Abundance and location of proteoglycans and hyaluronan within normal and myxomatous mitral valves.

INTRODUCTION: Extracellular matrix changes occur in many heart valve pathologies. For example, myxomatous mitral valves are reported to contain excess proteoglycans and hyaluronan. However, it is unknown which specific proteoglycans are altered in myxomatous valves. Because proteoglycans perform varied functions in connective tissues, this study was designed to identify and localize three matrix-associated proteoglycans, as well as hyaluronan and the hyaluronan receptor for endocytosis, within myxomatous and normal mitral valves. METHODS: Human mitral posterior leaflets (control, n=6-9; myxomatous, n=14-21; mean age, 61 years for all groups) were histochemically stained for proteoglycan core proteins, hyaluronan, and the hyaluronan receptor for endocytosis. Stain intensity was semiquantitatively graded to determine differences in marker abundance between normal and myxomatous valves. The proteoglycans were localized to different regions of the leaflet by correspondence to parallel Movat-stained sections. RESULTS: The proteoglycans decorin, biglycan, and versican were more abundant in myxomatous valves than in normal controls (P<.03). There was a gender effect on proteoglycan presence, but no age-related trends were observed. Hyaluronan and the hyaluronan receptor for endocytosis were distributed throughout all valves. There was no significant difference in hyaluronan between groups, but expression of the hyaluronan receptor for endocytosis was reduced in myxomatous valves compared to normal controls (P<.002). CONCLUSION: Excess decorin, biglycan, and versican may be associated with the remodeling of other matrix components in myxomatous mitral valves. Decreased expression of the hyaluronan receptor for endocytosis in myxomatous valves suggests that hyaluronan metabolism could be altered in myxomatous mitral valve disease. These findings contribute towards elucidating the pathogenesis of myxomatous mitral valve disease and developing potential new therapies.

Influence of cyclic strain and decorin deficiency on 3D cellularized collagen matrices.

Cyclic strain evokes the expression of the small leucine-rich proteoglycans decorin and biglycan in 2D cultures and native tissues. However, strain-dependent expression of these proteoglycans has not been demonstrated in engineered tissues. We hypothesized that the absence of decorin may compromise the effect of cyclic strain on the development of engineered tissues. Thus, we investigated the contribution of decorin to tissue organization in cyclically strained collagen gels relative to statically cultured controls. Decorin null (Dcn(-/-)) and wild-type murine embryonic fibroblasts were seeded within collagen gels and mechanically conditioned using a Flexcell Tissue Train culture system. After 8 days, the cyclically strained samples demonstrated greater collagen fibril density, proteoglycan content, and material strength for both cell types. On the other hand, increases in cell density, collagen fibril diameter, and biglycan expression were observed only in the cyclically strained gels seeded with Dcn(-/-) cells. Although cyclic strain caused an elevation in proteoglycan expression regardless of cell type, the type of proteoglycan differed between groups: the Dcn(-/-) cell-seeded gels produced an excess of biglycan not found in the wild-type controls. These results suggest that decorin-mediated tissue organization is strongly dependent upon tissue type and mechanical environment.

Endogenous overexpression of hyaluronan synthases within dynamically cultured collagen gels: Implications for vascular and valvular disease.

Hyaluronan is a ubiquitous component of the extracellular matrix with important roles in cell and tissue functions. Hyaluronan content is often elevated in cardiovascular diseases, such as mitral valve disease and atherosclerosis. The objective of this study was to determine the impact of endogenously produced hyaluronan dynamically cultured three-dimensional model of collagenous tissues. Collagen gels containing excess HA and hyaluronan synthase (has) overexpressing cells were grown in a cyclic strain environment to simulate cell-mediated matrix organization. Cyclic strain caused a significant elevation in the collagen fibril density, cell number, and hyaluronan content of the resulting collagen gels compared to those grown under a static strain regimen. The material behavior of collagen gels containing has overexpressing cells was also notably weakened compared to controls. Transmission electron microscopy and immunohistochemistry showed that proteoglycan distribution was influenced by both strain and has overexpression. The results were also dependent on the specific has isozyme overexpressed. This investigation helps to identify the mechanism by which hyaluronan acts in vivo to alter tissue material behavior in cardiovascular diseases such as myxomatous mitral valve disease and atherosclerosis.