Differential Effects of Fibronectin-DerivedOligopeptides on the Attachment of Rabbit Lens Epithelial Cells in vitro

Abstract

Among the Arg-Gly-Asp (RGD)-containing sequences that are known to be cell-binding domains of fibronectin, 500 µg/ml of Arg-Gly-Asp-Ser (RGDS) and Gly-Arg-Gly-Asp-Ser showed 100% inhibition of the attachment of cultured lens epithelial cells (TOTL-86 cells), when they were added to culture medium and coincubated with the cells for 24 h whereas RGD at concentrations of 500, 1,000 and 2,000 µg/ml had no such activity. After 48 h of cocultivation of 800, 400 or 200 µg/ml of RGDS with TOTL-86 cells, the percentage of floating cells was 100, 30.1 or 11.1%, respectively. After 144 h of cocultivation with RGDS, the percentage of floating cells was 1.6,2.4 or 1.9%, respectively, indicating that RGDS was not cytotoxic to lens epithelial cells. However, replacing the medium with fresh medium containing new RGDS peptide resulted in floating of cells. We also studied the inhibitory effect of two other amino acid sequences that are found in cell-binding sites of the fibronectin molecule: Glu-Ile-Leu-Asp-Val-Pro-Ser-Thr (EILDVPST) and Arg-Glu-Asp-Val (REDV). At 500 and 1,000 µg/ml, respectively, neither EILDVPST nor REDV has an inhibitory effect on the attachment of TOTL-86 cells, while RGDS at a concentration of 500 µg/ml completely inhibited the attachment of the cells in 24 h of incubation.

Control of fibrotic changes through the synergistic effects of anti-fibronectin antibody and an RGDS-tagged form of the same antibody

“…It is not surprising therefore, that fibronectin plays a critical role in mediating ECM deposition and inflammatory response in the development of fibrotic diseases. In vitro experiments with rabbit lens epithelial cells have demonstrated the role of fibronectin in promoting the adhesion and migration of these cells 10 21 , which contribute to the fibrotic pathology in posterior capsular opacification, through their differentiation into myofibroblasts in the presence of TGF-β2. Although fibronectin is present in minimal amounts in adult lens, exposure to plasma fibronectin and other growth factors, results in increased expression of fibronectin, following a cataract surgery 22 .…”

Inhibitory Effects of Arg-Gly-Asp (RGD) Peptide on Cell Attachment and Migration in a Human Lens Epithelial Cell Line

“…Palmade et al [24] reported that Ac-Arg-Gly-Asp-NH-CH2-Bzl (LCM 1910) which is an RGD-derived peptide signifi cantly decreased cell attachment on the lens capsule at a concentrationdependent effect, and cell proliferation was decreased on a plastic dish in the presence of LCM 1910 in bovine LEC culture. Sasabe et al [25] reported that 0.5 mg/ml of RGDS and GRGDS RGD peptide showed inhibitory effects on cell attachment in rabbit LEC culture, but 0.5, 1.0 and 2.0 mg/ml of the RGD sequence alone had no effect. Nishi et al [26] implanted a 4% RGD peptide or a combination of 10% RGD peptide and 23% EDTA containing polylactic-glycolic acid disk in the capsular bag in rabbit eyes.…”