Fabrication and Evaluation of Hyaluronidase-Responsive Scaffolds by Electrospinning with Antibacterial Properties for Tympanic Membrane Repair.

Tympanic membrane perforation (TMP) is prevalent in clinical settings. Patients with TMPs often suffer from infections caused by Staphylococcus aureus and Pseudomonas aeruginosa, leading to middle ear and external ear canal infections, which hinder eardrum healing. The objective of this study is to fabricate an enzyme-responsive antibacterial electrospun scaffold using poly(lactic-co-glycolic acid) and hyaluronic acid for the treatment of infected TMPs. The properties of the scaffold were characterized, including morphology, wettability, mechanical properties, degradation properties, antimicrobial properties, and biocompatibility. The results indicated that the fabricated scaffold had a core-shell structure and exhibited excellent mechanical properties, hydrophobicity, degradability, and cytocompatibility. Furthermore, in vitro bacterial tests and ex vivo investigations on eardrum infections suggested that this scaffold possesses hyaluronidase-responsive antibacterial properties. It may rapidly release antibiotics when exposed to the enzyme released by S. aureus and P. aeruginosa. These findings suggest that the scaffold has great potential for repairing TMPs with infections.

An Ion-Mediated Spiking Chemical Neuron based on Mott Memristor.

Artificial spiking neurons capable of interpreting ionic information into electrical spikes are critical to mimic biological signaling systems. Mott memristors are attractive for constructing artificial spiking neurons due to their simple structure, low energy consumption, and rich neural dynamics. However, challenges remain in achieving ion-mediated spiking and biohybrid-interfacing in Mott neurons. Here, a biomimetic spiking chemical neuron (SCN) utilizing an NbOx Mott memristor and oxide field-effect transistor-type chemical sensor is introduced. The SCN exhibits both excitation and inhibition spiking behaviors toward ionic concentrations akin to biological neural systems. It demonstrates spiking responses across physiological and pathological Na+ concentrations (1-200 × 10-3 m). The Na+-mediated SCN enables both frequency encoding and time-to-first-spike coding schemes, illustrating the rich neural dynamics of Mott neuron. In addition, the SCN interfaced with L929 cells facilitates real-time modulation of ion-mediated spiking under both normal and salty cellular microenvironments.

Characteristics of zero-valent iron surface oxide films under the catalytic interface reactions by assisting ligands in nitrate-contaminated groundwater.

The surface passivation layer coating on zero-valent iron (ZVI) particles impedes the electron transfer from ZVI to nitrate. To enhance the efficiency of nitrate reduction by Fe(0), we tested the chemical process and the thickness of the iron oxide film on the surface of Fe(0) particles, utilizing Fe2+aq in aqueous solution and wheat straw as ligands. A novel principal surface catalyzing reaction was formulated as follows: [Formula: see text] . When Fe2+aq concentration increased from 0 - 200 mg·L-1, the NO3- removal rate increased from 6.95% to 82.6% respectively during 12 h and it was 48%, 72%, 79% and 94% respectively in Fe0/WS ratio of 0, 0.25, 0.5 and 1 system. Uniform surface iron oxide films formed around the Fe(0) particles within 12 h after the adding Fe2+aq or wheat straw to the Fe(0) system. The composition and thickness of these films were dependent on the quantity of added materials. X-ray diffraction (XRD) analysis revealed that surface oxide iron mainly consisted of Fe2+ or Fe3+ oxides, with Fe3O4 being predominant. The X-ray photoelectron spectroscopy (XPS) etching indicated that the addition of Fe(0)/straw at mass ratios of 1 or system with 20 mg·L-1 Fe2+aq resulted in the thinnest surface iron oxide layer. The study demonstrated that reducing the oxide layer's thickness was achieved through partial catalysis and enhanced complexation capacity. This reduction was facilitated by the introduction of Fe2+aq or wheat straw into the Fe(0) system, potentially improving proton dissociation and promoting the ligand-assisted dissolution of Fe3+ oxides.

