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1.
Biomacromolecules ; 25(5): 3011-3017, 2024 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-38689515

RESUMO

Stabilization against the dilution-dependent disassembly of self-assembled nanoparticles is a requirement for in vivo application. Herein, we propose a simple and biocompatible cross-linking reaction for the stabilization of a series of nanoparticles formed by the self-assembly of amphiphilic HA-b-ELP block copolymers, through the alkylation of methionine residues from the ELP block with diglycidyl ether compounds. The core-cross-linked nanoparticles retain their colloidal properties, with a spherical core-shell morphology, while maintaining thermoresponsive behavior. As such, instead of a reversible disassembly when non-cross-linked, a reversible swelling of nanoparticles' core and increase of hydrodynamic diameter are observed with lowering of the temperature.


Assuntos
Reagentes de Ligações Cruzadas , Nanopartículas , Nanopartículas/química , Reagentes de Ligações Cruzadas/química , Temperatura , Polímeros/química , Elastina/química , Tamanho da Partícula
2.
J Colloid Interface Sci ; 668: 678-690, 2024 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-38710124

RESUMO

Aerogels, as a unique porous material, are expected to be used as insulation materials to solve the global environmental and energy crisis. Using chitosan, citric acid, pectin and phytic acid as raw materials, an all-biomass-based aerogel with high modulus was prepared by the triple strategy of ionic, physical and chemical cross-linking through directional freezing technique. Based on this three-dimensional network, the aerogel exhibited excellent compressive modulus (24.89 ± 1.76 MPa) over a wide temperature range and thermal insulation properties. In the presence of chitosan, citric acid and phytic acid, the aerogel obtained excellent fire safety (LOI value up to 31.2%) and antibacterial properties (antibacterial activity against Staphylococcus aureus and Escherichia coli reached 81.98% and 67.43%). In addition, the modified aerogel exhibited excellent hydrophobicity (hydrophobic angle of 146°) and oil-water separation properties. More importantly, the aerogel exhibited a biodegradation rate of up to 40.31% for 35 days due to its all-biomass nature. This work provides a green and sustainable strategy for the production of highly environmentally friendly thermal insulation materials with high strength, flame retardant, antibacterial and hydrophobic properties.


Assuntos
Antibacterianos , Quitosana , Ácido Cítrico , Escherichia coli , Géis , Staphylococcus aureus , Escherichia coli/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/química , Géis/química , Quitosana/química , Ácido Cítrico/química , Biomassa , Interações Hidrofóbicas e Hidrofílicas , Porosidade , Ácido Fítico/química , Pectinas/química , Reagentes de Ligações Cruzadas/química , Testes de Sensibilidade Microbiana , Propriedades de Superfície , Tamanho da Partícula , Temperatura
3.
AAPS PharmSciTech ; 25(5): 106, 2024 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-38724834

RESUMO

The primary factor underlying the virulence of Candida albicans is its capacity to form biofilms, which in turn leads to recurrent complications. Over-the-counter antifungal treatments have proven ineffective in eliminating fungal biofilms and the inflammatory cytokines produced during fungal infections. Chitosan nanoparticles offer broad and versatile therapeutic potential as both antifungal agents and carriers for antifungal drugs to combat biofilm-associated Candida infections. In our study, we endeavoured to develop chitosan nanoparticles utilising chitosan and the antifungal crosslinker phytic acid targeting C. albicans. Phytic acid, known for its potent antifungal and anti-inflammatory properties, efficiently crosslinks with chitosan. The nanoparticles were synthesised using the ionic gelation technique and subjected to analyses including Fourier transform infrared spectroscopy, dynamic light scattering, and zeta potential analysis. The synthesised nanoparticles exhibited dimensions with a diameter (Dh) of 103 ± 3.9 nm, polydispersity index (PDI) of 0.33, and zeta potential (ZP) of 37 ± 2.5 mV. These nanoparticles demonstrated an antifungal effect with a minimum inhibitory concentration (MIC) of 140 ± 2.2 µg/mL, maintaining cell viability at approximately 90% of the MIC value and reducing cytokine levels. Additionally, the nanoparticles reduced ergosterol content and exhibited a 62% ± 1.2 reduction in biofilm susceptibility, as supported by colony-forming unit (CFU) and XTT assays-furthermore, treatment with nanoparticles reduced exopolysaccharide production and decreased secretion of aspartyl protease by C. albicans. Our findings suggest that the synthesised nanoparticles effectively combat Candida albicans infections. In vivo studies conducted on a mouse model of vaginal candidiasis confirmed the efficacy of the nanoparticles in combating fungal infections in vivo.


