A ging is a major risk factor for the development of cardiovascular disease, and age-related changes in vascular function are hypothesized to influence the progression of atherosclerosis. One of the hallmarks of aging tissues is an impaired ability to regenerate, caused by the accumulation of senescent cells. Cellular senescence is a state of irreversible growth arrest 1 that can be triggered via the progressive shortening of telomeres (DNA sequence repeats that protect the ends of chromosomes) with successive rounds of cell division, known as replicative senescence.2 Senescence can also be induced by a variety of cellular stresses, independently of extensive proliferation, including oxidative stress 3,4 and activation of oncogenes. 5The signaling pathways that promote cellular senescence vary according to the initiating stimulus, cellular context, and other factors. Senescence can be induced by p16 INK4A (p16) and p14 ARF /p53 signaling pathways.5-8 p16 inhibits cyclin-dependent kinases 2 and 4, whereas p53 induces the cyclin-dependent © 2014 American Heart Association, Inc. Objective-Although atherosclerosis is associated with systemic risk factors such as age, high cholesterol, and obesity, plaque formation occurs predominately at branches and bends that are exposed to disturbed patterns of blood flow. The molecular mechanisms that link disturbed flow-generated mechanical forces with arterial injury are uncertain. To illuminate them, we investigated the effects of flow on endothelial cell (EC) senescence. Approach and Results-LDLR−/− (low-density lipoprotein receptor −/− ) mice were exposed to a high-fat diet for 2 to 12 weeks (or to a normal chow diet as a control) before the assessment of cellular senescence in aortic ECs. En face staining revealed that senescence-associated β-galactosidase activity and p53 expression were elevated in ECs at sites of disturbed flow in response to a high-fat diet. By contrast, ECs exposed to undisturbed flow did not express senescence-associated β-galactosidase or p53. Studies of aortae from healthy pigs (aged 6 months) also revealed enhanced senescence-associated β-galactosidase staining at sites of disturbed flow. These data suggest that senescent ECs accumulate at disturbed flow sites during atherogenesis. We used in vitro flow systems to examine whether a causal relationship exists between flow and EC senescence. Exposure of cultured ECs to flow (using either an orbital shaker or a syringe-pump flow bioreactor) revealed that disturbed flow promoted EC senescence compared with static conditions, whereas undisturbed flow reduced senescence. Gene silencing studies demonstrated that disturbed flow induced EC senescence via a p53-p21 signaling pathway. Disturbed flow-induced senescent ECs exhibited reduced migration compared with nonsenescent ECs in a scratch wound closure assay, and thus may be defective for arterial repair. However, pharmacological activation of sirtuin 1 (using resveratrol or SRT1720) protected ECs from disturbed flow-induced senescence. Conclusions-Distu...

Experimental data point to a determinant role for endothelial cell (EC) anionic sites in the regulation of vascular permeability. Previous studies have shown that EC anionic sites density is reduced in conditions of enhanced permeability. The pathophysiology of migraine and vascular headache encompasses dilatation of dural vessels and extravasation of plasma proteins. The current study was carried out to determine if the density of EC anionic sites is reduced in enhanced permeability of dural vessels. Enhanced permeability was chemically induced in rats by intravenous injection of substance P and was tested by assessing leakage of horseradish peroxidase (HRP). Anionic sites were labelled with cationic colloidal gold and their density was quantified from electron microscopy negatives. Experimental animals showed increased leakage of HRP from dural vessels. However, anionic sites in EC membranes (luminal and abluminal) showed no statistical differences when their mean densities in experimental and control animals were compared. The results indicate that in this model, factors other than the density of anionic sites may be important determinants in the permeability of dural vessels. Such factors may include structural alteration of ECs consistent with an increased permeability. In this study pronounced ultrastructural changes in ECs were noted in experimental animals including widening of intercellular junctions and an increase in the number of EC gaps and vesicles.

The scope of micro-CT imaging can be extended from current in vitro applications to establish post-extraction diagnosis of subtle odontogenic defects, in a manner similar to deriving histopathological diagnoses in extracted teeth. Ongoing technological advancements hold the promise for more widespread translatory applications.