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1.
Environ Monit Assess ; 194(12): 875, 2022 Oct 13.
Article in English | MEDLINE | ID: mdl-36227428

ABSTRACT

Polymers, such as partially hydrolyzed polyacrylamide (HPAM), are widely used in oil fields to enhance or improve the recovery of crude oil from the reservoirs. It works by increasing the viscosity of the injected water, thus improving its mobility and oil recovery. However, during such enhanced oil recovery (EOR) operations, it also produces a huge quantity of water alongside oil. Depending on the age and the stage of the oil reserve, the oil field produces ~ 7-10 times more water than oil. Such water contains various types of toxic components, such as traces of crude oil, heavy metals, and different types of chemicals (used during EOR operations such as HPAM). Thus, a huge quantity of HPAM containing produced water generated worldwide requires proper treatment and usage. The possible toxicity of HPAM is still ambiguous, but its natural decomposition product, acrylamide, threatens humans' health and ecological environments. Therefore, the main challenge is the removal or degradation of HPAM in an environmentally safe manner from the produced water before proper disposal. Several chemical and thermal techniques are employed for the removal of HPAM, but they are not so environmentally friendly and somewhat expensive. Among different types of treatments, biodegradation with the aid of individual or mixed microbes (as biofilms) is touted to be an efficient and environmentally friendly way to solve the problem without harmful side effects. Many researchers have explored and reported the potential of such bioremediation technology with a variable removal efficiency of HPAM from the oil field produced water, both in lab scale and field scale studies. The current review is in line with United Nations Sustainability Goals, related to water security-UNSDG 6. It highlights the scale of such HPAM-based EOR applications, the challenge of produced water treatment, current possible solutions, and future possibilities to reuse such treated water sources for other applications.


Subject(s)
Oil and Gas Fields , Petroleum , Acrylamides , Acrylic Resins/chemistry , Acrylic Resins/metabolism , Environmental Monitoring , Humans , Water Pollution
2.
Exp Eye Res ; 214: 108881, 2022 01.
Article in English | MEDLINE | ID: mdl-34871569

ABSTRACT

Corneal ulcers, a leading cause of blindness in the developing world are treated inappropriately without prior microbiology assessment because of issues related to availability or cost of accessing these services. In this work we aimed to develop a device for identifying the presence of Gram-positive or Gram-negative bacteria or fungi that can be used by someone without the need for a microbiology laboratory. Working with branched poly (N-isopropyl acrylamide) (PNIPAM) tagged with Vancomycin, Polymyxin B, or Amphotericin B to bind Gram-positive bacteria, Gram-negative bacteria and fungi respectively, grafted onto a single hydrogel we demonstrated specific binding of the organisms. The limit of detection of the microbes by these polymers was between 10 and 4 organisms per high power field (100X) for bacteria and fungi binding polymers respectively. Using ex vivo and animal cornea infection models infected with bacteria, fungi or both we than demonstrated that the triple functionalised hydrogel could pick up all 3 organisms after being in place for 30 min. To confirm the presence of bacteria and fungi we used conventional microbiology techniques and fluorescently labelled ligands or dyes. While we need to develop an easy-to-use either a colorimetric or an imaging system to detect the fluorescent signals, this study presents for the first time a simple to use hydrogel system, which can be applied to infected eyes and specifically binds different classes of infecting agents within a short space of time. Ultimately this diagnostic system will not require trained microbiologists for its use and will be used at the point-of-care.