Assuntos
Antifúngicos , Biofilmes , Candida albicans , Quitosana , Testes de Sensibilidade Microbiana , Nanopartículas , Ácido Fítico , Quitosana/química , Biofilmes/efeitos dos fármacos , Nanopartículas/química , Antifúngicos/farmacologia , Antifúngicos/administração & dosagem , Animais , Candida albicans/efeitos dos fármacos , Camundongos , Testes de Sensibilidade Microbiana/métodos , Ácido Fítico/farmacologia , Ácido Fítico/administração & dosagem , Ácido Fítico/química , Feminino , Candidíase/tratamento farmacológico , Tamanho da Partícula , Portadores de Fármacos/química , Reagentes de Ligações Cruzadas/química , Citocinas/metabolismo
4.
Invest Ophthalmol Vis Sci ; 65(5): 24, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38748430

RESUMO

Purpose: Hydrogels derived from decellularized tissues are promising biomaterials in tissue engineering, but their rapid biodegradation can hinder in vitro cultivation. This study aimed to retard biodegradation of a hydrogel derived from porcine decellularized lacrimal glands (dLG-HG) by crosslinking with genipin to increase the mechanical stability without affecting the function and viability of lacrimal gland (LG)-associated cells. Methods: The effect of different genipin concentrations on dLG-HG stiffness was measured rheologically. Cell-dependent biodegradation was quantified over 10 days, and the impact on matrix metalloproteinase (MMP) activity was quantified by gelatin and collagen zymography. The viability of LG epithelial cells (EpCs), mesenchymal stem cells (MSCs), and endothelial cells (ECs) cultured on genipin-crosslinked dLG-HG was assessed after 10 days, and EpC secretory activity was analyzed by ß-hexosaminidase assay. Results: The 0.5-mM genipin increased the stiffness of dLG-HG by about 46%, and concentrations > 0.25 mM caused delayed cell-dependent biodegradation and reduced MMP activity. The viability of EpCs, MSCs, and ECs was not affected by genipin concentrations of up to 0.5 mM after 10 days. Moreover, up to 0.5-mM genipin did not negatively affect EpC secretory activity compared to control groups. Conclusions: A concentration of 0.5-mM genipin increased dLG-HG stiffness, and 0.25-mM genipin was sufficient to prevent MMP-dependent degradation. Importantly, concentrations of up to 0.5-mM genipin did not compromise the viability of LG-associated cells or the secretory activity of EpCs. Thus, crosslinking with genipin improves the properties of dLG-HG for use as a substrate in LG tissue engineering.


Assuntos
Sobrevivência Celular , Reagentes de Ligações Cruzadas , Hidrogéis , Iridoides , Engenharia Tecidual , Animais , Iridoides/farmacologia , Iridoides/metabolismo , Suínos , Engenharia Tecidual/métodos , Reagentes de Ligações Cruzadas/farmacologia , Células Cultivadas , Células-Tronco Mesenquimais/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Células Endoteliais/efeitos dos fármacos , Materiais Biocompatíveis
5.
J Am Chem Soc ; 146(19): 13617-13628, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38695163

RESUMO

Here, we present a cross-linking approach to covalently functionalize and stabilize DNA origami structures in a one-pot reaction. Our strategy involves adding nucleotide sequences to adjacent staple strands, so that, upon assembly of the origami structure, the extensions form short hairpin duplexes targetable by psoralen-labeled triplex-forming oligonucleotides bearing other functional groups (pso-TFOs). Subsequent irradiation with UVA light generates psoralen adducts with one or both hairpin staples leading to site-specific attachment of the pso-TFO (and attached group) to the origami with ca. 80% efficiency. Bis-adduct formation between strands in proximal hairpins further tethers the TFO to the structure and generates "superstaples" that improve the structural integrity of the functionalized complex. We show that directing cross-linking to regions outside of the origami core dramatically reduces sensitivity of the structures to thermal denaturation and disassembly by T7 RNA polymerase. We also show that the underlying duplex regions of the origami core are digested by DNase I and thus remain accessible to read-out by DNA-binding proteins. Our strategy is scalable and cost-effective, as it works with existing DNA origami structures, does not require scaffold redesign, and can be achieved with just one psoralen-modified oligonucleotide.


Assuntos
Reagentes de Ligações Cruzadas , DNA , Conformação de Ácido Nucleico , Raios Ultravioleta , DNA/química , Reagentes de Ligações Cruzadas/química , Processos Fotoquímicos , Ficusina/química
6.
Carbohydr Polym ; 337: 122144, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38710569

RESUMO

In vivo, cells interact with the extracellular matrix (ECM), which provides a multitude of biophysical and biochemical signals that modulate cellular behavior. Inspired by this, we explored a new methodology to develop a more physiomimetic polysaccharide-based matrix for 3D cell culture. Maleimide-modified alginate (AlgM) derivatives were successfully synthesized using DMTMM to activate carboxylic groups. Thiol-terminated cell-adhesion peptides were tethered to the hydrogel network to promote integrin binding. Rapid and efficient in situ hydrogel formation was promoted by thiol-Michael addition "click" chemistry via maleimide reaction with thiol-flanked protease-sensitive peptides. Alginate derivatives were further ionically crosslinked by divalent ions present in the medium, which led to greater stability and allowed longer cell culture periods. By tailoring alginate's biofunctionality we improved cell-cell and cell-matrix interactions, providing an ECM-like 3D microenvironment. We were able to systematically and independently vary biochemical and biophysical parameters to elicit specific cell responses, creating custom-made 3D matrices. DMTMM-mediated maleimide incorporation is a promising approach to synthesizing AlgM derivatives that can be leveraged to produce ECM-like matrices for a broad range of applications, from in vitro tissue modeling to tissue regeneration.