Bioinspired Flexible Hydrogelation with Programmable Properties for Tactile Sensing.

Tactile sensing requires integrated detection platforms with distributed and highly sensitive haptic sensing capabilities along with biocompatibility, aiming to replicate the physiological functions of the human skin and empower industrial robotic and prosthetic wearers to detect tactile information. In this regard, short peptide-based self-assembled hydrogels show promising potential to act as bioinspired supramolecular substrates for developing tactile sensors showing biocompatibility and biodegradability. However, the intrinsic difficulty to modulate the mechanical properties severely restricts their extensive employment. Herein, by controlling the self-assembly of 9-fluorenylmethoxycarbonyl-modifid diphenylalanine (Fmoc-FF) through introduction of polyethylene glycol diacrylate (PEGDA), wider nanoribbons are achieved by untwisting from well-established thinner nanofibers, and the mechanical properties of the supramolecular hydrogels can be enhanced 10-fold, supplying bioinspired supramolecular encapsulating substrate for tactile sensing. Furthermore, by doping with poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) and 9-fluorenylmethoxycarbonyl-modifid 3,4-dihydroxy-l-phenylalanine (Fmoc-DOPA), the Fmoc-FF self-assembled hydrogels can be engineered to be conductive and adhesive, providing bioinspired sensing units and adhesive layer for tactile sensing applications. Therefore, the integration of these modules results in peptide hydrogelation-based tactile sensors, showing high sensitivity and sustainable responses with intrinsic biocompatibility and biodegradability. The findings establish the feasibility of developing programmable peptide self-assembly with adjustable features for tactile sensing applications.

Tuning the Biodegradation Rate of Silk Materials via Embedded Enzymes.

Conventional thinking when designing biodegradable materials and devices is to tune the intrinsic properties and morphological features of the material to regulate their degradation rate, modulating traditional factors such as molecular weight and crystallinity. Since regenerated silk protein can be directly thermoplastically molded to generate robust dense silk plastic-like materials, this approach afforded a new tool to control silk degradation by enabling the mixing of a silk-degrading protease into bulk silk material prior to thermoplastic processing. Here we demonstrate the preparation of these silk-based devices with embedded silk-degrading protease to modulate the degradation based on the internal presence of the enzyme to support silk degradation, as opposed to the traditional surface degradation for silk materials. The degradability of these silk devices with and without embedded protease XIV was assessed both in vitro and in vivo. Ultimately, this new process approach provides direct control of the degradation lifetime of the devices, empowered through internal digestion via water-activated proteases entrained and stabilized during the thermoplastic process.

Rapid UV Photo-Cross-Linking of α-Lactalbumin Hydrogel Biomaterial To Enable Wound Healing.

Although both the function and biocompatibility of protein-based biomaterials are better than those of synthetic materials, their usage as medical material is currently limited by their high costs, low yield, and low batch-to-batch reproducibility. In this article, we show how α-lactalbumin (α-LA), rich in tryptophan, was used to produce a novel type of naturally occurring, protein-based biomaterial suitable for wound dressing. To create a photo-cross-linkable polymer, α-LA was methacrylated at a 100-g batch scale with >95% conversion and 90% yield. α-LAMA was further processed using photo-cross-linking-based advanced processing techniques such as microfluidics and 3D printing to create injectable hydrogels, monodispersed microspheres, and patterned scaffolds. The obtained α-LAMA hydrogels show promising biocompatibility and degradability during in vivo testing. Additionally, the α-LAMA hydrogel can accelerate post-traumatic wound healing and promote new tissue regeneration. In conclusion, cheap and safe α-LAMA-based biomaterials could be produced, and they have a beneficial effect on wound healing. As a result, there may arise a potential partnership between the dairy industry and the development of pharmaceuticals.

Injectable cell-laden silk acid hydrogel.

