Tyrosine-based photoluminescent diketopiperazine supramolecular aggregates.

L-Tyrosine diketopiperazine (DKP) derivative 1 was synthesized, and the aggregation and photoluminescence behaviors were examined. A solution of 1 in tetrahydrofuran (THF) gradually became viscous at room temperature, and turned into the gel state 5 hours after preparation, as confirmed by dynamic viscoelasticity measurement. A solution of 1 in THF exhibited photoluminescence. Fibrous patterns were observed by transmission electron, atomic force and fluorescence microscopies. Dynamic light scattering, semiempirical molecular orbital and density functional theory calculations, as well as molecular dynamics simulations, indicated aggregate formation. This was attributed to intermolecular hydrogen bonding, mainly between the DKP moieties and partly between the urethane moieties, resulting in π-orbital overlap of the terminal phenyl groups leading to photoluminescence.

Three-dimensionally printable shear-thinning triblock copolypeptide hydrogels with antimicrobial potency.

Through rational design, block sequence controlled triblock copolypeptides comprising cysteine and tyrosine as well as a lysine or glutamic acid central block are devised. In these copolypeptides, each block contributes a specific property to the hydrogels to render them extrusion printable and antimicrobial. Three-dimensional (3D) printing of complex hydrogel structures with high shape retention is demonstrated. Moreover, composition dependent potent antimicrobial activity in contact-killing assays is elucidated.

Visualization of molecular binding sites at the nanoscale in the lift-up mode by amplitude-modulation atomic force microscopy.

We report a new approach to visualize the local distribution of molecular recognition sites with nanoscale resolution by amplitude-modulation atomic force microscopy. By integrating chemical modification of probes, photothermal excitation to drive a cantilever, and lift-up scanning over surface topography, we successfully visualized binding sites provided by streptavidin on a solid surface for biotin attached on an AFM probe. The optimization of measurement conditions was discussed in detail, and the application of the technique was verified with a different ligand-receptor system.

Study on Bacterial Antiadhesiveness of Stiffness and Thickness Tunable Cross-Linked Phospholipid Copolymer Thin-Film.

Bacterial adhesion on material surfaces is a significant problem in many areas, especially on medical devices. Upon colonizing a surface, bacteria tend to form biofilms and become difficult to eradicate. A multistep process is involved in bacterial biofilm formation, including primary adhesion to material surface and accumulation of bacterial cells. Controlling the primary adhesion of bacteria is an efficient way to manage biofilms. This study focused on the primary adhesion of bacteria on a copolymer thin-film composed of 2-methacryloyloxyethyl phosphorylcholine (MPC), 3-methacryloxypropyl trimethoxysilane (MPTMSi), and 3-(methacryloyloxy) propyl-tris(trimethylsilyloxy) silane (MPTSSi), which has anti-biofouling and thickness and stiffness tunable properties. We modulated the thickness (5-90 nm) and stiffness of the thin-film via changing the polymer concentration in the coating solution (dip coating). All polymer thin-films inhibited Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa primary adhesions. Interestingly, S. aureus adhesion was affected by the thickness and/or stiffness of the thin-film. We conclude that the mechanical property of the thin-film is one of the influential factors determining bacterial adhesion. These findings would be of significance in designing antibacterial materials.

3D Bioprinting in Skeletal Muscle Tissue Engineering.

Skeletal muscle tissue engineering (SMTE) aims at repairing defective skeletal muscles. Until now, numerous developments are made in SMTE; however, it is still challenging to recapitulate the complexity of muscles with current methods of fabrication. Here, after a brief description of the anatomy of skeletal muscle and a short state-of-the-art on developments made in SMTE with "conventional methods," the use of 3D bioprinting as a new tool for SMTE is in focus. The current bioprinting methods are discussed, and an overview of the bioink formulations and properties used in 3D bioprinting is provided. Finally, different advances made in SMTE by 3D bioprinting are highlighted, and future needs and a short perspective are provided.

A simple layer-stacking technique to generate biomolecular and mechanical gradients in photocrosslinkable hydrogels.

