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International Journal of Molecular... Jul 2023The outer membrane of Gram-negative bacteria contains a variety of pore-forming structures collectively referred to as porins. Some of these are voltage dependent, but...
The outer membrane of Gram-negative bacteria contains a variety of pore-forming structures collectively referred to as porins. Some of these are voltage dependent, but weakly so, closing at high voltages. Triplin, a novel bacterial pore-former, is a three-pore structure, highly voltage dependent, with a complex gating process. The three pores close sequentially: pore 1 at positive potentials, 2 at negative and 3 at positive. A positive domain containing 14 positive charges (the voltage sensor) translocates through the membrane during the closing process, and the translocation is proposed to take place by the domain entering the pore and thus blocking it, resulting in the closed conformation. This mechanism of pore closure is supported by kinetic measurements that show that in the closing process the voltage sensor travels through most of the transmembrane voltage before reaching the energy barrier. Voltage-dependent blockage of the pores by polyarginine, but not by a 500-fold higher concentrations of polylysine, is consistent with the model of pore closure, with the sensor consisting mainly of arginine residues, and with the presence, in each pore, of a complementary surface that serves as a binding site for the sensor.
Topics: Humans; Ion Channel Gating; Porins; Thiourea; Translocation, Genetic
PubMed: 37511231
DOI: 10.3390/ijms241411473 -
Advanced Science (Weinheim,... Nov 2023Modulating the inflammatory microenvironment can inhibit the process of inflammatory diseases (IDs). A tri-cross-linked inflammatory microenvironment-responsive hydrogel...
Triple Cross-linked Dynamic Responsive Hydrogel Loaded with Selenium Nanoparticles for Modulating the Inflammatory Microenvironment via PI3K/Akt/NF-κB and MAPK Signaling Pathways.
Modulating the inflammatory microenvironment can inhibit the process of inflammatory diseases (IDs). A tri-cross-linked inflammatory microenvironment-responsive hydrogel with ideal mechanical properties achieves triggerable and sustained drug delivery and regulates the inflammatory microenvironment. Here, this study develops an inflammatory microenvironment-responsive hydrogel (OD-PP@SeNPs) composed of phenylboronic acid grafted polylysine (PP), oxidized dextran (OD), and selenium nanoparticles (SeNPs). The introduction of SeNPs as initiators and nano-fillers into the hydrogel results in extra cross-linking of the polymer network through hydrogen bonding. Based on Schiff base bonds, Phenylboronate ester bonds, and hydrogen bonds, a reactive oxygen species (ROS)/pH dual responsive hydrogel with a triple-network is achieved. The hydrogel has injectable, self-healing, adhesion, outstanding flexibility, suitable swelling capacity, optimal biodegradability, excellent stimuli-responsive active substance release performance, and prominent biocompatibility. Most importantly, the hydrogel with ROS scavenging and pH-regulating ability protects cells from oxidative stress and induces macrophages into M2 polarization to reduce inflammatory cytokines through PI3K/AKT/NF-κB and MAPK pathways, exerting anti-inflammatory effects and reshaping the inflammatory microenvironment, thereby effectively treating typical IDs, including S. aureus infected wound and rheumatoid arthritis in rats. In conclusion, this dynamically responsive injectable hydrogel with a triple-network structure provides an effective strategy to treat IDs, holding great promise in clinical application.
Topics: Animals; Rats; NF-kappa B; Selenium; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Hydrogels; Reactive Oxygen Species; Staphylococcus aureus; MAP Kinase Signaling System; Nanoparticles
PubMed: 37740428
DOI: 10.1002/advs.202303167 -
Journal of Functional Biomaterials Dec 2023With the phase-out of amalgam and the increase in minimally invasive dentistry, there is a growing need for high-strength composite materials that can kill residual...
