Effective Treatment of Helicobacter pylori Infection Using Supramolecular Antimicrobial Peptide Hydrogels.

Helicobacter pylori can cause various gastric conditions including stomach cancer in an acidic environment. Although early H. pylori infections can be treated by antibiotics, prolonged antibiotic administrations may lead to the development of antimicrobial resistance, compromising the effectiveness of the treatments. Antimicrobial peptides (AMPs) have been reported to possess unique advantages against antimicrobial-resistant bacteria due to their rapid physical membrane disruptions and anti-inflammation/immunoregulation properties. Herein, we have developed an AMP hydrogel, which can be orally administered for the treatment of H. pylori infection. The hydrogel has potent antimicrobial activity against H. pylori, achieving bacterial eradication within minutes of action. Compared with the AMP solution, the hydrogel formulation significantly reduced the cytotoxicity and enhanced proteolytic stability. In vivo experiments suggested that the hydrogel formed at pH 4 had superior therapeutic effects to those at pH 7 and 10 hydrogels, attributed to its rapid release and bactericidal action within the acidic stomach environment. Compared to conventional antibiotic treatments, the AMP hydrogel had the advantages of fast bacterial killing in the gastric juice and obviated proton pump inhibitors during the treatment. Although both the AMP hydrogel and antibiotics suppressed the expression of pro-inflammatory cytokines, the former uniquely promoted inflammation resolution. These results indicate that the AMP hydrogels with effectiveness and biosafety may be potential candidates for the clinical treatment of H. pylori infections.

Peptide Self-assembly: From Ordered to Disordered.

ConspectusBiomolecular self-assembly is a ubiquitous occurrence in nature that gives rise to sophisticated superstructures that enable the implementation of complex biological functions. It encompasses both ordered structures, such as the DNA double helix, and disordered structures, such as the nucleolus and other nonmembranous organelles. In contrast to these highly organized ordered structures, which exhibit specific patterns or symmetry, disordered structures are characterized by their flexible and randomized molecular organization, which provides versatility, dynamicity, and adaptability to biological systems and contributes to the complexity and functionality of living organisms. However, these disordered structures usually exist in a thermodynamically metastable state. This means that these disordered structures are unstable and difficult to observe due to their short existence time. Achieving disordered structures through precise control of the assembly process and ensuring their stability and integrity pose significant challenges. Currently, ongoing research efforts are focused on the self-assembly of proteins with intrinsically disordered regions (IDRs). However, the structural complexity and instability of proteins present prohibitive difficulties in elucidating the multiscale self-assembly process. Therefore, simple peptides, as a segment of proteins, hold great promise in constructing self-assembly systems for related research. Since our finding on droplet-like disordered structures that occur transiently during the peptide self-assembly (PSA), our research is centered around the dynamic evolution of peptide supramolecular systems, particularly the modulation of a variety of assembled structures ranging from ordered to disordered.In this Account, we narrate our recent research endeavors on supramolecular structures formed by PSA, spanning from ordered structures to disordered structures. We delve into the mechanisms of structural regulation, shedding light on how these peptide-based structures can be controlled more precisely. Moreover, we emphasize the functional applications that arise from these structures. To begin, we conduct a comprehensive overview of various types of ordered structures that emerge from PSA, showcasing their diverse applications. Following, we elaborate on the discovery and development of droplet-like disordered structures that arise during PSA. A mechanistic study on multistep self-assembly processes mediated by liquid-liquid phase separation (LLPS) is critically emphasized. Ordered structures with different morphologies and functions can be obtained by subtly controlling and adjusting the metastable liquid droplets. In particular, we have recently developed solid glasses with long-range disorder, including noncovalent biomolecular glass based on amino acid and peptide derivatives, as well as high-entropy glass based on cyclic peptides. This demonstrates the great potential of using biologically derived molecules to create green and sustainable glassy materials.

Biomimetic Conductive Hydrogel Scaffolds with Anisotropy and Electrical Stimulation for In Vivo Skeletal Muscle Reconstruction.

