Tyrosinase-Modified UHMW SELP Polymers as Wet and Underwater Adhesives to Achieve Multi-interface Adhesion.

The presence of a hydration layer in humid and underwater environments challenges adhesive-substrate interactions and prevents effective bonding, which has become a significant obstacle to the development of adhesives in the industrial and biomedical fields. In this study, ultrahigh-molecular-weight (UHMW) silk-elastin-like proteins (SELP) with 3,4-dihydroxyphenylalanine (DOPA) converted from tyrosine residues by tyrosinase exhibited excellent adhesive properties on different interfaces, such as glass, aluminum, wood, polypropylene sheets, and pigskin, under both dry and wet conditions. Additionally, by incorporating trace amounts of cross-linking agents like Fe3+, NaIO4, and tris(hydroxymethyl) phosphine (THP), the mussel-inspired adhesives maintained a stable and excellent adhesion, broadening the conditions of application. Notably, the UHMW SELP adhesive exhibited remarkable underwater adhesion properties with a shear strength of 0.83 ± 0.17 MPa on glass. It also demonstrated good adhesion to biological tissues including the kidney, liver, heart, and lungs. In vitro cytocompatibility testing using L929 cells showed minimal toxicity, highlighting its potential application in the biomedical field. The sustainable, cytocompatible, cost-effective, and highly efficient adhesive provides valuable insights for the design and development of a new protein-based underwater adhesive for medical application.

Low-Temperature Extrusion of Waterborne Polyurethane-Polycaprolactone Composites for Multi-Material Bioprinting of Engineered Elastic Cartilage.

3D bioprinting of elastic cartilage tissues that are mechanically and structurally comparable to their native counterparts, while exhibiting favorable cellular behavior, is an unmet challenge. A practical solution for this problem is the multi-material bioprinting of thermoplastic polymers and cell-laden hydrogels using multiple nozzles. However, the processing of thermoplastic polymers requires high temperatures, which can damage hydrogel-encapsulated cells. In this study, the authors developed waterborne polyurethane (WPU)-polycaprolactone (PCL) composites to allow multi-material co-printing with cell-laden gelatin methacryloyl (GelMA) hydrogels. These composites can be extruded at low temperatures (50-60 °C) and high speeds, thereby reducing heat/shear damage to the printed hydrogel-capsulated cells. Furthermore, their hydrophilic nature improved the cell behavior in vitro. More importantly, the bioprinted structures exhibited good stiffness and viscoelasticity compared to native elastic cartilage. In summary, this study demonstrated low-temperature multi-material bioprinting of WPU-PCL-based constructs with good mechanical properties, degradation time-frames, and cell viability, showcasing their potential in elastic cartilage bio-fabrication and regeneration to serve broad biomedical applications in the future.

A potential bilayer skin substitute based on electrospun silk-elastin-like protein nanofiber membrane covered with bacterial cellulose.

Skin substitutes are designed to promote wound healing by replacing extracellular matrix. Silk-elastin-like protein is a renewable extracellular matrix-like material that integrated the advantages of silk and elastin-like protein. In this study, electrospun silk-elastin-like protein (SELP) nanofiber membrane covered with bacterial cellulose (BC) was created as a potential skin substitute to mimic gradient structure of epidermis and dermis of skin. The two layers were glued together using adhesive SELP containing 3,4-dihydroxyphenylalanine (DOPA) converted from tyrosine by tyrosinase. Skin topical drugs commonly used in clinical practice can penetrate through the SELP/BC barrier, and the rate of penetration is proportional to drug concentration. BC with dense fibrous structure can act as a barrier to preserve the inner SELP layer and prevent bacterial invasion, with a blocking permeation efficiency over 99% against four species of bacteria. Cell experiments demonstrated that the reticular fibers of SELP could provide an appropriate growth environment for skin cells proliferation and adhesion, which is considered to promote tissue repair and regeneration. The promising results support this strategy to fabricate a silk-elastin-like protein-based biomaterial for skin substitutes in the clinical treatment of full skin injuries and ulcers.

Laccase-mediated formation of hydrogels based on silk-elastin-like protein polymers with ultra-high molecular weight.

