Overcoming Endosomal Escape Barriers in Gene Drug Delivery Using De Novo Designed pH-Responsive Peptides.

A major challenge in using nanocarriers for intracellular drug delivery is their restricted capacity to escape from endosomes into the cytosol. Here, we significantly enhance the drug delivery efficiency by accurately predicting and regulating the transition pH (pH0) of peptides to modulate their endosomal escape capability. Moreover, by inverting the chirality of the peptide carriers, we could further enhance their ability to deliver nucleic acid drugs as well as antitumor drugs. The resulting peptide carriers exhibit versatility in transfecting various cell types with a high efficiency of up to 90% by using siRNA, pDNA, and mRNA. In vivo antitumor experiments demonstrate a tumor growth inhibition of 83.4% using the peptide. This research offers a potent method for the rapid development of peptide vectors with exceptional transfection efficiencies for diverse pathophysiological indications.

Rational design of antibodies and development of a novel method for (1-3)-ß-D glucan detection as an alternative to Limulus amebocyte lysate assay.

With advances in medicine, increasing medical interventions have increased the risk of invasive fungal disease development. (1-3)-ß-D glucan (BDG) is a common fungal biomarker in serological tests. However, the scarcity of Limulus resources for BDG detection poses a challenge. This study addresses the need for an alternative to Limulus amebocyte lysate by using BDG mutant antibody for chemiluminescence detection. The wild-type BDG antibody was obtained by immunizing rabbits. An optimal V52HI/N34L Y mutant antibody, which has increased 3.7-fold of the testing efficiency compared to the wild-type antibody, was first achieved by mutating "hot-spot" residues that contribute to strong non-covalent bonds, as determined by alanine scanning and molecular dynamics simulation. The mutant was then applied to develop the magnetic particle chemiluminescence method. 574 clinical samples were tested using the developed method, with a cutoff value of 95 pg/mL set by Limulus amebocyte lysate. The receiver operating characteristic curve demonstrated an area under the curve of 0.905 (95% CI: 0.880-0.929). Chemiluminescence detected an antigen concentration of 89.98 pg/mL, exhibiting a sensitivity of 83.33% and specificity of 89.76%. In conclusion, the results showed a good agreement with Limulus amebocyte lysate and demonstrated the feasibility of using BDG mutant antibodies for invasive fungal disease diagnosis. The new method based on chemiluminescence for detecting BDG could shorten the sample-to-result time to approximately 30 min, rescue Limulus from being endangered and is resource efficient in terms of equipment and the non-use of a skilled technician.

Computational Design of Peptide Assemblies.

With the ongoing development of peptide self-assembling materials, there is growing interest in exploring novel functional peptide sequences. From short peptides to long polypeptides, as the functionality increases, the sequence space is also expanding exponentially. Consequently, attempting to explore all functional sequences comprehensively through experience and experiments alone has become impractical. By utilizing computational methods, especially artificial intelligence enhanced molecular dynamics (MD) simulation and de novo peptide design, there has been a significant expansion in the exploration of sequence space. Through these methods, a variety of supramolecular functional materials, including fibers, two-dimensional arrays, nanocages, etc., have been designed by meticulously controlling the inter- and intramolecular interactions. In this review, we first provide a brief overview of the current main computational methods and then focus on the computational design methods for various self-assembled peptide materials. Additionally, we introduce some representative protein self-assemblies to offer guidance for the design of self-assembling peptides.

Algae-derived biochar nanozyme array for discrimination and detection of multiple pesticides in soil, water and food.

Despite the potential of nanozymes combined with sensor arrays for discriminating multiple pesticides simultaneously, they have few practical pesticide sensing uses due to the limited performance of existing nanozymes and the complexity of their preparation. Here, agricultural waste is utilized for the facile synthesis of high-performance biochar nanozymes and the fabrication of biochar nanozyme sensor arrays. The production of autogenous N-doped biochars with abundant surface functional groups and good peroxidase-like activities is achieved with different types of algae. High-performance biochar nanozyme sensor arrays can discriminate pesticides in a concentration range from 1 to 500 µM and in real samples from soil, lake water, seawater, apples, cucumbers, peaches, tomatoes and cabbages. Furthermore, pesticides can be quantified down to 1 µM. The development of high-performance nanozyme sensor arrays based on waste conversion could be a step toward pesticide discrimination and detection, which would improve human and environmental safety.

An ultra pH-responsive peptide nanocarrier for cancer gene therapy.

