The development of a substance or inhibitor-based treatment strategy for the prevention of aortic valve stenosis is a challenge and a main focus of medical research in this area. One strategy may be to use the tankyrase inhibitor XAV-939, which leads to Axin stabilisation and subsequent destruction of the β-catenin complex and dephosphorylation of β-catenin. The dephosphorylated active form of β-catenin (non-phospho-β-catenin) then promotes nuclear transcription that leads to osteogenesis. The aims of the present study were to develop an experimental system for inducing in vitro calcification of human aortic valvular interstitial cells (VICs) to investigate the potential anti-calcific effect of XAV-939 and to analyse expression of the Wnt signalling proteins and Sox9, a chondrogenesis regulator, in this model. Calcification of human VIC cultures was induced by cultivation in an osteogenic medium and the effect of co-incubation with 1μM XAV-939 was monitored. Calcification was quantified when mineral deposits were visible in culture and was histologically verified by von Kossa or Alizarin red staining and by IR-spectroscopy. Protein expression of alkaline phosphatase, Axin, β-catenin and Sox9 were quantified by western blotting. In 58% of the VIC preparations, calcification was induced in an osteogenic culture medium and was accompanied by upregulation of alkaline phosphatase. The calcification induction was prevented by the XAV-939 co-treatment and the alkaline phosphatase upregulation was suppressed. As expected, Axin was upregulated, but the levels of active non-phospho-β-catenin were also enhanced. Sox9 was induced during XAV-939 treatment but apparently not as a result of downregulation of β-catenin signalling. XAV-939 was therefore able to prevent calcification of human VIC cultures, and XAV-939 treatment was accompanied by upregulation of active non-phospho-β-catenin. Although XAV-939 does not downregulate active β-catenin, treatment with XAV-939 results in Sox9 upregulation that may prevent the calcification process.

Electroactive polymers (EAPs), especially dielectric elastomer actuators (DEAs), belong to a very promising and emerging class of functional materials. While DEAs are mostly utilized to rely on carbon-based electrodes, there are certain shortcomings of the use of carbon electrodes in the field of soft robotics. In this work we present a fish-like bending structure to serve as possible propulsion element, completely avoiding carbon-based electrodes. The presented robot is moving under water, using a particularly tailored conductive hydrogel as inner electrode and a highly anisotropic textile material to manipulate the bending behavior of the robot. The charge separation to drive two DEAs on the outsides of the robot is provided by the conductive hydrogel while the surrounding water serves as counter electrode. To characterize the hydrogel, tensile tests and impedance spectroscopy are used as measurement methods of choice. The performance of the robot was evaluated using a digital image correlation (DIC) measurement for its bending deflections under water. The developed fish-like robot was able to perform a dynamic bending movement, based on a tri-stable actuator setup. The performed measurements underpin the sufficient characteristics for an underwater application of conductive hydrogel electrodes as well as the applicability of the robotic concept for under water actuations.

Pathological ECM remodelling and biomineralization in human aortic valve and bioprosthesis tissue were investigated by Fourier transformed infrared (FT-IR) spectroscopic imaging and multivariate data analysis. Results of histological von Kossa staining to monitor hydroxyapatite biomineralization correlated to the definition of mineralized tissue using FT-IR spectroscopic imaging. Spectra exhibit signals of carbonate and phosphate groups of hydroxyapatite. Proteins could be identified by the amide I and amide II bands. Proteins were detected in the calcified human aortic valve tissue, but no absorption signals of proteins were observed in the mineralized bioprosthesis sample region. A shift of the amide I band from 1654 cm−1 to 1636 cm−1 was assumed to result from β-sheet structures. This band shift was observed in regions where the mineralization process had been identified but also in non-mineralized bioprosthesis tissue independent of prior implantation. The increased occurrence of β-sheet conformation is hypothesized to be a promoter of the biomineralization process. FT-IR spectroscopic imaging offers a wealth of chemical information. For example, slight variations in band position and intensity allow investigation of heterogeneity across aortic valve tissue sections. The exact evaluation of these properties and correlation with conventional histological staining techniques give insights into aortic valve tissue remodelling and calcific pathogenesis.