With the phase-out of amalgam and the increase in minimally invasive dentistry, there is a growing need for high-strength composite materials that can kill residual bacteria and promote tooth remineralization. This study quantifies how antibacterial polylysine (PLS) and re-mineralizing monocalcium phosphate monohydrate (MCPM) affect biofilms and the strength of dental composites. For antibacterial studies, the MCPM-PLS filler percentages were 0-0, 8-4, 12-6, and 16-8 wt% of the composite filler phase. Composite discs were immersed in 0.1% sucrose-supplemented broth containing (UA159) and incubated in an anaerobic chamber for 48 h. Surface biomass was determined by crystal violet (CV) staining. Growth medium pH was measured at 24 and 48 h. Biofilm bacterial viability (CFU), exo-polysaccharide (water-soluble glucan (WSG) and water-insoluble glucan (WIG)), and extracellular DNA (eDNA) were quantified. This was by serial dilution plate counting, phenol-sulfuric acid microassay, and fluorometry, respectively. The biaxial flexural strengths were determined after water immersion for 1 week, 1 month, and 1 year. The MCPM-PLS wt% were 8-4, 8-8, 16-4 and 16-8. The normalized biomass, WSG, and WIG showed a linear decline of 66%, 64%, and 55%, respectively, as the PLS level increased up to 8%. The surrounding media pH (4.6) was all similar. A decrease in bacterial numbers with the 12-6 formula and a significant reduction with 16-8 compared to the 0-0 formulation was observed. The eDNA concentrations in biofilms formed on 12-6 and 16-8 formulations were significantly less than the 0-0 control and 8-4 formulations. Doubling MCPM and PLS caused a 14 and 19% reduction in strength in 1 week, respectively. Average results were lower at 1 month and 1 year but affected less upon doubling MCPM and PLS levels. Moreover, a 4% PLS may help to reduce total biomass and glucan levels in biofilms on the above composites. Higher levels are required to reduce eDNA and provide bactericidal action, but these can decrease early strength.
PubMed: 38248680
DOI: 10.3390/jfb15010013 -
Advanced Science (Weinheim,... May 2024Non-antibiotic strategies are desperately needed to treat post-traumatic osteomyelitis (PTO) due to the emergence of superbugs, complex inflammatory microenvironments,...
Non-antibiotic strategies are desperately needed to treat post-traumatic osteomyelitis (PTO) due to the emergence of superbugs, complex inflammatory microenvironments, and greatly enriched biofilms. Previously, growing evidence indicated that quorum sensing (QS), a chemical communication signal among bacterial cells, can accelerate resistance under evolutionary pressure. This study aims to develop a medical dressing to treat PTO by inhibiting QS and regulating the inflammatory microenvironment, which includes severe oxidative stress and acid abscesses, through a reactive oxygen species (ROS)-responsive bond between N1- (4-borobenzoyl)-N3-(4-borobenzoyl)-the N1, the N1, N3, N3-tetramethylpropane-1,3-diamine (TSPBA) and polyvinyl alcohol (PVA), and the amino side chain of hyperbranched polylysine (HBPL). Physically enclosed QS inhibitors subsequently exerted the antibacterial effects. This hydrogel can scavenge hydrogen peroxide (HO), superoxide anion free radical (·O ), hydroxyl radicals (·OH) and 2,2-di(4-tert-octylphenyl)-1-picryl-hydrazyl (DPPH) to reduce oxidative stress and inhibit "bacteria-to-bacteria communication", thus clearing planktonic bacteria and biofilms, accelerating bacterial plasmolysis, reducing bacterial virulence and interfering with membrane transport. After in vivo treatment with hydrogel, nearly all bacteria are eliminated, inflammation is effectively inhibited, and osteogenesis and bone repair are promoted to facilitate recovery from PTO. The work demonstrates the clinical translational potential of the hydrogel in the treatment of drug-resistant bacteria induced PTO.
Topics: Quorum Sensing; Hydrogels; Osteomyelitis; Animals; Mice; Disease Models, Animal; Anti-Bacterial Agents; Oxidative Stress; Biofilms; Reactive Oxygen Species; Rats; Male
PubMed: 38482752
DOI: 10.1002/advs.202307969 -
Bioengineering & Translational Medicine May 2024Joint diseases, such as osteoarthritis, often require delivery of drugs to chondrocytes residing within the cartilage. However, intra-articular delivery of drugs to...
Joint diseases, such as osteoarthritis, often require delivery of drugs to chondrocytes residing within the cartilage. However, intra-articular delivery of drugs to cartilage remains a challenge due to their rapid clearance within the joint. This problem is further exacerbated by the dense and negatively charged cartilage extracellular matrix (ECM). Cationic nanocarriers that form reversible electrostatic interactions with the anionic ECM can be an effective approach to overcome the electrostatic barrier presented by cartilage tissue. For an effective therapeutic outcome, the nanocarriers need to penetrate, accumulate, and be retained within the cartilage tissue. Nanocarriers that adhere quickly to cartilage tissue after intra-articular administration, transport through cartilage, and remain within its full thickness are crucial to the therapeutic outcome. To this end, we used ring-opening polymerization to synthesize branched poly(l-lysine) (BPL) cationic nanocarriers with varying numbers of poly(lysine) branches, surface charge, and functional groups, while maintaining similar hydrodynamic diameters. Our results show that the multivalent BPL molecules, including those that are highly branched (i.e., generation two), can readily adhere and transport through the full thickness of cartilage, healthy and degenerated, with prolonged intra-cartilage retention. Intra-articular injection of the BPL molecules in mouse knee joint explants and rat knee joints showed their localization and retention. In summary, this study describes an approach to design nanocarriers with varying charge and abundant functional groups while maintaining similar hydrodynamic diameters to aid the delivery of macromolecules to negatively charged tissues.
