Escape Efficiency Research Articles

Steerable Microneedles Enabling Deep Delivery of Photosensitizers and CRISPR/Cas9 Systems for Effective Combination Cancer Therapy.

Although gene therapy has shown prospects in treating triple-negative breast cancer, it is insufficient to treat such a malignant tumor. Herein, nanoparticles (NPs)-embedded dissolving microneedles (IR780-PL/pFBXO44@MNs) with steerable and flectional property were developed to achieve the codelivery of FBXO44-targeted CRISPR/Cas9 plasmids (pFBXO44) and hydrophobic photosensitizers. For improved NP penetration in tumor tissue, collagenase@MNs were preapplied to degrade the tumor matrix. Under light irradiation, IR780 exhibited remarkable phototherapy, while the escape efficiency of NPs from lysosomes was improved. pFBXO44 was subsequently released in tumor cell cytoplasm via reducing the disulfide bonds of NPs, which could specifically knock out the FBXO44 gene to inhibit the migration and invasion of tumor cells. As a result, tumor cells were eradicated, and lung metastasis was effectively suppressed. This micelle-incorporated microneedle platform broadens the potential of combining gene editing and photo synergistic cancer therapy.

Nonviral In Vivo Delivery of CRISPR-Cas9 Using Protein-Agnostic, High-Loading Porous Silicon and Polymer Nanoparticles.

The complexity of CRISPR machinery is a challenge to its application for nonviral in vivo therapeutic gene editing. Here, we demonstrate that proteins, regardless of size or charge, efficiently load into porous silicon nanoparticles (PSiNPs). Optimizing the loading strategy yields formulations that are ultrahigh loading─>40% cargo by volume─and highly active. Further tuning of a polymeric coating on the loaded PSiNPs yields nanocomposites that achieve colloidal stability under cryopreservation, endosome escape, and gene editing efficiencies twice that of the commercial standard Lipofectamine CRISPRMAX. In a mouse model of arthritis, PSiNPs edit cells in both the cartilage and synovium of knee joints, and achieve 60% reduction in expression of the therapeutically relevant MMP13 gene. Administered intramuscularly, they are active over a broad dose range, with the highest tested dose yielding nearly 100% muscle fiber editing at the injection site. The nanocomposite PSiNPs are also amenable to systemic delivery. Administered intravenously in a model that mimics muscular dystrophy, they edit sites of inflamed muscle. Collectively, the results demonstrate that the PSiNP nanocomposites are a versatile system that can achieve high loading of diverse cargoes and can be applied for gene editing in both local and systemic delivery applications.

Construction of triphase interface for catalytic ozonation of polluted water

The utilization efficiency of ozone determines the cost of catalytic ozonation in water treatment. Herein, a triphase catalytic system was constructed by aerating ozone through a CeO2 loaded Al2O3 ceramic membrane (CeO2-CM) for disinfection and antibiotic degradation. Ozone aeration and a packed catalyst system (CeO2-Packing) were set as the controls. Results showed that CeO2-CM reduced the ozone escape by 34.6%–56.2%. The ozone utilization capacity of CeO2-CM for E. coli inactivation was 33.1% and 33.8% higher than those of CeO2-Packing and ozone aeration, respectively. The ozone utilization capacity of CeO2-CM for sulfamethoxazole degradation was 88.5% and 183.1% higher than those of CeO2-Packing and ozone aeration, respectively. CeO2-CM, with the lowest ozone escape and highest ozone utilization efficiency, significantly enhanced the performance of catalytic ozonation in disinfection and antibiotic degradation. This work proposes a feasible strategy for minimizing ozone consumption in water treatment.