As artificial extracellular matrix-like materials, silk-elastin-like protein (SELP) hydrogels, with excellent mechanical properties, high tunability, favorable biocompatibility, and controlled degradability, have become an important candidate in biomedical materials. In this study, SELP is composed of silk-like (GAGAGS) and elastin-like (GXGVP) tandem repeats, in which X residues are set as tyrosine and lysine. Furthermore, SELP polymers are prepared via SpyTag/SpyCatcher. To explore a gentler and more efficient enzymatic crosslinking method, an innovative method was invented to apply laccase to catalyze the formation of SELP hydrogels. Gelation could be successfully achieved in 2-5 min . SELP hydrogels mediated by laccase had the characteristic of low swelling rate, which could maintain a relatively stable shape even when immersed in water, and hence had the potential to be further developed into injectable biomaterials. Additionally, SELP hydrogels cross-linked by laccase showed excellent biocompatibility verified by L929 and HEK 293 T cells with cell viability >93.8 %. SELP hydrogels also exhibit good properties in sustained drug release and cell encapsulation in vitro. This study demonstrates a novel method to construct SELP hydrogels with excellent biocompatibility and expands the possibility of SELP-based material applications in biomedical fields.

Ultra-high molecular weight pullulan-based material with high deformability and shape-memory properties.

Most shape-memory polymers (SMPs) are derived from petroleum feedstocks, which have limitations due to their challenging manufacturing process. Accordingly, herein, a novel SMP based on microbially produced ultrahigh-molecular-weight (UHMW) pullulan was developed. UHMW pullulan cross-linked with 1,4-butanediol diglycidyl ether was wet-spun into fibers with high stretchability (1365 % strain) and excellent shape-memory properties. Furthermore, using three-dimensional (3D) printing, UHMW pullulan-based structures with highly complex shapes (for example, square, cruciform, pentagram and tubular structures), large deformability, and shape memory properties were fabricated. These 3D-printed structures exhibited four-dimensional (4D) programmable deformation under solvent stimulation, enabling the 4D printing of pullulan. The sustainable and eco-friendly approach proposed in this study for the production of pullulan-based SMPs promised to address the current limitations of petroleum-based SMPs.

Polylysine-decorated macroporous microcarriers laden with adipose-derived stem cells promote nerve regeneration in vivo.

Cell transplantation is an effective strategy to improve the repair effect of nerve guide conduits (NGCs). However, problems such as low loading efficiency and cell anoikis undermine the outcomes. Microcarriers are efficient 3D cell culture scaffolds, which can also prevent cell anoikis by providing substrate for adhesion during transplantation. Here, we demonstrate for the first time microcarrier-based cell transplantation in peripheral nerve repair. We first prepared macroporous chitosan microcarriers (CSMCs) by the emulsion-phase separation method, and then decorated the CSMCs with polylysine (pl-CSMCs) to improve cell affinity. We then loaded the pl-CSMCs with adipose-derived stem cells (ADSCs) and injected them into electrospun polycaprolactone/chitosan NGCs to repair rat sciatic nerve defects. The ADSCs-laden pl-CSMCs effectively improved nerve regeneration as demonstrated by evaluation of histology, motor function recovery, electrophysiology, and gastrocnemius recovery. With efficient cell transplantation, convenient operation, and the multiple merits of ADSCs, the ADSCs-laden pl-CSMCs hold good potential in peripheral nerve repair.

Hierarchical macro-microporous WPU-ECM scaffolds combined with Microfracture Promote in Situ Articular Cartilage Regeneration in Rabbits.

Tissue engineering provides a promising avenue for treating cartilage defects. However, great challenges remain in the development of structurally and functionally optimized scaffolds for cartilage repair and regeneration. In this study, decellularized cartilage extracellular matrix (ECM) and waterborne polyurethane (WPU) were employed to construct WPU and WPU-ECM scaffolds by water-based 3D printing using low-temperature deposition manufacturing (LDM) system, which combines rapid deposition manufacturing with phase separation techniques. The scaffolds successfully achieved hierarchical macro-microporous structures. After adding ECM, WPU scaffolds were markedly optimized in terms of porosity, hydrophilia and bioactive components. Moreover, the optimized WPU-ECM scaffolds were found to be more suitable for cell distribution, adhesion, and proliferation than the WPU scaffolds. Most importantly, the WPU-ECM scaffold could facilitate the production of glycosaminoglycan (GAG) and collagen and the upregulation of cartilage-specific genes. These results indicated that the WPU-ECM scaffold with hierarchical macro-microporous structures could recreate a favorable microenvironment for cell adhesion, proliferation, differentiation, and ECM production. In vivo studies further revealed that the hierarchical macro-microporous WPU-ECM scaffold combined with the microfracture procedure successfully regenerated hyaline cartilage in a rabbit model. Six months after implantation, the repaired cartilage showed a similar histological structure and mechanical performance to that of normal cartilage. In conclusion, the hierarchical macro-microporous WPU-ECM scaffold may be a promising candidate for cartilage tissue engineering applications in the future.