Assuntos
Alginatos , Química Click , Matriz Extracelular , Hidrogéis , Maleimidas , Compostos de Sulfidrila , Maleimidas/química , Alginatos/química , Compostos de Sulfidrila/química , Hidrogéis/química , Hidrogéis/síntese química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Humanos , Reagentes de Ligações Cruzadas/química , Adesão Celular/efeitos dos fármacos , Animais
7.
BMC Ophthalmol ; 24(1): 201, 2024 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-38698363

RESUMO

BACKGROUND: We aimed to employ Optical Coherence Tomography Angiography (OCTA) to comprehensively assess changes in the optic nerve head (ONH) and macular perfusion before and after the Corneal Collagen Cross-Linking (CCL) procedure in patients with keratoconus. METHODS: A total of 22 keratoconus patient's candidate for CCL procedures were included based on specific criteria, with meticulous exclusion criteria in place to minimize potential confounders. Participants underwent OCTA assessments of the ONH and macula using the Spectralis OCT (Heidelberg) before CCL, as well as at 1- and 3-months post-CCL. MATLAB software was utilized for image analysis. RESULTS: The mean age of the participants was 20.09 ± 6.11, including 59% male, and the mean intraocular pressure (IOP) before the surgery was 13.59 ± 2.85 mmHg. Peripapillary Retinal nerve fiber layer (ppRNFL) thickness and overall retinal thickness remained stable post-CCL. However, significant alterations were observed in macular vessel density, emphasizing regional variations in vascular response. For macular large vessel density (LVD), both superficial and deep vascular complex (SVC and DVC) demonstrated significant differences between before surgery and the 3 months post-surgery follow-up (p < 0.001 and p = 0.002, respectively). Optic nerve head markers demonstrated relative stability, except for changes in avascular complex density, which was 49.2 ± 2.2% before the surgery and decrease to 47.6 ± 1.7% three months after the operation (P-value = 0.005). CONCLUSION: While CCL appears to maintain the integrity of certain ocular structures, alterations in macular perfusion post-CCL suggest potential effects on retinal blood supply. Long-term monitoring is crucial to understand the implications of these changes, particularly in the context of conditions such as diabetes.


Assuntos
Colágeno , Reagentes de Ligações Cruzadas , Angiofluoresceinografia , Ceratocone , Disco Óptico , Vasos Retinianos , Tomografia de Coerência Óptica , Humanos , Tomografia de Coerência Óptica/métodos , Ceratocone/fisiopatologia , Ceratocone/diagnóstico , Masculino , Feminino , Colágeno/metabolismo , Adulto Jovem , Adulto , Angiofluoresceinografia/métodos , Vasos Retinianos/diagnóstico por imagem , Vasos Retinianos/fisiopatologia , Disco Óptico/irrigação sanguínea , Adolescente , Estudos Prospectivos , Fármacos Fotossensibilizantes/uso terapêutico , Fotoquimioterapia/métodos , Macula Lutea/diagnóstico por imagem , Macula Lutea/irrigação sanguínea
8.
Sci Rep ; 14(1): 10825, 2024 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-38734808

RESUMO

This study developed a kind of PEG-crosslinked O-carboxymethyl chitosan (O-CMC-PEG) with various PEG content for food packaging. The crosslinking agent of isocyanate-terminated PEG was firstly synthesized by a simple condensation reaction between PEG and excess diisocyanate, then the crosslink between O-carboxymethyl chitosan (O-CMC) and crosslinking agent occurred under mild conditions to produce O-CMC-PEG with a crosslinked structure linked by urea bonds. FT-IR and 1H NMR techniques were utilized to confirm the chemical structures of the crosslinking agent and O-CMC-PEGs. Extensive research was conducted to investigate the impact of the PEG content (or crosslinking degree) on the physicochemical characteristics of the casted O-CMC-PEG films. The results illuminated that crosslinking and components compatibility could improve their tensile features and water vapor barrier performance, while high PEG content played the inverse effects due to the microphase separation between PEG and O-CMC segments. The in vitro degradation rate and water sensitivity primarily depended on the crosslinking degree in comparison with the PEG content. Furthermore, caused by the remaining -NH2 groups of O-CMC, the films demonstrated antibacterial activity against Escherichia coli and Staphylococcus aureus. When the PEG content was 6% (medium crosslinking degree), the prepared O-CMC-PEG-6% film possessed optimal tensile features, high water resistance, appropriate degradation rate, low water vapor transmission rate and fine broad-spectrum antibacterial capacity, manifesting a great potential for application in food packaging to extend the shelf life.