The tumor microenvironment is a very complex and dynamic ecosystem. Although a variety of pH-responsive peptides have been reported to deliver nucleic acid drugs for cancer treatment, these responses typically only target the acidic microenvironment of the tumor or the lysosome, and the carrier suffers from issues such as low transfection efficiency and poor lysosomal escape within the cell. To address this problem, we have developed an ultra pH-responsive peptide nanocarrier that can efficiently deliver siRNA, pDNA, and mRNA into cancer cells by performing progressive dynamic assembly in response to pH changes in the acidic tumor microenvironment (pH 6.5-6.8) and the acidic intracellular lysosomal environment (pH 5.0-6.0). The maximum transfection efficiency was 87.1% for pDNA and 74.9% for mRNA, which is higher than that of peptide-based nanocarrier reported to date. In addition, the targeting sequence on the surface allows the peptide@siRNA complex to efficiently enter cancer cells, causing 96% of cancer cell mortality. The carrier has high biocompatibility and low cytotoxicity, making it highly promising for application in immunotherapy and gene therapy of tumors.

Efficient removal of chloroform from groundwater using activated percarbonate by cellulose nanofiber-supported Fe/Cu nanocomposites.

Chloroform (CF) is a recalcitrant halogenated methane (HM) that has received widespread attention due to its frequent detection in groundwater and its potential carcinogenic risk. In this study, TEMPO-oxidized cellulose nanofiber-supported iron/copper bimetallic nanoparticles (TOCNF-Fe/Cu), a novel composite catalyst, was synthesized to activate sodium percarbonate (SPC) for the removal of CF from groundwater. The results showed that over 96.3% of CF could be removed in a neutral reaction medium (pH 6.5-9) within 180 min using 0.66 g L-1 of TOCNF (0.32)-Fe/Cu (1) and 1 mM of SPC, which outperforms typical advanced oxidation processes. The reaction mechanism of the TOCNF-Fe/Cu-SPC system for the CF removal was elucidated. As demonstrated through electron paramagnetic resonance and quenching experiments, the TOCNF-Fe/Cu-SPC system was found to include •OH and O2•-, where the latter played a dominant role in the CF removal. DFT calculations indicated that TOCNF improved the electron transport capability of Fe/Cu and reduced the transition state energy. The Fe species on the surface of TOCNF-Fe/Cu were identified as the primary active sites for SPC activation, whereas the Cu species were beneficial to the regeneration of the Fe species. Additionally, TOCNF-Fe/Cu was found to have good recyclability and stability. The feasibility of the TOCNF-Fe/Cu-SPC system was further confirmed by applying it for the efficient removal of composite HMs from actually contaminated groundwater. Overall, the TOCNF-Fe/Cu-SPC system is an attractive candidate for the treatment of HM-contaminated groundwater.

Gold Nanoparticles Mineralized by Peptide Liquid Crystals with Dual-Functional Enzyme-like Activities: An Automatic Membrane Reactor for Glucose Detection.

The development of stable multifunctional enzyme mimics with tandem catalytic effects provides a great opportunity to construct economical and convenient bioassays. Inspired by biomineralization, in this work self-assembled N-(9-fluorenylmethoxycarbonyl)-protected tripeptide (Fmoc-FWK-NH2) liquid crystals were used as templates to in situ mineralize Au nanoparticles (AuNPs), and then a dual-functional enzyme-mimicking membrane reactor based on AuNPs and peptide-based hybrids was constructed. AuNPs with a uniform particle size and good dispersion were in situ reduced on the surface of the peptide liquid crystal due to the reduction of the indole group on the tryptophan residue, which exhibited excellent peroxidase-like and glucose oxidase-like activities simultaneously. Meanwhile, the oriented nanofibers aggregated into a three-dimensional network, which was further immobilized on the mixed cellulose membrane to form a membrane reactor. A biosensor was made to realize fast, low-cost, and automatic detection for glucose. This work represents a promising platform for the design and construction of novel multifunctional materials based on the biomineralization strategy.

Self-Assembly of Peptide-Lipid Nanoparticles for the Efficient Delivery of Nucleic Acids.

A transfection formulation is successfully developed to deliver nucleic acids by adding an auxiliary lipid (DOTAP) to the peptide, and the transfection efficiency of pDNA reaches 72.6%, which is close to Lipofectamine 2000. In addition, the designed KHL peptide-DOTAP complex exhibits good biocompatibility by cytotoxicity and hemolysis analysis. The mRNA delivery experiment indicates that the complex had a 9- or 10-fold increase compared with KHL or DOTAP alone. Intracellular localization shows that KHL/DOTAP can achieve good endolysosomal escape. Our design provides a new platform for improving the transfection efficiency of peptide vectors.