Design and characterization of plasticized bacterial cellulose/waterborne polyurethane composite with antibacterial function for nasal stenting.

A nasal stent capable of preventing adhesions and inflammation is of great value in treating nasal diseases. In order to solve the problems of tissue adhesion and inflammation response, we prepared plasticized bacterial cellulose (BCG) and waterborne polyurethane (WPU) composite with antibacterial function used as a novel nasal stent. The gelation behavior of BCG could contribute to protecting the paranasal sinus mucosa; meanwhile, the WPU with improved mechanical property was aimed at supporting the narrow nasal cavity. The thickness, size and the supporting force of the nasal stent could be adjusted according to the specific conditions of the nasal. Thermogravimetric analysis, contact angle and water absorption test were applied to investigate the thermal, hydrophilic and water absorption properties of the composite materials. The composite materials loaded with poly(hexamethylene biguanide) hydrochloride maintained well antibacterial activity over 12 days. Animal experiments further revealed that the mucosal epithelium mucosae damage of BCG-WPU composite was minor compared with that of WPU. This new type of drug-loaded nasal stent can effectively address the postoperative adhesions and infections while ensuring the health of nasal mucosal, and thus has an immense clinical application prospects in treating nasal diseases.

A novel waterborne polyurethane with biodegradability and high flexibility for 3D printing.

Three-dimensional (3D) printing provides a new approach of fabricating implantable products because it permits a flexible manner to extrude complex and customized shapes of the tissue scaffolds. Compared with other printable biomaterials, the polyurethane elastomer has several merits, including excellent mechanical properties and good biocompatibility. However, some intrinsic behavior, especially its high melting point and slow rate of degradation, hampered its application in 3D printed tissue engineering. Herein, we developed a 3D printable amino acid modified biodegradable waterborne polyurethane (WBPU) using a water-based green chemistry process. The flexibility of this material endows better compliance with tissue during implantation and prevents high modulus transplants from scratching surrounding tissues. The histocompatibility experiments show that the WBPU induces no apparent acute rejection or inflammation in vivo. We successfully fabricated a highly flexible WBPU scaffold by deposition 3D printing technology at a low temperature (50°C ~ 70 °C), and the printed products could support the adhesion and proliferation of chondrocytes and fibroblasts. The printed blocks possessed controllable degradability due to the different amounts of hydrophilic chain extender and did not cause accumulation of acidic products. In addition, we demonstrated that our WBPU is highly applicable for implantable tissue engineering because there is no cytotoxicity during its degradation. Taken together, we envision that this printable WBPU can be used as an alternative biomaterial for tissue engineering with low temperature printing, biodegradability, and compatibility.

Preparation of aminoalkyl-grafted bacterial cellulose membranes with improved antimicrobial properties for biomedical applications.

Bacterial cellulose (BC) membranes display special properties and structures, thus attracting much attention in application in the biomedical areas, for example, as implants for bone or cartilage tissue engineering, as substitutes for skin repairing, and as supports for controlled drug delivery. However, native BC lacks the activity to inhibit bacteria growth on its surface, which limits its applications in biomedical fields. There have been reports on chemical modification of BC membranes to endow them with antimicrobial properties needed for some special biomedical applications. In the present study, aminoalkyl-grafted BC membranes were prepared by alkoxysilane polycondensation using 3-aminopropyltriethoxysilane (APTES). The characterization for morphology and chemical composition showed that BC membranes were successfully grafted with aminoalkylsilane groups through covalent bonding. The surface morphology and roughness of the membranes changed after chemical grafting. Furthermore, after grafting with APTES, the membranes got less hydrophilic than native BC. The aminoalkyl-grafted BC membranes showed strong antibacterial properties against Staphylococcus aureus and Escherichia coli and moreover, they were nontoxic to normal human dermal fibroblasts. These results indicate that aminoalkyl-grafted BC membranes are potential to be used for biomedical applications.

