Biological Applications Research Articles

Bioelectret Materials and Their Bioelectric Effects for Tissue Repair: A Review.

Biophysical and clinical medical studies have confirmed that biological tissue lesions and trauma are related to the damage of an intrinsic electret (i.e., endogenous electric field), such as wound healing, embryonic development, the occurrence of various diseases, immune regulation, tissue regeneration, and cancer metastasis. As exogenous electrical signals, such as conductivity, piezoelectricity, ferroelectricity, and pyroelectricity, bioelectroactives can regulate the endogenous electric field, thus controlling the function of cells and promoting the repair and regeneration of tissues. Materials, once polarized, can harness their inherent polarized static electric fields to generate an electric field through direct stimulation or indirect interactions facilitated by physical signals, such as friction, ultrasound, or mechanical stimulation. The interaction with the biological microenvironment allows for the regulation and compensation of polarized electric signals in damaged tissue microenvironments, leading to tissue regeneration and repair. The technique shows great promise for applications in the field of tissue regeneration. In this paper, the generation and change of the endogenous electric field and the regulation of exogenous electroactive substances are expounded, and the latest research progress of the electret and its biological effects in the field of tissue repair include bone repair, nerve repair, drug penetration promotion, wound healing, etc. Finally, the opportunities and challenges of electret materials in tissue repair were summarized. Exploring the research and development of new polarized materials and the mechanism of regulating endogenous electric field changes may provide new insights and innovative methods for tissue repair and disease treatment in biological applications.

Mitochondria-targeted and photo-activated CO release for synergistic photodynamic therapy

In photodynamic therapy, organelle-targeted strategies are allowed to selectively impair specific cell functions, leading to efficiently eliminating cancer cells. Carbon monoxide (CO), as a highly toxic gaseous signaling molecule, often plays an active role in enhancing the sensitivity of cancer cells during cancer treatment, making cells prone to apoptosis. Therefore, we report a photosensitizer Cy-Mn, which possesses the capability of photoactivatable CO releasing and 1O2 generation. Furthermore, during the treatment, Cy-Mn has perfect synergistic effect on the cancer cells for enhanced photodynamic therapy. Under 760nm laser irradiation, Cy-Mn released manganese ions, which altered the permeability transition pore of the mitochondria membrane, reduced the mitochondrial membrane potential, and exhibited severe phototoxicity. Additionally, we investigated the imaging of Cy-Mn in vivo, and found Cy-Mn displayed excellent fluorescence imaging capabilities, which further promotes its biological applications in imaging-guided PDT.

An instructional design strategy for optimization of GelMA hydrogels material properties

Gelatin methacryloyl (GelMA) hydrogels have become a key biomaterial in various biomedical fields, such as tissue regeneration engineering and drug delivery, due to their exceptional biocompatibility and customizable physical and chemical characteristics. The resemblance of GelMA to the extracellular matrix and its ability to be cured with light make it a popular choice for researchers. This research delves into how the material properties of GelMA hydrogels, specifically the density of the molecular network (DMN), impact their performance in different applications. By preparing GelMA hydrogels with different degrees of substitution (DS) and concentrations, and utilizing these attributes as key measurable parameters for determining DMN, we conducted a comprehensive evaluation of properties relevant to biological applications. We also suggest strategies for optimizing these properties. Through the application of the DMN theory, we show that precise control over material properties allows for tailored customization of GelMA hydrogels for specific tissue engineering and drug delivery purposes. This study offers valuable insights into the optimization of GelMA hydrogel properties to advance the field of tissue engineering and improve customization of cellular microenvironments.

