Fluorinated Lipid Nanoparticles for Enhancing mRNA Delivery Efficiency.

Lipid nanoparticles (LNPs), a nonviral nucleic acid delivery system, have shown vast potential for vaccine development and disease treatment. LNPs assist mRNA to cross physiological barriers such as cell membranes and endosomes/lysosomes, promoting the intracellular presentation of mRNA. However, the endosome escape efficiency and biosafety of currently commercialized LNPs are still unsatisfactory, resulting in underutilization of mRNA. Herein, we report that fluorinated modification of the 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol)-2000 (PEG-DSPE), termed as FPD, in the LNPs can improve the delivery efficiency of mRNA. FPD accounts for only 1.5% of lipids in LNPs but could mediate a 5-fold and nearly 2-fold enhancement of mRNA expression efficiency in B16F10 tumor cells and primary dendritic cells, respectively. Mechanism studies reveal that FPD promotes the cellular internalization of LNPs as well as endosome escape. In vivo studies substantiate that FPD can augment overall mRNA expression at least 3-fold, either by intravenous or intraperitoneal injection, compared to LNPs prepared with nonfluorinated PEG-lipids at a relatively low mRNA dose. Besides, with the introduction of FPD, mRNA expression in the spleen augmented compared to that of the DMG-PEG commercial formulations. Benefiting from a prudent dosage of fluorine, the fluorinated LNPs display favorable biosafety profiles at cellular and zoological levels.

RCN1 deficiency inhibits oral squamous cell carcinoma progression and THP-1 macrophage M2 polarization.

Reticulocalbin 1 (RCN1), a calcium-binding protein located in the endoplasmic reticulum (ER) lumen, contains six conserved regions. Its main functions include maintaining intracellular homeostasis and regulating cell proliferation and apoptosis, and it plays an important role in the development of various tumours. However, the exact function of RCN1 in oral squamous cell carcinoma (OSCC) is not fully understood. Therefore, the aim of this study was to investigate the effects of RCN1 on the biological behaviour of OSCC and the regulation of tumour-associated macrophage (TAM) polarization. The expression of RCN1 in OSCC and normal oral mucosa was evaluated through bioinformatics analysis and immunohistochemical staining. The growth, migration, and invasion of OSCC cells were observed after knockdown of RCN1 using CCK-8 and Transwell assays. Apoptosis was detected by flow cytometry. The effect of tumour cell-derived RCN1 on the polarization of THP-1 macrophages was investigated by establishing a coculture model of THP-1 macrophages and OSCC cells. Additionally, changes in the expression levels of relevant proteins were detected using Western blotting. The upregulation of RCN1 in tumour tissues compared to normal oral mucosal tissues is associated with a poor prognosis and can be utilized as a prognostic indicator for OSCC. Knockdown of RCN1 inhibited the proliferation, migration, and invasion of OSCC cells. Additionally, knockdown of RCN1 in Cal-27 and SCC-25 cells resulted in inhibition of the M2 polarization of THP-1 macrophages. RCN1 knockdown inhibits OSCC progression and M2 macrophage polarization. Targeting RCN1 may be a promising approach for OSCC treatment.

Ultrarapid-Acting Microneedles for Immediate Delivery of Biotherapeutics.

Subcutaneous (SC) injection is a common administration route for rapid and efficient delivery of biotherapeutics. However, syringe-based injections usually require professional assistance and are associated with pain and potential risks of infections, thus leading to undesired patient compliance and poor life quality. Herein, this work presents an ultrarapid-acting microneedle (URA-MN) patch for immediate transdermal delivery of therapeutics in a minimally invasive manner. Effervescent agents are incorporated into the tip of URA-MN for rapid generation of CO2 bubbles upon insertion into the skin, immediately powering the biotherapeutics release within a few minutes. The release kinetics of diverse agents including liraglutide (LRT), insulin, and heparin from the URA-MN patches are evaluated in three different mouse models, and the rapid release of biotherapeutics and potent therapeutic effects are achieved with only 5 min administration. Noteworthily, attributed to the short application duration and negligible residuals of MN matrix remaining in the skin, the URA-MN patch shows desirable biocompatibility after six-week administration.

TGF-ßRII/EP300/SMAD4 cascade signaling pathway promotes invasion and glycolysis in oral squamous cell carcinoma.