Ionizable Lipid Nanoparticles for mRNA Delivery

Therapeutics based on messenger RNA (mRNA) have become a promising class of methods for various diseases’ treatments in recent years. The safe and effective delivery of mRNA into target cells is the premise of the clinical translation of mRNA. Due to the protonation of ionizable lipids in acidic conditions, the ionizable lipid nanoparticles (iLNPs) have lower toxicity and higher endosomal escape efficiency than permanent cationic lipid nanoparticles (cLNPs). They not only exhibit excellent delivery ability in commercial COVID‐19 mRNA vaccines but also are highly potential carriers for delivering mRNA drugs. This review summarizes the composition, preparation, stability of mRNA iLNPs, and the recent progress in the mRNA delivery by iLNPs. According to the structure characteristics of ionizable heads, the ionizable lipids are classified into tertiary amine, quaternary amine, imidazole core, piperidine core, piperazine core, and diketopiperazine core‐based head lipids. Their design, optimization, and applications on mRNA delivery are discussed. The current status of representative clinical trials based on mRNA iLNPs delivery is also analyzed. The challenges and prospects of mRNA iLNPs delivery in clinical applications are proposed at the end.

Functional L-Arginine Derivative as an Efficient Vector for Intracellular Protein Delivery for Potential Cancer Therapy.

The utilization of cytosolic protein delivery is a promising approach for treating various diseases by replacing dysfunctional proteins. Despite the development of various nanoparticle-based intracellular protein delivery methods, the complicated chemical synthesis of the vector, loading efficiency and endosomal escape efficiency of proteins remain a great challenge. Recently, 9-fluorenylmethyloxycarbonyl (Fmoc)-modified amino acid derivatives have been used to self-assemble into supramolecular nanomaterials for drug delivery. However, the instability of the Fmoc group in aqueous medium restricts its application. To address this issue, the Fmoc ligand neighboring arginine was substituted for dibenzocyclooctyne (DBCO) with a similar structure to Fmoc to obtain stable DBCO-functionalized L-arginine derivative (DR). Azide-modified triethylamine (crosslinker C) was combined with DR to construct self-assembled DRC via a click chemical reaction for delivering various proteins, such as BSA and saporin (SA), into the cytosol of cells. The hyaluronic-acid-coated DRC/SA was able to not only shield the cationic toxicity, but also enhance the intracellular delivery efficiency of proteins by targeting CD44 overexpression on the cell membrane. The DRC/SA/HA exhibited higher growth inhibition efficiency and lower IC50 compared to DRC/SA toward various cancer cell lines. In conclusion, DBCO-functionalized L-arginine derivative represents an excellent potential vector for protein-based cancer therapy.

SARS-CoV-2 omicron BA.1.1 is highly resistant to antibody neutralization of convalescent serum from the origin strain

The emergence and rapid spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant (BA.1.1) has attracted global attention. The numerous mutations in the spike protein suggest that it may have altered susceptibility to immune protection elicited by the existing coronavirus disease 2019 (COVID-19) infection. We used a live virus neutralization test and SARS-CoV-2 pseudotype vesicular stomatitis virus vector-based neutralization assay to assess the degree of immune escape efficiency of the original, Delta (B1.617.2), and Omicron strains against the serum antibodies from 64 unvaccinated patients who had recovered from COVID-19 and the results were strongly correlated. The convalescent serum neutralization was more markedly reduced against the Omicron variant (9.4–57.9-fold) than the Delta variant (2.0–4.5-fold) as compared with the original strain. Our results demonstrate the reduced fusion and notable immune evasion capabilities of the Omicron variants, highlighting the importance of accelerating the development of vaccines targeting them.

Light-Activated siRNA Endosomal Release (LASER) by Porphyrin Lipid Nanoparticles.