Gelatin and lipidoid integrate to create gelasomes to enhance siRNA delivery with low toxicity

RNAi therapeutics possess the potential to cure many uncurable human diseases. For instance, RNAi therapeutics using liposomes showed remarkable survival benefits in patients with liver diseases. However, the extension of liposomes to deliver RNA to cure other ailments has largely been unsuccessful. Therefore, researchers are focusing on designing and testing different combinations of materials for versatile RNA delivery applications. Yet, an efficient and safe RNA delivery platform has not been identified. In this work, we have developed a new class of RNA-delivery vehicle called “Gelasomes,” using an incongruous combination of gelatin and lipidoid to exploit each material's unique properties while overcoming their inherent limitations. The low in vivo toxicity of Gelasomes is attributed to the exterior gelatin layers that shield the exposure of cationic lipidoid-siRNA clusters and yet present a biocompatible surface. Indeed, toxicity studies in mice indicate that repeated administration of Gelasomes (up to 48 mg/kg BW) is well-tolerated with no notable changes in body weight, hematology, or serum chemistry. Interestingly, the gelatin outer layer efficiently protects siRNA from serum degradation (48 h), preserving its functionality beyond two months of storage. Notably, Gelasomes possess dual siRNA conjugation modes, i.e., electrostatic binding with lipidoid core and covalent attachment to gelatin surface. The bivalency coupled with lipidoids' high transfection efficiency rendered Gelasomes with remarkably high gene silencing efficiency (>90 %) at very low treatment doses in vitro (40 μg/mL). In vivo studies further confirmed the high gene silencing ability of Gelasomes in non-small cell lung tumor mouse models. This new platform is tunable on all fronts: size, degree of surface coating, and biomolecule functionalization. Truncating the lipidoid C14-tail to a C8-tail yielded Gelasomes of reduced size. As lipidoids with different carbon lengths are synthesizable, we can develop a library of Gelasomes with different sizes. The surface coating with less gelatin resulted in high transfection efficiency at low doses of Gelasomes. The structure of Gelasomes offers chemical handles to couple target-specific molecules like antibodies to tune their properties for efficient biological application.

Research Progress of Cyanine-based Near-Infrared Fluorescent Probes for Biological Application.

Cyanine-based near-infrared (NIR) fluorescent probes have played vital roles in biological application due to their low interference from background fluorescence, deep tissue penetration, high sensitivity, and minimal photodamage to biological samples. They are widely utilized in molecular recognition, medical diagnosis, biomolecular detection, and biological imaging. Herein, we provide a review of recent advancements in cyanine-based NIR fluorescent probes for the detection of pH, cells, tumor as well as their application in photothermal therapy (PTT) and photodynamic therapy (PDT).

Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.

Genome editing technologies based on DNA-dependent polymerases (DDPs) could offer several benefits compared with other types of editors to install diverse edits. Here, we develop click editing, a genome writing platform that couples the advantageous properties of DDPs with RNA-programmable nickases to permit the installation of a range of edits, including substitutions, insertions and deletions. Click editors (CEs) leverage the 'click'-like bioconjugation ability of HUH endonucleases with single-stranded DNA substrates to covalently tether 'click DNA' (clkDNA) templates encoding user-specifiable edits at targeted genomic loci. Through iterative optimization of the modular components of CEs and their clkDNAs, we demonstrate the ability to install precise genome edits with minimal indels in diverse immortalized human cell types and primary fibroblasts with precise editing efficiencies of up to ~30%. Editing efficiency can be improved by rapidly screening clkDNA oligonucleotides with various modifications, including repair-evading substitutions. Click editing is a precise and versatile genome editing approach for diverse biological applications.

Exploring Nanohydrogels: Crafting Innovations for Biomedical Advancements

AbstractHere in, we discuss different types of nanohydrogels in the field of biomedicine, highlighting their unique properties such as size, biocompatibility, and tuneable drug release capabilities. Nanohydrogels are considered versatile tools for applications in tissue engineering, drug delivery, and diagnostics, with the ability to target specific sites and release therapeutic agents in a controlled manner. The controlled‐release characteristics of nanohydrogels are particularly emphasized for targeted drug delivery, potentially transforming therapies for conditions like cancer by maximizing treatment efficacy while minimizing damage to healthy tissues. Several research articles published on nanohydrogels has been steadily increasing over the years, indicating growing interest and research activity in this field. This review provides an overview of contemporary methodologies employed in nanohydrogel design, their utilization in biomedical contexts, and their pivotal role in shaping advanced therapeutics. Recent investigations on nanohydrogels are scrutinized for their extensive utility in devising strategies conducive to finely tuned biological applications. Challenges and obstacles associated with the nanohydrogel technology in biomedicine also has been discussed. Cautious handling of the interaction of nanohydrogels with complex biological systems is also emphasized for the successful integration of this technology into medical applications.

