RESUMEN
Here, we demonstrate the concerted inhibition of different influenza A virus (IAV) strains using a low-molecular-weight dual-action linear polymer. The 6'-sialyllactose and zanamivir conjugates of linear polyglycerol are optimized for simultaneous targeting of hemagglutinin and neuraminidase on the IAV surface. Independent of IAV subtypes, hemagglutination inhibition data suggest better adsorption of the heteromultivalent polymer than homomultivalent analogs onto the virus surface. Cryo-TEM images imply heteromultivalent compound-mediated virus aggregation. The optimized polymeric nanomaterial inhibits >99.9% propagation of various IAV strains 24 h postinfection in vitro at low nM concentrations and is up to 10000× more effective than the commercial zanamivir drug. In a human lung ex vivo multicyclic infection setup, the heteromultivalent polymer outperforms the commercial drug zanamivir and homomultivalent analogs or their physical mixtures. This study authenticates the translational potential of the dual-action targeting approach using small polymers for broad and high antiviral efficacy.
Asunto(s)
Alphainfluenzavirus , Glicosilación , Polímeros/química , Polímeros/farmacología , Alphainfluenzavirus/efectos de los fármacos , Gripe Humana/tratamiento farmacológico , Antivirales/química , Antivirales/farmacología , Humanos , Zanamivir/química , Zanamivir/farmacologíaRESUMEN
As virus outbreaks continue to pose a challenge, a nonspecific viral inhibitor can provide significant benefits, especially against respiratory viruses. Polyglycerol sulfates recently emerge as promising agents that mediate interactions between cells and viruses through electrostatics, leading to virus inhibition. Similarly, hydrophobic C60 fullerene can prevent virus infection via interactions with hydrophobic cavities of surface proteins. Here, two strategies are combined to inhibit infection of SARS-CoV-2 variants in vitro. Effective inhibitory concentrations in the millimolar range highlight the significance of bare fullerene's hydrophobic moiety and electrostatic interactions of polysulfates with surface proteins of SARS-CoV-2. Furthermore, microscale thermophoresis measurements support that fullerene linear polyglycerol sulfates interact with the SARS-CoV-2 virus via its spike protein, and highlight importance of electrostatic interactions within it. All-atom molecular dynamics simulations reveal that the fullerene binding site is situated close to the receptor binding domain, within 4 nm of polyglycerol sulfate binding sites, feasibly allowing both portions of the material to interact simultaneously.
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COVID-19 , Fulerenos , Humanos , SARS-CoV-2 , Fulerenos/farmacología , Unión ProteicaRESUMEN
Mucins are the key component of the defensive mucus barrier. They are extended fibers of very high molecular weight with diverse biological functions depending strongly on their specific structural parameters. Here, we present a mucin-inspired nanostructure, produced via a synthetic methodology to prepare methacrylate-based dendronized polysulfates (MIP-1) on a multi gram-scale with high molecular weight (MW=450â kDa) and thiol end-functionalized mucin-inspired polymer (MIP) via RAFT polymerization. Cryo-electron tomography (Cryo-ET) analysis of MIP-1 confirmed a mucin-mimetic wormlike single-chain fiber structure (length=144±59â nm) in aqueous solution. This biocompatible fiber showed promising activity against SARS-CoV-2 and its mutant strain, with a remarkable low half maximal (IC50 ) inhibitory concentration (IC50 =10.0â nM). Additionally, we investigate the impact of fiber length on SARS-CoV-2 inhibition by testing other functional polymers (MIPs) of varying fiber lengths.
Asunto(s)
COVID-19 , Impresión Molecular , Humanos , Mucinas , SARS-CoV-2 , Polímeros/farmacología , Polímeros/química , Impresión Molecular/métodosRESUMEN
Evidence is strengthening to suggest that the novel SARS-CoV-2 mutant Omicron, with its more than 60 mutations, will spread and dominate worldwide. Although the mutations in the spike protein are known, the molecular basis for why the additional mutations in the spike protein that have not previously occurred account for Omicron's higher infection potential, is not understood. We propose, based on chemical rational and molecular dynamics simulations, that the elevated occurrence of positively charged amino acids in certain domains of the spike protein (Delta: +4; Omicron: +5 vs. wild type) increases binding to cellular polyanionic receptors, such as heparan sulfate due to multivalent charge-charge interactions. This observation is a starting point for targeted drug development.