Anisotropic cellulose nanocrystalline sponge sheets with ultrahigh water fluxes and oil/water selectivity.

Oily sewage caused by oil spill accidents has become a severe problem in the last decades. Hence, two-dimensional sheet-like filter materials for oil/water separation have received widespread attention. Porous sponge materials were developed using cellulose nanocrystals (CNCs) as raw materials. They are environmentally friendly and easy to prepare, with high flux and separation efficiency. The 1,2,3,4-butane tetracarboxylic acid cross-linked anisotropic cellulose nanocrystalline sponge sheet (B-CNC) exhibited ultrahigh water fluxes driven by gravity alone, depending on the aligned structure of channels and the rigidity of CNCs. Meanwhile, the sponge gained superhydrophilic/underwater superhydrophobic wettability with an underwater oil contact angle of up to 165.7° due to its ordered micro/nanoscale structure. B-CNC sheets displayed high oil/water selectivity without additional material doping or chemical modification. For oil/water mixtures, high separation fluxes of approximately 100,000 L·m-2·h-1 and separation efficiencies of up to 99.99 % were obtained. For a Tween 80-stabilized toluene-in-water emulsion, the flux reached >50,000 L·m-2·h-1, and the separation efficiency was above 99.7 %. B-CNC sponge sheets showed significantly higher fluxes and separation efficiencies than other bio-based two-dimensional materials. This research provides a facile and straightforward fabrication method of environmental-friendly B-CNC sponges for rapid, selective oil/water separation.

Glutathione peroxidase-like nanozymes: mechanism, classification, and bioapplication.

The field of nanozymes is developing rapidly. In particular, glutathione peroxidase (GPx)-like nanozymes, which catalytically reduce H2O2/organic hydroperoxides to H2O/alcohols, have attracted considerable attention. GPx-like nanozymes are powerful antioxidant enzymes known to combat oxidative stress. They have broad applications, including cytoprotection, anti-inflammation, neuroprotection, tumor therapy, and anti-aging. Although much progress has been made, GPx-like nanozymes have not been well discussed or fully reviewed as other nanozymes. This review aims to summarize recent advances on GPx-like nanozymes from the vantage point of mechanism, classification, and bioapplication. Future prospects for advancing their design and application are also discussed.

Biomimetic mineralization based on self-assembling peptides.

Biomimetic science has attracted great interest in the fields of chemistry, biology, materials science, and energy. Biomimetic mineralization is the process of synthesizing inorganic minerals under the control of organic molecules or biomolecules under mild conditions. Peptides are the motifs that constitute proteins, and can self-assemble into various hierarchical structures and show a high affinity for inorganic substances. Therefore, peptides can be used as building blocks for the synthesis of functional biomimetic materials. With the participation of peptides, the morphology, size, and composition of mineralized materials can be controlled precisely. Peptides not only provide well-defined templates for the nucleation and growth of inorganic nanomaterials but also have the potential to confer inorganic nanomaterials with high catalytic efficiency, selectivity, and biotherapeutic functions. In this review, we systematically summarize research progress in the formation mechanism, nanostructural manipulation, and applications of peptide-templated mineralized materials. These can further inspire researchers to design structurally complex and functionalized biomimetic materials with great promising applications.

Ternary Synergy of Lys, Dopa, and Phe Results in Strong Cohesion of Peptide Films.

Synergistic interactions between 3,4-dihydroxyphenylalanine (Dopa, Y*), cationic residues, and the aromatic rings have been recently highlighted as influential factors that enhance the underwater adhesion strength of mussel foot proteins and their derivatives. In this study, we report the first ever evidence of a cation-catechol-benzene ternary synergy between Y*, lysine (Lys, K), and phenylalanine (Phe, F) in adhesive peptides. We synthesized three hexapeptides containing a different combination of Y*, K, and F, i.e., (KY*)3, (KF)3, and (KY*F)2, respectively, exploring the relationship between the cohesive performance and molecular architecture of peptides. The peptide with the (KY*F)2 sequence displays the strongest underwater cohesion energy of 10.3 ± 0.3 mJ m-2 from direct nanoscale surface force measurements. Combined with molecular dynamics simulation, we demonstrated that there are more bonding interactions (including cation-π, π-π, and hydrogen bond interactions) in (KY*F)2 compared to the other two peptides. In addition, peptide (KY*F)2 still shows the strongest cohesive energies of 7.6 ± 0.7 and 3.7 ± 0.5 mJ m-2 in acidic and high-ionic strength environments, respectively, although the cohesive energy decreases compared to the value in pure water. Our results further explain the underwater cohesion mechanisms combining multiple interactions and offer insights on designing Dopa containing underwater adhesives.

