Purchase this article with an account.
C Latz, G Pavon-Djavid, F El Khadali, V Migonney; Increased Activation of FAK, ERK1/2 in Human Fibroblasts and Rabbit Lens Epithelial Cells Upon Adhesion to Modified Acrylic Polymers in Comparison to PMMA . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4003.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate the adhesion inhibiting effect of modified acrylic polymers (EF35) in comparison to polymethylmetacrylate (PMMA) Methods: Cell culture: Human fibroblasts McCoy and primary rabbit lens epithelial cells (LEC) were serum-starved overnight and harvested by limited trypsin/EDTA treatment; cells were held in suspension for 1h and seeded onto fibronectin-coated PMMA and EF35 discs. At various time points following plating the attached cells were either counted, labeled for F-actin, or lysed. Whole cell lysates were separated by SDS-PAGE and blotted against P-ERK1/2 , FAK, or phosphotyrosine. Blots were scanned and analyzed with an image analysis program. Polymer synthesis: EF35 was synthesized by radical polymerization and characterized by NMR and infrared analysis. Commercially available PMMA served as control. Results: Adhesion: McCoy cells adhered to PMMA and EF35 similarly. The number of cells that adhered to EF35 was lower than that of PMMA. Adhesion to PMMA was faster than to EF35. Western blot of whole cell lysates revealed increased and more sustained activation levels of FAK in cells seeded on EF35. Additionally, activation of ERK1/2 was found for a longer period in EF35 seeded cells. Morphologic studies on actin stained cells showed decreased adhesion to EF35 in rabbit LECs and decreased F-Actin formation near the outer membranes of McCoy cells. Conclusion: Integrin- mediated activation of FAK and ERK1/2 is more pronounced in cells plated onto substituted polymers. Our results indicate a different adhesion mechanism on substituted polymers leading to an increased and sustained activation of integrin mediated kinases in human fibroblasts. F-actin labeling demonstrates changes in the cytosceletons of human fibroblasts and rabbit LECs.
This PDF is available to Subscribers Only