INTRODUCTION: EP300 is considered to be a cancer suppressor gene that plays a role in tumor development, but some studies have reported that it is not an oral squamous cell carcinoma suppressor gene, because there was neither epigenetic inactivation of the gene nor a mutation resulting in functional impairment. However, there is no relevant study on whether EP300 is the exact carcinogenic effect and its mechanisms of carcinogenic effects in oral squamous cell carcinoma. METHODS: Western blot analysis and quantitative real time polymerase chain reaction experiments verified the protein and mRNA expression of EP300 in oral squamous cell carcinoma; The effects of EP300 knockout on glucose consumption and lactic acid production were detected by glycolysis experiments; The relationship between pathway-related proteins and EP300 was verified by bioinformatics analysis and co-immunoprecipitation experiment. RESULTS: Our experimental results confirm that the protein and mRNA of EP300 are highly expressed in oral squamous cell carcinoma, and after knocking out the EP300, the glycolysis ability, invasion, migration, and other biological functions of oral squamous cell carcinoma, are inhibited at the same time. Pathway-related experiments have confirmed that EP300 plays a role in promoting cancer through the transforming growth factor-beta receptor II (TGF-ßRII)/EP300/Smad4 cascade pathway. CONCLUSION: EP300 plays a carcinogenic role in OSCC showed that the TGF-ßRII/EP300/Smad4 cascade pathway is involved in oral squamous cell carcinoma.

Bioresponsive Immunotherapeutic Materials.

The human immune system is an interaction network of biological processes, and its dysfunction is closely associated with a wide array of diseases, such as cancer, infectious diseases, tissue damage, and autoimmune diseases. Manipulation of the immune response network in a desired and controlled fashion has been regarded as a promising strategy for maximizing immunotherapeutic efficacy and minimizing side effects. Integration of "smart" bioresponsive materials with immunoactive agents including small molecules, biomacromolecules, and cells can achieve on-demand release of agents at targeted sites to reduce overdose-related toxicity and alleviate off-target effects. This review highlights the design principles of bioresponsive immunotherapeutic materials and discusses the critical roles of controlled release of immunoactive agents from bioresponsive materials in recruiting, housing, and manipulating immune cells for evoking desired immune responses. Challenges and future directions from the perspective of clinical translation are also discussed.

Sustainable Bioplastic Made from Biomass DNA and Ionomers.

Plastics play important roles in modern life and currently the development of plastic recycling is highly demanding and challenging. To relieve this dilemma, one option is to develop new sustainable bioplastics that are compatible with the environment over the whole material life cycle. We report a sustainable bioplastic made from natural DNA and biomass-derived ionomers, termed as DNA plastics. The sustainability involves all aspects of the production, use, and end-of-life options of DNA plastics: (1) the raw materials are derived from biorenewable resources; (2) the water-processable strategy is environmentally friendly, not involving high-energy consumption, the use of organic solvents, and the production of byproducts; (3) recyclable and nondestructive use is achieved to significantly prolong the service lifetime of the plastics; and (4) the disposal of waste plastics follows two green routes including the recycling of waste plastics and enzyme-triggered controllable degradation under mild conditions. Besides, DNA plastics can be "aqua-welded" to form arbitrary designed products such as a plastic cup. This work provides a solution to transform biobased hydrogel to bioplastic and demonstrates the closed-loop recycling of DNA plastics, which will advance the development of sustainable materials.

Controllable assembly/disassembly of polyphenol-DNA nanocomplex for cascade-responsive drug release in cancer cells.

Developing nanocarrier systems with sufficient drug loading ability and efficient drug release behavior in cells is a powerful strategy to maximize therapeutic efficacies and minimize side effects of administered drugs. However, the two aspects are usually contradictory in a single nanocarrier. Herein, polyphenol-DNA nanocomplex with controllable assembly/disassembly behaviors is developed for responsive and sequential drug release in cancer cells. Programmable assembly of branched-DNA achieves multiple-gene loading, afterwards tannic acid (TA), plant-derived polyphenols as drugs mediate assembly of branched-DNA to form nanocomplex. Intracellularly, two-step disassembly process of nanocomplex enables efficient gene/drug release. Lysosomal acidic microenvironment induces the disassembly of nanocomplex to release TA and branched-DNA. Glutathione and DNase I in cytoplasm trigger the precise release of genes from branched-DNA. The efficacy of multiple-gene/chemo-therapy is demonstrated using in vitro and in vivo models. This work provides a controllable assembly/disassembly route to resolve the conflict between sufficient drug loading and efficient drug release in cells for therapeutics.

Super-Soft DNA/Dopamine-Grafted-Dextran Hydrogel as Dynamic Wire for Electric Circuits Switched by a Microbial Metabolism Process.