Biospecific cation-π interaction by modulating molecular hydration and supramolecular structure of short peptides.

This study presents novel adhesive materials that use cation-π interactions to achieve highly specific cohesive interaction under water. The materials are short length peptides based on the FKF motif flanked by different side groups. Using the surface forces apparatus, we show that the composition of the side group allows to finely tune the strength of the cohesive and adhesive energies of the peptide and its specificity, meaning its capacity to bind strongly only to substrates bearing the same peptide. The interfacial properties of these adhesive peptides are shown to strongly depend on the composition of the deposition solvent, with DMSO being the solvent of choice to achieve high cohesive and adhesive energies. This result was correlated with the supramolecular structure of the peptide film and confirmed that needle-like structures can significantly enhance the adhesion of the material. Altogether, we showed that cation-π interaction can be used efficiently to create adhesive materials that incorporate features already known for underwater adhesives such as activation via solvent displacement, as well as new ones such as specificity and supramolecular structure enhanced adhesion.

Peptidyl Virus-Like Nanovesicles as Reconfigurable "Trojan Horse" for Targeted siRNA Delivery and Synergistic Inhibition of Cancer Cells.

The self-assembly of peptidyl virus-like nanovesicles (pVLNs) composed of highly ordered peptide bilayer membranes that encapsulate the small interfering RNA (siRNA) is reported. The targeting and enzyme-responsive sequences on the bilayer's surface allow the pVLNs to enter cancer cells with high efficiency and control the release of genetic drugs in response to the subcellular environment. By transforming its structure in response to the highly expressed enzyme matrix metalloproteinase 7 (MMP-7) in cancer cells, it helps the siRNA escape from the lysosomes, resulting in a final silencing efficiency of 92%. Moreover, the pVLNs can serve as reconfigurable "Trojan horse" by transforming into membranes triggered by the MMP-7 and disrupting the cytoplasmic structure, thereby achieving synergistic anticancer effects and 96% cancer cell mortality with little damage to normal cells. The pVLNs benefit from their biocompatibility, targeting, and enzyme responsiveness, making them a promising platform for gene therapy and anticancer therapy.

One-Step Synthesis of Peptide-Gold Nanoclusters with Tunable Fluorescence and Enhanced Gene Delivery Efficiency.

In this study, peptide-gold nanoclusters with tunable fluorescence were prepared by a simple "one-pot" method, which were used for gene localization and delivery in vivo to achieve efficient intracellular colocalization, uptake, and transfection. The efficiency of pDNA transfection was up to 70.6%, and there was no obvious cytotoxicity. This study proves that the simple-composition and bio-friendly peptide-gold nanoclusters are promising gene delivery carriers and can provide a powerful theoretical and experimental basis for the application of peptide-metal nanocomplexes in gene delivery and other biomedicine fields.

Bioinspired porphyrin-peptide supramolecular assemblies and their applications.

Inspired by the hierarchical chiral assembly of porphyrin-proteins in photosynthetic systems, the hierarchical self-assembly of porphyrin-amino acids/peptides provides a novel strategy for constructing functional materials. How to artificially simulate the assembly of porphyrins, proteins, and other cofactors in the photosynthesis system to obtain persistent strong light capture, charge separation and catalytic reactions has become an important concern in the construction of biomimetic photosynthesis systems. This paper summarizes the different assembly strategies adopted in recent years, the effects of driving forces on self-assembly, and the application of porphyrin-peptides in catalysis and biomedicine, and briefly discusses the challenges and prospects for future research.

Coordination-Induced Self-Assembly of a Dipeptide into Multifunctional Chiral Nanostructures.

Short peptides could be used as chiral motifs to self-assemble into various artificial nanostructures with supramolecular or nanoscale chirality, but their applications still need to be expanded. Here, under the mediation of metal ions, the ferrocene-diphenylalanine (Fc-LFLF) peptide can self-assemble into various chiral nanostructures, including right-handed helical microflowers mediated by Cu2+, left-handed nanofibers mediated by Ag+, and right-handed nanofibers mediated by Zn2+ and Cd2+. Meanwhile, the gold nanoparticles could be mineralized and deposited on Cu2+/Fc-LFLF microflowers to form AuNPs@Cu2+/Fc-LFLF, which showed significantly improved catalytic activity. The Ag+ could be further mineralized on the peptide nanofibers to form AgNPs@Fc-LFLF, showing an excellent antibacterial effect. Overall, this study provides new insights into the chiral self-assembly of short peptides and demonstrates that the chiral peptide-metal assemblies may have broad prospects in the fields of biocatalysis and antimicrobials.

