Polydopamine-Induced Multilevel Engineering of Regenerated Silk Fibroin Fiber for Photothermal Conversion.

Solid photothermal materials with favorable biocompatibility and modifiable mechanical properties demonstrate obvious superiority and growing demand. In this work, polydopamine (PDA) induced functionalization of regenerated silk fibroin (RSF) fibers has satisfactory photothermal conversion ability and flexibility. Based on multilevel engineering, RSF solution containing PDA nanoparticles is wet spun to PDA-incorporating RSF (PDA@RSF) fibers, and then the fibers are coated with PDA via oxidative self-polymerization of dopamine to form PDA@RSF-PDA (PRP) fibers. During the wet spinning process, PDA is to adjust the mechanical properties of RSF by affecting its hierarchical structure. Meanwhile, coated PDA gives the PRP fibers extensive absorption of near-infrared light and sunlight, which is further fabricated into PRP fibrous membranes. The temperature of PRP fibrous membranes can be adjusted and increases to about 50 °C within 360 s under 808 nm laser irradiation with a power density of 0.6 W cm-2 , and PRP fibrous membranes exhibit effective photothermal cytotoxicity both in vitro and in vivo. Under the simulated sunlight, the temperature of PRP fiber increases to more than 200 °C from room temperature and the material can generate 4.5 V voltage when assembled with a differential thermal battery, which means that the material also has the potential for flexible wearable electronic devices.

Coupling of Silk Fibroin Nanofibrils Enzymatic Membrane with Ultra-Thin PtNPs/Graphene Film to Acquire Long and Stable On-Skin Sweat Glucose and Lactate Sensing.

The applications of enzymatic biosensors are largely limited by their relatively poor stability and short lifespan. Herein, a bio-active porous enzymatic nanofiber (PEN) membrane composed of silk fibroin nanofibrils (SFNFs) and enzymes is developed to effectively retain the enzymes in the 3D space. The 3D functional scaffolds formed by SFNFs can immobilize enzymes and provide a large surface area for molecular/ion diffusion and biochemical reactions. The PEN membrane is subsequently attached to an ultra-thin PtNPs/graphene (Pt-G) nanocomposite film to facilitate the electron transport between the enzymes and electrodes, permitting highly effective glucose and lactate sensing with long and stable performance. The as-assembled glucose and lactate sensors demonstrate high sensitivity, good cyclic reproducibility, and in particular long-term stability of up to 25 and 23.6 h, respectively. These glucose sensors have a working life that is ≈1.25-times longer than that of the best available sensors reported so far. Moreover, a wearable platform based on the sensors is developed for real-time analysis of sweat during outdoor exercising to transmit signals to a mobile handset. The high sensitivity, comfort and long-term stability of the device can benefit for long-term in-line surveillance of physiological conditions.

Reinforcement of Silk Microneedle Patches for Accurate Transdermal Delivery.

Microneedles (MNs) have attracted considerable attention in the pharmaceutical field as a minimally invasive delivery alternative to hypodermic needles. Current material systems of MNs have gradually shifted from metals, ceramics, and silicon to polymer in consideration of toughness and drug loading capacity. Silk fibroin (SF) is considered one of the most promising alternatives because it combines the ability to maintain the activity of biomolecules, adjustable mechanical strength, and excellent biocompatibility. However, the strength and hardness of SF MNs need to be carefully optimized to ensure skin epidermis penetration and controlled drug release, which are rarely explored in reported works. Here, the synergistic effect of glutaraldehyde-based cross-linking and water vapor annealing post-treatment is presented as an effective method to promote the formation of SF molecular networks and the mechanical strength of SF MNs. Moreover, the reinforced MN substrate is coated with a drug-loaded SF layer with low crystallinity. The drug release experiments demonstrate the successful controlled release of rhodamine B, horseradish peroxidase, and tetracycline, which suggests the great potential in the application of vaccine, antibiosis, cosmetology, and so forth.

Biomimetic Salinity Power Generation Based on Silk Fibroin Ion-Exchange Membranes.