Engineering dynamic systems or materials to respond to biological process is one of the major tasks in synthetic biology and will enable wide promising applications, such as robotics and smart medicine. Herein, a super-soft and dynamic DNA/dopamine-grafted-dextran hydrogel, which shows super-fast volume-responsiveness with high sensitivity upon solvents with different polarities and enables creation of electric circuits in response to microbial metabolism is reported. Synergic permanent and dynamic double networks are integrated in this hydrogel. A serials of dynamic hydrogel-based electric circuits are fabricated: 1) triggered by using water as switch, 2) triggered by using water and petroleum ether as switch pair, 3) a self-healing electric circuit; 4) remarkably, a microbial metabolism process which produces ethanol triggering electric circuit is achieved successfully. It is envisioned that the work provides a new strategy for the construction of dynamic materials, particularly DNA-based biomaterials; and the electric circuits will be highly promising in applications, such as soft robotics and intelligent systems.

Encapsulating Microorganisms inside Electrospun Microfibers as a Living Material Enables Room-Temperature Storage of Microorganisms.

Room-temperature storage and transportation of microorganisms maximize the power of microorganisms in healthcare, energy, and environment. Recently, paper-based biotechnologies have been developed to enable room-temperature storage of a variety of nonliving biosystems such as diagnostic devices and cell-free systems. Herein, room-temperature storage of living microorganisms is realized by an electrospun nonwoven paper containing convex region, which is composed of coiled microfibers with dense distribution of microorganisms. Microorganisms are encapsulated into the microfibers and remain intact after electrospinning. Poly(ethylene oxide) is used as polymer matrix, and glycerol and dextran are used as additives. When the contents of glycerol and dextran are optimized as 5 and 0.4%, the room-temperature time is prolonged to 2 days, more than 8 folds as compared with the control group. Upon demand, the microorganisms can be activated by adding water and used for culturing microorganisms directly. Furthermore, mechanisms which account for microbial activity and storage are studied. Our microfiber-based strategy is universal for the room-temperature storage of prokaryotic and eukaryotic microorganisms in the solid formulation. Besides, our microorganism/polymer complex structures represent novel living materials via a bottom-up strategy, which are of great potential for new biomedical applications.

Comprehensively durable superhydrophobic metallic hierarchical surfaces via tunable micro-cone design to protect functional nanostructures.

Superhydrophobic surfaces have been intensively investigated in recent years. However, their durability remains a major challenge before superhydrophobic surfaces can be employed in practice. Although various works have focused on overcoming this bottleneck, no single surface has ever been able to achieve the comprehensive durability (including tangential abrasion durability, dynamic impact durability and adhesive durability) required by stringent industrial requirements. Within the hierarchical structures developed for superhydrophobicity in typical plants or animals by natural evolution, microstructures usually provide mechanical stability, strength and flexibility to protect functional nanostructures to enable high durability. However, this mechanism for achieving high durability is rarely studied or reported. We employed an ultrafast laser to fabricate micro/nanohierarchical structures on metal surfaces with tunable micro-cones and produced abundant nanostructures. We then systematically investigated their comprehensive mechanical durability by fully utilizing the protective effect of the microstructures on the functional nanostructures via the tunable design of micro-cones. We confirm that the height and spatial period of the microstructures were crucial for the tangential abrasion durability and dynamic impact durability, respectively. We finally fabricated optimized superhydrophobic tungsten hierarchical surfaces, which could withstand 70 abrasion cycles, 28 min of solid particle impact or 500 tape peeling cycles to retain contact angles of greater than 150° and sliding angles of less than 20°, which demonstrated exceptional comprehensive durability. The comprehensive durability, in particular the dynamic impact durability and adhesive durability, are among the best published results. This research clarifies the mechanism whereby the microstructures effectively protected the functional nanostructures to achieve high durability of the superhydrophobic surfaces and is promising for improving the durability of superhydrophobic surfaces and thus for practical applications.

General Strategy toward Dual-Scale-Controlled Metallic Micro-Nano Hybrid Structures with Ultralow Reflectance.