Asunto(s)
COVID-19 , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , COVID-19/virología , Humanos , Mutación , SARS-CoV-2/genética , Glicoproteína de la Espiga del Coronavirus/química , Glicoproteína de la Espiga del Coronavirus/genéticaRESUMEN
2D nanomaterials have garnered widespread attention in biomedicine and bioengineering due to their unique physicochemical properties. However, poor functionality, low solubility, intrinsic toxicity, and nonspecific interactions at biointerfaces have hampered their application in vivo. Here, biocompatible polyglycerol units are crosslinked in two dimensions using a graphene-assisted strategy leading to highly functional and water-soluble polyglycerols nanosheets with 263 ± 53 nm and 2.7 ± 0.2 nm average lateral size and thickness, respectively. A single-layer hyperbranched polyglycerol containing azide functional groups is covalently conjugated to the surface of a functional graphene template through pH-sensitive linkers. Then, lateral crosslinking of polyglycerol units is carried out by loading tripropargylamine on the surface of graphene followed by lifting off this reagent for an on-face click reaction. Subsequently, the polyglycerol nanosheets are detached from the surface of graphene by slight acidification and centrifugation and is sulfated to mimic heparin sulfate proteoglycans. To highlight the impact of the two-dimensionality of the synthesized polyglycerol sulfate nanosheets at nanobiointerfaces, their efficiency with respect to herpes simplex virus type 1 and severe acute respiratory syndrome corona virus 2 inhibition is compared to their 3D nanogel analogs. Four times stronger in virus inhibition suggests that 2D polyglycerols are superior to their current 3D counterparts.
RESUMEN
Search of new strategies for the inhibition of respiratory viruses is one of the urgent health challenges worldwide, as most of the current therapeutic agents and treatments are inefficient. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic and has taken lives of approximately two million people to date. Even though various vaccines are currently under development, virus, and especially its spike glycoprotein can mutate, which highlights a need for a broad-spectrum inhibitor. In this work, inhibition of SARS-CoV-2 by graphene platforms with precise dual sulfate/alkyl functionalities is investigated. A series of graphene derivatives with different lengths of aliphatic chains is synthesized and is investigated for their ability to inhibit SARS-CoV-2 and feline coronavirus. Graphene derivatives with long alkyl chains (>C9) inhibit coronavirus replication by virtue of disrupting viral envelope. The ability of these graphene platforms to rupture viruses is visualized by atomic force microscopy and cryogenic electron microscopy. A large concentration window (10 to 100-fold) where graphene platforms display strongly antiviral activity against native SARS-CoV-2 without significant toxicity against human cells is found. In this concentration range, the synthesized graphene platforms inhibit the infection of enveloped viruses efficiently, opening new therapeutic and metaphylactic avenues against SARS-CoV-2.
Asunto(s)
Grafito/química , SARS-CoV-2/química , Antivirales/farmacología , COVID-19/epidemiología , COVID-19/virología , Microscopía por Crioelectrón , Humanos , Microscopía de Fuerza Atómica , Pandemias , SARS-CoV-2/efectos de los fármacosRESUMEN
BACKGROUND: Traditional medicines based on herbal extracts have been proposed as affordable treatments for patients suffering from coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Teas and drinks containing extracts of Artemisia annua and Artemisia afra have been widely used in Africa in efforts to prevent SARS-CoV-2 infection and fight COVID-19. METHODS: The plant extracts and Covid-Organics drink produced in Madagascar were tested for plaque reduction using both feline coronavirus and SARS-CoV-2 in vitro. Their cytotoxicities were also investigated. RESULTS: Several extracts as well as Covid-Organics inhibited SARS-CoV-2 and FCoV infection at concentrations that did not affect cell viability. CONCLUSIONS: Some plant extracts show inhibitory activity against FCoV and SARS-CoV-2. However, it remains unclear whether peak plasma concentrations in humans can reach levels needed to inhibit viral infection following consumption of teas or Covid-Organics. Clinical studies are required to evaluate the utility of these drinks for COVID-19 prevention or treatment of patients.