Subject(s)
Acrylic Resins/metabolism , Corneal Ulcer/diagnosis , Eye Infections, Bacterial/diagnosis , Eye Infections, Fungal/diagnosis , Hydrogels/metabolism , Ligands , Acrylic Resins/chemistry , Amphotericin B/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Bacteria/metabolism , Candidiasis/diagnosis , Candidiasis/microbiology , Corneal Ulcer/microbiology , Eye Infections, Bacterial/microbiology , Eye Infections, Fungal/microbiology , Fungi/drug effects , Fungi/metabolism , Humans , Hydrogels/chemistry , Microbial Sensitivity Tests , Polymyxin B/pharmacology , Pseudomonas Infections/diagnosis , Pseudomonas Infections/microbiology , Rabbits , Staphylococcal Infections/diagnosis , Staphylococcal Infections/microbiology , Vancomycin/pharmacology
3.
Chembiochem ; 22(24): 3452-3461, 2021 12 10.
Article in English | MEDLINE | ID: mdl-34596326

ABSTRACT

Smart hydrogels hold much potential for biocatalysis, not only for the immobilization of enzymes, but also for the control of enzyme activity. We investigated upper critical solution temperature-type poly N-acryloyl glycinamide (pNAGA) hydrogels as a smart matrix for the amine transaminase from Bacillus megaterium (BmTA). Physical entrapment of BmTA in pNAGA hydrogels results in high immobilization efficiency (>89 %) and high activity (97 %). The temperature-sensitiveness of pNAGA is preserved upon immobilization of BmTA and shows a gradual deswelling upon temperature reduction. While enzyme activity is mainly controlled by temperature, deactivation tended to be higher for immobilized BmTA (≈62-68 %) than for free BmTA (≈44 %), suggesting a deactivating effect due to deswelling of the pNAGA gel. Although the deactivation in response to hydrogel deswelling is not yet suitable for controlling enzyme activity sufficiently, it is nevertheless a good starting point for further optimization.


Subject(s)
Acrylic Resins/metabolism , Bacillus megaterium/enzymology , Hydrogels/metabolism , Temperature , Transaminases/metabolism , Acrylic Resins/chemistry , Hydrogels/chemistry , Molecular Structure
4.
Yakugaku Zasshi ; 141(9): 1079-1086, 2021.
Article in Japanese | MEDLINE | ID: mdl-34471009

ABSTRACT

Protein affinity reagents that specifically and strongly bind to target molecules are widely used in disease detection, diagnosis, and therapy. Although antibodies and their fragments are the gold standard in protein-protein inhibitors (PPIs), synthetic polymers such as linear polymers, dendrimers, and nanoparticles as cost-effective PPIs have attracted great attention as alternatives to antibodies. These polymers exhibit high affinity to the target by imitating natural protein-protein interactions. However, only a few in vivo applications have been reported. Here, our recent advances in the development of synthetic polymers for in vivo application are reviewed. Poly(N-isopropylacrylamide) (pNIPAm) was used as a model of synthetic affinity reagents. Incorporation of both sulfated carbohydrate and hydrophobic monomers into lightly crosslinked pNIPAm nanoparticles (NPs) captured and neutralized vascular endothelial growth factor (VEGF) and inhibited tumor growth upon intravenous injection into tumor-bearing mice. Modification of a liposome with the pNIPAm-based linear polymer increased the polymer circulation time after intravenous injection and improved the affinity for the target. The pNIPAm-based NPs delivered by oral administration captured the target small molecules and inhibited their absorption from the intestine. Our recent findings provide useful information for the design of synthetic polymers that capture target molecules in vivo.


Subject(s)
Acrylic Resins , Drug Design/methods , Molecular Targeted Therapy/methods , Nanoparticles , Polymers , Acrylic Resins/administration & dosage , Acrylic Resins/chemistry , Acrylic Resins/metabolism , Acrylic Resins/pharmacology , Administration, Oral , Animals , Humans , Hydrophobic and Hydrophilic Interactions , Indicators and Reagents , Injections, Intravenous , Liposomes , Mice , Nanoparticles/chemistry , Neoplasms/drug therapy , Neoplasms/pathology , Polymers/administration & dosage , Polymers/chemistry , Polymers/metabolism , Polymers/pharmacology , Vascular Endothelial Growth Factor A/administration & dosage , Vascular Endothelial Growth Factor A/metabolism , Vascular Endothelial Growth Factor A/pharmacology
5.
Carbohydr Polym ; 255: 117332, 2021 Mar 01.
Article in English | MEDLINE | ID: mdl-33436175