The nature of the hydrogel scaffold mimicking extracellular matrix plays a crucial role in tissue engineering like skeletal muscle repair. Herein, an anisotropic and conductive hydrogel scaffold is fabricated using gelatin methacryloyl (GelMA) as the matrix hydrogel and silver nanowire (AgNW) as the conductive dopant, through a directional freezing technique for muscle defect repair. The scaffold has an anisotropic structure composed of a directional longitudinal section and a honeycomb cross-section, with high mechanical strength of 10.5 kPa and excellent conductivity of 0.26 S m-1 . These properties are similar to native muscle extracellular matrix (ECM) and allow for cell orientation under the guidance of contact cues and electrical stimulation synergistically. In vitro experiments show that the scaffold's oriented structure combined with electrical stimulation results in enhanced myotube formation, with a length of up to 863 µm and an orientation rate of 81%. Furthermore, the electrically stimulated scaffold displays a promoted muscle reconstruction ability when transplanted into rats with muscle defects, achieving a muscle mass and strength restoration ratio of 95% and 99%, respectively, compared to normal levels. These findings suggest that the scaffold has great potential in muscle repair applications.

Molecular Engineering of Ordered Piezoelectric Sulfonic Acid-Containing Assemblies.

Sulfonic acid-containing bioorganic monomers with wide molecular designability and abundant hydrogen bonding sites hold great potential to design diverse functional biocrystals but have so far not been explored for piezoelectric energy harvesting applications due to the lack of strategies to break the centrosymmetry of their assemblies. Here, a significant molecular packing transformation from centrosymmetric into non-centrosymmetric conformation by the addition of an amide terminus in the sulfonic acid-containing bioorganic molecule is demonstrated, allowing a high electromechanical response. The amide-functionalized molecule self-assembles into a polar supramolecular parallel ß-sheet-like structure with a high longitudinal piezoelectric coefficient d11 = 15.9 pm V-1 that produces the maximal open-circuit voltage of >1 V and the maximal power of 18 nW in nanogenerator devices pioneered. By contrast, molecules containing an amino or a cyclohexyl terminus assemble into highly symmetric 3D hydrogen bonding diamondoid-like networks or 2D double layer structures that show tunable morphologies, thermostability, and mechanical properties but non-piezoelectricity. This work not only presents a facile approach to achieving symmetry transformation of bioorganic assemblies but also demonstrates the terminal group and the property correlation for tailor-made design of high-performance piezoelectric biomaterials.

Biosynthesis of Multifunctional Transformable Peptides for Inducing Tumor Cell Apoptosis.

Engineered nanomaterials hold great promise to improve the specificity of disease treatment. Herein, a fully protein-based material is obtained from nonpathogenic Escherichia coli (E. coli), which is capable of morphological transformation from globular to fibrous in situ for inducing tumor cell apoptosis. The protein-based material P1 is comprised of a ß-sheet-forming peptide KLVFF, pro-apoptotic protein BAK, and GFP along with targeting moieties. The self-assembled nanoparticles of P1 transform into nanofibers in situ in the presence of cathepsin B, and the generated nanofibrils favor the dimerization of functional BH3 domain of BAK on the mitochondrial outer membrane, leading to efficient anticancer activity both in vitro and in vivo via mitochondria-dependent apoptosis through Bcl-2 pathway. To precisely manipulate the morphological transformation of biosynthetic molecules in living cells, a spatiotemporally controllable anticancer system is constructed by coating P1-expressing E. coli with cationic conjugated polyelectrolytes to release the peptides in situ under light irradiation. The biosynthetic peptide-based enzyme-catalytic transformation strategy in vivo would offer a novel perspective for targeted delivery and shows great potential in precision disease therapeutics.

Two-photon nanoprobes based on bioorganic nanoarchitectonics with a photo-oxidation enhanced emission mechanism.