This study presents an injectable cell-laden hydrogel system based on silk acid, a carboxylated derivative of natural silk fibroin, which exhibits promising applications in biomedicine. The hydrogel is produced under physiological conditions (37 °C and pH 7.4) via physical crosslinking. Notably, the hydrogel demonstrates remarkable cytocompatibility, enabling efficient cell encapsulation, and exhibits good injectability. These promising results strongly indicate the potential of silk acid hydrogel for transformative applications, including 3D cell culture, targeted cell delivery, and tissue engineering.

Achieving Reliable and Ultrafast Memristors via Artificial Filaments in Silk Fibroin.

The practical implementation of memristors in neuromorphic computing and biomimetic sensing suffers from unexpected temporal and spatial variations due to the stochastic formation and rupture of conductive filaments (CFs). Here, the biocompatible silk fibroin (SF) is patterned with an on-demand nanocone array by using thermal scanning probe lithography (t-SPL) to guide and confine the growth of CFs in the silver/SF/gold (Ag/SF/Au) memristor. Benefiting from the high fabrication controllability, cycle-to-cycle (temporal) standard deviation of the set voltage for the structured memristor is significantly reduced by ≈95.5% (from 1.535 to 0.0686 V) and the device-to-device (spatial) standard deviation is also reduced to 0.0648 V. Besides, the statistical relationship between the structural nanocone design and the resultant performance is confirmed, optimizing at the small operation voltage (≈0.5 V) and current (100 nA), ultrafast switching speed (sub-100 ns), large on/off ratio (104 ), and the smallest switching slope (SS < 0.01 mV dec-1 ). Finally, the short-term plasticity and leaky integrated-and-fire behavior are emulated, and a reliable thermal nociceptor system is demonstrated for practical neuromorphic applications.

Silk Acid as an Implantable Biomaterial for Tissue Regeneration.

Silk fibroin derived from the domesticated silkworm Bombyx mori is a protein-based biopolymer with low immunogenicity, intrinsic biodegradability, and tunable mechanical properties, showing great potential in biomedical applications. Using chemical modification to alter the primary structure of silk fibroin enables the expanded generation of new silk-based biomaterials. Inspired by the molecular structure of hyaluronic acid, which is enriched in carboxyl groups, an efficient method with scaling-up potential to achieve controlled carboxylation of silk fibroin to prepare silk acid (SA) is reported, and the biological properties of SA are further studied. The SA materials show tunable hydrophilicity and enzymatic degradation properties at different carboxylation degrees (CDs). Subcutaneous implantation in mice for up to 1 month reveals that the SA materials with a high CD present enhanced degradation while causing a mild foreign-body response, including a low inflammatory response and reduced fibrotic encapsulation. Immunofluorescence analysis further indicates that the SA materials show pro-angiogenesis properties and promote M2-type macrophage polarization to facilitate tissue regeneration. This implies great promise for SA materials as a new implantable biomaterial for tissue regeneration.

White-light crosslinkable milk protein bioadhesive with ultrafast gelation for first-aid wound treatment.

BACKGROUND: Post-traumatic massive hemorrhage demands immediately available first-aid supplies with reduced operation time and good surgical compliance. In-situ crosslinking gels that are flexibly adapting to the wound shape have a promising potential, but it is still hard to achieve fast gelation, on-demand adhesion, and wide feasibility at the same time. METHODS: A white-light crosslinkable natural milk-derived casein hydrogel bioadhesive is presented for the first time. Benefiting from abundant tyrosine residues, casein hydrogel bioadhesive was synthesized by forming di-tyrosine bonds under white light with a ruthenium-based catalyst. We firstly optimized the concentration of proteins and initiators to achieve faster gelation and higher mechanical strength. Then, we examined the degradation, cytotoxicity, tissue adhesion, hemostasis, and wound healing ability of the casein hydrogels to study their potential to be used as bioadhesives. RESULT: Rapid gelation of casein hydrogel is initiated with an outdoor flashlight, a cellphone flashlight, or an endoscopy lamp, which facilitates its usage during first-aid and minimally invasive operations. The rapid gelation enables 3D printing of the casein hydrogel and excellent hemostasis even during liver hemorrhage due to section injury. The covalent binding between casein and tissue enables robust adhesion which can withstand more than 180 mmHg blood pressure. Moreover, the casein-based hydrogel can facilitate post-traumatic wound healing caused by trauma due to its biocompatibility. CONCLUSION: Casein-based bioadhesives developed in this study pave a way for broad and practical application in emergency wound management.

