plate at 700 x g for 3 minutes. 5.2.2 Remove 100 mL of media from each well and transfer it to another plate, maintaining plate format. Use this plate immediately for the LDH assay (see Section 5.5). 5.2.3 Remove remaining media from original plate and discard. 5.2.4 Add 200 mL of fresh media to all wells. 5.2.5 Add 50 mL of MTT to all wells. 5.2.6 Cover in aluminum foil and incubate at 37°C for 4 hours. 5.2.7 Remove plate from incubator and spin at 700 x g for 3 minutes. 5.2.8 Aspirate media and MTT. 5.2.9 Add 200 mL of DMSO to all wells. 5.2.10 Add 25 mL of glycine buffer to all wells. Place on shaker to mix. 5.2.11 Read absorbance at 570 nm on plate reader.5.1.1 Harvest cryopreserved cells from prepared flasks (limit to 20 passages). 5.1.2 Count cell concentration using a coulter counter or hemocytometer. 5.1.3 Dilute cells to a density of 2.5 x 105 cells/mL in M199 (3% FBS) cell culture media. 5.1.4 Plate 100 mL cells/well as per plate format (Appendix) for four 96-well plates (time zero, 6 hour sample exposure, 24 hour sample exposure, 48 hour sample exposure). The format indicates no cells in rows D and E as they serve as particle blanks to be subtracted from cell treatment wells. Each plate accommodates two samples (Rows A–C and F–H). Each nanoparticle is tested at nine dilutions. Column 11 receives the APAP positive control and column 12 receives Triton X-100. 5.1.5 Incubate plates for 24 hours at 5% CO2, 37°C and 95% humidity.4.1.1 Acetaminophen (APAP) positive control: Add 19 mg to a total volume of 5 mL M199 Cell Culture Media (with 3% FBS) to make a 25 mM solution. Sterile filter using a 0.2 mm filter.4.1.2 1% Triton-X-100 positive control: Add 1 mL of Triton-X-100 to 99 mL of media. Sterile filter using a 0.2 mm filter.4.2.1 MTT solution: 5 mg/mL MTT in PBS, store for up to one month at 4°C in dark 4.2.2 Glycine Buffer: 0.1 M glycine (MW 75.07), 0.1 M NaCl (MW 58.44), pH 10.5, store at room temperature.5.3.1 The highest concentration of nanoparticle tested should be at the limit of solubility. The test sample should be at physiological pH. Neutralization of acidic/basic test samples may be required. 5.3.2 Dilute test compound in media, making a total of 9 1:4 dilutions. 5.3.3 Add 100 mL of each sample dilution and positive control to 6 hour, 24 hour and 48 hour exposure plates as per the plate format (Appendix).4.3.1 Reconstitute catalyst in 1 mL dH20 for 10 min with occasional vortexing (stable for 2 weeks at 4°C). 4.3.2 Reaction mixture (for one 96-well plate): Add 250 mL of reconstituted catalyst solution to 11.25 mL of dye solution (stable for 2 weeks at 4°C).
Study Protocol URINCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdf
Study Protocol Version1.11.11.11.11.11.11.11.11.1
Study Protocol Parameters Name
Study Protocol Parameters Name Term Accession Number
Study Protocol Parameters Name Term Source REF
Study Protocol Components NameMTT; acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakeracetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; biovision LDH-cytotoxicity assay kit; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakerMTT; acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakerM199 cell culture media; fetal bovine serum; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakeracetaminophen; M199 cell culture media; fetal bovine serumtriton-X-100; M199 cell culture media; fetal bovine serumMTT; glycine; sodium chlorideacetaminophen; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakerbiovision LDH-cytotoxicity assay kit
Study Protocol Components Typereagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrumentreagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrumentreagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrumentreagent; reagent; material; instrument; instrument; instrumentreagent; reagent; reagentreagent; reagent; reagentreagent; reagent; reagentreagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrumentreagent
Study Protocol Components Type Term Accession NumberNPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290;NPO_290; ; NPO_1436; NPO_1436; NPO_1436NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290;NPO_290; ; NPO_1436; NPO_1436; NPO_1436NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290;NPO_290; ; NPO_1436; NPO_1436; NPO_1436NPO_290; NPO_290; ; NPO_1436; NPO_1436; NPO_1436NPO_290; NPO_290; NPO_290NPO_290; NPO_290; NPO_290NPO_290; NPO_290; NPO_290NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; ; NPO_1436; NPO_1436; NPO_1436NPO_290
Study Protocol Components Type Term Source REFNPO; NPO; NPO; NPO; NPO; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO; NPO; NPO; NPO; NPO; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO; NPO; NPO; NPO; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO; NPO; ; NPO; NPO; NPONPO; NPO; NPONPO; NPO; NPONPO; NPO; NPONPO; NPO; NPO; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO
STUDY CONTACTS
Study Person Last NameSmith
Study Person First NameJane
Study Person Mid InitialsK
Study Person Emailsmithj@mail.nih.gov
Study Person Phone1231231235
Study Person Fax
Study Person AddressLaboratory Street, City, State 111111
Study Person AffiliationDoe Laboratories
Study Person Rolesinvestigator
Study Person Roles Term Accession Number
Study Person Roles Term Source REFMO