Assuntos
Antibacterianos , Quitosana , Escherichia coli , Embalagem de Alimentos , Polietilenoglicóis , Quitosana/química , Quitosana/análogos & derivados , Quitosana/farmacologia , Embalagem de Alimentos/métodos , Antibacterianos/química , Antibacterianos/farmacologia , Polietilenoglicóis/química , Escherichia coli/efeitos dos fármacos , Reagentes de Ligações Cruzadas/química , Espectroscopia de Infravermelho com Transformada de Fourier , Staphylococcus aureus/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Resistência à Tração
9.
Int J Mol Sci ; 25(9)2024 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-38732231

RESUMO

Regenerative medicine is an interdisciplinary field aiming at restoring pathologically damaged tissues and whole organs by cell transplantation in combination with proper supporting scaffolds. Gelatine-based ones are very attractive due to their biocompatibility, rapid biodegradability, and lack of immunogenicity. Gelatine-based composite hydrogels, containing strengthening agents to improve their modest mechanical properties, have been demonstrated to act as extracellular matrices (ECMs), thus playing a critical role in "organ manufacturing". Inspired by the lysyl oxidase (LO)-mediated process of crosslinking, which occurs in nature to reinforce collagen, we have recently developed a versatile protocol to crosslink gelatine B (Gel B) in the presence or absence of LO, using properly synthesized polystyrene- and polyacrylic-based copolymers containing the amine or aldehyde groups needed for crosslinking reactions. Here, following the developed protocol with slight modifications, we have successfully crosslinked Gel B in different conditions, obtaining eight out of nine compounds in high yield (57-99%). The determined crosslinking degree percentage (CP%) evidenced a high CP% for compounds obtained in presence of LO and using the styrenic amine-containing (CP5/DMAA) and acrylic aldehyde-containing (CPMA/DMAA) copolymers as crosslinking agents. ATR-FTIR analyses confirmed the chemical structure of all compounds, while optical microscopy demonstrated cavernous, crater-like, and labyrinth-like morphologies and cavities with a size in the range 15-261 µm. An apparent density in the range 0.10-0.45 g/cm3 confirmed the aerogel-like structure of most samples. Although the best biodegradation profile was observed for the sample obtained using 10% CP5/DMAA (M3), high swelling and absorption properties, high porosity, and good biodegradation profiles were also observed for samples obtained using the 5-10% CP5/DMAA (M4, 5, 6) and 20% CPMA/DMAA (M9) copolymers. Collectively, in this work of synthesis and physicochemical characterization, new aerogel-like composites have been developed and, based on their characteristics, which fit well within the requirements for TE, five candidates (M3, M4, M5, M6, and M9) suitable for future biological experiments on cell adhesion, infiltration and proliferation, to confirm their effective functioning, have been identified.


Assuntos
Materiais Biocompatíveis , Gelatina , Hidrogéis , Medicina Regenerativa , Alicerces Teciduais , Gelatina/química , Alicerces Teciduais/química , Medicina Regenerativa/métodos , Materiais Biocompatíveis/química , Hidrogéis/química , Hidrogéis/síntese química , Humanos , Engenharia Tecidual/métodos , Reagentes de Ligações Cruzadas/química
10.
Anal Chem ; 96(19): 7506-7515, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38690851

RESUMO

Alzheimer's disease (AD) is a progressive neurological disorder featuring abnormal protein aggregation in the brain, including the pathological hallmarks of amyloid plaques and hyperphosphorylated tau. Despite extensive research efforts, understanding the molecular intricacies driving AD development remains a formidable challenge. This study focuses on identifying key protein conformational changes associated with the progression of AD. To achieve this, we employed quantitative cross-linking mass spectrometry (XL-MS) to elucidate conformational changes in the protein networks in cerebrospinal fluid (CSF). By using isotopically labeled cross-linkers BS3d0 and BS3d4, we reveal a dynamic shift in protein interaction networks during AD progression. Our comprehensive analysis highlights distinct alterations in protein-protein interactions within mild cognitive impairment (MCI) states. This study accentuates the potential of cross-linked peptides as indicators of AD-related conformational changes, including previously unreported site-specific binding between α-1-antitrypsin (A1AT) and complement component 3 (CO3). Furthermore, this work enables detailed structural characterization of apolipoprotein E (ApoE) and reveals modifications within its helical domains, suggesting their involvement in MCI pathogenesis. The quantitative approach provides insights into site-specific interactions and changes in the abundance of cross-linked peptides, offering an improved understanding of the intricate protein-protein interactions underlying AD progression. These findings lay a foundation for the development of potential diagnostic or therapeutic strategies aimed at mitigating the negative impact of AD.


