Document Detail


Evaluation of a range of anti-proliferative assays for the preclinical screening of anti-psoriatic drugs: a comparison of colorimetric and fluorimetric assays with the thymidine incorporation assay.
MedLine Citation:
PMID:  20482335     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Established treatments for psoriasis are generally based on antiproliferative, anti-inflammatory, or differentiation-modifying activity, or a combination of these effects. New agents for the treatment of psoriasis could be identified by high-throughput screening (HTS) of large compound libraries using keratinocyte proliferation models. Although several new proliferation assays have been developed, the radioactive [(3)H]-thymidine incorporation assay is still considered to be the gold standard for the evaluation of keratinocyte proliferation in vitro. In this study, we compare a number of simple, and reliable, colorimetric (MTT, NRU, SRB, and CVS), and fluorimetric (CAM and AB) methods with the [(3)H]-thymidine incorporation assay for the measurement of keratinocyte proliferation in the exponential growth phase in 96-well formats. The concentrations that induced 50% growth inhibition (GI(50)) were determined by each assay for the established antipsoriatics, dithranol, and methotrexate. Strong correlations were observed between the percentage growth inhibitions determined by the radioactive and the colorimetric assays, with no significant differences (P > 0.05) between their GI(50) values. The colorimetric assays are thus suitable alternatives to the radioactive assay for quantifying keratinocyte growth inhibition. We have also validated the use of the HaCaT cell line as a representative of the hyperproliferative psoriatic epidermis, in the preclinical screening of experimental anti-psoriatic agents.
Authors:
Suja Elizabeth George; Rosaleen J Anderson; Anne Cunningham; Michael Donaldson; Paul W Groundwater
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Assay and drug development technologies     Volume:  8     ISSN:  1557-8127     ISO Abbreviation:  Assay Drug Dev Technol     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-06-28     Completed Date:  2010-10-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101151468     Medline TA:  Assay Drug Dev Technol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  389-400     Citation Subset:  IM    
Affiliation:
Sunderland Pharmacy School, University of Sunderland, Wharncliffe Street, Sunderland, United Kingdom.
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MeSH Terms
Descriptor/Qualifier:
Anthralin / pharmacology
Anti-Infective Agents, Local / pharmacology
Caspase 3 / metabolism
Caspase 7 / metabolism
Cell Line
Cell Proliferation / drug effects*
Colorimetry
Dermatologic Agents / pharmacology
Dose-Response Relationship, Drug
Drug Evaluation, Preclinical / methods*
Fluorescent Dyes
Fluorometry
Humans
Keratinocytes / drug effects
Methotrexate / pharmacology
Psoriasis / drug therapy*
Reproducibility of Results
Tetrazolium Salts
Thiazoles
Thymidine / metabolism*
Chemical
Reg. No./Substance:
0/Anti-Infective Agents, Local; 0/Dermatologic Agents; 0/Fluorescent Dyes; 0/Tetrazolium Salts; 0/Thiazoles; 298-93-1/thiazolyl blue; 480-22-8/Anthralin; 50-89-5/Thymidine; 59-05-2/Methotrexate; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 7

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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