Molecular Design and Preparation of Protein-Based Soft Ionic Conductors with Tunable Properties.

Protein-based soft ionic conductors have attracted considerable research interest in recent years with great potential in applications at the human-machine interfaces. However, a fundamental mechanistic understanding of the ionic conductivity of silk-based ionic conductors is still unclear. Here, we first developed an environmental-friendly and scalable method to fabricate silk-based soft ionic conductors using silk proteins and calcium chloride. The mechanistic understanding of the ion transport and molecular interactions between calcium ions and silk proteins at variable water contents was investigated in-depth by combining experimental and simulation approaches. The results show that calcium ions primarily interact with amide groups in proteins at a low water content. The ionic conductivity is low since the calcium ions are confined around silk proteins within 2.0-2.6 Å. As water content increases, the calcium ions are hydrated with the formation of water shells, leading to the increased distance between calcium ions and silk proteins (3.3-6.0 Å). As a result, the motion of the calcium ions increased to achieve a higher ionic conductivity. By optimizing the ratio of the silk proteins, calcium ions, and water, silk-based soft ionic conductors with good stretchability and self-healing properties can be obtained. Such protein-based soft ionic conductors can be further used to fabricate smart devices such as electrochromic devices.

Supertough and highly stretchable silk protein-based films with controlled biodegradability.

Naturally derived protein-based biopolymers are considered potential biomaterials in biomedical applications and eco-friendly materials for replacing current petroleum-based polymers due to their good biocompatibility, low environmental impact, and tunable degradability. However, current strategies for fabricating protein-based materials with superior properties and tailored functionality in a scalable manner are still lacking. Here, we demonstrate an aqueous-based scalable approach for fabricating silk protein-based films through controlled molecular self-assembly (CMS) of silk proteins with plasticizers and salt ions. The films fabricated using this method can achieve a toughness of up to 64±5 MJ/m3 with a stretchability of up to 574±31%. We also demonstrate the tunable enzymatic degradability, low in vitro cytotoxicity, and good in vivo biocompatibility of the films. Furthermore, the films can be patterned with predesigned complex structures through laser cutting and functionalized with bioactive components. The functional silk protein-based films show great potential in various applications, including flexible electronics, bioelectronics, tissue engineering, and bioplastic packaging. STATEMENT OF SIGNIFICANCE: Inspired by the naturally optimized multi-scale self-assembly of silk proteins in natural silks, we develop an aqueous-based approach for scalable production of superior protein-based films through controlled molecular self-assembly (CMS) of silk proteins with glycerol and calcium ions. The prepared silk films present outstanding mechanical properties, controlled enzymatic biodegradability, low in vitro cytotoxicity, and good in vivo biocompatibility. Notably, the films fabricated using this method can achieve a high toughness of 64±5 MJ/m3 with a stretchability of 594±31%. The approach introduced in this work provides a facile route toward making silk-based materials with superior properties. It also paves new avenues for developing functional protein-based materials with precisely controlled structures and properties for various applications.

Degradable silk-based soft actuators with magnetic responsiveness.