Lipid nanoparticles (LNPs) have achieved clinical success in delivering small interfering RNAs (siRNAs) for targeted gene therapy. However, endosomal escape of siRNA into the cytosol remains a fundamental challenge for LNPs. Herein, we report a strategy termed light-activated siRNA endosomal release (LASER) to address this challenge. We established a porphyrin-LNP by incorporating porphyrin-lipids into the clinically approved Onpattro formulation. The porphyrin-LNP maintained the physical properties of an LNP and generated reactive oxygen species (ROS) when irradiated with near-infrared (NIR) light. Using confocal microscopy, we revealed that porphyrin-lipids within the LNP translocate to endosomal membranes during endocytosis. The translocated porphyrin-lipids generated ROS under light irradiation and enabled LASER through endosomal membranes disruption as observed through GAL-9 recruitment and transmission electron microscopy (TEM). By establishing a quantitative confocal imaging method, we confirmed that porphyrin-LNPs can increase siRNA endosomal escape efficiency by up to 2-fold via LASER and further enhance luciferase target knockdown by 4-fold more in luciferase-transfected prostate cancer cells. Finally, we formulated porphyrin-LNPs encapsulated with gold nanoparticles (GNP) and visualized the LASER effect within prostate tumors via TEM, confirming the light-activated endosomal membrane disruption and subsequent GNP release into cytosols in vivo. Overall, porphyrin-LNPs and the LASER approach enhanced siRNA endosomal escape and significantly improved knockdown efficacy. We believe the versatility of this technology could be applied to various LNP-based RNA therapeutics.

Escape panels in trawls: does placement matter when every individual contacting the panel can escape?

Escape panels are one of the bycatch reduction devices most used in trawl fisheries but their efficiency rely on fish actively contacting the panel to escape. To investigate if contact behaviour changes at different panel placements, we tested a 300 mm square mesh panel placed in the upper panel of the codend at 3, 4 and 7 m from the codline. Seven competing models of contact probability were fitted to the empirical data. Based on the results, we inferred that panel placement significantly affects escape efficiency due to a change in type of contact behaviour. Cod ( Gadus morhua) showed a contact increasing with length when the panel was closest to the codline, while contact probability decreased with length at the other placements. Similarly, contact probability for plaice ( Pleuronectes platessa) was found to increase with length at 3 and 4 m, whereas a length-independent contact best represented the data at 7 m. Finally, Nephrops ( Nephrops norvegicus) had in general low contact probability. The results provide new knowledge regarding species and placement-dependent panel escape.

Cell penetrating peptide: A potent delivery system in vaccine development.

One of the main obstacles to most medication administrations (such as the vaccine constructs) is the cellular membrane's inadequate permeability, which reduces their efficiency. Cell-penetrating peptides (CPPs) or protein transduction domains (PTDs) are well-known as potent biological nanocarriers to overcome this natural barrier, and to deliver membrane-impermeable substances into cells. The physicochemical properties of CPPs, the attached cargo, concentration, and cell type substantially influence the internalization mechanism. Although the exact mechanism of cellular uptake and the following processing of CPPs are still uncertain; but however, they can facilitate intracellular transfer through both endocytic and non-endocytic pathways. Improved endosomal escape efficiency, selective cell targeting, and improved uptake, processing, and presentation of antigen by antigen-presenting cells (APCs) have been reported by CPPs. Different in vitro and in vivo investigations using CPP conjugates show their potential as therapeutic agents in various medical areas such as infectious and non-infectious disorders. Effective treatments for a variety of diseases may be provided by vaccines that can cooperatively stimulate T cell-mediated immunity (T helper cell activity or cytotoxic T cell function), and immunologic memory. Delivery of antigen epitopes to APCs, and generation of a potent immune response is essential for an efficacious vaccine that can be facilitated by CPPs. The current review describes the delivery of numerous vaccine components by various CPPs and their immunostimulatory properties.

Cytoplasmic delivery of siRNA using human-derived membrane penetration-enhancing peptide