Powering implanted medical devices (IMDs) is a long-term challenge since their use in biological environments requires a long-term and stable supply of power and a biocompatible and biodegradable battery system. Here, silk fibroin-based ion-exchange membranes are developed using bionics principles for reverse electrodialysis devices (REDs). Silk fibroin nanofibril (SNF) membranes are negatively and positively modified, resulting in strong cation and anion selectivity that regulates ion diffusion to generate electric power. These oppositely charged SNF membranes are assembled with Ag/AgCl electrodes into a multicompartment RED. By filling them with 10 and 0.001 mM NaCl solutions, a maximum output power density of 0.59 mW/m2 at an external loading resistance of 66 kΩ is obtained. In addition, 10 pairs of SNF membranes produce a considerable voltage of 1.58 V. This work is a proof of concept that key components of battery systems can be fabricated with protein materials. Combined with the emergence of water-based battery technologies, the findings in this study provide insights for the construction of tissue-integrated batteries for the next generation of IMDs.

Programing Performance of Silk Fibroin Superstrong Scaffolds by Mesoscopic Regulation among Hierarchical Structures.

To design higher-strength natural scaffold materials, wool keratin (WK) rich in α-helix structures is used as a well-defined foreign substrate, which induces the formation of ß-crystallites in silk fibroin (SF). Consequently, the macroscopic properties of silk materials (such as the rheological properties of SF hydrogels and the mechanical properties of stents) can be manipulated by governing the change in the hierarchical mesoscopic structure of silk materials. In this work, by monitoring the structure and morphology in the SF gel process, the mechanism of the effect of keratin on SF network formation was speculated, which was further used to design ultra-high-strength protein scaffolds. It has been confirmed that WK accelerates the gelation of SF by reducing the multistep nucleation barrier and increasing the primary nucleation sites, and then establishing a high-density SF domain network. The modulus of the protein composite scaffold prepared by this facile strategy can reach 11.55 MPa, and the MC-3T3 cells can grow well on the scaffold surface. The results suggest that freeze-dried biocompatible SF-based scaffolds are potential candidates for bone tissue engineering.

Zwitterionic PMCP-Modified Polycaprolactone Surface for Tissue Engineering: Antifouling, Cell Adhesion Promotion, and Osteogenic Differentiation Properties.

Biodegradable polycaprolactone (PCL) has been widely applied as a scaffold material in tissue engineering. However, the PCL surface is hydrophobic and adsorbs nonspecific proteins. Some traditional antifouling modifications using hydrophilic moieties have been successful but inhibit cell adhesion, which is not ideal for tissue engineering. The PCL surface is modified with bioinspired zwitterionic poly[2-(methacryloyloxy)ethyl choline phosphate] (PMCP) via surface-initiated atom transfer radical polymerization to improve cell adhesion through the unique interaction between choline phosphate (CP, on PMCP) and phosphate choline (PC, on cell membranes). The hydrophilicity of the PCL surface is significantly enhanced after surface modification. The PCL-PMCP surface reduces nonspecific protein adsorption (e.g., up to 91.7% for bovine serum albumin) due to the zwitterionic property of PMCP. The adhesion and proliferation of bone marrow mesenchymal stem cells on the modified surface is remarkably improved, and osteogenic differentiation signs are detected, even without adding any osteogenesis-inducing supplements. Moreover, the PCL-PMCP films are more stable at the early stage of degradation. Therefore, the PMCP-functionalized PCL surface promotes cell adhesion and osteogenic differentiation, with an antifouling background, and exhibits great potential in tissue engineering.

Thermoresponsive hydrogels based on a phosphorylated star-shaped copolymer: mimicking the extracellular matrix for in situ bone repair.

A type of four-arm star-shaped copolymer (star-PAA(PEA)-PNIPAM), consisting of the thermoresponsive block (poly(N-isopropylacrylamide), PNIPAM) and the phosphorylated functional block O-phosphoethanolamine (PEA) grafted poly(acrylic acid) (PAA(PEA)), is synthesized by atom transfer radical polymerization (ATRP) and subsequent modification. Owing to the unique superiority of the star-shaped structure, star-PAA(PEA)-PNIPAM can transform from the sol to gel state in response to the physiological temperature (37 °C) at a relatively low polymer concentration (>0.5 wt%). In addition, because of the enriched phosphorylated functional groups, the hydrogel formed by star-PAA(PEA)-PNIPAM can mimic the acidic extracellular matrix protein to adsorb calcium ions and mineralize in situ, both in in vitro and in in vivo experiments. Meanwhile, it is favorable for cell adhesion and proliferation due to its appropriate three-dimensional interspace. Thus, the biocompatible star-PAA(PEA)-PNIPAM hydrogel has great potential for bone repair applications.

Glucose-Responsive Micelles for Controlled Insulin Release Based on Transformation from Amphiphilic to Double Hydrophilic.