ABSTRACT

To fabricate a mucoadhesive hydrogel with superior properties for local delivery of cisplatin (CDDP) to colorectal cancer, a hardcore bottle-brush polymer (HCBBP) was developed through grafting of poly(acrylic acid) (PAA) on cellulose nanocrystals (CNC) at 6, 9 and 12 CNC:PAA w/w ratios. The developed materials were characterized by acid-base titrations, FT-IR, electron microscopy, muco-rheological behaviour in the presence of mucin, in vitro drug release and anticancer activity against human HCT-116 colorectal cancer cells. The results showed CNC-g-PAA9 to have superior rheological behavior in the presence of mucin compared to CNC and other gels under study indicating beneficial mucoadhesive characteristics. CNC-g-PAA9:CDDP complex showed slow CDDP release causing a significant increase in IC 50 of the drug (> 3-fold) against HCT116 cells. The developed CNC-PAA9 hydrogel showed no intrinsic cytotoxicity on its own. The results point to a great promise for CNC-g-PAA9 as mucoadhesive hydrogels for local platinum delivery in colorectal cancer.


Subject(s)
Acrylic Resins/chemistry , Antineoplastic Agents/pharmacology , Cellulose/chemistry , Cisplatin/pharmacology , Drug Carriers , Hydrogels/chemical synthesis , Acrylic Resins/metabolism , Antineoplastic Agents/metabolism , Cell Adhesion , Cell Survival/drug effects , Cellulose/metabolism , Cisplatin/metabolism , Drug Compounding/methods , Drug Liberation , HCT116 Cells , Humans , Hydrogels/metabolism , Inhibitory Concentration 50 , Kinetics , Mucins/metabolism , Nanoparticles/chemistry , Nanoparticles/ultrastructure
6.
J Biomed Mater Res B Appl Biomater ; 109(1): 92-101, 2021 01.
Article in English | MEDLINE | ID: mdl-32627383

ABSTRACT

Like many other cell types, neuroblastoma cells are also known to respond to mechanical cues in their microenvironment in vitro. They were shown to have mechanotransduction pathways, which result in enhanced neuronal morphology on stiff substrates. However, in previous studies, the differentiation process was monitored only by morphological parameters. Motivated by the lack of comprehensive studies that investigate the effects of mechanical cues on neuroblastoma differentiation, we used SH-SY5Y cells differentiated on polyacrylamide (PA) gels as a model. Cells differentiated on the surface of PA hydrogels with three different elastic moduli (0.1, 1, and 50 kPa) were morphologically evaluated and their electrophysiological responsiveness was probed using calcium imaging. Immunodetection of neural marker TUJ1 and p-FAK was used for biochemical characterization. Groups with defined stiffness that are matching and nonmatching to neural tissue extracellular matrix were used to distinguish biomimetic results from other effects. Results show that while cells display morphologies that do not resemble neurons on soft substrates, they are in fact electrophysiologically more responsive and abundant in neuronal marker TUJ1. Our findings suggest that while neuronal differentiation occurs more efficiently in microenvironments mechanically mimicking neural tissue, the SH-SY5Y model demonstrates morphologies that conflict with neuronal behavior under these conditions. These results are expected to contribute considerable input to researchers that use SH-SY5Y as a neuron model.


Subject(s)
Cell Differentiation/physiology , Mechanotransduction, Cellular/physiology , Neuroblastoma/metabolism , Neurons/cytology , Acrylic Resins/chemistry , Acrylic Resins/metabolism , Biomarkers/metabolism , Focal Adhesion Kinase 1/metabolism , Humans , Hydrogels/chemistry , Hydrogels/metabolism , Mechanical Phenomena , Surface Properties , Tubulin/metabolism
7.
Water Res ; 190: 116704, 2021 Feb 15.
Article in English | MEDLINE | ID: mdl-33279745