BackgroundAlthough protein-based methods using cell-penetrating peptides such as TAT have been expected to provide an alternative approach to siRNA delivery, the low efficiency of endosomal escape of siRNA/protein complexes taken up into cells by endocytosis remains a problem. Here, to overcome this problem, we adopted the membrane penetration-enhancing peptide S19 from human syncytin 1 previously identified in our laboratory.ResultsWe prepared fusion proteins in which the S19 and TAT peptides were fused to the viral RNA-binding domains (RBDs) as carrier proteins, added the RBD-S19-TAT/siRNA complex to human cultured cells, and investigated the cytoplasmic delivery of the complex and the knockdown efficiency of target genes. We found that the intracellular uptake of the RBD-S19-TAT/siRNA complex was increased compared to that of the RBD-TAT/siRNA complex, and the expression level of the target mRNA was decreased. Because siRNA must dissociate from RBD and bind to Argonaute 2 (Ago2) to form the RNA-induced silencing complex (RISC) after the protein/siRNA complex is delivered into the cytoplasm, a dilemma arises: stronger binding between RBD and siRNA increases intracellular uptake but makes RISC formation more difficult. Thus, we next prepared fusion proteins in which the S19 and TAT peptides were fused with Ago2 instead of RBD and found that the efficiencies of siRNA delivery and knockdown obtained using TAT-S19-Ago2 were higher than those using TAT-Ago2. In addition, we found that the smallest RISC delivery induced faster knockdown than traditional siRNA lipofection, probably due to the decreased time required for RISC formation in the cytoplasm.ConclusionThese results indicated that S19 and TAT-fused siRNA-binding proteins, especially Ago2, should be useful for the rapid and efficient delivery of siRNA without the addition of any endosome-disrupting agent.

Cancerous pH-responsive polycarboxybetaine-coated lipid nanoparticle for smart delivery of siRNA against subcutaneous tumor model in mice.

Lipid nanoparticles (LNPs) have been commonly used as a vehicle for nucleic acids, such as small interfering RNA (siRNA); the surface modification of LNPs is one of the determinants of their delivery efficiency especially in systemic administration. However, the applications of siRNA-encapsulated LNPs are limited due to a lack effective systems to deliver to solid tumors. Here, we report a smart surface modification using a charge-switchable ethylenediamine-based polycarboxybetaine for enhancing tumor accumulation via interaction with anionic tumorous tissue constituents due to selective switching to cationic charge in response to cancerous acidic pH. Our polycarboxybetaine-modified LNP could enhance cellular uptake in cancerous pH, resulting in facilitated endosomal escape and gene knockdown efficiency. After systemic administration, the polycarboxybetaine-modified LNP accomplished high tumor accumulation in SKOV3-luc and CT 26 subcutaneous tumor models. The siPLK-1-encapsulated LNP thereby accomplished significant tumor growth inhibition. This study demonstrates a promising potential of the pH-responsive polycarboxybetaine as a material for modifying the surface of LNPs for efficient nucleic acid delivery.

Construction of hierarchical nanostructures and NiO nanosheets@nanorods for efficient urea electrooxidation

Hierarchical NiO nanosheets@nanorods have been rationally designed and constructed for efficient urea electrooxidation in an alkaline solution. The critical synthetic strategy, engaging the one-step anion-competitive reaction, precisely integrates two nickel-based materials into a heterostructure with Ni(OH)2 nanosheets and NiC2O4 nanorods. Benefiting from the hierarchically porous structure and high specific surface area, the NiO NNs can improve the escape efficiency of gas in electrochemical reactions and maintain sustainability. Furthermore, this distinctive structure can expose highly dispersed active sites for enhancing urea molecules' adsorption, surface-dependent redox reactions, and electrical conductivities. As a result, these hierarchical NiO nanosheets@nanorods exhibit superior activity with a low overpotential of 156 mV at 10 mA/cm2, and a slight Tafel slope of 40.7 mV/dec, and high stability with almost no decay of 12,000 s for urea electrooxidation. This work promotes the application of well-designed hierarchical structure in electrooxidizing urea and provides a possibility for highly efficient electrolysis of alkaline urea wastewater.

Modified Co electronic states in double-anionic CoPS nanocrystals induce highly efficient electrocatalytic hydrogen evolution

Pyrite-type CoS2 with half-metal properties has attracted more attention in the energy field because it is a kind of non-precious metal electrocatalyst with inexpensive, environmentally friendly, and sustainable merits. Particularly, it can produce significant spin-polarized electrons from Co electronic states to run efficient hydrogen evolution reactions (HER). In this work, we focus attention on double-anionic CoPS nanocrystals (NCs) and explore electrocatalytic hydrogen evolution performance. Theoretical calculations indicate that phosphorus introduction into CoS2 shifts the d-band of Co atom towards the Fermi surface, making the surface activation barrier of CoPS (0.98 eV) far lower than that of CoS2 (1.41 eV). By growing the rod-like Co3(PO4)2 with surface decoration of CoPS NCs on the CoS2 wrapped carbon fiber paper as electrochemical cathode, we show that such a composite structure can effectively improve the escape efficiency of H2 due to large specific surface area. The current density reaches 10 mA cm−2 at 47 mV. The tafel slope is only 52 mV dec−1, which is far smaller than that of CoS2. Spectroscopic characterization suggests that the modified electronic states of CoPS NCs and good charge transport kinetics in the compound are responsible for the observed high-efficiency HER performance. This study provides a useful strategy for designing and fabricating efficient electrocatalysts with synergistic effect of double anions and hierarchical activity.