Asunto(s)
Antivirales/farmacología , Artemisia/química , Extractos Vegetales/farmacología , SARS-CoV-2/efectos de los fármacos , Animales , Antivirales/química , Línea Celular , Supervivencia Celular/efectos de los fármacos , Coronavirus Felino/efectos de los fármacos , Coronavirus Felino/crecimiento & desarrollo , Extractos Vegetales/química , SARS-CoV-2/crecimiento & desarrollo , Ensayo de Placa ViralRESUMEN
Inhibition of herpes simplex virus type 1 (HSV-1) binding to the host cell surface by highly sulfated architectures is among the promising strategies to prevent virus entry and infection. However, the structural flexibility of multivalent inhibitors plays a major role in effective blockage and inhibition of virus receptors. In this study, we demonstrate the inhibitory effect of a polymer scaffold on the HSV-1 infection by using highly sulfated polyglycerols with different architectures (linear, dendronized, and hyperbranched). IC50 values for all synthesized sulfated polyglycerols and the natural sulfated polymer heparin were determined using plaque reduction infection assays. Interestingly, an increase in the IC50 value from 0.03 to 374 nM from highly flexible linear polyglycerol sulfate (LPGS) to less flexible scaffolds, namely, dendronized polyglycerol sulfate and hyperbranched polyglycerol sulfate was observed. The most potent LPGS inhibits HSV-1 infection 295 times more efficiently than heparin, and we show that LPGS has a much reduced anticoagulant capacity when compared to heparin as evidenced by measuring the activated partial thromboplastin time. Furthermore, prevention of infection by LPGS and the commercially available drug acyclovir were compared. All tested sulfated polymers do not show any cytotoxicity at concentrations of up to 1 mg/mL in different cell lines. We conclude from our results that more flexible polyglycerol sulfates are superior to less flexible sulfated polymers with respect to inhibition of HSV-1 infection and may constitute an alternative to the current antiviral treatments of this ubiquitous pathogen.
Asunto(s)
Herpesvirus Humano 1 , Antivirales/farmacología , Glicerol , Polímeros , SulfatosRESUMEN
Geometry-matching has been known to benefit the formation of stable biological interactions in natural systems. Herein, we report that the spiky nanostructures with matched topography to the influenza A virus (IAV) virions could be used to design next-generation advanced virus inhibitors. We demonstrated that nanostructures with spikes between 5 and 10 nm bind significantly better to virions than smooth nanoparticles, due to the short spikes inserting into the gaps of glycoproteins of the IAV virion. Furthermore, an erythrocyte membrane (EM) was coated to target the IAV, and the obtained EM-coated nanostructures could efficiently prevent IAV virion binding to the cells and inhibit subsequent infection. In a postinfection study, the EM-coated nanostructures reduced >99.9% virus replication at the cellular nontoxic dosage. We predict that such a combination of geometry-matching topography and cellular membrane coating will also push forward the development of nanoinhibitors for other virus strains, including SARS-CoV-2.
Asunto(s)
Betacoronavirus/ultraestructura , Infecciones por Coronavirus/virología , Nanoestructuras/ultraestructura , Neumonía Viral/virología , Antivirales/farmacología , Betacoronavirus/efectos de los fármacos , Sitios de Unión , COVID-19 , Infecciones por Coronavirus/tratamiento farmacológico , Diseño de Fármacos , Humanos , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza A/ultraestructura , Microscopía Electrónica , Modelos Biológicos , Nanotecnología , Pandemias , Neumonía Viral/tratamiento farmacológico , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus/efectos de los fármacos , Glicoproteína de la Espiga del Coronavirus/ultraestructura , Internalización del Virus/efectos de los fármacosRESUMEN
Here we report that negatively charged polysulfates can bind to the spike protein of SARS-CoV-2 via electrostatic interactions. Using a plaque reduction assay, we compare inhibition of SARS-CoV-2 by heparin, pentosan sulfate, linear polyglycerol sulfate (LPGS) and hyperbranched polyglycerol sulfate (HPGS). Highly sulfated LPGS is the optimal inhibitor, with an IC50 of 67â µg mL-1 (approx. 1.6â µm). This synthetic polysulfate exhibits more than 60-fold higher virus inhibitory activity than heparin (IC50 : 4084â µg mL-1 ), along with much lower anticoagulant activity. Furthermore, in molecular dynamics simulations, we verified that LPGS can bind more strongly to the spike protein than heparin, and that LPGS can interact even more with the spike protein of the new N501Y and E484K variants. Our study demonstrates that the entry of SARS-CoV-2 into host cells can be blocked via electrostatic interactions, therefore LPGS can serve as a blueprint for the design of novel viral inhibitors of SARS-CoV-2.