Two-photon absorption (TPA) fluorescence imaging holds great promise in diagnostics and biomedicine owing to its unparalleled spatiotemporal resolution. However, the adaptability and applicability of currently available TPA probes, which act as a critical element for determining the imaging contrast effect, is severely challenged by limited photo-luminescence in vivo. This is particularly a result of uncontrollable aggregation that causes fluorescence quenching, and inevitable photo-oxidation in harsh physiological milieu, which normally leads to bleaching of the dye. Herein, we describe the remarkably enhanced TPA fluorescence imaging capacity of self-assembling near-infrared (NIR) cyanine dye-based nanoprobes (NPs), which can be explained by a photo-oxidation enhanced emission mechanism. Singlet oxygen generated during photo-oxidation enables chromophore dimerization to form TPA intermediates responsible for enhanced TPA fluorescence emission. The resulting NPs possess uniform size distribution, excellent stability, more favorable TPA cross-section and anti-bleaching ability than a popular TPA probe rhodamine B (RhB). These properties of cyanine dye-based TPA NPs promote their applications in visualizing blood circulation and tumoral accumulation in real-time, even to cellular imaging in vivo. The photo-oxidation enhanced emission mechanism observed in these near-infrared cyanine dye-based nanoaggregates opens an avenue for design and development of more advanced TPA fluorescence probes.

Supramolecular Nanoarchitectonics Based on Antagonist Peptide Self-Assembly for Treatment of Liver Fibrosis.

Therapeutic peptides have attracted increasing attention as anti-fibrotic drug candidates. However, the rapid degradation and insufficient liver accumulation of therapeutic peptides have seriously hampered their clinical translation. Here, the use of supramolecular nanoarchitectonics is reported to fabricate nanodrugs from therapeutic peptides for treating liver fibrosis. Self-assembling antagonist peptides are rationally designed and manipulated into uniform peptide nanoparticles with well-defined nanostructures and uniform sizes. Significantly, the peptide nanoparticles show enhanced accumulation in liver sites and limited distribution in other tissues. In vivo results show that the peptide nanoparticles exhibit greatly enhanced anti-fibrotic activity compared to the pristine antagonist along with good biocompatibility. These results indicate that self-assembly is a promising nanoarchitectonics approach to enhance the anti-fibrotic activity of therapeutic peptides for treating liver fibrosis.

Bubble wall confinement-driven molecular assembly toward sub-12 nm and beyond precision patterning.

Patterning is attractive for nanofabrication, electron devices, and bioengineering. However, achieving the molecular-scale patterns to meet the demands of these fields is challenging. Here, we propose a bubble-template molecular printing concept by introducing the ultrathin liquid film of bubble walls to confine the self-assembly of molecules and achieve ultrahigh-precision assembly up to 12 nanometers corresponding to the critical point toward the Newton black film limit. The disjoining pressure describing the intermolecular interaction could predict the highest precision effectively. The symmetric molecules exhibit better reconfiguration capacity and smaller preaggregates than the asymmetric ones, which are helpful in stabilizing the drainage of foam films and construct high-precision patterns. Our results confirm the robustness of the bubble template to prepare molecular-scale patterns, verify the criticality of molecular symmetry to obtain the ultimate precision, and predict the application potential of high-precision organic patterns in hierarchical self-assembly and high-sensitivity sensors.

Functional chromopeptide nanoarchitectonics: molecular design, self-assembly and biological applications.

Chromoproteins are a class of delicate natural compounds that elegantly complex photosensitive species with proteins and play a central role in important life processes, such as photosynthesis. Inspired by chromoproteins, researchers integrate simple peptides and photosensitive molecular motifs to generate chromopeptides. Compared with chromoproteins, chromopeptides exhibit a relatively simple molecular structure, flexible and adjustable photophysical properties, and a capability of programmable self-assembly. Chromopeptide self-assembly has attracted great attention as the resultant high-level architectures exhibit an ingenious combination of photofunctions and biofunctions. This review systematically summarizes recent advances in chromopeptide nanoarchitectonics with particular focus on the design strategy, assembly mechanism, and structure-function relationship. Among them, the effect of peptide sequences and the variation in photophysical performance are critically emphasized. On this basis, various applications, including biomedicine and artificial photosynthesis, are discussed together with the future prospects of chromopeptide nanoarchitectonics. This review will provide insights into chromopeptide nanoarchitectonics and corresponding materials with precise designs, flexible nanostructures and versatile functions. In addition, knowledge involving chromopeptide nanoarchitectonics may aid in the development of many other kinds of supramolecular biological materials and bioengineering techniques.

Biomolecular glass with amino acid and peptide nanoarchitectonics.