Endosomal Escape of Bioactives Deployed via Nanocarriers: Insights Into the Design of Polymeric Micelles

Cytoplasmic delivery of bioactives requires the use of strategies such as active transport, electroporation, or the use of nanocarriers such as polymeric nanoparticles, liposomes, micelles, and dendrimers. It is essential to deliver bioactive molecules in the cytoplasm to achieve targeted effects by enabling organelle targeting. One of the biggest bottlenecks in the successful cytoplasmic delivery of bioactives through nanocarriers is their sequestration in the endosomes that leads to the degradation of drugs by progressing to lysosomes. In this review, we discussed mechanisms by which nanocarriers are endocytosed, the mechanisms of endosomal escape, and more importantly, the strategies that can be and have been employed for their escape from the endosomes are summarized. Like other nanocarriers, polymeric micelles can be designed for endosomal escape, however, a careful control is needed in their design to balance between the possible toxicity and endosomal escape efficiency. Keeping this in view, polyion complex micelles, and polymers that have the ability to escape the endosome, are fully discussed. Finally, we provided some perspectives for designing the polymeric micelles for efficient cytoplasmic delivery of bioactive agents through endosomal escape.

Experimental investigation on the air-cushion effect during free fall of a trimaran section using an air escape control method

Slamming is common when a trimaran sails in rough seas. Owing to the semiclosed structural shape of a trimaran, an air cushion can easily form, reducing the slamming pressure. Therefore, the air-cushion effect during water entry of a trimaran section is investigated in this work. To study the air-cushion effect quantitively, an experimental procedure together with a novel model design is presented. Partly opened acrylic plates are pasted to a trimaran section model and the opening size is changed to control the efficiency of air escape from the front and rear of the model. The formation of the air cushion and the transformation of the water surface are captured by a high-speed camera. The influences of the opening size and the initial release height of the model on the slamming pressure are analyzed to investigate the air-cushion effect. The most novel idea of the present paper is the design of the asymmetrical openings on the front and rear acrylic plates pasted to the symmetrical model. Through this design, the significant difference produced by the air-cushion effect can be clearly distinguished by comparing the left and right halves of the model. Results show that the air cushion can postpone the emergence time of the pressure peak, decrease the magnitude of the pressure peak, and magnify the pressure fluctuation phenomenon.

Discovery of a Cyclic Cell-Penetrating Peptide with Improved Endosomal Escape and Cytosolic Delivery Efficiency.

Cyclic cell-penetrating peptide 12 (CPP12) is highly efficient for the cytosolic delivery of a variety of cargo molecules into mammalian cells in vitro and in vivo. However, its cytosolic entry efficiency is substantially reduced at lower concentrations or in the presence of serum proteins. In this study, CPP12 analogs were prepared by replacing its hydrophobic residues with amino acids of varying hydrophobicity and evaluated for cellular entry. Substitution of l-3-benzothienylalanine (Bta) for l-2-naphthylalanine (Nal) resulted in CPP12-2, which exhibits up to 3.8-fold higher cytosolic entry efficiency than CPP12, especially at low CPP concentrations; thanks to improved endosomal escape efficiency. CPP12-2 is well suited for the cytosolic delivery of highly potent cargos to achieve biological activity at low concentrations.

Resonant Tunneling Diodes: Mid-Infrared Sensing at Room Temperature.