Assuntos
Doença de Alzheimer , Apolipoproteínas E , Espectrometria de Massas , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Doença de Alzheimer/diagnóstico , Humanos , Apolipoproteínas E/química , Apolipoproteínas E/metabolismo , Reagentes de Ligações Cruzadas/química , Conformação Proteica , alfa 1-Antitripsina/química , alfa 1-Antitripsina/metabolismo , Disfunção Cognitiva/metabolismo
11.
Anal Chem ; 96(19): 7373-7379, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38696819

RESUMO

Cross-linking mass spectrometry (XL-MS) has evolved into a pivotal technique for probing protein interactions. This study describes the implementation of Parallel Accumulation-Serial Fragmentation (PASEF) on timsTOF instruments, enhancing the detection and analysis of protein interactions by XL-MS. Addressing the challenges in XL-MS, such as the interpretation of complex spectra, low abundant cross-linked peptides, and a data acquisition bias, our current study integrates a peptide-centric approach for the analysis of XL-MS data and presents the foundation for integrating data-independent acquisition (DIA) in XL-MS with a vendor-neutral and open-source platform. A novel workflow is described for processing data-dependent acquisition (DDA) of PASEF-derived information. For this, software by Bruker Daltonics is used, enabling the conversion of these data into a format that is compatible with MeroX and Skyline software tools. Our approach significantly improves the identification of cross-linked products from complex mixtures, allowing the XL-MS community to overcome current analytical limitations.


Assuntos
Reagentes de Ligações Cruzadas , Espectrometria de Massas , Software , Fluxo de Trabalho , Reagentes de Ligações Cruzadas/química , Peptídeos/química , Peptídeos/análise , Humanos
12.
ACS Biomater Sci Eng ; 10(5): 3343-3354, 2024 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-38695560

RESUMO

Moldable tissue-sealant hydrogels were developed herein by combining the yield stress fluidity of a Carbomer and in situ cross-linking of 3-arm PEG-thiol (PEG-SH) and 4-arm PEG-acrylate (PEG-AC). The Carbomer was mixed with each PEG oligomer to form two aqueous precursors: Carbomer/PEG-SH and Carbomer/PEG-AC. The two hydrogel precursors exhibited sufficient yield stress (>100 Pa) to prevent dripping from their placement on the tissue surface. Moreover, these hydrogel precursors exhibited rapid restructuring when the shear strain was repeatedly changed. These rheological properties contribute to the moldability of these hydrogel precursors. After mixing these two precursors, they were converted from yield-stress fluids to chemically cross-linked hydrogels, Carbomer/PEG hydrogel, via thiol-Michael addition. The gelation time was 5.0 and 11.2 min at 37 and 25 °C, respectively. In addition, the Carbomer/PEG hydrogels exhibited higher cellular viability than the pure Carbomer. They also showed stable adhesiveness and burst pressure resistance to various tissues, such as the skin, stomach, colon, and cecum of pigs. The hydrogels showed excellent tissue sealing in a cecum ligation and puncture model in mice and improved the survival rate due to their tissue adhesiveness and biocompatibility. The Carbomer/PEG hydrogel is a potential biocompatible tissue sealant that surgeons can mold. It was revealed that the combination of in situ cross-linkable PEG oligomers and yield stress fluid such as Carbomer is effective for developing the moldable tissue sealant without dripping of its hydrogel precursors.


Assuntos
Hidrogéis , Polietilenoglicóis , Compostos de Sulfidrila , Hidrogéis/química , Hidrogéis/farmacologia , Polietilenoglicóis/química , Animais , Camundongos , Compostos de Sulfidrila/química , Adesivos Teciduais/química , Adesivos Teciduais/farmacologia , Suínos , Reagentes de Ligações Cruzadas/química , Reologia , Humanos , Resinas Acrílicas
13.
Biomacromolecules ; 25(5): 3169-3177, 2024 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-38684138

RESUMO

Achieving precise control over gelator alignment and morphology is crucial for crafting tailored materials and supramolecular structures with distinct properties. We successfully aligned the self-assembled micelles formed by a functionalized dipeptide 2NapFF into long 1-D "gel noodles" by cross-linking with divalent metal chlorides. We identify the most effective cross-linker for alignment, enhancing mechanical stability, and imparting functional properties. Our study shows that Group 2 metal ions are particularly suited for creating mechanically robust yet flexible gel noodles because of their ionic and nondirectional bonding with carboxylate groups. In contrast, the covalent nature and high directional bonds of d-block metal ions with carboxylates tend to disrupt the self-assembly of 2NapFF. Furthermore, the 2NapFF-Cu noodles demonstrated selective antibacterial activity, indicating that the potent antibacterial property of the copper(II) ion is preserved within the cross-linked system. By merging insights into molecular alignment, gel extrusion processing, and integrating specific functionalities, we illustrate how the versatility of dipeptide-based gels can be utilized in creating next-generation soft materials.