Glass is ubiquitous in life and widely used in various fields. However, there is an urgent need to develop biodegradable and biorecyclable glasses that have a minimal environmental footprint toward a sustainable society and a circular materials economy. Here, we report a family of eco-friendly glasses of biological origin fabricated using biologically derived amino acids or peptides through the classic heating-quenching procedure. Amino acids and peptides with chemical modification at their ends are found able to form a supercooled liquid before decomposition and eventually glass upon quenching. These developed glasses exhibit excellent glass-forming ability and optical characteristics and are amenable to three-dimensional-printed additive manufacturing and mold casting. Crucially, the glasses show biocompatibility, biodegradability, and biorecyclability beyond the currently used commercial glasses and plastic materials.

Hydrogel Nanoarchitectonics of a Flexible and Self-Adhesive Electrode for Long-Term Wireless Electroencephalogram Recording and High-Accuracy Sustained Attention Evaluation.

Hydrogels are ideal building blocks to fabricate the next generation of electrodes for acquiring high-quality physiological electrical signals, for example, electroencephalography (EEG). However, collection of EEG signals still suffers from electrode deformation, sweating, extensive body motion and vibration, and environmental interference. Herein, polyvinyl alcohol and polyvinylpyrrolidone are selected to prepare a hydrogel network with tissue-like modulus and excellent flexibility. Additionally, polydopamine nanoparticles, obtained by polydopamine peroxidation, are integrated into the hydrogel to endow them with higher transparency, higher self-adhesion, and lower impedance. Consequently, a multichannel and wirelessly operated hydrogel electrode can establish a conformal and stable interface with tissue and illustrate high channel uniformity, low interfacial contact impedance, low power noise, long-term stability, and a tolerance to sweat and motion. Furthermore, the hydrogel electrode shows the unprecedented ability to classify the recorded high-quality prefrontal EEG signals into seven-category sustained attention with high accuracy (91.5%), having great potential applications in the assessment of human consciousness and in multifunctional diagnoses.

Covalently triggered self-assembly of peptide-based nanodrugs for cancer theranostics.

Covalently triggered peptide self-assembly is achieved through sequential integration of spontaneous covalent reaction and noncovalent interactions, thus both enhancing the physiological stability and extending unexpected functionality of the resulting peptide-based assemblies, different from popular supramolecular peptide self-assembly merely associated with noncovalent interactions. This review summarizes the recent progress on the development of covalently triggered peptide self-assembly for cancer theranostics. Especially, we propose the fundamental design principle of covalently triggered peptide self-assembly for constructing a variety of peptide-based assemblies including nanoparticles, nanofibers, hollow nanospheres, and other nanoarchitectures. Subsequently, the discussion is anchored in an overview of representative covalently assembled peptide-based nanodrugs for the cancer theranostics. Finally, the challenges and perspectives on the clinical potential of the covalently assembled peptide-based nanodrugs are highlighted. This review will provide new insights into construction of peptide-based nanodrugs through combination of covalent reaction and noncovalent self-assembly and prompt their clinical applications in cancer diagnosis and therapeutics.

Bioactive Peptide Nanodrugs Based on Supramolecular Assembly for Boosting Immunogenic Cell Death-Induced Cancer Immunotherapy.

Immunogenic cell death (ICD)-induced immunotherapy holds promise for complete elimination and long-term protective immune responses against cancer by combining direct tumor cell killing and antitumor immune response. Some therapeutic approaches (such as hyperthermia, photodynamic therapy, or radiotherapy) and inducers (certain chemotherapy drugs, oncolytic viruses) have been devoted to initiating and/or boosting ICD, leading to the activation of tumor-specific immune responses. Recently, supramolecular assembled bioactive peptide nanodrugs have been employed to improve the efficacy of ICD-induced cancer immunotherapy by increasing tumor targeted accumulation as well as responsive release of ICD inducers, directly inducing high levels of ICD and realizing the simultaneous enhancement of immune response through the immune function of the active peptide itself. Here, the authors review bioactive peptide nanodrugs based on supramolecular assembly, mainly as an intelligent delivery system, a direct ICD inducer and an immune response enhancer, for boosting ICD induced cancer immunotherapy. The functions of diverse bioactive peptides used in the construction of nanodrugs are described. The design of a supramolecular assembly, the mechanism of boosting ICD, and synergetic effects of bioactive peptides combined immunotherapy are critically emphasized.

