Variations in mechanical properties of the extracellular matrix occurs in various processes, such as tissue fibrosis. The impact of changes in tissue stiffness on cell behaviour are studied in vitro using various types of biomaterials and methods. Stiffness patterning of hydrogel scaffolds, through the use of stiffness gradients for instance, allows the modelling and studying of cellular responses to fibrotic mechanisms. Gelatine methacryloyl (GelMA) has been used extensively in tissue engineering for its inherent biocompatibility and the ability to precisely tune its mechanical properties. Visible light is now increasingly employed for crosslinking GelMA hydrogels as it enables improved cell survival when performing cell encapsulation. We report here, the photopatterning of mechanical properties of GelMA hydrogels with visible light and eosin Y as the photoinitiator using physical photomasks and projection with a digital micromirror device. Using both methods, binary hydrogels with areas of different stiffnesses and hydrogels with stiffness gradients were fabricated. Their mechanical properties were characterised using force indentation with atomic force microscopy, which showed the efficiency of both methods to spatially pattern the elastic modulus of GelMA according to the photomask or the projected pattern. Crosslinking through projection was also used to build constructs with complex shapes. Overall, this work shows the feasibility of patterning the stiffness of GelMA scaffolds, in the range from healthy to pathological stiffness, with visible light. Consequently, this method could be used to build in vitro models of healthy and fibrotic tissue and study the cellular behaviours involved at the interface between the two.
An integrated instrument is being developed to study live cardiac trabeculae, which is capable of stimulating the muscle under controlled conditions while measuring the heat production, force, and sarcomere length distribution. To improve the accuracy of estimation of stress, strain, and volumetric heat production, the geometry of the muscle must be known. A spectral domain optical coherence tomography system (SD-OCT) has been constructed and calibrated to image the trabecula mounted inside the instrument. This system was mounted above the muscle chamber and a series of equally-spaced cross-sectional images were obtained. These were processed using a workflow developed to extract cross-sectional area and volume. The initial results have demonstrated the feasibility of using OCT to capture the overall geometry of cardiac trabecula mounted in the instrument. Further work will be directed to improve the image quality for larger samples and apply meshing algorithms to the acquired data.
. Significance: A non-destructive technique for accurately characterizing the spatial distribution of optical properties of soft tissue membranes may give improved outcomes in many tissue engineering applications. Aim: This study aimed to develop a non-destructive macroscopic imaging technique that is sensitive to optical anisotropy, typical of fibrous components in soft tissue membranes, and can address some of the difficulties caused by the complex turbid nature of these tissues. Approach: A near-infrared Mueller matrix imaging polarimeter employing logarithm decomposition was developed and used to conduct transmission measurements of all the polarization properties across the full thickness of bovine pericardium tissue. Results: The full Mueller matrix was measured across a sample of calf bovine pericardium and revealed significant retardance (linear and circular) and depolarization in this tissue. Regions with a uniform axis of optical anisotropy were identified. Mueller matrix imaging demonstrated that the exhibited circular retardance was sufficient to lead to possible misinterpretation of apparent fiber orientation when using conventional polarization imaging techniques for such tissues. Conclusions: Mueller matrix imaging can identify regional distributions of optical anisotropy in calf bovine pericardium. This new capability is a promising development in non-destructive imaging for tissue selection.
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