Resonant tunneling diode photodetectors appear to be promising architectures with a simple design for mid-infrared sensing operations at room temperature. We fabricated resonant tunneling devices with GaInAsSb absorbers that allow operation in the μm range with significant electrical responsivity of 0.97 A/W at 2004 nm to optical readout. This paper characterizes the photosensor response contrasting different operational regimes and offering a comprehensive theoretical analysis of the main physical ingredients that rule the sensor functionalities and affect its performance. We demonstrate how the drift, accumulation, and escape efficiencies of photogenerated carriers influence the electrostatic modulation of the sensor’s electrical response and how they allow controlling the device’s sensing abilities.

Collective response of microrobotic swarms to external threats

Many animal species organize within groups to achieve advantages compared to being isolated. Such advantages can be found e.g. in collective responses which are less prone to individual failures or noise and thus provide better group performance. Inspired by social animals, here we demonstrate with a swarm of microrobots made from programmable active colloidal particles (APs) that their escape from a hazardous area can originate from a cooperative group formation. As a consequence, the escape efficiency remains almost unchanged even when half of the APs are not responding to the threat. Our results not only confirm that incomplete or missing individual information in robotic swarms can be compensated by other group members but also suggest strategies to increase the responsiveness and fault-tolerance of robotic swarms when performing tasks in complex environments.

Drastic decline in sera neutralization against SARS-CoV-2 Omicron variant in Wuhan COVID-19 convalescents

ABSTRACT Global concern has been raised by the emergence and rapid transmission of the heavily mutated SARS-CoV-2 Omicron variant (B.1.1.529). So far, the infection features and immune escape ability of the Omicron variant have not been extensively studied. Here, we produced the Omicron pseudovirus and compared its entry, membrane fusion, and immune escape efficiency with the original strain and the dominating Delta variant. We found the Omicron variant showed slightly higher infectivity than the Delta variant and a similar ability to compete with the Delta variant in using Angiotensin-converting enzyme 2 (ACE2) in a BHK21-ACE2 cell line. However, the Omicron showed a significantly reduced fusogenicity than the original strain and the Delta variant in both BHK21-ACE2 and Vero-E6 cells. The neutralization assay testing the Wuhan convalescents’ sera one-year post-infection showed a more dramatic reduction (10.15 fold) of neutralization against the Omicron variant than the Delta variant (1.79 fold) compared with the original strain with D614G. Notably, immune-boosting through three vaccine shots significantly improved the convalescents’ immunity against the Omicron variants. Our results reveal a reduced fusogenicity and a striking immune escape ability of the Omicron variant, highlighting the importance of booster shots against the challenge of the SARS-CoV-2 antigenic drift.

Hyaluronic acid-modified redox-sensitive hybrid nanocomplex loading with siRNA for non-small-cell lung carcinoma therapy

A novel hyaluronic acid (HA)-modified hybrid nanocomplex HA-SeSe-COOH/siR-93C@PAMAM, which could efficiently deliver siRNA into tumor cells via a redox-mediated intracellular disassembly, was constructed for enhanced antitumor efficacy. Thereinto, siR-93C (siRNA) and positive PAMAM were firstly mixed into the electrostatic nano-intermediate, and then diselenide bond (-SeSe-)-modified HA was coved to shield excessive positive charges. This hybrid nanocomplex displayed uniform dynamic sizes, high stability, controlled zeta potential and narrow PDI distribution. Moreover, the -SeSe- linkage displayed GSH/ROS dual responsive properties, improving intracellular trafficking of siRNA. In vitro assays in A549 cell line presented that HA-SeSe-COOH/siR-93C@PAMAM has low cytotoxicity, rapid lysosomal escape and significant transfection efficiency; besides, an efficient proliferation inhibition ability and enhanced apoptosis. Furthermore, in animal studies, this negative-surfaced hybrid nanocomplex showed a prolonged circulation in blood and improved inhibition of tumor growth. All these results verified our hypothesis in this study that diselenide bonds-modified HA could promote not only stability and safety of nanoparticles in vivo but also intracellular behavior of siRNA via redox-dual sensitive properties; furthermore, this hybrid nanocomplex provided a visible potential approach for siRNA delivery in the antitumor field.