DNAzymes as Catalysts for l-Tyrosine and Amyloid ß Oxidation.

Single-stranded deoxyribonucleic acids have an enormous potential for catalysis by applying tailored sequences of nucleotides for individual reaction conditions and substrates. If such a sequence is guanine-rich, it may arrange into a three-dimensional structure called G-quadruplex and give rise to a catalytically active DNA molecule, a DNAzyme, upon addition of hemin. Here, we present a DNAzyme-mediated reaction, which is the oxidation of l-tyrosine toward dityrosine by hydrogen peroxide. With an optimal stoichiometry between DNA and hemin of 1:10, we report an activity of 101.2 ± 3.5 µUnits (µU) of the artificial DNAzyme Dz-00 compared to 33.0 ± 1.8 µU of free hemin. Exemplarily, DNAzymes may take part in neurodegeneration caused by amyloid beta (Aß) aggregation due to l-tyrosine oxidation. We show that the natural, human genome-derived DNAzyme In1-sp is able to oxidize Aß peptides with a 4.6% higher yield and a 33.3% higher velocity of the reaction compared to free hemin. As the artificial DNAzyme Dz-00 is even able to catalyze Aß peptide oxidation with a 64.2% higher yield and 337.1% higher velocity, an in-depth screening of human genome-derived DNAzymes may identify further candidates with similarly high catalytic activity in Aß peptide oxidation.

Cytocompatible, Injectable, and Electroconductive Soft Adhesives with Hybrid Covalent/Noncovalent Dynamic Network.

Synthetic conductive biopolymers have gained increasing interest in tissue engineering, as they can provide a chemically defined electroconductive and biomimetic microenvironment for cells. In addition to low cytotoxicity and high biocompatibility, injectability and adhesiveness are important for many biomedical applications but have proven to be very challenging. Recent results show that fascinating material properties can be realized with a bioinspired hybrid network, especially through the synergy between irreversible covalent crosslinking and reversible noncovalent self-assembly. Herein, a polysaccharide-based conductive hydrogel crosslinked through noncovalent and reversible covalent reactions is reported. The hybrid material exhibits rheological properties associated with dynamic networks such as self-healing and stress relaxation. Moreover, through fine-tuning the network dynamics by varying covalent/noncovalent crosslinking content and incorporating electroconductive polymers, the resulting materials exhibit electroconductivity and reliable adhesive strength, at a similar range to that of clinically used fibrin glue. The conductive soft adhesives exhibit high cytocompatibility in 2D/3D cell cultures and can promote myogenic differentiation of myoblast cells. The heparin-containing electroconductive adhesive shows high biocompatibility in immunocompetent mice, both for topical application and as injectable materials. The materials could have utilities in many biomedical applications, especially in the area of cardiovascular diseases and wound dressing.

Reversibly Assembled Electroconductive Hydrogel via a Host-Guest Interaction for 3D Cell Culture.

The study of cells responding to an electroconductive environment is impeded by the lack of a method, which would allow the encapsulation of cells in an extracellular matrix-like 3D electroactive matrix, and more challengingly, permit a simple mechanism to release cells for further characterization. Herein, we report a polysaccharide-based conductive hydrogel system formed via a ß-cyclodextrin-adamantane host-guest interaction. Oxidative polymerization of 3,4-ethylenedioxythiophene (EDOT) in the presence of adamantyl-modified sulfated alginate (S-Alg-Ad) results in bio-electroconductive polymer PEDOT:S-Alg-Ad, which can form hydrogel with poly-ß-cyclodextrin (Pß-CD). The PEDOT:S-Alg-Ad/Pß-CD hydrogels can be tuned on aspects of mechanical and electrical properties, exhibit self-healing feature, and are injectable. Electron microscopy suggested that the difference in stiffness and conductivity is associated with the nacre-like layered nanostructures when different sizes of PEDOT:S-Alg-Ad nanoparticles were used. Myoblast C2C12 cells were encapsulated in the conductive hydrogel and exhibited proliferation rate comparable to that in nonconductive S-Alg-Ad/Pß-CD hydrogel. The cells could be released from the hydrogels by adding the ß-CD monomer. Astonishingly, the conductive hydrogel can dramatically promote myotube-like structure formation, which is not in the non-electroconductive hydrogel. The ability to embed and release cells in an electroconductive environment will open new doors for cell culture and tissue engineering.

Noncovalently Assembled Electroconductive Hydrogel.

Cross-linking biomolecules with electroconductive nanostructures through noncovalent interactions can result in modular networks with defined biological functions and physical properties such as electric conductivity and viscoelasticity. Moreover, the resulting matrices can exhibit interesting features caused by the dynamic assembly process, such as self-healing and molecular ordering. In this paper, we present a physical hydrogel system formed by mixing peptide-polyethylene glycol and poly(3,4-ethylenedioxythiophene):polystyrene sulfonate. This combinatorial approach, which uses different modular building blocks, could lead to high tunability on aspects of rheology and electrical impedance. The proposed physical hydrogel system is characterized by both a self-healing ability and injectability. Interestingly, the formation of hydrogels at relatively low concentrations led to a network of closer molecular packing of poly(3,4-ethylenedioxythiophene) nanoparticles, reflected by the enhanced conductivity. The biopolymer system can be used to develop three-dimensional cell cultures with incorporated electric stimuli, as evidenced by its contribution to the survival and proliferation of encapsulated mesenchymal stromal cells and their differentiation upon electrical stimulation.