Asunto(s)
Antivirales/metabolismo , Heparina/metabolismo , Poliéster Pentosan Sulfúrico/metabolismo , SARS-CoV-2/química , Glicoproteína de la Espiga del Coronavirus/metabolismo , Internalización del Virus/efectos de los fármacos , Células A549 , Animales , Antivirales/química , Chlorocebus aethiops , Heparina/química , Humanos , Simulación de Dinámica Molecular , Poliéster Pentosan Sulfúrico/química , Polímeros/química , Polímeros/metabolismo , Unión Proteica , Glicoproteína de la Espiga del Coronavirus/química , Electricidad Estática , Células VeroRESUMEN
Multivalent binding inhibitors are a promising new class of antivirals that prevent virus infections by inhibiting virus binding to cell membranes. The design of these inhibitors is challenging as many properties, for example, inhibitor size and functionalization with virus attachment factors, strongly influence the inhibition efficiency. Here, virus binding inhibitors are synthesized, the size and functionalization of which are inspired by mucins, which are naturally occurring glycosylated proteins with high molecular weight (MDa range) and interact efficiently with various viruses. Hyperbranched polyglycerols (hPGs) with molecular weights ranging between 10 and 2600 kDa are synthesized, thereby hitting the size of mucins and allowing for determining the impact of inhibitor size on the inhibition efficiency. The hPGs are functionalized with sialic acids and sulfates, as suggested from the structure of mucins, and their inhibition efficiency is determined by probing the inhibition of influenza A virus (IAV) binding to membranes using various methods. The largest, mucin-sized inhibitor shows potent inhibition at pm concentrations, while the inhibition efficiency decreases with decreasing the molecular weight. Interestingly, the concentration-dependent IAV inhibition shows a biphasic behavior, which is attributed to differences in the binding affinity of the inhibitors to the two IAV envelope proteins, neuraminidase, and hemagglutinin.
Asunto(s)
Glicerol , Virus de la Influenza A , Mucinas , Polímeros , Acoplamiento Viral , Animales , Antivirales/farmacología , Membrana Celular/metabolismo , Membrana Celular/virología , Perros , Glicerol/síntesis química , Glicerol/metabolismo , Glicerol/farmacología , Hemaglutininas Virales/metabolismo , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza A/metabolismo , Células de Riñón Canino Madin Darby , Peso Molecular , Mucinas/química , Neuraminidasa/metabolismo , Polímeros/síntesis química , Polímeros/metabolismo , Polímeros/farmacología , Acoplamiento Viral/efectos de los fármacosRESUMEN
The topographic features of an implant, which mechanically regulate cell behaviors and functions, are critical for the clinical success in tissue regeneration. How cells sense and respond to the topographical cues, e.g., interfacial roughness, is yet to be fully understood and even debatable. Here, the mechanotransduction and fate determination of human mesenchymal stem cells (MSCs) on surface roughness gradients are systematically studied. The broad range of topographical scales and high-throughput imaging is achieved based on a catecholic polyglycerol coating fabricated by a one-step-tilted dip-coating approach. It is revealed that the adhesion of MSCs is biphasically regulated by interfacial roughness. The cell mechanotransduction is investigated from focal adhesion to transcriptional activity, which explains that cellular response to interfacial roughness undergoes a direct force-dependent mechanism. Moreover, the optimized roughness for promoting cell fate specification is explored.