Assuntos
Antibacterianos , Cobre , Géis , Antibacterianos/química , Antibacterianos/farmacologia , Cobre/química , Cobre/farmacologia , Géis/química , Reagentes de Ligações Cruzadas/química , Dipeptídeos/química , Dipeptídeos/farmacologia , Micelas , Testes de Sensibilidade Microbiana , Staphylococcus aureus/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos
14.
Proc Natl Acad Sci U S A ; 121(19): e2317703121, 2024 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-38687792

RESUMO

Fluorescence labeling of chemically fixed specimens, especially immunolabeling, plays a vital role in super-resolution imaging as it offers a convenient way to visualize cellular structures like mitochondria or the distribution of biomolecules with high detail. Despite the development of various distinct probes that enable super-resolved stimulated emission depletion (STED) imaging of mitochondria in live cells, most of these membrane-potential-dependent fluorophores cannot be retained well in mitochondria after chemical fixation. This lack of suitable mitochondrial probes has limited STED imaging of mitochondria to live cell samples. In this study, we introduce a mitochondria-specific probe, PK Mito Orange FX (PKMO FX), which features a fixation-driven cross-linking motif and accumulates in the mitochondrial inner membrane. It exhibits high fluorescence retention after chemical fixation and efficient depletion at 775 nm, enabling nanoscopic imaging both before and after aldehyde fixation. We demonstrate the compatibility of this probe with conventional immunolabeling and other strategies commonly used for fluorescence labeling of fixed samples. Moreover, we show that PKMO FX facilitates correlative super-resolution light and electron microscopy, enabling the correlation of multicolor fluorescence images and transmission EM images via the characteristic mitochondrial pattern. Our probe further expands the mitochondrial toolkit for multimodal microscopy at nanometer resolutions.


Assuntos
Aldeídos , Corantes Fluorescentes , Microscopia de Fluorescência , Mitocôndrias , Mitocôndrias/metabolismo , Humanos , Corantes Fluorescentes/química , Aldeídos/metabolismo , Aldeídos/química , Microscopia de Fluorescência/métodos , Células HeLa , Reagentes de Ligações Cruzadas/química , Animais , Membranas Mitocondriais/metabolismo
15.
Jpn J Ophthalmol ; 68(3): 225-232, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38557918

RESUMO

PURPOSE: We aimed to compare the efficacy and safety of accelerated contact lens-assisted cross-linking (CA-CXL) with Lotrafilcon B and Comfilcon A lenses in keratoconus (KC) patients with thin corneas. STUDY DESIGN: Retrospective, single-center study. MATERIALS AND METHODS: We retrospectively included 51 eyes of 39 KC patients with corneal thickness <400µm after epithelial scraping (Epi-off), who underwent accelerated CA-CXL treatment with Lotrafilcon B (n=20) and Comfilcon A (n=31). Uncorrected and corrected distance visual acuity (UDVA and CDVA), manifest refraction values, corneal topographic data and endothelial cell density were recorded at preoperative and postoperative 1st, 3rd and 6th month controls. RESULTS: CDVA in the Comfilcon A group was higher than CDVA before surgery at 6 months postoperatively (p<0.001). When the two lenses were compared, CDVA was found to be significantly higher in the Lotrafilcon B group in the preoperative, postoperative 1st month and 3rd month values, but there was no significant difference between the postoperative 6th month values (p=0.028, p=0.018, p=0.044, p=0.181, respectively). The maximum keratometry (Kmax) value at the 6th month after surgery in the Comfilcon A group was significantly lower than in the Lotrafilcon B group (p=0,009). There was no significant difference between the endothelial cell density values between the groups (p=0.623, p=0.609, p=0.794, p=0.458, respectively). There was no significant difference between the progression, regression, and stability rates of the two groups (p=0.714). CONCLUSIONS: Accelerated CA-CXL with Lotrafilcon B and Comfilcon A silicone hydrogel lenses is a safe and effective method to stop progression in patients with thin corneas.


Assuntos
Colágeno , Topografia da Córnea , Reagentes de Ligações Cruzadas , Ceratocone , Fotoquimioterapia , Fármacos Fotossensibilizantes , Refração Ocular , Riboflavina , Acuidade Visual , Humanos , Ceratocone/diagnóstico , Ceratocone/fisiopatologia , Ceratocone/tratamento farmacológico , Ceratocone/terapia , Ceratocone/metabolismo , Feminino , Masculino , Estudos Retrospectivos , Acuidade Visual/fisiologia , Fármacos Fotossensibilizantes/uso terapêutico , Adulto , Riboflavina/uso terapêutico , Fotoquimioterapia/métodos , Adulto Jovem , Refração Ocular/fisiologia , Colágeno/metabolismo , Resultado do Tratamento , Córnea/patologia , Raios Ultravioleta , Seguimentos , Adolescente , Contagem de Células , Substância Própria/metabolismo , Endotélio Corneano/patologia , Lentes de Contato Hidrofílicas , Crosslinking Corneano
16.
Int J Biol Macromol ; 267(Pt 1): 131432, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38583849

RESUMO

The potential applications of cellulose nanofibril-based foam materials can be expanded by their enhanced water durability. This study proposes two crosslinking methods to improve the water durability of the oven-dried carboxymethylated cellulose nanofibril (CMCNF) foam. The first method involves the addition of a crosslinker, polyamideamine epichlorohydrin. The second method is the self-crosslinking of CMCNFs via heat treatment at 140 °C for less than an hour, which is a simple way to crosslink CMCNF-based materials. Both crosslinking methods resulted in excellent water durability and wet resilience of the foams, which also exhibited high water absorbency. Furthermore, neither method affected the structural nor mechanical properties of the oven-dried CMCNF foams. In particular, self-crosslinking by heat treatment proved to be as effective as using a crosslinking agent. Compared to the freeze-dried foam, the oven-dried foam exhibited slower methylene blue (MB) dye adsorption but a higher maximum adsorption capacity (238-250 mg/g), attributed to the closed pore structure and a larger specific surface area. In addition, the isotherm and reusability of the foam for MB adsorption were investigated. These crosslinking processes expanded the potential use of oven-dried CMCNF foams as adsorbents for cationic dyes.