A biocompatible bacterial cellulose/tannic acid composite with antibacterial and anti-biofilm activities for biomedical applications.

Biofilm-associated infections are in a high rate of recurrence and biofilms show formidable resistance to current antibiotics, making them a growing challenge in biomedical field. In this study, a biocompatible composite was developed by incorporating tannic acid (TA) and MgCl2 to bacterial cellulose (BC) for antimicrobial and anti-biofilm purposes. The morphology was investigated by scanning electron microscopy (SEM), and chemical structure were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectra (XPS). In vitro release profiles of tannic acid revealed that the Mg2+ cross-links help impede the release of TA from BC matrix, while composite BC-TA lacked Mg2+ ionic cross-links, thus more TA was released from the hydrogel. The BC-TA-Mg composites also displayed strong antibacterial activity against S. aureus, E. coli and P. aeruginosa. Moreover, the composites significantly reduced biofilm formation of S. aureus and P. aeruginosa after 24 h incubation by ∼80% and ∼87%, respectively. As a consequence, the BC-TA-Mg composites are a very promising material for combating biofilm-associated infections in biomedical and public health fields.

PCL-MECM-Based Hydrogel Hybrid Scaffolds and Meniscal Fibrochondrocytes Promote Whole Meniscus Regeneration in a Rabbit Meniscectomy Model.

Regeneration of an injured meniscus continues to be a scientific challenge due to its poor self-healing potential. Tissue engineering provides an avenue for regenerating a severely damaged meniscus. In this study, we first investigated the superiority of five concentrations (0%, 0.5%, 1%, 2%, and 4%) of meniscus extracellular matrix (MECM)-based hydrogel in promoting cell proliferation and the matrix-forming phenotype of meniscal fibrochondrocytes (MFCs). We found that the 2% group strongly enhanced chondrogenic marker mRNA expression and cell proliferation compared to the other groups. Moreover, the 2% group showed the highest glycosaminoglycan (GAG) and collagen production by day 14. We then constructed a hybrid scaffold by 3D printing a wedge-shaped poly(ε-caprolactone) (PCL) scaffold as a backbone, followed by injection with the optimized MECM-based hydrogel (2%), which served as a cell delivery system. The hybrid scaffold (PCL-hydrogel) clearly yielded favorable biomechanical properties close to those of the native meniscus. Finally, PCL scaffold, PCL-hydrogel, and MFCs-loaded hybrid scaffold (PCL-hydrogel-MFCs) were implanted into the knee joints of New Zealand rabbits that underwent total medial meniscectomy. Six months postimplantation we found that the PCL-hydrogel-MFCs group exhibited markedly better gross appearance and cartilage protection than the PCL scaffold and PCL-hydrogel groups. Moreover, the regenerated menisci in the PCL-hydrogel-MFCs group had similar histological structures, biochemical contents, and biomechanical properties as the native menisci in the sham operation group. In conclusion, PCL-MECM-based hydrogel hybrid scaffold seeded with MFCs can successfully promote whole meniscus regeneration, and cell-loaded PCL-MECM-based hydrogel hybrid scaffold may be a promising strategy for meniscus regeneration in the future.

An ultrasound-controllable release system based on waterborne polyurethane/chitosan membrane for implantable enhanced anticancer therapy.