Asunto(s)
Diferenciación Celular , Células Madre Mesenquimatosas , Adhesión Celular , Adhesiones Focales , Humanos , Mecanotransducción Celular/fisiología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/ultraestructura , Microscopía Electrónica de Rastreo , Regeneración/fisiología , Propiedades de SuperficieRESUMEN
Recently emerging graphene-based 2D nanoplatforms with multiple therapeutic modalities provide enormous opportunities to combat pathogenic bacterial infections. However, because these materials suffer from complicated synthesis, massive dosage requirements, and abundant nonlocalized heat, much more simplified, tunable, and localized eradication approaches are urgently required. Herein, we report on the fabrication of the metal-organic-framework (MOF)-derived 2D carbon nanosheets (2D-CNs) with phase-to-size transformation and localized bacterial eradication capabilities for augmented anti-infective therapy. The MOF-derived, ZnO-doped carbon on graphene (ZnO@G) is first synthesized and then anchored with phase transformable thermally responsive brushes (TRB) by in situ polymerization to yield the TRB-ZnO@G. The TRB-ZnO@G exhibits flexible 2D nanostructures, high photothermal activities, sustained Zn2+ ions release, and ON-OFF switchable phase-to-size transformation abilities. Notably, the near-infrared-triggered formation of TRB-ZnO@G-bacteria aggregations enables localized massive Zn2+ ions penetration, physical cutting, and hyperthermia killing, which synergistically enhance the disruption of bacterial membranes and intracellular substances. The obtained novel 2D-CNs not only present robust and localized multiple bacterial eradication capabilities with nearly 100% bactericidal efficiency at low concentrations but also possess rapid and safe skin wound disinfection via a short-time photothermal treatment without damaging normal skin tissues or causing accumulative toxicities, thus presenting great potential for broad-spectrum eradication of pathogenic bacteria.
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Antiinfecciosos/farmacología , Infecciones Bacterianas/tratamiento farmacológico , Estructuras Metalorgánicas/química , Nanoestructuras/química , Antiinfecciosos/química , Bacterias/efectos de los fármacos , Infecciones Bacterianas/microbiología , Carbono/química , Desinfección , Grafito/química , Humanos , Estructuras Metalorgánicas/uso terapéutico , Óxido de Zinc/químicaRESUMEN
In this study, we demonstrate the concept of "topology-matching design" for virus inhibitors. With the current knowledge of influenzaâ A virus (IAV), we designed a nanoparticle-based inhibitor (nano-inhibitor) that has a matched nanotopology to IAV virions and shows heteromultivalent inhibitory effects on hemagglutinin and neuraminidase. The synthesized nano-inhibitor can neutralize the viral particle extracellularly and block its attachment and entry to the host cells. The virus replication was significantly reduced by 6 orders of magnitude in the presence of the reverse designed nano-inhibitors. Even when used 24â hours after the infection, more than 99.999 % inhibition is still achieved, which indicates such a nano-inhibitor might be a potent antiviral for the treatment of influenza infection.
Asunto(s)
Antivirales/farmacología , Diseño de Fármacos , Virus de la Influenza A/efectos de los fármacos , Gripe Humana/tratamiento farmacológico , Nanopartículas/química , Zanamivir/farmacología , Animales , Antivirales/síntesis química , Antivirales/química , Perros , Glicerol/química , Glicerol/farmacología , Humanos , Lactosa/análogos & derivados , Lactosa/química , Lactosa/farmacología , Células de Riñón Canino Madin Darby/efectos de los fármacos , Células de Riñón Canino Madin Darby/virología , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Tamaño de la Partícula , Polímeros/química , Polímeros/farmacología , Ácidos Siálicos/química , Ácidos Siálicos/farmacología , Propiedades de Superficie , Replicación Viral/efectos de los fármacos , Zanamivir/síntesis química , Zanamivir/químicaRESUMEN
Flexible multivalent 3D nanosystems that can deform and adapt onto the virus surface via specific ligand-receptor multivalent interactions can efficiently block virus adhesion onto the cell. We here report on the synthesis of a 250â nm sized flexible sialylated nanogel that adapts onto the influenza A virus (IAV) surface via multivalent binding of its sialic acid (SA) residues with hemagglutinin spike proteins on the virus surface. We could demonstrate that the high flexibility of sialylated nanogel improves IAV inhibition by 400 times as compared to a rigid sialylated nanogel in the hemagglutination inhibition assay. The flexible sialylated nanogel efficiently inhibits the influenza A/X31 (H3N2) infection with IC50 values in low picomolar concentrations and also blocks the virus entry into MDCK-II cells.
