Restoration of epidermal organization and function in response to a variety of pathophysiological insults is critically dependent on coordinated keratinocyte migration, proliferation, and stratification during the process of wound healing. These processes are mediated by the reconfiguration of both cell-cell (desmosomes, adherens junctions) and cell-matrix (focal adhesions, hemidesmosomes) junctions and the cytoskeletal filament networks that they serve to interconnect. In this study, we investigated the role of substrate elasticity (stiffness) on keratinocyte colony formation in vitro during the process of nascent epithelial sheet formation as triggered by the calcium switch model of keratinocyte culture. Keratinocytes cultured on pepsin digested type I collagen coated soft (nominal E = 1.2 kPa) polyacrylamide gels embedded with fluorescent microspheres exhibited (i) smaller spread contact areas, (ii) increased migration velocities, and (iii) increased rates of colony formation with more cells per colony than did keratinocytes cultured on stiff (nominal E = 24 kPa) polyacrylamide gels. As assessed by tracking of embedded microsphere displacements, keratinocytes cultured on soft substrates generated large local substrate deformations that appeared to recruit adjacent keratinocytes into joining an evolving colony. Together with the observed differences in keratinocyte kinematics and substrate deformations, we developed two ad hoc analyses, termed distance rank (DR) and radius of cooperativity (RC), that help to objectively ascribe what we perceive as increasingly cooperative behavior of keratinocytes cultured on soft versus stiff gels during the process of colony formation. We hypothesize that the differences in keratinocyte colony formation observed in our experiments could be due to cell-cell mechanical signaling generated via local substrate deformations that appear to be correlated with the increased expression of β4 integrin within keratinocytes positioned along the periphery of an evolving cell colony.

For children born with a single functional ventricle, the Fontan operation bypasses the right ventricle by forming a four-way total cavopulmonary connection adapting the existing ventricle for the systemic circulation. However, upon adulthood, many Fontan patients exhibit low cardiac output and elevated venous pressure, eventually requiring a heart transplantation. Despite efforts to develop a Fontan pump or use an existing ventricular assist device for failing Fontan support, there is still no device designed or tested for subpulmonary support. Penn State University is developing a hydrodynamically levitated Fontan circulatory assist device (FCAD) for bridge-to-transplant or destination therapy. The FCAD hemodynamics, at both steady and pulsatile conditions for three pump operating conditions, were quantified using particle image velocimetry to determine the velocity magnitudes and Reynolds normal and shear stresses. Data were acquired at three planes (0 mm and ±25% of the radius) for the inferior and superior vena cavae inlets and the pulmonary artery outlet. The inlets had a blunt velocity profile that became skewed towards the collecting volute as fluid approached the rotor. At the outlet, regardless of the flow condition, a high-velocity jet exited the volute and moved downstream in a helical pattern. Turbulent stresses observed at the volute exit were influenced by the rotor's rotation. Regardless of inlet conditions, the pump demonstrated advantageous behavior for clinical use with a predictable flow field and a low risk of platelet adhesion and hemolysis based on calculated wall shear rates and turbulent stresses, respectively.

Supraphysiologic high shear stresses created in calcific aortic stenosis (AS) are known to cause hemostatic abnormalities, however, the relationship between the complex blood flows over the severity of AS and hemostatic abnormalities still remains unclear. This study systematically characterized the blood flow in mild, moderate, and severe AS. A series of large eddy simulations (LES) validated by particle image velocimetry were performed on physiologically representative AS models with a peak physiologic flow condition of 18 liter per minute. Time-accurate velocity fields, transvalvular pressure gradient, and laminar viscous—and turbulent (or Reynolds) shear stresses (RSSmax) were evaluated for each degree of severity. The peak velocities of mild, moderate, and severe AS were on the order of 2.0, 4.0, and 8.0 m/s, respectively. Jet velocity in severe AS was highly skewed with extremely high velocity (as high as 8 m/s) and mainly traveled through the posterior aortic wall up to the aortic arch while still carrying a relatively high velocity, that is, >4 m/s. The mean laminar viscous wall shear stresses (WSS) for mild, moderate, and severe AS were on the order of 40, 100, and 180 Pa, respectively. The RSSmax were on the order of 260, 490, and 2,500 Pa for mild, moderate, and severe AS, respectively. This study may provide a link between altered flows in AS and hemostatic abnormalities such as acquired von Willebrand syndrome and hemolysis, thus, help diagnosing and timing of the treatment.

No abstract