A hierarchal model for bacterial cell inactivation in solution by direct and indirect treatment using cold atmospheric plasmas

Cold atmospheric plasma devices have shown promise for a variety of plasma medical applications, including wound healing and bacterial inactivation often performed in liquids. In the latter application, plasma-produced reactive oxygen and nitrogen species (RONS) interact with and damage bacterial cells, though the exact mechanism by which cell damage occurs is unclear. Computational models can help elucidate relationships between plasma-produced RONS and cell killing by enabling direct comparison between dissimilar plasma devices and by examining the effects of changing operating parameters in these devices. In biological applications, computational models of plasma-liquid interactions would be most effective in design and optimization of plasma devices if there is a corresponding prediction of the biological outcome. In this work, we propose a hierarchal model for planktonic bacterial cell inactivation by plasma produced RONS in liquid. A previously developed reaction mechanism for plasma induced modification of cysteine was extended to provide a basis for cell killing by plasma-produced RONS. Results from the model are compared to literature values to provide proof of concept. Differences in time to bacterial inactivation as a function of plasma operating parameters including gas composition and plasma source configuration are discussed. Results indicate that optimizing gas-phase reactive nitrogen species production may be key in the design of plasma devices for disinfection.

Multi-resonance plasmonic refractive index sensor based on maze-shaped resonators for biological applications

Multi-resonance plasmonic refractive index sensor based on maze-shaped resonators for biological applications

Posttranscriptional tuning of gene expression over a large dynamic range in synthetic tobacco chloroplast operons.

Achieving optimally balanced gene expression within synthetic operons requires regulatory elements capable of providing a spectrum of expression levels. In this study, we investigate the expression of gfp reporter gene in tobacco chloroplasts, guided by variants of the plastid atpH 5' UTR, which harbors a binding site for PPR10, a protein that activates atpH at the posttranscriptional level. Our findings reveal that endogenous tobacco PPR10 confers distinct levels of reporter activation when coupled with the tobacco and maize atpH 5' UTRs in different design contexts. Notably, high GFP expression was not coupled to the stabilization of monocistronic gfp transcripts in dicistronic reporter lines, adding to the evidence that PPR10 activates translation via a mechanism that is independent of its stabilization of monocistronic transcripts. Furthermore, the incorporation of a tRNA upstream of the UTR nearly abolishes gfp mRNA (and GFP protein), presumably by promoting such rapid RNA cleavage and 5' exonucleolytic degradation that PPR10 had insufficient time to bind and protect gfp RNA, resulting in a substantial reduction in GFP accumulation. When combined with a mutant atpH 5' UTR, the tRNA leads to an exceptionally low level of transgene expression. Collectively, this approach allows for tuning of reporter gene expression across a wide range, spanning from a mere 0.02-25% of the total soluble cellular protein. These findings highlight the potential of employing cis-elements from heterologous species and expand the toolbox available for plastid synthetic biology applications requiring multigene expression at varying levels.

Se-functionalized ZIF-8 nanoparticles:synthesis, characterization and disruption of biofilms and quorum sensing inSerratia marcescens.