ABSTRACT

Cationic polyacrylamide (cPAM) is a widely used flocculant to dewater sewage sludge (SS) for high-solids anaerobic digestion (AD), and its degradation is crucial since it would release toxic acrylamide (AM) once entering environment. Hydrothermal treatment (HTT) is an efficient method to enhance the AD efficiency of SS. However, the effects of cPAM on AD of SS and the degradation of cPAM during HTT-AD process have not be studied. The study showed cPAM at 20 mg/g TS increased methane yield of SS from 127.0 to 138.9 ml CH4/g TS in HTT-AD process, and the biodegradability of cPAM was 76.3%, which was much higher than that (7.4%) without HTT. In HTT-AD process, the enrichment of certain microbes (e.g. Gelria sp.) was observed, which might be related with cPAM degradation. HTT decreased the molecular weight (MW) of cPAM, and resulted in the production of 2-hydroxy-ethyl-trimethylammonium, ammonia, trimethylamine, and ethanol. Methane potential tests of the main HTT products also showed they were easily to be degraded. Overall, HTT-AD integrated process was an efficient method to reduce environmental risk of cPAM as well as increase energy output (biogas), and the study also provided insights into the degradation mechanism of cPAM during HTT.


Subject(s)
Acrylic Resins , Sewage , Acrylic Resins/metabolism , Anaerobiosis , Bioreactors , Methane
8.
Drug Dev Ind Pharm ; 46(5): 706-716, 2020 May.
Article in English | MEDLINE | ID: mdl-32266837

ABSTRACT

The aim of the present work was to develop a promising drug delivery system of oxiconazole nitrate-loaded solid lipid nanoparticles (SLNs) topical gel to enhance the drug effectiveness for the treatment of Tinea infection. SLNs were prepared by emulsification-solvent evaporation method. Particle size and entrapment efficiency of the prepared SLNs were investigated. An appropriate formulation was selected and examined for morphology and physicochemical characterization adopting Scanning electron microscope and Differential scanning colorimetry. In-vitro drug release was also investigated. The selected SLNs were loaded into 1% Carbopol 934 gel that was investigated for homogeneity, pH, grittiness, spreadability, viscosity and in vitro drug release. Clinical study for the developed gel system compared to the corresponding marketed product was conducted on 28 patients. The results revealed that the prepared oxiconazole nitrate SLNs had drug entrapment efficiency ranging from 41.34% to 75.07% and zeta potential lying between -13 and -50. Physicochemical characterization revealed a decrease in the drug crystallinity in the prepared SLNs. The gel formulation showed appropriate physical characteristics and sustained in-vitro drug release. Clinical study for the prepared oxiconazole nitrate SLNs gel showed significantly less side effects, better patient satisfaction and superior clinical improvement compared with the corresponding marketed product.


Subject(s)
Acrylic Resins/administration & dosage , Antifungal Agents/administration & dosage , Imidazoles/administration & dosage , Nanoparticles/administration & dosage , Skin Absorption/drug effects , Tinea/drug therapy , Acrylic Resins/chemical synthesis , Acrylic Resins/metabolism , Adult , Animals , Antifungal Agents/chemical synthesis , Antifungal Agents/metabolism , Drug Carriers/administration & dosage , Drug Carriers/chemical synthesis , Drug Carriers/metabolism , Drug Compounding/methods , Female , Gels , Humans , Imidazoles/chemical synthesis , Imidazoles/metabolism , Lipids , Male , Middle Aged , Nanoparticles/chemistry , Nanoparticles/metabolism , Organ Culture Techniques , Rats , Rats, Wistar , Skin Absorption/physiology , Tinea/metabolism , Tinea/pathology , Young Adult
9.
Int J Pharm ; 581: 119292, 2020 May 15.
Article in English | MEDLINE | ID: mdl-32243967