PubMed: 38818117
DOI: 10.1002/btm2.10612 -
Journal of Nanobiotechnology May 2024Approximately 80 percent of the total RNA in cells is ribosomal RNA (rRNA), making it an abundant and inexpensive natural source of long, single-stranded nucleic acid,...
Approximately 80 percent of the total RNA in cells is ribosomal RNA (rRNA), making it an abundant and inexpensive natural source of long, single-stranded nucleic acid, which could be used as raw material for the fabrication of molecular origami. In this study, we demonstrate efficient and robust construction of 2D and 3D origami nanostructures utilizing cellular rRNA as a scaffold and DNA oligonucleotide staples. We present calibrated protocols for the robust folding of contiguous shapes from one or two rRNA subunits that are efficient to allow folding using crude extracts of total RNA. We also show that RNA maintains stability within the folded structure. Lastly, we present a novel and comprehensive analysis and insights into the stability of RNA:DNA origami nanostructures and demonstrate their enhanced stability when coated with polylysine-polyethylene glycol in different temperatures, low Mg concentrations, human serum, and in the presence of nucleases (DNase I or RNase H). Thus, laying the foundation for their potential implementation in emerging biomedical applications, where folding rRNA into stable structures outside and inside cells would be desired.
Topics: RNA, Ribosomal; Nucleic Acid Conformation; Nanostructures; Humans; RNA Folding; DNA; Polylysine; Polyethylene Glycols
PubMed: 38698435
DOI: 10.1186/s12951-024-02489-2 -
Journal of Food Protection Jul 2023The purpose of the study was to investigate the mechanism of inactivation of Serratia liquefaciens by different treatments, namely corona discharge plasma (CDP),...
The purpose of the study was to investigate the mechanism of inactivation of Serratia liquefaciens by different treatments, namely corona discharge plasma (CDP), ε-polylysine (ε-PL), and corona discharge plasma combined with ε-polylysine (CDP plus ε-PL). The results showed that the combined treatment of CDP and ε-PL exhibited significant antibacterial effects. The total number of colonies of S. liquefaciens dropped by 0.49 log CFU/mL following 4 min of CDP treatment, 4MIC ε-PL treatment for 6 h alone decreased the amounts of colonies by 2.11 log CFU/mL, and 6 h of treatment with 4MIC ε-PL after the bacterium was treated with CDP could decrease the number of colonies by 6.77 log CFU/mL. Scanning electron microscopy images showed that the combined treatment of CDP and ε-PL caused the most serious damage to the cell morphology. Electrical conductivity, nucleic acid, and PI staining indicated that the combined treatment dramatically enhanced the permeability of the cell membrane. In addition, the combined treatment led to a significant decrease in SOD and POD enzyme activities in S. liquefaciens, which prevented energy metabolism. Finally, the determination of free and intracellular ε-PL concentrations confirmed that the treatment of CDP could cause the bacteria to bind more ε-PL and exert more significant bacterial inhibition. Therefore, CDP and ε-PL had a synergistic effect in the inhibition of S. liquefaciens.
Topics: Polylysine; Serratia liquefaciens; Anti-Bacterial Agents; Cell Membrane; Microscopy, Electron, Scanning
PubMed: 37295216
DOI: 10.1016/j.jfp.2023.100078 -
Polymers Nov 2023The aim of this investigation was to scrutinize the effects of a thermal treatment on the electrostatic complex formed between gum arabic (GA) and ε-polylysine (ε-PL),...
