Impact of interstitial flow on cartilage matrix synthesis and NF-kB transcription factor mRNA expression in a novel perfusion bioreactor.

This work is focused on designing an easy-to-use novel perfusion system for articular cartilage (AC) tissue engineering and using it to elucidate the mechanism by which interstitial shear upregulates matrix synthesis by articular chondrocytes (AChs). Porous chitosan-agarose (CHAG) scaffolds were synthesized and compared to bulk agarose (AG) scaffolds. Both scaffolds were seeded with osteoarthritic human AChs and cultured in a novel perfusion system with a medium flow velocity of 0.33 mm/s corresponding to 0.4 mPa surfice shear and 40 mPa CHAG interstitial shear. While there were no statistical differences in cell viability for perfusion versus static cultures for either scaffold type, CHAG scaffolds exhibited a 3.3-fold higher (p < 0.005) cell viability compared to AG scaffold cultures. Effects of combined superficial and interstitial perfusion for CHAG showed 150- and 45-fold (p < 0.0001) increases in total collagen (COL) and 13- and 2.2-fold (p < 0.001) increases in glycosaminoglycans (GAGs) over AG non-perfusion and perfusion cultures, respectively, and a 1.5-fold and 3.6-fold (p < 0.005) increase over non-perfusion CHAG cultures. Contrasting CHAG perfusion and static cultures, chondrogenic gene comparisons showed a 3.5-fold increase in collagen type II/type I (COL2A1/COL1A1) mRNA ratio (p < 0.05), and a 1.3-fold increase in aggrecan mRNA. Observed effects are linked to NF-κB signal transduction pathway inhibition as confirmed by a 3.2-fold (p < 0.05) reduction of NF-κB mRNA expression upon exposure to perfusion. Our results demonstrate that pores play a critical role in improving cell viability and that interstitial flow caused by medium perfusion through the porous scaffolds enhances the expression of chondrogenic genes and extracellular matrix through downregulating NF-κB1.

Uniformly valid solution for acoustic propagation in weakly tapered circular waveguides: liquid jet example.

The propagation of ultrasound down laminar liquid jets has potential applications to the stimulation of liquid drop production [J. B. Lonzaga, C. F. Osterhoudt, D. B. Thiessen, and P. L. Marston, J. Acoust. Soc. Am. 121, 3323-3330 (2007)] as well as to the coupling of ultrasound to objects through contact with a jet. In normal gravity, a jet issuing from a nozzle becomes tapered as the jet accelerates downward. A uniformly valid solution for the acoustic propagation in a weakly tapered, liquid jet waveguide in air with a turning point is derived using Langer's transformation and the method of multiple scales. The loss of energy from transmission into the air and from thermal viscous absorption is neglected. A solvability condition is used to obtain the leading-order correction due to the taper of the waveguide. This asymptotic solution is validated using finite-element numerical calculations. The ultrasonic wave amplitude is enhanced in the region of the jet close to the cutoff of the excited mode.

Liquid jet response to internal modulated ultrasonic radiation pressure and stimulated drop production.

Experimental evidence shows that a liquid jet in air is an acoustic waveguide having a cutoff frequency inversely proportional to the jet diameter. Ultrasound applied to the jet supply liquid can propagate within the jet when the acoustic frequency is near to or above the cutoff frequency. Modulated radiation pressure is used to stimulate large amplitude deformations and the breakup of the jet into drops. The jet response to the modulated internal ultrasonic radiation pressure was monitored along the jet using (a) an optical extinction method and (b) images captured by a video camera. The jet profile oscillates at the frequency of the radiation pressure modulation and where the response is small, the amplitude was found to increase in proportion to the square of the acoustic pressure amplitude as previously demonstrated for oscillating drops [P.L. Marston and R.E. Apfel, J. Acoust. Soc. Am. 67, 27-37 (1980)]. Small amplitude deformations initially grow approximately exponentially with axial distance along the jet. Though aspects of the perturbation growth can be approximated from Rayleigh's analysis of the capillary instability, some detailed features of the observed jet response to modulated ultrasound are unexplained neglecting the effects of gravity.

Hysteresis and mode coupling in capillary bridge oscillations: observations.

We investigate nonlinear axisymmetric oscillations of capillary bridges in a Plateau tank of density-matched liquids. The liquids are selected to have unusually small kinematic viscosities. Large amplitude oscillations are excited by applying oscillating Maxwell stresses. The modal frequency response is measured by incrementing the excitation frequency. In a narrow range of frequencies the response depends on the direction (downward or upward) of the increments in a way consistent with a lumped-parameter model of hysteresis for weakly damped oscillators having a mode-softening nonlinearity. The bridge length is selected so that the third harmonic is the natural frequency of a higher-order capillary mode and that mode also exhibits hysteresis.

Manipulation of fluid objects with acoustic radiation pressure.

Conditions are summarized for manipulating and stabilizing fluid objects based on the acoustic radiation pressure of standing waves. Examples include (but are not limited to) liquid drops, gas bubbles in liquids, and cylindrical liquid bridges. The emphasis is on situations where the characteristic wavelength of the acoustic field is large in comparison to the relevant dimension of the fluid object. Tables are presented for ease of comparing the signs of qualitatively different radiation force parameters for a variety of fluid objects.

Active electrostatic control of liquid bridge dynamics and stability.

Stabilization of cylindrical liquid bridges beyond the Rayleigh-Plateau limit has been demonstrated in both Plateau-tank experiments and in short-duration low gravity on NASA KC-135 aircraft using an active electrostatic control method. The method controls the (2,0) capillary mode using an optical modal-amplitude detector and mode-coupled electrostatic feedback stress. The application of mode-coupled stresses to a liquid bridge is also a very useful way to study mode dynamics. A pure (2,0)-mode oscillation can be excited by periodic forcing and then the forcing can be turned off to allow for a free decay from which the frequency and damping of the mode is measured. This can be done in the presence or absence of feedback control. Mode-coupled feedback stress applied in proportion to modal amplitude with appropriate gain leads to stiffening of the mode allowing for stabilization beyond the Rayleigh-Plateau limit. If the opposite sign of gain is applied the mode frequency is reduced. It has also been demonstrated that, by applying feedback in proportion to the modal velocity, the damping of the mode can be increased or decreased depending on the velocity gain. Thus, both the mode frequency and damping can be independently controlled at the same time and this has been demonstrated in Plateau-tank experiments. The International Space Station (ISS) has its own modes of oscillation, some of which are in a low frequency range comparable to the (2,0)-mode frequency of typical liquid bridges. In the event that a vibration mode of the ISS were close to the frequency of a capillary mode it would be possible, with active electrostatic control, to shift the capillary-mode frequency away from that of the disturbance and simultaneously add artificial damping to further reduce the effect of the g-jitter. In principle, this method could be applied to any fluid configuration with a free surface.

Observation of the enhanced backscattering of light by the end of a tilted dielectric cylinder owing to the caustic merging transition.

The scattering of light by obliquely illuminated circular dielectric cylinders was previously demonstrated to be enhanced by a merger of Airy caustics at a critical tilt angle. [Appl. Opt. 37, 1534 (1998)]. A related enhancement is demonstrated here for backward and near-backward scattering for cylinders cut with a flat end perpendicular to the cylinder's axis. It is expected that merged caustics will enhance the backscattering by clouds of randomly oriented circular cylinders that have appropriately flat ends.