Asunto(s)
Antivirales/farmacología , Virus de la Influenza A/efectos de los fármacos , Ácido N-Acetilneuramínico/química , Nanogeles/química , Animales , Antivirales/química , Perros , Virus de la Influenza A/fisiología , Concentración 50 Inhibidora , Células de Riñón Canino Madin Darby , Microscopía de Fuerza Atómica , Microscopía Fluorescente , Internalización del Virus/efectos de los fármacosRESUMEN
Nanointerfacial decoration of silver nanoparticles (AgNPs) is an ideal protocol to improve the antibacterial efficiency of diverse nanomaterials, including carbon nanotube (CNT), graphene, and many other intensively studied nanoarchitectures, which provides a tremendous possibility for designing advanced antibacterial biomaterials and biomedical devices. However, the direct exposure of AgNPs will lead to potential mammalian cell apoptosis and death, which significantly limits their biological applications. In this study, we demonstrated a green and one-step approach to achieve robust antibacterial and highly biocompatible AgNP-CNT composites. AgNPs were produced via mussel-inspired "one-step" in situ reduction and coating process and were anchored onto the surface of a CNT. Simultaneously, protective polymer layers were formed to shield the AgNPs to improve their biocompatibility. Because of the bactericidal efficiency of AgNPs, the composites showed robust antibacterial efficiency in terms of both inhibition of bacterial cell growth and bacterial killing activity. Moreover, owing to the shielding effects of the polymer coatings, the nanocomposites exhibited much improved compatibility with human umbilical vein endothelial cells compared with bare AgNP-CNTs. Furthermore, the nanocomposites exhibited good stability in psychological solutions. With integrated excellent antibacterial activity, cell compatibility, and long-term stability, it is believed that the synthesized AgNP-CNT composites will be of promising potential in antibacterial applications. Meanwhile, the proposed strategies can also be applied to fabricate many other kinds of AgNP-based composites because of the versatile functionality of catecholic polymers.
Asunto(s)
Antibacterianos , Materiales Biocompatibles , Bivalvos , Nanopartículas del Metal , Nanotubos de Carbono , Plata , Animales , Antibacterianos/química , Antibacterianos/farmacología , Apoptosis/efectos de los fármacos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Escherichia coli/crecimiento & desarrollo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Nanopartículas del Metal/química , Nanopartículas del Metal/uso terapéutico , Plata/química , Plata/farmacología , Staphylococcus aureus/crecimiento & desarrolloRESUMEN
Surface modification has long been of great interest to impart desired functionalities to the bioimplants. However, due to the limitations of recent technologies in surface modification, it is highly desirable to explore novel protocols, which can advantageously and efficiently endow the inert material surfaces with versatile biofunctionalities. Herein, to achieve versatile and rapid postfunctionalization of polymeric membrane, we demonstrate a new strategy for the fabrication of ß-cyclodextrin (ß-CD) modified host membrane substrate that can recognize a series of well-designed guest macromolecules. The surface assembly procedure was driven by the host-guest interaction between adamantane (Ad) and ß-CD. ß-CD immobilized host membrane was fabricated via two steps: (1) epoxy groups enriched poly(ether sulfone) (PES) membrane was first prepared via in situ cross-linking polymerization and subsequently phase separation; (2) mono-6-deoxy-6-ethylenediamine-ß-CD (EDA-ß-CD) was then anchored onto the surface of the epoxy functionalized PES membrane to obtain PES-CD. Subsequently, three types of Ad-terminated polymers, including Ad-poly(styrenesulfonate-co-sodium acrylate) (Ad-PSA), Ad-methoxypoly(ethylene glycol) (Ad-PEG), and Ad-poly(methyl chloride-quaternized 2-(dimethylamino)ethyl methacrylate (Ad-PMT), were separately assembled onto the ß-CD immobilized surfaces to endow the membranes with anticoagulant, antifouling, and antibacterial capability, respectively. Activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT) measurements were carried out to explore the anticoagulant activity. The antifouling capability was evaluated via protein adsorption and platelet adhesion measurements. Moreover, Staphyllococcous aureus (S. aureus) was selected as model bacteria to evaluate the antibacterial ability of the functionalized membranes. The results indicated that well-regulated blood compatibility, antifouling capability, and bactericidal activity could be achieved by the proposed rapid postfunctionalization on polymeric membranes. This approach of versatile and rapid postfunctionalization is promising for the preparation of multifunctional polymeric membrane materials to meet with various demands for the further applications.