The aim of this investigation was to scrutinize the effects of a thermal treatment on the electrostatic complex formed between gum arabic (GA) and ε-polylysine (ε-PL), with the goal of improving the antibacterial properties and reducing the hygroscopicity of ε-PL. The heated complex with a ratio of 1:4 exhibited an encapsulation efficiency of 93.3%. Additionally, it had an average particle size of 350.3 nm, a polydispersity index of 0.255, and a zeta potential of 18.9 mV. The formation of the electrostatic complex between GA and ε-PL was confirmed through multispectral analysis, which demonstrated the participation of hydrogen bonding and hydrophobic and electrostatic interactions, as well as the enhanced effect of heat treatment on these forces within the complex. The complex displayed a core-shell structure, with a regular distribution and a shape that was approximately spherical, as observed in the transmission electron microscopy images. Additionally, the heated GA-ε-PL electrostatic composite exhibited favorable antibacterial effects on Salmonella enterica and Listeria monocytogenes, with reduced minimum inhibitory concentrations (15.6 μg/mL and 62.5 μg/mL, respectively) and minimum bactericidal concentrations (31.3 μg/mL and 156.3 μg/mL, respectively) compared to free ε-PL or the unheated electrostatic composite. Moreover, the moisture absorption of ε-PL reduced from 92.6% to 15.0% in just 48 h after being incorporated with GA and subsequently subjected to heat. This research showed a way to improve the antibacterial efficiency and antihygroscopicity of ε-PL, reducing its application limitations as an antimicrobial substance to some extent.
PubMed: 38231920
DOI: 10.3390/polym15234517 -
Frontiers in Nutrition 2024This study investigated the effects of nisin combined with ε-polylysine on microorganisms and the refrigerated quality of fresh-cut jackfruit. After being treated with...
This study investigated the effects of nisin combined with ε-polylysine on microorganisms and the refrigerated quality of fresh-cut jackfruit. After being treated with distilled water (control), nisin (0.5 g/L), ε-polylysine (0.5 g/L), and the combination of nisin (0.1 g/L) and ε-polylysine (0.4 g/L), microporous modified atmosphere packaging (MMAP) was carried out and stored at 10 ± 1°C for 8 days. The microorganisms and physicochemical indexes were measured every 2 days during storage. The results indicated that combined treatment (0.1 g/L nisin, 0.4 g/L ε-polylysine) had the best preservation on fresh-cut jackfruit. Compared with the control, combined treatment inhibited microbial growth (total bacterial count, mold and yeast), reduced the weight loss rate, respiratory intensity, polyphenol oxidase and peroxidase activities, and maintained higher sugar acid content, firmness, and color. Furthermore, it preserved higher levels of antioxidant compounds, reduced the accumulation of malondialdehyde and hydrogen peroxide, thereby reducing oxidative damage and maintaining high nutritional and sensory qualities. As a safe application of natural preservatives, nisin combined with ε-polylysine treatment has great application potential in the fresh-cut jackfruit industry.
PubMed: 38419851
DOI: 10.3389/fnut.2024.1299810 -
Journal of Controlled Release :... Dec 2023Adipose-derived mesenchymal stem cell-derived small extracellular vesicles (Ad-MSC-sEVs/AMEs) combined with scaffold materials are used in tissue-engineered bladders;...
Negatively charged bladder acellular matrix loaded with positively charged adipose-derived mesenchymal stem cell-derived small extracellular vesicles for bladder tissue engineering.
Adipose-derived mesenchymal stem cell-derived small extracellular vesicles (Ad-MSC-sEVs/AMEs) combined with scaffold materials are used in tissue-engineered bladders; however, the lack of retention leads to limited distribution of AMEs in the scaffold areas and low bioavailability of AMEs after bladder reconstruction. To improve retention of AMEs, we developed a novel strategy that modifies the surface charge of the bladder acellular matrix (BAM) via oxidative self-polymerization of dopamine-reducing graphene oxide (GO) and AMEs using ε-polylysine-polyethylene-distearyl phosphatidylethanolamine (PPD). We evaluated two BAM surface modification methods and evaluated the biocompatibility of materials and PPD and electrostatic adherence effects between PPD-modified AMEs and rGO-PDA/BAM in vivo and in vitro. Surface modification increased retention of AMEs, enhanced regeneration of bladder structures, and increased electrical conductivity of rGO-PDA/BAM, thereby improving bladder function recovery. RNA-sequencing revealed 543 miRNAs in human AMEs and 514 miRNAs in rat AMEs. A Venn diagram was used to show target genes of miRNA with the highest proportion predicted by the four databases; related biological processes and pathways were predicted by KEGG and GO analyses. We report a strategy for improving bioavailability of AMEs for bladder reconstruction and reveal that enriched miR-21-5p targets PIK3R1 and activates the PI3K/Akt pathway to promote cell proliferation and migration.
Topics: Rats; Humans; Animals; Tissue Engineering; Urinary Bladder; Phosphatidylinositol 3-Kinases; Extracellular Matrix; Mesenchymal Stem Cells; MicroRNAs; Extracellular Vesicles
PubMed: 37944669
DOI: 10.1016/j.jconrel.2023.10.048