Surface Self-Assembly of Dipeptides on Porous CaCO3 Particles Promoting Cell Internalization.

The self-assembling behavior of peptides and derivatives is crucial in the natural process to construct various architectures and achieve specific functions. However, the surface or interfacial self-assembly, in particular, on the surface of micro- or nanoparticles is even less systematically investigated. Here, uniform porous CaCO3 microparticles were prepared with different charged, hydrophobic and hydrophilic surfaces to assess the self-assembling behavior of dipeptides composed of various sequences. Experimental results indicate that dipeptides with a negative charge in an aqueous solution preferred to self-assemble on the hydrophobic and positively charged surface of CaCO3 particles, which can be ascribed to the electrostatic and hydrophobic interaction between dipeptides and CaCO3 particles. Meanwhile, the Log p (lipid-water partition coefficient) of dipeptides has a significant effect on the self-assembling behavior of dipeptides on the surface of porous CaCO3; dipeptides with high Log p preferred to self-assemble on the surface of CaCO3 particles, resulting in the improved cell internalization efficiency of particles with low cytotoxicity. After loading with a model drug (doxorubicin), the particles show obvious antitumor activity in animal experiments and can reduce Dox side effects effectively.

Long-range ordered amino acid assemblies exhibit effective optical-to-electrical transduction and stable photoluminescence.

Bio-endogenous peptide molecules are ideal components for fabrication of biocompatible and environmentally friendly semiconductors materials. However, to date, their applications have been limited due to the difficulty in obtaining stable, high-performance devices. Herein, simple amino acid derivatives fluorenylmethoxycarbonyl-leucine (Fmoc-L) and fluorenylmethoxycarbonyl-tryptophan (Fmoc-W) are utilized to form long-range ordered supramolecular nanostructures by tight aromatic stacking and extensive hydrogen bonding with mechanical, electrical and optical properties. For the first time, without addition of any photosensitizers, pure Fmoc-L microbelts and Fmoc-W microwires exhibit Young's modulus up to 28.79 and 26.96 GPa, and unprecedently high values of photocurrent responses up to 2.2 and 2.3 µA/cm2, respectively. Meanwhile, Fmoc-W microwires with stable blue fluorescent emission under continuous excitation are successfully used as LED phosphors. Mechanism analysis shows that these two amino acids derivatives firstly formed dimers to reduce the bandgap, then further assemble into bioinspired semiconductor materials using the dimers as the building blocks. In this process, aromatic residues of amino acids are more conducive to the formation of semiconducting characteristics than fluorenyl groups. STATEMENT OF SIGNIFICANCE: Long-range ordered amino acid derivative assemblies with mechanical, electrical and optical properties were fabricated by a green and facile biomimetic strategy. These amino acid assemblies have Young's modulus comparable to that of concrete and exhibit typical semiconducting characteristics. Even without the addition of any photosensitizer, pure amino acid assemblies can still produce a strong photocurrent response and an unusually stable photoluminescence. The results suggest that amino acid structures with hydrophilic C-terminal and aromatic residues are more conducive to the formation of semiconducting characteristics. This work unlocks the potential for amino acid molecules to self-assemble into high-performance bioinspired semiconductors, providing a reference for customized development of biocompatible and environmentally friendly semiconductor materials through rational molecular design.

Orally administered covalently-assembled antioxidative peptide nanoparticles for inflammatory bowel disease therapy.