The majority of research on nanomaterials has been concentrated on metal nanoparticles since they are easily made and manipulated. Nanomaterials have shown a wide range of applications in biology. Nevertheless, their bioactivity declines due to their extreme susceptibility to chemical and physical stimuli. The goal of encapsulating these nanomaterials in a matrix is gradually being pursued, which boosts their affordability, stability, and usability. Metal-organic frameworks, often known as MOFs, have the potential to be the best platforms for encapsulating metal nanoparticles due to their well-defined frameworks, persistent porosity, and flexibility in modification. In this investigation, we report the synthesis and optimization of polyvinylpyrrolidone (PVP)-stabilized Se (0) nanoparticles and novel Se@ZIF-8 by chemical method. The sizes and morphologies of Se (0) and Se@ZIF-8 were affected by the ratios of Se/Zn2+ and [hmim]/Zn2+ used. The optimized Se@ZIF-8 nanoparticles exhibited a particle size and zeta potential of 319 nm and -34 mv respectively. Transmission electron microscopy displayed spherical morphology for Se(0) nanoparticles, whereas the surface morphology of novel Se@ZIF-8 nanoparticles was drastically changed to hexagonal shaped structures with smooth surface morphologies in scanning electron microscopy. The DTA, TG/DTG, XRD analysis confirmed the presence of novel Se incorporated ZIF-8 nanoparticulate framework. The synthesized novel Se@ZIF-8 nanoparticles showed efficient antibacterial activity as evidenced by low MIC values. Interestingly, these Se@ZIF-8 NPs not only inhibited biofilm formation in S. marcescens, but also effectively eradicated mature biofilms by degrading the eDNA of the EPS layer. It was observed that Se@ZIF-8 targeted the Quroum Sensing pathway and reduced its associated virulence factors production. This work opens up a different approach of Se@ZIF-8 nanoparticles as novel antibiotics to treat biofilm-associated infections caused by S. marcescens and offer a solution for antimicrobial resistance.&#xD.

Tailor-Made Gold Nanomaterials for Applications in Soft Bioelectronics and Optoelectronics.

In modern nanoscience and nanotechnology, gold nanomaterials are indispensable building blocks that have demonstrated a plethora of applications in catalysis, biology, bioelectronics, and optoelectronics. Gold nanomaterials possess many appealing material properties, such as facile control over their size/shape and surface functionality, intrinsic chemical inertness yet with high biocompatibility, adjustable localized surface plasmon resonances, tunable conductivity, wide electrochemical window, etc. Such material attributes have been recently utilized for designing and fabricating soft bioelectronics and optoelectronics. This motivates to give a comprehensive overview of this burgeoning field. The discussion of representative tailor-made gold nanomaterials, including gold nanocrystals, ultrathin gold nanowires, vertically aligned gold nanowires, hard template-assisted gold nanowires/gold nanotubes, bimetallic/trimetallic gold nanowires, gold nanomeshes, and gold nanosheets, is begun. This is followed by the description of various fabrication methodologies for state-of-the-art applications such as strain sensors, pressure sensors, electrochemical sensors, electrophysiological devices, energy-storage devices, energy-harvesting devices, optoelectronics, and others. Finally, the remaining challenges and opportunities are discussed.

Liquid Gallium-Assisted Pyrolysis of MOF Affording CNT Non-Hollow Frameworks in High Yields for High-Performance Sodium-Ion Battery Anode.

Carbon materials have great potential for applications in energy, biology, and environment due to their excellent chemical and physical properties. Their preparation by carbonization methods encounters limitations and the carbon loss during pyrolysis in the form of gaseous molecules results in low yield of carbon materials. Herein a low-energy (600°C) and high-yield (82wt.%) carbonization strategy is developed using liquid gallium-assisted pyrolysis of metal-organic frameworks (MOFs) affording the N-doped carbon nanotube (CNT) non-hollow frameworks encapsulating Co nanoparticles. The liquid gallium layer offers protection against air, promotes heat transfer, and limits the escape of small carbonaceous gaseous molecules, which greatly improve the yields of the pyrolysis reaction. Experimental and theoretical results reveal that the synergistic interaction between CNTs and N/O-containing groups gives a non-hollow framework composed of N/O-enriched and open CNTs (NOCNTF-15, 15 denotes the 15mm thickness of the liquid gallium layer during the pyrolysis) with high specific capacity (185mAhg-1 at 10Ag-1) and ultra-stable cyclability (stable operation at 10Ag-1 and 50°C for 20000 cycles). This study provides a unique approach to carbonization that facilitates the practical application of low-cost CNTs and other MOFs-derived carbon materials in high-performance sodium-ion batteries (SIBs).

