In cardiac tissue engineering, the use of bioreactors is fundamental for applying controlled mechanical stimuli on cells and recreate a physiological environment for cardiomyocyte cultures. This work is focused on the design of a sensorized Squeeze Pressure bioreactor (SQPR 3.0) able to apply a periodic contactless hydrodynamic pressure on tissue constructs. This system was then tested with H2c9, a murine cardiomyoblast cell line, to investigate the effect of different stimulation times (2h, 24h, 30h) on cell shape and cardiotypic marker expression.
In this paper, we discuss the basic design requirements for the development of physiologically meaningful in vitro systems comprising cells, scaffolds and bioreactors, through a bottom up approach. Very simple micro- and milli-fluidic geometries are first used to illustrate the concepts, followed by a real device case-study. At each step, the fluidic and mass transport parameters in biological tissue design are considered, starting from basic questions such as the minimum number of cells and cell density required to represent a physiological system and the conditions necessary to ensure an adequate nutrient supply to tissues. At the next level, we consider the use of three-dimensional scaffolds, which are employed both for regenerative medicine applications and for the study of cells in environments which better recapitulate the physiological milieu. Here, the driving need is the rate of oxygen supply which must be maintained at an appropriate level to ensure cell viability throughout the thickness of a scaffold. Scaffold and bioreactor design are both critical in defining the oxygen profile in a cell construct and are considered together. We also discuss the oxygen-shear stress trade-off by considering the levels of mechanical stress required for hepatocytes, which are the limiting cell type in a multi-organ model. Similar considerations are also made for glucose consumption in cell constructs. Finally, the allometric approach for generating multi-tissue systemic models using bioreactors is described.
Permeability studies across epithelial barriers are of primary importance in drug delivery as well as in toxicology. However, traditional in vitro models do not adequately mimic the dynamic environment of physiological barriers. Here, we describe a novel two-chamber modular bioreactor for dynamic in vitro studies of epithelial cells. The fluid dynamic environment of the bioreactor was characterized using computational fluid dynamic models and measurements of pressure gradients for different combinations of flow rates in the apical and basal chambers. Cell culture experiments were then performed with fully differentiated Caco-2 cells as a model of the intestinal epithelium, comparing the effect of media flow applied in the bioreactor with traditional static transwells. The flow increases barrier integrity and tight junction expression of Caco-2 cells with respect to the static controls. Fluorescein permeability increased threefold in the dynamic system, indicating that the stimulus induced by flow increases transport across the barrier, closely mimicking the in vivo situation. The results are of interest for studying the influence of mechanical stimuli on cells, and underline the importance of developing more physiologically relevant in vitro tissue models. The bioreactor can be used to study drug delivery, chemical, or nanomaterial toxicity and to engineer barrier tissues.
Patients affected by motor disorders of the hand and having residual voluntary movements of fingers or wrist can benefit from self-rehabilitation exercises performed with so-called dynamic hand splints. These systems consist of orthoses equipped with elastic cords or springs, which either provide a sustained stretch or resist voluntary movements of fingers or wrist. These simple systems are limited by the impossibility of modulating the mechanical stiffness. This limitation does not allow for customizations and real-time control of the training exercise, which would improve the rehabilitation efficacy. To overcome this limitation, 'active' orthoses equipped with devices that allow for electrical control of the mechanical stiffness are needed. Here, we report on a solution that relies on compact and light-weight electroactive elastic transducers that replace the passive elastic components. We developed a variable-stiffness transducer made of dielectric elastomers, as the most performing types of electromechanically active polymers. The transducer was manufactured with a silicone film and tested with a purposely-developed stiffness control strategy that allowed for electrical modulations of the force-elongation response. Results showed that the proposed new technology is a promising and viable solution to develop electrically controllable dynamic hand orthoses for hand rehabilitation. ? 2013 IPEM.
We describe here a wearable, wireless, compact, and lightweight tactile display, able to mechanically stimulate the fingertip of users, so as to simulate contact with soft bodies in virtual environments. The device was based on dielectric elastomer actuators, as high-performance electromechanically active polymers. The actuator was arranged at the user’s fingertip, integrated within a plastic case, which also hosted a compact high-voltage circuitry. A custom-made wireless control unit was arranged on the forearm and connected to the display via low-voltage leads. We present the structure of the device and a characterization of it, in terms of electromechanical response and stress relaxation. Furthermore, we present results of a psychophysical test aimed at assessing the ability of the system to generate different levels of force that can be perceived by users.