HYPOTHESIS: Inflammatory bowel disease (IBD) is a chronic inflammation disease and still faces many therapeutic challenges, such as ineffective treatments, antibiotic resistance, and systematic toxicity. In order to improve the therapeutic efficacy of IBD, it is thus urgent to develop efficient, non-toxic and conveniently-administrated nanoagents to replace the currently used medicines. Casein phosphopeptide (CPP) has been found capable of chelating transition metal ions to suppress reactive oxygen species (ROS) generation, showing the potential for the treatment of IBD. However, CPP easily suffers from hydrolysis and enzymatic degradation, which limits its further clinical application. Covalent assembly of CPP to form nanoparticles (GCPP NPs) may be an efficient way to enhance the CPP stability in physiological environment and finally improve its capability of in vivo antioxidation and IBD treatment. EXPERIMENTS: We synthesized GCPP NPs through covalent assembly of Genipin and CPP, followed by characterizing their physicochemical properties. GCPP NPs were incubated under different physiological conditions including phosphate buffered saline, cell culture media, simulated gastrointestinal fluid for evaluation of stability. Cytotoxicity and antioxidation activities of GCPP NPs were tested in vitro under the 3T3 cell line using the ABTS and MTT assays, respectively. Finally, a DSS-induced mouse colitis model was established to assess specific accumulation and good therapeutic efficacy of GCPP NPs via an oral administration strategy. FINDINGS: GCPP NPs are robust and stable to overcome easy degradation of CPP even under the harsh gastrointestinal environments, which are adapted for oral administration. As-prepared GCPP NPs show benign antioxidant activity to scavenge ROS. Meanwhile, nanoscale GCPP NPs can passively accumulate and maintain at inflamed sites. The body weight and colon length of DSS-induced colitis mice treated by GCPP NPs perform a rehabilitation trend. These results indicate that GCPP NPs, as peptide-based therapeutic nanoagents, have great potential in the anti-inflammatory treatment of IBD by oral administration.

Correction: Coordination-assembled myricetin nanoarchitectonics for sustainably scavenging free radicals.

[This corrects the article DOI: 10.3762/bjnano.13.23.].

Schiff Base Mediated Dipeptide Assembly toward Nanoarchitectonics.

Dynamic covalent chemistry (DCC) is fascinating because of its dual nature. It perfectly combines the reversible nature of noncovalent bonds with the robustness of covalent bonds, effectively enhancing the stability of assemblies and meanwhile giving rise to unprecedented properties. Therefore, integration of DCC with supramolecular chemistry has emerged as a versatile and an extraordinarily useful approach in directing peptide assembly. This Minireview focuses on a recent strategy, which exploits dynamic Schiff base chemistry in combination with supramolecular chemistry, to mediate dipeptide assembly toward nanoarchitectonics. Diversified structures, new emergent properties, and their related applications are highlighted. Lastly, the opportunities and prospects in this exciting field are also introduced.

Albumin-mediated "Unlocking" of supramolecular prodrug-like nanozymes toward selective imaging-guided phototherapy.

Construction of an activatable photosensitizer and integration into an adaptive nanozyme during phototherapy without producing off-target toxicity remains a challenge. Herein, we have fabricated a prodrug-like supramolecular nanozyme based on a metallic-curcumin and cyanine co-assembly. The albumin-mediated phenol AOH group transformation of nanozyme changes its adjustable oxygen stress from negative superoxide dismutase-like activity of ROS-scavenging to positive photo oxidase activity with an ROS-amplifying capacity. It further increases the depth penetration of a nanozyme in a tumor spheroid, selectively targeting tumorous phototherapy. It also triggers a signal in targeted tumor cells and helps increase cancer cell ablation. This work suggests new options for development of activatable supramolecular nanozymes and provides a synergetic prodrug-like nanozyme strategy for early diagnosis and preclinical phototherapeutics.

Assessment of the Characteristics of Waxy Rice Mutants Generated by CRISPR/Cas9.

The cooking and eating quality of rice grains is a major focus from a consumer's perspective and is mainly determined by the apparent amylose content (AAC) of the starch. Waxy rice, a type of rice with an AAC of less than 2%, is an important goal for the breeding of high-quality rice. In recent years, the cloning of the Waxy (Wx) gene has revealed the molecular mechanism of the formation of waxy traits in rice. However, there have been limited studies on the physicochemical properties, such as gelatinization temperature, rapid viscosity analyzer profile, and amylopectin fine structure of wx mutants. In the current study, a rapid and highly efficient strategy was developed through the CRISPR/Cas9 gene-editing system for generating wx mutants in the background of five different rice varieties. The wx mutation significantly reduced the AAC and starch viscosity but did not affect the major agronomic traits (such as plant height, panicle number per plant, grain number per panicle, and seed-setting frequency). Incorporation of the wx mutation into varieties with low initial AAC levels resulted in further reduction in AAC, but without significantly affecting the original, desirable gelatinization traits and amylopectin structure types, suggesting that parents with low initial AAC should be preferred in breeding programs.