Soft actuators with stimuli-responsiveness have great potential in biomedical applications such as drug delivery and minimally invasive surgery. In this study, protein-based soft actuators with magnetic actuation are fabricated using naturally occurring silk proteins and synthesized Fe3O4 magnetic nanoparticles (NPs). Briefly, magnetic silk films are first prepared by solution casting of a mixture containing silk proteins, synthesized Fe3O4 NPs, and glycerol. The molecular structures of the magnetic silk films are characterized by FTIR spectroscopy, which show that the ß-sheet content in the films is about 20%. The mechanical tests show that the magnetic silk films can be stretched to over 200% under wet conditions and Young's modulus is estimated to be 4.89 ± 0.69 MPa, matching the stiffness of soft tissues. Furthermore, the enzymatic degradability, good biocompatibility, and in vivo X-ray visibility of the films are demonstrated by the in vitro enzymatic degradation test, in vivo biocompatibility test, and micro-CT imaging, respectively. Degradable silk-based soft actuators with magnetic responsiveness are successfully prepared by thermal forming or plastic molding of the magnetic silk films. The fabricated soft actuators can be actuated and move with precise locomotive gaits in solutions using a magnet. In addition, the retention of the soft actuators and localized drug delivery in gastrointestinal tracts by attaching a magnet to the abdominal skin are demonstrated using model systems. The degradable silk-based soft actuators provide many opportunities for improving current therapeutic strategies in biomedicine.

Natural polymer-based scaffolds for soft tissue repair.

Soft tissues such as skin, muscle, and tendon are easily damaged due to injury from physical activity and pathological lesions. For soft tissue repair and regeneration, biomaterials are often used to build scaffolds with appropriate structures and tailored functionalities that can support cell growth and new tissue formation. Among all types of scaffolds, natural polymer-based scaffolds attract much attention due to their excellent biocompatibility and tunable mechanical properties. In this comprehensive mini-review, we summarize recent progress on natural polymer-based scaffolds for soft tissue repair, focusing on clinical translations and materials design. Furthermore, the limitations and challenges, such as unsatisfied mechanical properties and unfavorable biological responses, are discussed to advance the development of novel scaffolds for soft tissue repair and regeneration toward clinical translation.

Scalable Milk-Derived Whey Protein Hydrogel as an Implantable Biomaterial.

There are limited naturally derived protein biomaterials for the available medical implants. High cost, low yield, and batch-to-batch inconsistency, as well as intrinsically differing bioactivity in some of the proteins, make them less beneficial as common implant materials compared to their synthetic counterparts. Here, we present a milk-derived whey protein isolate (WPI) as a new kind of natural protein-based biomaterial for medical implants. The WPI was methacrylated at 100 g bench scale, >95% conversion, and 90% yield to generate a photo-cross-linkable material. WPI-MA was further processed into injectable hydrogels, monodispersed microspheres, and patterned scaffolds with photo-cross-linking-based advanced processing methods including microfluidics and 3D printing. In vivo evaluation of the WPI-MA hydrogels showed promising biocompatibility and degradability. Intramyocardial implantation of injectable WPI-MA hydrogels in a model of myocardial infarction attenuated the pathological changes in the left ventricle. Our results indicate a possible therapeutic value of WPI-based biomaterials and give rise to a potential collaboration between the dairy industry and the production of medical therapeutics.

Processing-Structure-Properties Relationships of Glycerol-Plasticized Silk Films.

Silk possesses excellent mechanical properties and biocompatibility due to its unique protein sequences and hierarchical structures. Thus, it has been widely used as a biomaterial in a broad spectrum of biomedical applications. In this study, an in-depth investigation of glycerol-plasticized silk films was carried out to understand the processing-structure-properties relationships. A series of glycerol-plasticized silk films with glycerol contents in the range of 0 to 30% (w/w) were prepared. The molecular structures and organizations of silk proteins and the interactions between glycerol and proteins were studied using FTIR, XRD, and DSC. At a low glycerol content (<12%), DSC revealed that the glass transition temperature and thermally induced crystallization temperature decreased as the glycerol content increased, implying that glycerol mainly interacts with silk proteins through hydrogen bonding. As the glycerol content further increased, the chain mobility of the silk proteins was promoted, leading to the formation of ß-sheet structures, water insolubility, and increased crystallinity. In addition, the stretchability and toughness of the films were significantly enhanced. The role of glycerol as a plasticizer in regulating the silk protein structures and determining the properties of the films was thoroughly discussed.

Emerging Biopolymer-Based Bioadhesives.

