Document Detail


Differences in the TGF-{beta}1-induced profibrotic response of anterior and posterior corneal keratocytes in vitro.
MedLine Citation:
PMID:  19907023     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Purpose. To characterize phenotypic differences between anterior and posterior corneal keratocytes after stimulation with the profibrotic agent transforming growth factor-beta1 (TGF-beta1) in vitro. Methods. Sixteen corneas from healthy felines were obtained immediately after death. Lamellar dissection was performed to separate the anterior and posterior stroma at approximately 50% depth either manually (n = 2) or with a Moria microkeratome (300-mum head; n = 14). Cells from the anterior and posterior stroma were cultured separately but under identical conditions. Using immunohistochemistry and Western blot techniques, Ki-67 staining and relative expression of Thy-1, alpha smooth muscle actin (alpha-SMA), and fibronectin were assessed after stimulation with different TGF-beta1 concentrations. In addition, anterior and posterior cells cultured in different concentrations of TGF-beta1 were wounded with a razor blade, and the wound area and time to closure were determined. Results. Stimulation by all concentrations of TGF-beta1 increased the proportion of Ki-67-positive cells in anterior and posterior cell cultures, but this increase was noted earlier in posterior cells than in anterior cells. Increasing TGF-beta1 concentration also increased the relative expression of Thy-1, alpha-SMA, and fibronectin in anterior and posterior fibroblasts. However, anterior cells expressed these fibrotic markers at lower TGF-beta1 concentrations than did posterior keratocytes. After mechanical wounding, posterior cells closed the wound area faster than did anterior cells at all concentrations of TGF-beta1. Conclusions. The present experiments show that anterior and posterior corneal keratocytes exhibit different sensitivities to the profibrotic growth factor TGF-beta1. This heterogeneity of keratocyte response may impact wound closure after mechanical wounding.
Authors:
Holly B Hindman; Jennifer N Swanton; Richard P Phipps; Patricia J Sime; Krystel R Huxlin
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2009-11-11
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  51     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-03-26     Completed Date:  2010-04-26     Revised Date:  2014-09-21    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1935-42     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Actins / metabolism
Animals
Antigens, Thy-1 / metabolism
Blotting, Western
Cats
Cells, Cultured
Cornea / injuries
Corneal Stroma / cytology,  drug effects*,  metabolism
Fibroblasts / drug effects*,  metabolism
Fibronectins / metabolism
Immunohistochemistry
Ki-67 Antigen / metabolism
Phenotype
Transforming Growth Factor beta1 / pharmacology*
Grant Support
ID/Acronym/Agency:
ES01247/ES/NIEHS NIH HHS; EY015836/EY/NEI NIH HHS; EY017123/EY/NEI NIH HHS; HL095402/HL/NHLBI NIH HHS; HL75432/HL/NHLBI NIH HHS; K23 EY019353/EY/NEI NIH HHS; K23 EY019353/EY/NEI NIH HHS; K23 EY019353-01/EY/NEI NIH HHS; KL2 RR024136/RR/NCRR NIH HHS; KL2 RR024136/RR/NCRR NIH HHS; KL2 RR024136-016493/RR/NCRR NIH HHS; L30 EY019138/EY/NEI NIH HHS; L30 EY019138/EY/NEI NIH HHS; L30 EY019138-01/EY/NEI NIH HHS; P0EY01319F/EY/NEI NIH HHS; P30 ES001247/ES/NIEHS NIH HHS; P30 ES001247-250109/ES/NIEHS NIH HHS; P30 ES001247-25S10109/ES/NIEHS NIH HHS; P30 EY001319/EY/NEI NIH HHS; P30 EY001319-35/EY/NEI NIH HHS; R01 EY015836/EY/NEI NIH HHS; R01 EY015836-04/EY/NEI NIH HHS; R01 EY015836-05/EY/NEI NIH HHS; R01 EY017123/EY/NEI NIH HHS; R01 EY017123-04/EY/NEI NIH HHS; R01 HL075432/HL/NHLBI NIH HHS; R01 HL075432-04/HL/NHLBI NIH HHS; R01 HL075432-04S2/HL/NHLBI NIH HHS; R03 HL095402/HL/NHLBI NIH HHS; R03 HL095402-02/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Actins; 0/Antigens, Thy-1; 0/Fibronectins; 0/Ki-67 Antigen; 0/Transforming Growth Factor beta1
Comments/Corrections

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