Functional metal surfaces with minimum optical reflection over a broadband spectrum have essential importance for optical and optoelectronic devices. However, the intrinsically large optical impedance mismatch between metals and the free space causes a huge obstacle in achieving such a purpose. We propose and experimentally demonstrate a general pulse injection controlled ultrafast laser direct writing strategy for fabricating highly effective antireflection structures on metal surfaces. The presented strategy can implement separate and flexible modifications on both microscale frame structures and nanoscale particles, a benefit from which is that optimized geometrical light trapping and enhanced effective medium effect reducing the surface reflection can be simultaneously achieved within one hybrid structure. Thus, comprehensively improved antireflection performances can be realized. Hybrid structures with substantial nanoparticles hierarchically attached on regularly arrayed microcones are generally constructed on different metal surfaces, achieving highly efficient light absorption over ultraviolet to near-infrared broadband spectrum regions. Reflectance minimums of 1.4%, 0.29%, and 2.5% are reached on Cu, Ti, and W surfaces, respectively. The presented strategy is simple in process, adaptable for different kinds of metals, reproduceable in dual-scale structural features, and feasible for large-area production. All these advantages make the strategy as well as the prepared antireflection structures excellent candidates for practical applications.

Large-Scale Tunable 3D Self-Supporting WO3 Micro-Nano Architectures as Direct Photoanodes for Efficient Photoelectrochemical Water Splitting.

Hydrogen production from water based on photoelectrochemical (PEC) reactions is feasible to solve the urgent energy crisis. Herein, hierarchical 3D self-supporting WO3 micro-nano architectures in situ grown on W plates are successfully fabricated via ultrafast laser processing hybrid with thermal oxidation. Owing to the large surface area and efficient interface charge transfer, the W plate with hierarchical porous WO3 nanoparticle aggregates has been directly employed as the photoanode for excellent PEC performance, which exhibits a high photocurrent density of 1.2 mA cm-2 at 1.0 V vs Ag/AgCl (1.23 V vs RHE) under AM 1.5 G illumination and reveals excellent structural stability during long-term PEC water splitting reactions. The nanoscale and microscale features can be facilely tuned by controlling the laser processing parameters and the thermal oxidation conditions to achieve improved PEC activity. The presented hybrid method is simple, cost-effective, and controllable for large-scale fabrication, which should provide a new and general route that how the properties of conventional metal oxides can be improved via hierarchical 3D micro-nano configurations.

Precise Control of the Number of Layers of Graphene by Picosecond Laser Thinning.

The properties of graphene can vary as a function of the number of layers (NOL). Controlling the NOL in large area graphene is still challenging. In this work, we demonstrate a picosecond (ps) laser thinning removal of graphene layers from multi-layered graphene to obtain desired NOL when appropriate pulse threshold energy is adopted. The thinning process is conducted in atmosphere without any coating and it is applicable for graphene films on arbitrary substrates. This method provides many advantages such as one-step process, non-contact operation, substrate and environment-friendly, and patternable, which will enable its potential applications in the manufacturing of graphene-based electronic devices.

Porous nitrogen-doped carbon microspheres derived from microporous polymeric organic frameworks for high performance electric double-layer capacitors.

This research presents a simple and efficient method to synthesize porous nitrogen-doped carbon microspheres (PNCM) by the carbonization of microporous poly(terephthalaldehyde-pyrrole) organic frameworks (PtpOF). The common KOH activation process is used to tune the porous texture of the PNCM and produce an activated-PNCM (A-PNCM). The PNCM and A-PNCM with specific surface area of 921 and 1303 m(2) g(-1) , respectively, are demonstrated as promising candidates for EDLCs. At a current density of 0.5 A g(-1) , the specific capacitances of the PNCM and A-PNCM are 248 and 282 F g(-1) , respectively. At the relatively high current density of 20 A g(-1) , the capacitance remaining is 95 and 154 F g(-1) , respectively. Capacity retention of the A-PNCM is more than 92% after 10000 charge/discharge cycles at a current density of 2 A g(-1) .

Anti-Recombinant Gametocyte 56 Protein IgY Protected Chickens from Homologous Coccidian Infection.

Coccidiosis is caused by intra-cellular infection of Eimeria spp., which goes through a complex life cycle in the intestinal mucosa of infected hosts. Specific immunoglobulins (IgY) could be produced in egg yolk by immunizing hens with specific antigens. In the present study, we cloned the E. maxima gam56 gene, expressed the GST-GAM56 fusion protein and raised IgY to GST-GAM56 in hens. The anti-GST-GAM56 IgY antibody was isolated and used to treat chickens infected with E. maxima oocysts. Intramuscular injection of the antibodies provided minimal protection against parasite infection. However, oral dosing of the IgY 3 or 5 d after oocyst inoculation significantly improved body weight gain, reduced oocyst output and intestinal lesion score were reduced at 3 or 5 d after oocyst challenging, compared to the untreated control group. Our findings suggest that the IgY to gam56 could be an effective prophylactic or therapeutic agent against E. maxima infection in chickens and should have a practical application value.