A facile method to fabricate versatile keratin cryogels for tissue engineering applications.

Human hair keratin (HHK) has been extensively explored as a biomaterial for soft tissue regeneration due to their excellent bioactivity and biocompatibility. The possibility to fabricate HHK into three-dimensional (3D) hydrogels with physical properties resembling soft tissues has been well demonstrated. However, conventional keratin hydrogels often exhibit a dense architecture that could hinder cell filtration. In the present study, HHK-based cryogels were fabricated using a freeze-thaw (FT) method, where oxidized dopamine (ODA) was employed to covalently crosslink thiol/amine rich-keratin molecules at sub-zero temperatures. The obtained HHK-ODA cryogels have micron-sized pores ranging between 100 and 200 µm and mechanical properties that can be tuned by varying the crosslinking density between ODA and HHK. Through optimization of the weight content of ODA and the number of FT cycles, the compressive strengths and stiffnesses of these cryogels achieved 15-fold increments from ∼1.5 kPa to ∼22 kPa and ∼300 Pa to ∼5000 Pa, respectively. The HHK-ODA cryogels competently supported human dermal fibroblast spreading and proliferation. Overall, this study exhibited a facile method to fabricate mechanically superior keratin-based cryogels with cell compatible microarchitecture, circumventing the need for complicated chemical modifications and the use of cytotoxic crosslinkers.

Ion-Mediated Recombination Dynamics in Perovskite-Based Memory Light-Emitting Diodes for Neuromorphic Control Systems.

Neuromorphic devices can help perform memory-heavy tasks more efficiently due to the co-localization of memory and computing. In biological systems, fast dynamics are necessary for rapid communication, while slow dynamics aid in the amplification of signals over noise and regulatory processes such as adaptation- such dual dynamics are key for neuromorphic control systems. Halide perovskites exhibit much more complex phenomena than conventional semiconductors due to their coupled ionic, electronic, and optical properties which result in modulatable drift, diffusion of ions, carriers, and radiative recombination dynamics. This is exploited to engineer a dual-emitter tandem device with the requisite dual slow-fast dynamics. Here, a perovskite-organic tandem light-emitting diode (LED) capable of modulating its emission spectrum and intensity owing to the ion-mediated recombination zone modulation between the green-emitting quasi-2D perovskite layer and the red-emitting organic layer is introduced. Frequency-dependent response and high dynamic range memory of emission intensity and spectra in a LED are demonstrated. Utilizing the emissive read-out, image contrast enhancement as a neuromorphic pre-processing step to improve pattern recognition capabilities is illustrated. As proof of concept using the device's slow-fast dynamics, an inhibition of the return mechanism is physically emulated.

Influence of Ionic Additives in the PEDOT:PSS Hole Transport Layers for Efficient Blue Perovskite Light Emitting Diodes.

Ruddlesden-Popper (RP) perovskites have been gaining traction in the development of high-efficiency or blue-emitting perovskite light emitting diodes (PeLEDs) due to the unique energy funneling mechanism, which enhances photoluminescence intensity, and dimensional control, which enables spectral tuning. In a conventional p-i-n device structure, the quality of RP perovskite films, including grain morphology and defects, as well as device performance can be significantly influenced by the underlying hole-transport layer (HTL). Poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) is commonly used in several PeLEDs as an HTL because of its high electrical conductivity and optical transparency. Nonetheless, the energy level mismatch and exciton quenching caused by PEDOT:PSS often compromises PeLED performance. Herein, we investigate the mitigation of these effects through addition of work-function-tunable PSS Na to the PEDOT:PSS HTL and assess the impact on blue PeLED performance. Surface analysis of the modified PEDOT:PSS HTLs reveals a PSS-rich layer that alleviates exciton quenching at the HTL/perovskite interface. At an optimal concentration of 6% PSS Na addition, an improvement in the external quantum efficiency is observed, with champion blue and sky-blue PeLEDs achieving 4% (480 nm) and 6.36% (496 nm), respectively, while operation stability is prolonged by fourfold.

Human hair proteins as natural reactive oxygen species scavengers for in vitro applications.

Human hair proteins are recognized for their intrinsically high cysteine content. They can be solubilized while preserving their highly reductive thiol groups for free radical scavenging applications. The presence of aromatic and nucleophilic amino acids such as methionine, serine, phenylalanine, and threonine further contribute to the antioxidative potential of this material. Herein, utilizing the DPPH (2,2-diphenyl-1-picrylhydrazyl) and acellular 2',7'-dichlorodihydrofluorescein diacetate (H2 DCFDA) assays, keratins are demonstrated to possess the highest radical scavenging activity among the studied hair proteins. Consequently, protection against hydrogen peroxide-induced oxidative stress in human dermal fibroblasts (HDFs) cultured in human hair keratin supplemented media is demonstrated. Quenching of reactive oxygen species in the HDF is observed using the CellROX Green dye and the expression levels of antioxidant (HMOX1, SOD2, GPX1) and tumor suppressor (TP53) genes is analyzed using qPCR. Collectively, this study presents further evidence and demonstrates the in vitro application potential of hair proteins, especially keratins, as an antioxidizing supplement.

Anisotropic hair keratin-dopamine composite scaffolds exhibit strain-stiffening properties.