Anti-relapse therapy after surgery plays a critical role in cancer therapy. New strategies maximizing the delivery of drugs to tumor cells while reducing toxic side effects on normal tissues and organs are still urgently required. In order to solve the problems of the poor delivery and inadequate distribution of cytotoxic chemotherapeutic drugs in the clinical application, an ultrasound-controllable and implantable release-system that utilized waterborne polyurethane (WPU) and chitosan (CS) composite membrane as drug carrier with wide flexible loading capacity for doxorubicin (DOX) was described in present work. Benefiting from the hydrophilic segment in WPU and bioactivity of amino groups on side chains of CS, the resulting composite films exhibited fine biodegradability, favorable cytocompatibility and excellent blood compatibility. The in vitro release studies illustrated that the drug-loading membranes displayed a well sustained release effect manifested in slow release, stability and no sudden release, and the DOX was able to release in an ultrasound-controlled manner. Cellular uptake assay and CCK 8 assay showed that the DOX can be released efficiently from the drug-loading matrix and taken up by tumor cells. As a means of adjuvant local treatment, this work provided a facile approach to the design of ultrasound-regulated membrane matrix that is highly beneficial not only due to the higher and long-term therapeutic efficiency, and improvement of utilization efficiency of chemotherapeutic drugs but also the low toxicity to normal cells.

Enhanced Neurite Outgrowth on a Multiblock Conductive Nerve Scaffold with Self-Powered Electrical Stimulation.

Electrical stimulation (ES) is widely applied to promote nerve regeneration. Currently, metal needles are used to exert external ES, which may cause pain and risk of infection. In this work, a multiblock conductive nerve scaffold with self-powered ES by the consumption of glucose and oxygen is prepared. The conductive substrate is prepared by in situ polymerization of polypyrrole (PPy) on the nanofibers of bacterial cellulose (BC). Platinum nanoparticles are electrodeposited on the anode side for glucose oxidation, while nitrogen-doped carbon nanotubes (N-CNTs) are loaded on the cathode side for oxygen reduction. The scaffold shows good mechanical property, flexibility and conductivity. The scaffold can form a potential difference of above 300 mV between the anode and the cathode in PBS with 5 × 10-3 m glucose. Dorsal root ganglions cultured on the Pt-BC/PPy-N-CNTs scaffold are 55% longer in mean neurite length than those cultured on BC/PPy. In addition, in vivo study indicates that the Pt-BC/PPy-N-CNTs scaffold promotes nerve regeneration compared with the BC/PPy group. This paper presents a novel design of a nerve scaffold with self-powered ES. In the future, it can be combined with other features to promote nerve regeneration.

Recyclable Fully Biobased Chitosan Adsorbents Spray-Dried in One Pot to Microscopic Size and Enhanced Adsorption Capacity.

A facile one-pot spray-drying process was developed for fabrication and in situ crosslinking of chitosan microspheres to improve the adsorption capacity by microscopic design. A fully biobased nature was achieved by utilizing genipin (GP) as a crosslinking agent and chitosan-derived nanographene oxide (nGO) as a property tuner. The produced chitosan microspheres were further proven as powerful adsorbents for common wastewater contaminants such as anionic dyes and pharmaceutical contaminants, here modeled by methyl orange (MO) and diclofenac sodium (DCF). By regulating the amount of GP and nGO, as well as by controlling the process parameters including the spray-drying inlet temperature and postheat treatment, the surface morphology, size, zeta potential, and adsorption efficiency of the microspheres could be tuned accordingly. The adsorption efficiency for MO and DCF reached 98.9 and 100%, respectively. The microspheres retained high DCF adsorption efficiency after six adsorption and desorption cycles, and the recyclability was improved by the incorporated nGO. The fabricated microspheres, thus, have great potential as reusable and eco-friendly adsorbents.

Tunable chitosan hydrogels for adsorption: Property control by biobased modifiers.

A sustainable strategy to fabricate chitosan-based composite hydrogels with tunable properties and controllable adsorption capacity of trace pharmaceuticals was demonstrated. Two biobased modifiers were utilized to tune the properties, nano-graphene oxide (nGO) derived from chitosan via microwave-assisted carbonization and oxidation, and genipin as the crosslinking agent. An increase in genipin content facilitated an increase in the degree of crosslinking as shown by improved storage modulus and decreased swelling ratio. Increasing nGO content changed the surface microtopography of the hydrogel which correlated with the surface wettability. nGO also catalyzed the genipin-crosslinking reaction. The hydrogel was further shown to be an effective adsorbent for a common anti-inflammatory drug, diclofenac sodium (DCF), with the removal efficiency ranging from 91 to 100% after 48 h. DCF adsorption efficiency could be tuned through simple alteration of nGO and genipin concentration, which provides promising potential for this environmental-friendly adsorbent in removal of DCF from pharmaceutical waste water.