Recently, stimuli-responsive carriers have been paid much attention to control cargo release due to their obvious advantages such as targeted delivery, reduced systematic cytotoxicity and enhanced therapeutic efficiency. In this study, a well-defined block copolymer synthesized via ATRP, i.e., poly(ethylene glycol)-b-poly(2-diisopropylaminoethyl methacrylate) (PEG-b-PDPA), has been used to investigate the insulin release behavior in response to glucose changes for potential diabetes mellitus (DM) therapy. Based on the enzymatic catalytic reaction of glucose and glucose oxidase (GOD), the acidic product (gluconic acid) can reduce the micro-environmental pH value. Thereby, the hydrophobic PDPA block with pH sensitivity can rapidly be protonated in response to the decrease of pH value. Due to the partial protonated PDPA block undergoing a variation from hydrophobic to hydrophilic, the self-assembled nanomicelle can gradually release loaded insulin in a regulated model. According to the characterizations of size, morphology, drug loading efficiency, controlled insulin release behavior, glucose sensitivity and cytotoxicity, we conclude that this delicately designed glucose-responsive nanomicelle would be an efficient self-regulated carrier for controlled insulin release for potential DM therapy.

Engineering Porous Poly(lactic acid) Scaffolds with High Mechanical Performance via a Solid State Extrusion/Porogen Leaching Approach.

A knotty issue concerning the poor mechanical properties exists in the porogen leaching approach to porous scaffolds, despite its advantage in tuning pore structure. To address this hurdle, solid state extrusion (SSE) combined with porogen leaching was utilized to engineer porous scaffolds of poly(lactic acid) (PLA). Advances introduced by poly(ethylene glycol) (PEG) caused the PLA ductile to be processed and, on the other hand, enabled the formation of interconnected pores. Thus, a well-interconnected porous architecture with high connectivity exceeding 97% and elevated porosity over 60% was obtained in the as-prepared PLA scaffolds with the composition of NaCl higher than 75.00 wt % and PEG beyond 1.25 wt %. More strikingly, the pore walls of macropores encompassed countless micropores and rough surface topography, in favor of transporting nutrients and metabolites as well as cell attachment. The prominent compressive modulus of the PLA scaffolds was in the range of 85.7⁻207.4 MPa, matching the normal modulus of human trabecular bone (50⁻250 MPa). By means of alkaline modification to improve hydrophilicity, biocompatible porous PLA scaffolds exhibited good cell attachment. These results suggest that the SSE/porogen leaching approach provides an eligible clue for fabricating porous scaffolds with high mechanical performance for use as artificial extracellular matrices.

Choline phosphate functionalized surface: protein-resistant but cell-adhesive zwitterionic surface potential for tissue engineering.

A choline phosphate (CP) modified surface is designed to resist protein adsorption due to its zwitterionic properties and simultaneously promote cell adhesion though its universal interaction with phosphate choline (PC) headgroups of the cell membrane. This work provides a new approach to obtain a cell-adhesive surface with a non-biofouling 'background', which has a potential for tissue engineering.

Tumor-acidity activated surface charge-conversion of polymeric nanocarriers for enhanced cell adhesion and targeted drug release.

The development of stimuli-responsive polymeric nanocarriers could significantly enhance drug bioavailability due to improved pharmacokinetics and biodistribution. However, in the drug delivery process, the poor cell uptake of drug-loaded carriers has greatly limited the therapeutic efficiency for anti-cancer applications. Herein, 2,3-dimethylmaleic anhydride (DMMA) is engineered into the well-defined biodegradable amphiphilic block copolymer poly(D,L-lactide)-block-poly(2-aminoethyl methacrylate) (PLA-b-PAEMA) to construct a tumor-acidity activated nanocarrier (PLA-b-PAEMA/DMMA) for potential tumor therapy. After the loading of positively charged DOX·HCl into the negatively charged corona structure through electrostatic attraction, this carrier is expected to prolong the blood circulation time and smartly convert surface charge from negative to positive for enhanced tumor cell uptake and targeted drug release. Furthermore, this carrier exhibits additional cytotoxicity for tumor cells after the tumor-acidity activated surface charge-conversion from negative to positive. Thus, this smart carrier is a feasible candidate for potential cancer therapy.

Modulated regeneration of acid-etched human tooth enamel by a functionalized dendrimer that is an analog of amelogenin.