Human hair keratin (HHK) has been successfully explored as raw materials for three-dimensional scaffolds for soft tissue regeneration due to its excellent biocompatibility and bioactivity. However, none of the reported HHK based scaffolds is able to replicate the strain-stiffening capacity of living tissues when responding to large deformations. In the present study, strain-stiffening property was achieved in scaffolds fabricated from HHK via a synergistic effect of well-defined, aligned microstructure and chemical crosslinking. Directed ice-templating method was used to fabricate HHK-based scaffolds with highly aligned (anisotropic) microstructure while oxidized dopamine (ODA) was used to crosslink covalently to HHKs. The resultant HHK-ODA scaffolds exhibited strain-stiffening behavior characterized by the increased gradient of the stress-strain curve after the yield point. Both ultimate tensile strength and the elongation at break were enhanced significantly (~700 kPa, ~170%) in comparison to that of HHK scaffolds lacking of aligned microstructure or ODA crosslinking. In vitro cell culture studies indicated that HHK-ODA scaffolds successfully supported human dermal fibroblasts (HDFs) adhesion, spreading and proliferation. Moreover, anisotropic HHK-ODA scaffolds guided cell growth in alignment with the defined microstructure as shown by the highly organized cytoskeletal networks and nuclei distribution. The findings suggest that HHK-ODA scaffolds, with strain-stiffening properties, biocompatibility and bioactivity, have the potential to be applied as biomimetic matrices for soft tissue regeneration.

Liposome interaction with macrophages and foam cells for atherosclerosis treatment: effects of size, surface charge and lipid composition.

Liposomes are potential drug carriers for atherosclerosis therapy due to low immunogenicity and ease of surface modifications that allow them to have prolonged circulation half-life and specifically target atherosclerotic sites to increase uptake efficiency. However, the effects of their size, charge, and lipid compositions on macrophage and foam cell behaviour are not fully understood. In this study, liposomes of different sizes (60 nm, 100 nm and 180 nm), charges (-40 mV, -20 mV, neutral, +15 mV and +30 mV) and lipid compositions (1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, L-a-phosphatidylcholine, and egg sphingomyelin) were synthesized, characterized and exposed to macrophages and foam cells. Compared to 100 nm neutral 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) liposomes, flow cytometry and confocal imaging indicated that cationic liposomes and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DSPC) liposomes were internalized more by both macrophages and foam cells. Through endocytosis inhibition, phagocytosis and clathrin-mediated endocytosis were identified as the dominant mechanisms of uptake. Anionic and DSPC liposomes induced more cholesterol efflux capacity in foam cells. These results provide a guide for the optimal size, charge, and lipid composition of liposomes as drug carriers for atherosclerosis treatment.

Self-Assembly of Solubilized Human Hair Keratins.

Human hair keratins have proven to be a viable biomaterial for diverse regenerative applications. However, the most significant characteristic of this material, the ability to self-assemble into nanoscale intermediate filaments, has not been exploited. Herein, we successfully demonstrated the induction of hair-extracted keratin self-assembly in vitro to form dense, homogeneous, and continuous nanofibrous networks. These networks remain hydrolytically stable in vitro for up to 5 days in complete cell culture media and are compatible with primary human dermal fibroblasts and keratinocytes. These results enhance the versatility of human hair keratins for applications where structured assembly is of benefit.

Characterization of Anisotropic Human Hair Keratin Scaffolds Fabricated via Directed Ice Templating.

Human hair keratin (HHK) is successfully exploited as raw materials for 3D scaffolds for soft tissue regeneration owing to its excellent biocompatibility and bioactivity. However, most HHK scaffolds are not able to achieve the anisotropic mechanical properties of soft tissues such as tendons and ligaments due to lack of tunable, well-defined microstructures. In this study, directed ice templating method is used to fabricate anisotropic HHK scaffolds that are characterized by aligned pores (channels) in between keratin layers in the longitudinal plane. In contrast, pores in the transverse plane maintain a homogenous rounded morphology. Channel widths throughout the scaffolds range from ≈5 to ≈15 µm and are tunable by varying the freezing temperature. In comparison with HHK scaffolds with random, isotropic pore structures, the tensile strength of anisotropic HHK scaffolds is enhanced significantly by up to fourfolds (≈200 to ≈800 kPa) when the tensile load is applied in the direction parallel to the aligned pores. In vitro results demonstrate that the anisotropic HHK scaffolds are able to support human dermal fibroblast adhesion, spreading, and proliferation. The findings suggest that HHK scaffolds with well-defined, aligned microstructure hold promise as templates for soft tissues regeneration by mimicking their anisotropic properties.

Cryogelation of Human Hair Keratins.

Human hair keratins (HHK) are known for their biocompatibility and potential to regulate cell response, possibly due to the presence of the leucine-aspartic-valine cell adhesion and signaling motifs. Together with the abundance of cysteine residues in HHK, 3D HHK scaffolds are fabricated through cryogelation based on spontaneous disulfide crosslinks and noncovalent interactions. Herein, the molecular mechanism of HHK self-assembly during cryogelation is interrogated and the influence of cryogelation parameters on the properties of the resultant scaffolds is studied. With successive freeze-thaw cycles, the storage modulus (G') of HHK cryogels substantially improves from 116.4 Pa at freeze-thaw cycle 3 (FT3) to 1908.7 Pa at freeze-thaw cycle 10 (FT10). Meanwhile, it is found that complete thiol-capping of HHK samples significantly inhibits cryogel formation as compared to partially or uncapped HHK samples, suggesting the dominant role of disulfide stabilization in cryogelation. Finally, uniaxial compression tests on HHK sponges demonstrate that FT cycling, from 0 to 10, is able to improve the compression modulus of sponges by ≈12-folds. These findings show that macroscale properties of HHK cryogels can be conveniently modulated by physical parameters of cryogelation and that disulfide bonding is the main stabilizing force in HHK cryogels.