The safe Laccase@ZIF-8-prodrug system with GSH redox cycle for effective targeted cancer therapy with low off-target toxicity.

Nanotechnology has been widely used in cancer treatment but only a small fraction (0.7 %) of the administered nanoparticle was delivered into a solid tumor, while the remaining fraction causes off-target toxicity in healthy tissues. The activation of prodrugs by exogenous enzymes can be used as an effective anticancer strategy to reduce systemic toxicity. In this study, the laccase@zeolitic imidazolate framework-8 (LA@ZIF-8) enzyme-activated prodrug system was created based on the high catalytic activity of LA in an acidic environment and the pH response (pH∼5.5) of ZIF-8. Quercetin (QU) was selected as the prodrug, which is non-toxic and even beneficial without being activated by LA. LA could be precisely released in the acidic tumor microenvironment for activating nontoxic QU successfully to produce toxic oxidized quercetin (OQU), and the LA was steady loaded on the LA@ZIF-8 under physiological conditions (pH∼7.4). Especially, the high concentrations of glutathione (GSH) in tumors could accelerate the oxidation of QU by LA. Meanwhile, GSH could be consumed continuously by the reduction of OQU for regenerating QU, thus a LA-QU-GSH redox was formed ingeniously. Therefore, the synergistic effect of OQU toxicity and GSH depletion induced reactive oxygen species (ROS) accumulation and tumor cell apoptosis. Overall, the LA@ZIF-8-QU prodrug system provides ideas for safe and effective cancer treatment with low off-target toxicity.

Pervaporation Polyvinyl Alcohol Membranes Modified with Zr-Based Metal Organic Frameworks for Isopropanol Dehydration.

Metal-organic frameworks (MOFs) are perceptive modifiers for the creation of mixed matrix membranes to improve the pervaporation performance of polymeric membranes. In this study, novel membranes based on polyvinyl alcohol (PVA) modified with Zr-MOFs (MIL-140A, MIL-140A-AcOH, and MIL-140A-AcOH-EDTA) particles were developed for enhanced pervaporation dehydration of isopropanol. Two membrane types (substrateless-freestanding; and formed on polyacrylonitrile support-composite) were prepared. The additional cross-linking of membranes with glutaraldehyde was carried out to circumvent membrane stability in pervaporation dehydration of diluted solutions. The synthesized Zr-MOFs were characterized by scanning electron microscopy, X-ray powder diffraction analysis, and specific surface area measurement. The structure and physicochemical properties of the developed membranes were investigated by Fourier-transform infrared spectroscopy, scanning electron and atomic force microscopies, thermogravimetric analysis, swelling experiments, and contact angle measurements. The PVA and PVA/Zr-MOFs membranes were evaluated in pervaporation dehydration of isopropanol in a wide concentration range. It was found that the composite cross-linked PVA membrane with 10 wt% MIL-140A had optimal pervaporation performance in the isopropanol dehydration (12-100 wt% water) at 22 °C: 0.15-1.33 kg/(m2h) permeation flux, 99.9 wt% water in the permeate, and is promising for the use in the industrial dehydration of alcohols.

Study of the kinetics and mechanism of ferrocene-tripeptide inhibiting insulin aggregation.

Peptides are gaining popularity as neurodegenerative disease-targeting drugs due to their medicinal value and simplicity in the biomedical and pharmaceutical industry fields. Herein, based on previously studied ferrocene-modified dipeptide (Fc-dipeptide) ferrocene-L-Phe-L-Phe (Fc-FF), we developed four Fc-tripeptides, i.e., Fc-FFY, Fc-FFF, Fc-FFD, and Fc-FFK, to further study the anti-amyloid effects of peptide-based inhibitors. The results showed that all Fc-tripeptides inhibited the formation of insulin fibrils in a dose-dependent manner more responsive than Fc-FF. Meanwhile, Fc-FFY and Fc-FFF had a more significant inhibitory effect on insulin amyloid fibrillation than Fc-FFD and Fc-FFK. In addition, molecular dynamics simulations indicated that Fc-FFY and Fc-FFF were contacted mainly with the hydrophobic core residues of insulin chain A and chain B, respectively. The research indicated that Fc-tripeptides have potential effects in preventing protein misfolding diseases and could be further used to design effective anti-amyloidosis compounds.