In the bioinspired repair process of tooth enamel, it is important to simultaneously mimic the organic-matrix-induced biomineralization and increase the binding strength at the remineralization interface. In this work, a fourth-generation polyamidoamine dendrimer (PAMAM) is modified by dimethyl phosphate to obtain phosphate-terminated dendrimer (PAMAM-PO3H2) since it has a similar dimensional scale and peripheral functionalities to that of amelogenin, which plays important role in the natural development process of enamel. Its phosphate group has stronger affinity for calcium ion than carboxyl group and can simultaneously provide strong hydroxyapatite (HA)-binding capability. The MTT assay demonstrates the low cytotoxicity of PAMAM-PO3H2. Adsorption tests indicate that PAMAM-PO3H2 can be tightly adsorbed on the human tooth enamel. Scanning electron microscopy and X-ray diffraction are used to analyze the remineralization process. After being incubated in artificial saliva for 3weeks, there is a newly generated HA layer of 11.23µm thickness on the acid-etched tooth enamel treated by PAMAM-PO3H2, while the thickness for the carboxyl-terminated one (PAMAM-COOH) is only 6.02µm. PAMAM-PO3H2 can regulate the remineralization process to form ordered new crystals oriented along the Z-axis and produce an enamel prism-like structure that is similar to that of natural tooth enamel. The animal experiment also demonstrates that PAMAM-PO3H2 can induce significant HA regeneration in the oral cavity of rats. Thus PAMAM-PO3H2 shows great potential as a biomimetic restorative material for human tooth enamel.

Phosphorylated dendronized poly(amido amine)s as protein analogues for directing hydroxylapatite biomineralization.

Dendronized poly(amido amine)s (DPs) bearing tri-phosphate or bis-phosphonate peripheral groups are synthesized. These worm-like DPs can template the formation of BMSCs adhesive hydroxylapatite (HA) on the nano-scale, or self-assemble into mineral-collecting microfibers on the micro-scale, exhibiting similar functions of non-collagenous proteins (NCPs) in the natural biomineralization process of HA.

Macroscopic supramolecular assembly of rigid building blocks through a flexible spacing coating.

Macroscopic supramolecular assembly is a promising method for manufacturing macroscopic, ordered structures for tissue-engineering scaffolds. A flexible spacing coating is shown to overcome undesired surface and size effects and to enable assembly of macroscopic cubes with host/guest groups. The assembled pairs disassembled upon introduction of competitive guest molecules, thereby demonstrating a multivalent assembly mechanism.

Triclosan-loaded poly(amido amine) dendrimer for simultaneous treatment and remineralization of human dentine.

In order to treat dental caries of damaged dentine, triclosan-loaded carboxyl-terminated poly(amido amine) dendrimer (PAMAM-COOH) is prepared and characterized. While being incubated in artificial saliva, triclosan-loaded PAMAM-COOH formulation can induce in situ remineralization of hydroxyapatite (HA) on etched dentine, and the regenerated HA has a similar crystal structure with natural dentine. It can also release the encapsulated triclosan for a long period. The interesting drug release profiles are controlled by both dendrimer encapsulation capability and the mineralization degree, which are ideal to obtain multifunctional properties of long-term release of anti-bacterial drug for local treatment during the remineralization process. The triclosan-loaded G4-COOH provides a general strategy to cure dental caries and repair damaged dentine at the same time, which forms a potential restorative material for dental repair.

Thermoresponsive hydrogels from phosphorylated ABA triblock copolymers: a potential scaffold for bone tissue engineering.

A series of thermoresponsive and biocompatible ABA triblock copolymers in which the outer A blocks comprise poly(N-isopropylacrylamide) and the central B block consists of O-phosphoethanolamine (PEA) grafted poly(acrylic acid) (PAA(PEA)) are achieved by atom transfer radical polymerization (ATRP) and subsequent modification. At a relatively low concentration (2 w/v% in phosphate buffered saline), the triblock copolymers can form free-standing gels at 37 °C. Using a combination of variable-temperature 1H NMR, dynamic light scattering, and rheological measurements, it is demonstrated that the gelation behavior is highly dependent on both the length of A blocks and the substitution degree of phosphate group. To examine the potential application as scaffold for bone tissue engineering, the physical gels are incubated in the simulated body fluid (SBF) for 2 weeks. Obvious nucleation and growth of hydroxyapatite are found in the gels, as indicated by the scanning electron microscope, energy dispersive spectroscopy, and X-ray diffraction measurements. The triblock copolymers also exhibit low cytotoxicity in cell viability test. Thus the triblock copolymers have great potential for bone tissue engineering.