Influence of laser power and beam path under nonuniform AC electric fields on 3D microvortex flow

This paper describes the effect of optical light on the generation and manipulation of microvortex flow named ‘twin opposing microvortex’ (TOMV) flow. This opto-electrohydrodynamic (OEHD) technique combines optical light, i.e. infrared (IR) laser (1064 nm), with non-uniform AC electric fields generated from a pair of indium tin oxide (ITO) electrodes. When the IR laser beam passes through the electric fields, a rapid and three-dimensional (3D) vortex flow is generated in a microchamber. When the laser beam passes through the electric fields, especially the exposed ITO electrode, the direction of the TOMV flow as well as its strength are controlled. With an AC signal of 107 kHz and various voltages below a peak-to-peak voltage of 10 V, laser power is varied up to 1.5 W and the path of a laser beam relative to the electrode (300 μm long and 16 μm wide) is manipulated. The maximum in-plane velocity outside the electrode region was obtained by micron-resolution particle image velocimetry (μPIV). When the laser beam passes through the left or right side of the lower electrode, the TOMV flow field rotates counterclockwise or clockwise, respectively. Applying optical light on an ITO electrode creates in situ and on-demand microvortex flow, which increases the feasibility of OEHD technique in various biological and chemical applications (e.g., mixing and delivering nanofluids in microfluidic devices).

Multicolor Tuning of Perylene Diimides Dyes for Targeted Organelle Imaging In Vivo.

The adjustment of the emission wavelengths and cell permeability of the perylene diimides (PDI) for multicolor cell imaging is a great challenge. Herein, based on a bay-region substituent engineering strategy, multicolor perylene diimides (MCPDI) were rationally designed and synthesized by introducing azetidine substituents on the bay region of PDIs. With the fine-tuned electron-donating ability of the azetidine substituents, these MCPDI showed high brightness, orange, red, and near infrared (NIR) fluorescence along with Stokes shifts increasing from 35 to 110 nm. Interestingly, azetidine substituents distorted to the plane of the MCPDI dyes, and the twist angle of monosubstituted MCPDI was larger than that of disubstituted MCPDI, which might efficiently decrease their π-π stacking. Moreover, all of these MCPDI dyes were cell-permeable and selectively stained various organelles for multicolor imaging of multiple organelles in living cells. Two-color imaging of lipid droplets (LDs) and other organelles stained with MCPDI dyes was performed to reveal the interaction between the LDs and other organelles in living cells. Furthermore, a NIR-emitting MCPDI dye with a mitochondria-targeted characteristic was successfully applied for tumor-specific imaging. The facile synthesis, excellent stability, high brightness, tunable fluorescence emission, and Stokes shifts make these MCPDI promising fluorescent probes for biological applications.

Streamflow in the United States: Characteristics, trends, regime shifts, and extremes

Long-term streamflow observations contain essential information for understanding hydrological changes and managing water resources. A continental-scale dataset or analysis of temporal streamflow change is still lacking across hydrologic gauges in the Conterminous United States (CONUS). Here, we compiled 70 years of streamflow records from 1951 to 2021 at ~ 8000 hydrologic stations across the CONUS and characterized temporal trends, regime shifts, and extreme events using a Bayesian time series analysis algorithm. We found that the occurrences of sudden streamflow changes (e.g., regime shifts and extreme events) have been increasing with time across the CONUS. In addition, we derived 181 streamflow indicators that are valuable for hydrological and biological applications, such as the duration and frequency of high or low streamflow events. The Mississippi River Basin, especially the middle and lower parts, was a hot spot of high-frequency high-flow events. Overall, we anticipate the dataset generated here offers valuable information for understanding and quantifying changes in water resources across the CONUS.

Biological Applications, In Vitro Cytotoxicity, Cellular Uptake, and Apoptotic Pathway Studies Induced by Ternary Cu (II) Complexes Involving Triflupromazine with Biorelevant Ligands.