Asunto(s)
Antibacterianos/química , Ciclodextrinas/química , Polímeros/química , Sulfonas/química , Antibacterianos/síntesis química , Antibacterianos/farmacología , Incrustaciones Biológicas/prevención & control , Relación Dosis-Respuesta a Droga , Interacciones Hidrofóbicas e Hidrofílicas , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Polimerizacion , Polímeros/síntesis química , Polímeros/farmacología , Staphylococcus aureus/citología , Staphylococcus aureus/efectos de los fármacos , Relación Estructura-Actividad , Sulfonas/síntesis química , Sulfonas/farmacología , Propiedades de SuperficieRESUMEN
Combining the advantages of the fibrous nanostructure of carbon nanotubes (CNTs) and the bioactivities of heparin/heparin-mimicking polyanions, functional nanofibrous heparin or heparin-mimicking multilayers were constructed on PVDF membrane with highly promoted endothelialization and antithrombogenic activities. Oxidized CNT (oCNT) was first functionalized with water-soluble chitosan (polycation), then enwrapped with heparin or a typical sulfonated heparin-mimicking polymers (poly(sodium 4-styrenesulfonate-co-sodium methacrylate)) to construct the multilayers. Then, the surface-deposited multilayers were constructed via electrostatic layer-by-layer assembly of the functionalized oCNTs. The scanning electron microscope and atom force microscope images confirmed that the coated multilayers exhibited nanofibrous and porous structure. The live/dead cell staining and cell viability assay results indicated that the coated nanofibrous multilayers had excellent compatibility with endothelial cells. The cell morphology observation further confirmed the promotion ability of surface endothelialization due to the coated heparin/heparin-mimicking multilayers. Further systematical evaluation on blood compatibility revealed that the surface heparin/heparin-mimicking multilayer-coated membranes also had significantly improved blood compatibility including restrained platelet adhesion and activation, prolonged blood clotting times, and inhibited activation of coagulation and complement factors. In summary, the proposed nanofibrous multilayers integrated endothelialization and antithrombogenic properties; meanwhile, the heparin-mimicking coating validated comparable performances as heparin coating. Herein, it is expected that the surface coating of nanofibrous multilayers, especially the facilely constructed heparin-mimicking coating, may have great application potential in biomedical fields.
Asunto(s)
Materiales Biocompatibles Revestidos/química , Fibrinolíticos/química , Heparina/química , Células Endoteliales de la Vena Umbilical Humana/fisiología , Nanofibras/química , Adsorción , Supervivencia Celular , Células Cultivadas , Materiales Biocompatibles Revestidos/farmacología , Activación de Complemento , Fibrinógeno/química , Fibrinolíticos/farmacología , Heparina/farmacología , Humanos , Ensayo de Materiales , Imitación Molecular , Tiempo de Tromboplastina Parcial , Adhesividad Plaquetaria , Polivinilos/química , Unión Proteica , Albúmina Sérica Bovina/químicaRESUMEN
In this study, we proposed a catechol chemistry inspired approach to construct surface self-cross-linked polymer nanolayers for the design of versatile biointerfaces. Several representative biofunctional polymers, P(SS-co-AA), P(SBMA-co-AA), P(EGMA-co-AA), P(VP-co-AA), and P(MTAC-co-AA), were first synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, and then the catecholic molecules (dopamine, DA) were conjugated to the acrylic acid (AA) units by the facile carbodiimide chemistry. Then, the catechol (Cat) group conjugated biofunctional polymers, named PSS-Cat, PSBMA-Cat, PEGMA-Cat, PVP-Cat, and PMTAC-Cat, were applied for the construction of self-cross-linked nanolayers on polymeric substrates via the pH induced catechol cross-linking and immobilization. The XPS spectra, surface morphology, and wettability gave robust evidence that the catechol conjugated polymers were successfully coated, and the coated substrates possessed increased surface roughness and hydrophilicity. Furthermore, the systematic in vitro investigation of protein adsorption, platelet adhesion, activated partial thromboplastin time (APTT), thrombin time (TT), cell viability, and antibacterial ability confirmed that the coated nanolayers conferred the substrates with versatile biological performances. The PSS-Cat coated substrate had low blood component activation and excellent anticoagulant activity; while the PEGMA-Cat and PSBMA-Cat showed ideal resistance to protein fouling and inhibition of platelet activation. The PSS-Cat and PVP-Cat coated substrates exhibited promoted endothelial cell proliferation and viability. The PMTAC-Cat coated substrate showed an outstanding activity on bacterial inhibition. In conclusion, the catechol chemistry inspired approach allows the self-cross-linked nanolayers to be easily immobilized on polymeric substrates with the stable conformation and multiple biofunctionalities. It is expected that this low-cost and facile bioinspired coating system will present great potential in creating novel and versatile biointerfaces.