Warp-Knitted Spacer Fabrics: A Versatile Platform to Generate Fiber-Reinforced Hydrogels for 3D Tissue Engineering.

Mesenchymal stem cells (MSCs) possess huge potential for regenerative medicine. For tissue engineering approaches, scaffolds and hydrogels are routinely used as extracellular matrix (ECM) carriers. The present study investigated the feasibility of using textile-reinforced hydrogels with adjustable porosity and elasticity as a versatile platform for soft tissue engineering. A warp-knitted poly (ethylene terephthalate) (PET) scaffold was developed and characterized with respect to morphology, porosity, and mechanics. The textile carrier was infiltrated with hydrogels and cells resulting in a fiber-reinforced matrix with adjustable biological as well as mechanical cues. Finally, the potential of this platform technology for regenerative medicine was tested on the example of fat tissue engineering. MSCs were seeded on the construct and exposed to adipogenic differentiation medium. Cell invasion was detected by two-photon microscopy, proliferation was measured by the PrestoBlue assay. Successful adipogenesis was demonstrated using Oil Red O staining as well as measurement of secreted adipokines. In conclusion, the given microenvironment featured optimal mechanical as well as biological properties for proliferation and differentiation of MSCs. Besides fat tissue, the textile-reinforced hydrogel system with adjustable mechanics could be a promising platform for future fabrication of versatile soft tissues, such as cartilage, tendon, or muscle.

Identification of aurintricarboxylic acid as a potent allosteric antagonist of P2X1 and P2X3 receptors.

The homotrimeric P2X3 receptor, one of the seven members of the ATP-gated P2X receptor family, plays a crucial role in sensory neurotransmission. P2X3 receptor antagonists have been identified as promising drugs to treat chronic cough and are suggested to offer pain relief in chronic pain such as neuropathic pain. Here, we analysed whether compounds affect P2X3 receptor activity by high-throughput screening of the Spectrum Collection of 2000 approved drugs, natural products and bioactive substances. We identified aurintricarboxylic acid (ATA) as a nanomolar-potency antagonist of P2X3 receptor-mediated responses. Two-electrode voltage clamp electrophysiology-based concentration-response analysis and selectivity profiling revealed that ATA strongly inhibits the rP2X1 and rP2X3 receptors (with IC50 values of 8.6 nM and 72.9 nM, respectively) and more weakly inhibits P2X2/3, P2X2, P2X4 or P2X7 receptors (IC50 values of 0.76 µM, 22 µM, 763 µM or 118 µM, respectively). Patch-clamp analysis of mouse DRG neurons revealed that ATA inhibited native P2X3 and P2X2/3 receptors to a similar extent than rat P2X3 and P2X2/3 receptors expressed in Xenopus oocytes. In a radioligand binding assay, up to 30 µM ATA did not compete with [3H]-ATP for rP2X3 receptor binding, indicating a non-competitive mechanism of action. Molecular docking studies, site-directed mutagenesis and concentration-response analysis revealed that ATA binds to the negative allosteric site of the hP2X3 receptor. In summary, ATA as a drug-like pharmacological tool compound is a nanomolar-potency, allosteric antagonist with selectivity towards αß-methylene-ATP-sensitive P2X1 and P2X3 receptors.