The novel mixed-ligand complexes derived from the parent antidepressant phenothiazine drug triflupromazine (TFP) were synthesized along with the secondary ligands glycine and histidine. [Cu(TFP)(Gly)Cl]·2H2O (1) and [Cu(TFP)(His)Cl]·2H2O (2) were examined for their in vitro biological properties. Cyclic voltammetry was used to study the binding of both complexes to CT-DNA. The two complexes were examined for antiviral, antiparasite, and anti-inflammatory applications. An in vitro cytotoxicity study on two different cancer cell lines, MCF-7, HepG2, and a normal cell line, HSF, shows promising selective cytotoxicity for cancer cells. An investigation of the cell cycle and apoptosis rates was evaluated by flow cytometry with Annexin V-FITC/Propidium Iodide (PI) staining of the treated cells. Gene expression and western blotting were carried out to determine the expression levels of the pro-apoptotic markers and the anti-apoptotic marker Bcl2. The tested complexes decreased cell viability and triggered apoptosis in human tumor cell lines. Molecular docking was also used to simulate Bcl2 inhibition. Finally, complex (2) has potent antitumor effects on human tumor cells, especially against HepG2 cells, as seen in the cellular drug uptake assay. Consequently, complex (2) may prove useful against cancer, especially liver cancer. For further understanding, it needs to be explored in vivo.

Engineering a Self-Assembled Protein Cage for Targeted Dual Functionalization.

Self-assembled protein cages are attractive scaffolds for organizing various proteins of interest (POIs) toward applications in synthetic biology and medical science. However, specifically attaching multiple POIs to a single protein cage remains challenging, resulting in diversity among the functionalized particles. Here, we present the engineering of a self-assembled protein cage, DTMi3ST, capable of independently recruiting two different POIs using SpyCatcher (SC)/SpyTag (ST) and DogCatcher (DC)/DogTag (DT) chemistries, thereby reducing variability between assemblies. Using fluorescent proteins as models, we demonstrate controlled targeting of two different POIs onto DTMi3ST protein cages both in vitro and inside living cells. Furthermore, dual functionalization of the DTMi3ST protein cage with a membrane-targeting peptide and β-galactosidase resulted in the construction of membrane-bound enzyme assemblies in Escherichia coli, leading to a 69.6% enhancement in substrate utilization across the membrane. This versatile protein cage platform provides dual functional nanotools for biological and biomedical applications.

Fabrication of heterocellular spheroids with controllable core-shell structure using inertial focusing effect for scaffold-free 3D cell culture models.

Three-dimensional (3D) cell culture models capable of emulating the biological functions of natural tissues are pivotal in tissue engineering and regenerative medicine. Despite progress, the fabrication of in vitro heterocellular models that mimic the intricate structures of natural tissues remains a significant challenge. In this study, we introduce a novel, scaffold-free approach leveraging the inertial focusing effect in rotating hanging droplets for the reliable production of heterocellular spheroids with controllable core-shell structures. Our method offers precise control over the core-shell spheroid's size and geometry by adjusting the cell suspension density and droplet morphology. We successfully applied this technique to create hair follicle organoids, integrating dermal papilla cells within the core and epidermal cells in the shell, thereby achieving markedly enhanced hair inducibility compared to mixed-structure models. Furthermore, we have developed melanoma tumor spheroids that accurately mimic the dynamic interactions between tumor and stromal cells, showing increased invasion capabilities and altered expressions of cellular adhesion molecules and proteolytic enzymes. These findings underscore the critical role of cellular spatial organization in replicating tissue functionality in vitro. Our method represents a significant advancement towards generating heterocellular spheroids with well-defined architectures, offering broad implications for biological research and applications in tissue engineering.

Progress in Preparation and Application of Fucoxanthin from Marine Algae

This paper reviews the preparation and application of fucoxanthin from marine algae. It details the progress in various aspects, including biological origin, chemical structure, production process, structure identification, physiological functions, and applications, based on experimental data and clinical applications. The paper also highlights technical challenges and issues in fucoxanthin research, such as improving yield, purity, and structure modification. Finally, it summarizes the research findings and discusses future research directions and prospects for fucoxanthin from marine algae, offering suggestions for further studies.