Physicochemical and biological gradients are desirable features for hydrogels to enhance their relevance to biological environments for three-dimensional (3D) cell culture. Therefore, simple and efficient techniques to generate chemical, physical and biological gradients within hydrogels are highly desirable. This work demonstrates a technique to generate biomolecular and mechanical gradients in photocrosslinkable hydrogels by stacking and crosslinking prehydrogel solution in a layer by layer manner. Partial crosslinking of the hydrogel allows mixing of prehydrogel solution with the previous hydrogel layer, which makes a smooth gradient profile, rather than discrete layers. This technique enables the generation of concentration gradients of bovine serum albumin in both gelatin methacryloyl (GelMA) and poly(ethylene glycol) diacrylate hydrogels, as well as mechanical gradients across a hydrogel containing varying gel concentrations. Fluorescence microscopy, mechanical testing, and scanning electron microscopy show that the gradient profiles can be controlled by changing both the volume and concentration of each layer as well as intensity of UV exposure. GelMA hydrogel gradients with different Young's moduli were successfully used to culture human fibroblasts. The fibroblasts migrated along the gradient axis and showed different morphologies. In general, the proposed technique provides a rapid and simple approach to design and fabricate 3D hydrogel gradients for in vitro biological studies and potentially for in vivo tissue engineering applications.

Minimally Invasive and Regenerative Therapeutics.

Advances in biomaterial synthesis and fabrication, stem cell biology, bioimaging, microsurgery procedures, and microscale technologies have made minimally invasive therapeutics a viable tool in regenerative medicine. Therapeutics, herein defined as cells, biomaterials, biomolecules, and their combinations, can be delivered in a minimally invasive way to regenerate different tissues in the body, such as bone, cartilage, pancreas, cardiac, skeletal muscle, liver, skin, and neural tissues. Sophisticated methods of tracking, sensing, and stimulation of therapeutics in vivo using nano-biomaterials and soft bioelectronic devices provide great opportunities to further develop minimally invasive and regenerative therapeutics (MIRET). In general, minimally invasive delivery methods offer high yield with low risk of complications and reduced costs compared to conventional delivery methods. Here, minimally invasive approaches for delivering regenerative therapeutics into the body are reviewed. The use of MIRET to treat different tissues and organs is described. Although some clinical trials have been performed using MIRET, it is hoped that such therapeutics find wider applications to treat patients. Finally, some future perspective and challenges for this emerging field are highlighted.

Advances and Future Perspectives in 4D Bioprinting.

Three-dimensionally printed constructs are static and do not recapitulate the dynamic nature of tissues. Four-dimensional (4D) bioprinting has emerged to include conformational changes in printed structures in a predetermined fashion using stimuli-responsive biomaterials and/or cells. The ability to make such dynamic constructs would enable an individual to fabricate tissue structures that can undergo morphological changes. Furthermore, other fields (bioactuation, biorobotics, and biosensing) will benefit from developments in 4D bioprinting. Here, the authors discuss stimuli-responsive biomaterials as potential bioinks for 4D bioprinting. Natural cell forces can also be incorporated into 4D bioprinted structures. The authors introduce mathematical modeling to predict the transition and final state of 4D printed constructs. Different potential applications of 4D bioprinting are also described. Finally, the authors highlight future perspectives for this emerging technology in biomedicine.

Damage-free tip-enhanced Raman spectroscopy for heat-sensitive materials.

We report a method to establish experimental conditions for tip-enhanced Raman spectroscopy (TERS) with low thermal and mechanical damage to samples. In this method, we monitor the thermal desorption of thiol molecules from a gold-coated probe of an atomic force microscope (AFM) via TERS spectra. Temperatures for desorption of thiol molecules (60-100 °C) from gold surfaces cover the temperature range for degradation of heat-sensitive biomaterials (e.g. proteins). By monitoring the desorption of the thiols on the probe, we can estimate the power of an excitation laser for the samples to reach their critical temperatures for thermal degradation. Furthermore, we also found that an active oscillation of AFM cantilevers significantly promotes the heat transfer from the probe to the surrounding medium. This enables us to employ a higher power density of the excitation laser, resulting in a stronger Raman signal compared with the signal obtained with a contact mode. We propose that this combinatory method is effective in acquiring strong TERS signals while suppressing thermal and mechanical damage to soft and heat-sensitive samples.