Ultraviolet light-irradiated collagen III modulates expression of cytoskeletal and surface adhesion molecules in rat aortic smooth muscle cells in vitro

Abstract

Systemic and pulmonary hypertension is characterised by structural reconstruction of the vascular wall which includes hypertrophy and hyperplasia of vascular smooth muscle cells (VSMCs) and fibroproduction. We hypothesise that these changes are stimulated by non-enzymatic modification of collagen molecules in the injured vascular wall by radicals. We exposed collagen III to ultraviolet (UV) light irradiation which, as indicated by fluorescence and electrophoretic analyses, resulted in its fragmentation. Both irradiated and control unmodified collagen were adsorbed on culture dishes and seeded with VSMCs derived from the rat thoracic aorta. During the first week after seeding, the cells on the modified collagen attained significantly higher population density (by 15-83%), higher mitotic index (by 31-135%) and higher BrdU labelling index (by 32%). However, these cells were less resistant to spontaneous and trypsin-mediated detachment from the growth support. As revealed using enzyme-linked immunosorbent assay in 3-day-old cultures, the cells growing on the irradiated collagen exhibited a lower concentration of beta-1 integrins (-10%, measured per milligram of protein), vinculin (-18%), talin (-6%) and vimentin (-15%). Immunofluorescence staining showed that these molecules were distributed more diffusely and less organised into focal adhesion plaques or cytoskeletal fibres. The concentration of two adhesion molecules of immunoglobulin type, ICAM-1 and VCAM-1, was increased by 11% and 16%, respectively. The concentration of alpha-v integrins and alpha-actin was unchanged; the latter, however, formed fewer distinct microfilament bundles in cells on the modified collagen. Our results suggest that the VSMCs growing on UV-modified collagen are more prone to escape the growth control mediated by cell-extracellular matrix contact and can bind the cells of the immune system.

Enhanced Growth and Osteogenic Differentiation of Human Osteoblast-Like Cells on Boron-Doped Nanocrystalline Diamond Thin Films

“…Nevertheless, as shown in our earlier studies and in studies by other authors, even weak changes in the production of adhesion, cytoskeletal, differentiation and immunomodulatory molecules can significantly influence the morphology and behaviour of cells, e.g., vascular smooth muscle cells on collagen III substrates treated by UV light [56]. Similarly, in a study performed by Jeng et al (1993) [57] on cultured human umbilical vein endothelial cells, even a small increase in the expression of immunoglobuline and selectin adhesion molecules, induced by treatment with oxidized human plasma low density lipids (LDL), resulted in a significant increase in the number and adhesion strength of monocytes bound to the endothelial cells.…”

Adhesion and Growth of Vascular Smooth Muscle Cells on Nanostructured and Biofunctionalized Polyethylene

“…The non-specific binding of the secondary antibody on samples grafted with Fn and BSA was significantly reduced due to the presence of these biomolecules. In this context it should be pointed out that in studies using immunocytochemical methods, BSA [25,26,29], the whole blood serum [30] or solutions of other biomolecules, e.g., gelatin [12] are commonly used for blocking non-specific binding sites for antibodies. In addition, the fluorescence intensity on HDPE grafted with Fn or BSA was higher than on LDPE, which showed that Fn and BSA were grafted in a higher concentration on the HDPE surface than on LDPE [Figure 2(A,B)].…”

In vitro Proliferation and Osteogenic Differentiation of Human Bone Marrow-derived Mesenchymal Stem Cells Cultured with Hardystonite (Ca2ZnSi 2O7) and β-TCP Ceramics

“…Cell proliferation and differentiation are two key points in the application of MSCs in bone regeneration. Osteogenic differentiated cells generally show a low level of proliferation [30,31]. It has been reported that plasma-implanted titanium surfaces support osteogenic differentiation of bone marrow cells at the expense of reduced proliferation [32].…”