Fitzroy J Byfield | PubMed Health

PubMed2006

OxLDL increases endothelial stiffness, force generation, and network formation.

This study investigates the effect of oxidatively modified low density lipoprotein (OxLDL) on the biomechanical properties of human aortic endothelial cells (HAECs). We show that treatment with OxLDL results in a 90% decrease in the membrane deformability of HAECs, as determined by micropipette aspiration. Furthermore, aortic endothelial cells freshly isolated from hypercholesterolemic pigs were significantly stiffer than cells isolated from healthy animals. Interestingly, OxLDL had no effect on membrane cholesterol of HAECs but caused the disappearance of a lipid raft marker, GM1, from the plasma membrane. Both an increase in membrane stiffness and a disappearance of GM1 were also observed in cells that were cholesterol-depleted by methyl-beta-cyclodextrin. Additionally, OxLDL treatment of HAECs embedded within collagen gels resulted in increased gel contraction, indicating an increase in force generation by the cells. This increase in force generation correlated with an increased ability of HAECs to elongate and form networks in a three-dimensional environment. Increased force generation, elongation, and network formation were also observed in cholesterol-depleted cells. We suggest, therefore, that exposure to OxLDL results in the disruption or redistribution of lipid rafts, which in turn induces stiffening of the endothelium, an increase in endothelial force generation, and the potential for network formation.

Animals↗

PubMed2004

Cholesterol depletion increases membrane stiffness of aortic endothelial cells.

This study has investigated the effect of cellular cholesterol on membrane deformability of bovine aortic endothelial cells. Cellular cholesterol content was depleted by exposing the cells to methyl-beta-cyclodextrin or enriched by exposing the cells to methyl-beta-cyclodextrin saturated with cholesterol. Control cells were treated with methyl-beta-cyclodextrin-cholesterol at a molar ratio that had no effect on the level of cellular cholesterol. Mechanical properties of the cells with different cholesterol contents were compared by measuring the degree of membrane deformation in response to a step in negative pressure applied to the membrane by a micropipette. The experiments were performed on substrate-attached cells that maintained normal morphology. The data were analyzed using a standard linear elastic half-space model to calculate Young elastic modulus. Our observations show that, in contrast to the known effect of cholesterol on membrane stiffness of lipid bilayers, cholesterol depletion of bovine aortic endothelial cells resulted in a significant decrease in membrane deformability and a corresponding increase in the value of the elastic coefficient of the membrane, indicating that cholesterol-depleted cells are stiffer than control cells. Repleting the cells with cholesterol reversed the effect. An increase in cellular cholesterol to a level higher than that of normal cells, however, had no effect on the elastic properties of bovine aortic endothelial cells. We also show that although cholesterol depletion had no apparent effect on the intensity of F-actin-specific fluorescence, disrupting F-actin with latrunculin A abrogated the stiffening effect. We suggest that cholesterol depletion increases the stiffness of the membrane by altering the properties of the submembrane F-actin and/or its attachment to the membrane.

Animals↗