Assuntos
Celulose , Corantes , Reagentes de Ligações Cruzadas , Azul de Metileno , Nanofibras , Água , Nanofibras/química , Adsorção , Água/química , Celulose/química , Celulose/análogos & derivados , Corantes/química , Azul de Metileno/química , Reagentes de Ligações Cruzadas/química , Poluentes Químicos da Água/química , Purificação da Água/métodos , Epicloroidrina/química
17.
Indian J Ophthalmol ; 72(5): 712-717, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38648433

RESUMO

PURPOSE: To compare the changes encountered in corneal biomechanics and aberration profile following accelerated corneal collagen cross-linking (CXL) using hypo-osmolar and iso-osmolar riboflavin in corneal thicknesses of <400 and >400 microns, respectively. METHODS: This is a prospective, interventional, comparative study involving 100 eyes of 75 patients with progressive keratoconus. Eyes were divided into two groups based on corneal thickness: group 1 included eyes with a corneal thickness of <400 microns who underwent hypo-osmolar CXL, and group 2 included eyes with a corneal thickness of >400 microns who underwent iso-osmolar CXL. Corneal biomechanical and aberration profiles were evaluated and compared between groups. RESULTS: In group 1, all higher-order aberrations (HOA) except secondary astigmatism significantly decreased from baseline; however, in group 2, only coma and trefoil decreased. The corneal resistance factor and corneal hysteresis significantly improved in both groups, which was significantly greater in group 2 than in group 1. The change in inverse radius, deformation amplitude, and tomographic biomechanical index was significantly improved in group 2 as compared to group 1. CONCLUSION: Improvement in corrected distance visual acuity and decrease in HOA were significantly better in the hypo-osmolar CXL group; however, the improvement in biomechanical strength of the cornea was significantly better in the iso-osmolar group.


Assuntos
Colágeno , Córnea , Topografia da Córnea , Reagentes de Ligações Cruzadas , Ceratocone , Fármacos Fotossensibilizantes , Riboflavina , Raios Ultravioleta , Acuidade Visual , Adolescente , Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Fenômenos Biomecânicos , Colágeno/metabolismo , Córnea/diagnóstico por imagem , Córnea/fisiopatologia , Córnea/efeitos dos fármacos , Substância Própria/metabolismo , Substância Própria/efeitos dos fármacos , Aberrações de Frente de Onda da Córnea/fisiopatologia , Reagentes de Ligações Cruzadas/uso terapêutico , Seguimentos , Ceratocone/tratamento farmacológico , Ceratocone/fisiopatologia , Ceratocone/diagnóstico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Estudos Prospectivos , Refração Ocular/fisiologia , Riboflavina/uso terapêutico , Acuidade Visual/fisiologia , Criança
18.
Oral Health Prev Dent ; 22: 171-180, 2024 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-38687029

RESUMO

PURPOSE: To investigate the microbiological outcomes obtained with either subgingival debridement (SD) in conjunction with a gel containing sodium hypochlorite and amino acids followed by subsequent application of a cross-linked hyaluronic acid gel (xHyA) gel, or with SD alone. MATERIALS AND METHODS: Forty-eight patients diagnosed with stages II-III (grades A/B) generalised periodontitis were randomly treated with either SD (control) or SD plus adjunctive sodium hypochlorite/amino acids and xHyA gel (test). Subgingival plaque samples were collected from the deepest site per quadrant in each patient at baseline and after 3 and 6 months. Pooled sample analysis was performed using a multiplex polymerase chain reaction (PCR)-based method for the identification of detection frequencies and changes in numbers of the following bacteria: Aggregatibacter actinomycetemcomitans (A.a), Porphyromonas gingivalis (P.g), Tannerella forsythia (T.f), Treponema denticola (T.d), and Prevotella intermedia (P.i). RESULTS: In terms of detection frequency, in the test group, statistically significant reductions were found for P.g, T.f, T.d and P.i (p < 0.05) after 6 months. In the control group, the detection frequencies of all investigated bacterial species at 6 months were comparable to the baseline values (p > 0.05). The comparison of the test and control groups revealed statistically significant differences in detection frequency for P.g (p = 0.034), T.d (p < 0.01) and P.i (p = 0.02) after 6 months, favouring the test group. Regarding reduction in detection frequency scores, at 6 months, statistically significant differences in favour of the test group were observed for all investigated bacterial species: A.a (p = 0.028), P.g (p = 0.028), T.f (p = 0.004), T.d (p <0.001), and P.i (p = 0.003). CONCLUSIONS: The present microbiological results, which are related to short-term outcomes up to 6 months post-treatment, support the adjunctive subgingival application of sodium hypochlorite/amino acids and xHyA to subgingival debridement in the treatment of periodontitis.