Bioadhesives have been widely used in healthcare and biomedical applications due to their ease-of-operation for wound closure and repair compared to conventional suturing and stapling. However, several challenges remain for developing ideal bioadhesives, such as unsatisfied mechanical properties, non-tunable biodegradability, and limited biological functions. Considering these concerns, naturally derived biopolymers have been considered good candidates for making bioadhesives owing to their ready availability, facile modification, tunable mechanical properties, and desired biocompatibility and biodegradability. Over the past several years, remarkable progress has been made on biopolymer-based adhesives, covering topics from novel materials designs and advanced processing to clinical translation. The developed bioadhesives have been applied for diverse applications, including tissue adhesion, hemostasis, antimicrobial, wound repair/tissue regeneration, and skin-interfaced bioelectronics. Here in this comprehensive review, recent progress on biopolymer-based bioadhesives is summarized with focuses on clinical translations and multifunctional bioadhesives. Furthermore, challenges and opportunities such as weak adhesion strength at the hydrated state, mechanical mismatch with tissues, and unfavorable immune responses are discussed with an aim to facilitate the future development of high-performance biopolymer-based bioadhesives.

Radially Aligned Porous Silk Fibroin Scaffolds as Functional Templates for Engineering Human Biomimetic Hepatic Lobules.

Bioengineering functional hepatic tissue constructs that physiologically replicate the human native liver tissue in vitro is sought for clinical research and drug discovery. However, the intricate architecture and specific biofunctionality possessed by the native liver tissue remain challenging to mimic in vitro. In the present study, a versatile strategy to fabricate lobular-like silk protein scaffolds with radially aligned lamellar sheets, interconnected channels, and a converging central cavity was designed and implemented. A proof-of-concept study to bioengineer biomimetic hepatic lobules was conducted through coculturing human hepatocytes and primary endothelial cells on these lobular-like scaffolds. Relatively long-term viability of hepatocyte/endothelial cells was found along with cell alignment and organization in vitro. The hepatocytes showed special epithelial polarity and bile duct formation, similar to the liver plate, while the aligned endothelial cells generated endothelial networks, similar to natural hepatic sinuses. This endowed the three-dimensional (3D) tissue constructs with the capability to recapitulate hepatic-like parenchymal-mesenchymal growth patterns in vitro. More importantly, the cocultured hepatocytes outperformed monocultures or monolayer cultures, displaying significantly enhanced hepatocyte functions, including functional gene expression, albumin (ALB) secretion, urea synthesis, and metabolic activity. Thus, this functional unit with a biomimetic phenotype provides a novel technology for bioengineering biomimetic hepatic lobules in vitro, with potential utility as a building block for bioartificial liver (BAL) engineering or as a robust tool for drug metabolism investigation.

Geochemical stability of zero-valent iron modified raw wheat straw innovatively applicated to in situ permeable reactive barrier: N2 selectivity and long-term denitrification.

The zero-valent iron (ZVI) modified wheat straw materials are widely used for treating groundwater by permeable reactive barrier (PRB). We report the performance of a field-scale PRB filled with ZVI modified wheat straw materials for nitrate (NO3-)-contaminated groundwater. In lab-scale PRB filled with ZVI modified wheat straw material, NO3- concentration entering the PRB was varied (27.80-59.86 mg L-1) according to the in situ NO3- contamination. A stable NO3- removal rate of 90% was achieved at a controlled hydraulic retention time of 22 days, together with a proportion of denitrifying bacteria up to 34.37%. The field-scale PRB filled with ZVI modified wheat straw material was successful at removing NO3- from groundwater (removal percentages ≥60%) at a groundwater flow rate of 0.01 m3 d-1. Monitoring of groundwater within this PRB provided evidences that the nitrogen gas (N2) selectivity increased with lower ammonia (NH4+) generated from ZVI reduction of NO3-, and few emission of NO2- present due to denitrification capacity in this PRB. The results are finally compared with the few others reported existing PRBs for nitrate-contaminated groundwater worldwide, and demonstrated that the ZVI modified wheat straw material would be an effective fillings for field PRB to remediate groundwater.