Assuntos
Aggregatibacter actinomycetemcomitans , Aminoácidos , Placa Dentária , Ácido Hialurônico , Porphyromonas gingivalis , Prevotella intermedia , Hipoclorito de Sódio , Tannerella forsythia , Treponema denticola , Humanos , Ácido Hialurônico/uso terapêutico , Hipoclorito de Sódio/uso terapêutico , Aggregatibacter actinomycetemcomitans/efeitos dos fármacos , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Porphyromonas gingivalis/efeitos dos fármacos , Feminino , Pessoa de Meia-Idade , Masculino , Prevotella intermedia/efeitos dos fármacos , Tannerella forsythia/efeitos dos fármacos , Treponema denticola/efeitos dos fármacos , Adulto , Placa Dentária/microbiologia , Aminoácidos/uso terapêutico , Desbridamento Periodontal/métodos , Carga Bacteriana/efeitos dos fármacos , Géis , Terapia Combinada , Seguimentos , Reagentes de Ligações Cruzadas/uso terapêutico , Bolsa Periodontal/microbiologia , Bolsa Periodontal/terapia , Periodontite/microbiologia , Periodontite/terapia , Periodontite/tratamento farmacológico
19.
Int J Biol Macromol ; 267(Pt 1): 131135, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38574914

RESUMO

The study involves the preparation and characterization of crosslinked-carboxymethyl cellulose (CMC) films using varying amounts of citric acid (CA) within the range 5 %-20 %, w/w, relative to the dry weight of CMC. Through techniques such as Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, carbonyl content analysis, and gel fraction measurements, the successful crosslinking between CMC and CA is confirmed. The investigation includes an analysis of chemical structure, physical and optical characteristics, swelling behavior, water vapor transmission rate, moisture content, and surface morphologies. The water resistance of the cross-linked CMC films exhibited a significant improvement when compared to the non-crosslinked CMC film. The findings indicated that films crosslinked with 10 % CA demonstrated favorable properties for application as edible coatings. These transparent films, ideal for packaging, prove effective in preserving the quality and sensory attributes of fresh bananas, including color retention, minimized weight loss, slowed ripening through inhibiting amyloplast degradation, and enhanced firmness during storage.


Assuntos
Carboximetilcelulose Sódica , Ácido Cítrico , Filmes Comestíveis , Embalagem de Alimentos , Musa , Carboximetilcelulose Sódica/química , Ácido Cítrico/química , Embalagem de Alimentos/métodos , Musa/química , Vapor , Reagentes de Ligações Cruzadas/química , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química , Conservação de Alimentos/métodos
20.
Chem Pharm Bull (Tokyo) ; 72(5): 454-470, 2024 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-38644216

RESUMO

This study investigates the efficacy of modified Albizia procera gum as a release-retardant polymer in Diltiazem hydrochloride (DIL) matrix tablets. Carboxymethylated Albizia procera gum (CAP) and ionically crosslinked carboxymethylated Albizia procera gum (Ca-CAP) were utilized, with Ca-CAP synthesized via crosslinking CAP with calcium ions (Ca2+) using calcium chloride (CaCl2). Fourier Transform (FT) IR analysis affirmed polymer compatibility, while differential scanning calorimetry (DSC) and X-ray diffraction (XRD) assessed thermal behavior and crystallinity, respectively. Zeta potential analysis explored surface charge and electrostatic interactions, while rheology examined flow and viscoelastic properties. Swelling and erosion kinetics provided insights into water penetration and stability. CAP's carboxymethyl groups (-CH2-COO-) heightened divalent cation reactivity, and crosslinking with CaCl2 produced Ca-CAP through -CH2-COO- and Ca2+ interactions. Structural similarities between the polymers were revealed by FTIR, with slight differences. DSC indicated modified thermal behavior in Ca-CAP, while Zeta potential analysis showcased negative charges, with Ca-CAP exhibiting lower negativity. XRD highlighted increased crystallinity in Ca-CAP due to calcium crosslinking. Minimal impact on RBC properties was observed with both polymers compared to the positive control as water for injection (WFI). Ca-CAP exhibited improved viscosity, strength, controlled swelling, and erosion, allowing prolonged drug release compared to CAP. Stability studies confirmed consistent six-month drug release, emphasizing Ca-CAP's potential as a stable, sustained drug delivery system over CAP. Robustness and accelerated stability tests supported these findings, underscoring the promise of Ca-CAP in controlled drug release applications.


Assuntos
Diltiazem , Gomas Vegetais , Comprimidos , Diltiazem/química , Gomas Vegetais/química , Comprimidos/química , Albizzia/química , Liberação Controlada de Fármacos , Reagentes de Ligações Cruzadas/química , Reagentes de Ligações Cruzadas/síntese química
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