ABSTRACT

The use of water-insoluble carriers for amorphous solid dispersions (ASDs) has attracted more recent interest as the kinetic solubility profiles (KSP) from these systems can achieve a more sustained level of supersaturation when compared with ASDs based on water-soluble polymers. However, the effect of swelling capacity of water-insoluble carriers on the resulting KSP of ASDs has not been fully explored in terms of their achievable degree and extent of drug supersaturation. Thus, the objective of this study is to compare kinetic solubility profiles of ASDs based on commercially available water-insoluble carriers in order to bridge this knowledge gap and provide fundamental information important to the design of ASDs based on water-insoluble carriers. This was achieved by comparing the KSP from non-sink dissolution studies of ASDs of two model poorly-water soluble drugs, indomethacin (IND) and posaconazole (PCZ) based on commercially available water-insoluble carriers with different equilibrium water swelling such as fully hydrolyzed (physically crosslinked) poly (vinyl alcohol) (PVA), Eudragit RS PO, as well as chemically crosslinked PHEMA hydrogels . Our results show that the higher the swelling capacity of the water-insoluble carrier, the faster the rate of supersaturation generation, and the shorter the sustained supersaturation due to drug precipitation. The interplay of particle size, total dose and the swelling capacity was also shown to be an essential aspect when tailoring the supersaturation generation from water-insoluble polymer-based ASDs. The importance of the swelling feature was confirmed using firstly different polymer carriers (PVA, Eudragit RS PO, and PHEMA) and then polymer samples of identical composition and drug loading but with different swelling capacities (e.g., PVA, physically crosslinked to different degrees). Furthermore, a large drug partitioning value between the polymer carrier and dissolution medium was found to limit the extent of drug release or supersaturation buildup from these ASDs. Finally, the existence of electrostatic polymer-drug interactions realized from our molecular dynamic simulations supports the observed impact of the large partitioning of the model drug IND between the polymer ED RS PO and the dissolution medium, thereby leading to a lower degree of supersaturation generation (or slower drug release) from this ASD.


Subject(s)
Acrylic Resins/metabolism , Drug Liberation , Polyhydroxyethyl Methacrylate/metabolism , Water/metabolism , Acrylic Resins/chemistry , Calorimetry, Differential Scanning/methods , Drug Carriers/chemistry , Drug Carriers/metabolism , Polyhydroxyethyl Methacrylate/chemistry , Solubility , Water/chemistry
10.
Hernia ; 24(6): 1293-1306, 2020 12.
Article in English | MEDLINE | ID: mdl-32006122

ABSTRACT

PURPOSE: Two innovative reinforced biologic materials were studied in a non-human primate hernia repair model. The test articles, which combine layers of ovine decellularized extracellular matrix with minimal amounts of synthetic polymer, were evaluated for their biologic performance as measured by inflammatory response, healing kinetics, integration, and remodeling into functional host tissue. For comparison, seven clinically used biologic and synthetic meshes were also studied. METHODS: Animals were implanted with test articles in surgically created full-thickness midline abdominal wall defects, and evaluated macroscopically and histologically at 4, 12, and 24 weeks. RESULTS: Macroscopically, biologics resorbed and remodeled into naturally appearing tissue; the reinforced biologics appeared similar, but remodeled earlier and were less prone to stretch. Synthetics developed a layer of reactive tissue above and separate from the contracted mesh structure. At early time points, the collagen networks of biologics and reinforced biologics were infiltrated by host cells primarily as a peripheral layer on the biologics. As early as 12 weeks, the collagen networks associated with the reinforced biologics remodeled into organized host collagen. By 24 weeks, both reinforced biologics and biologics had low levels of inflammation. In contrast, a foreign body response persisted at 24 weeks with the synthetics, which had developed less organized collagen, separate in space from the actual mesh. CONCLUSIONS: The current study shows a favorable response to reinforced biologics, which were associated with an initial inflammatory response, resolving by later time points, followed by active remodeling, and the formation of new morphologically functional collagen.


Subject(s)
Acrylic Resins/metabolism , Biocompatible Materials/chemistry , Herniorrhaphy/methods , Surgical Mesh/standards , Animals , Disease Models, Animal , Humans , Sheep
11.
Biomacromolecules ; 21(2): 892-902, 2020 02 10.
Article in English | MEDLINE | ID: mdl-31895978

ABSTRACT

Here, we report the striking properties such as high stretchability, self-healing, and adhesiveness of an amphiphilic copolymeric hydrogel (poly(acrylic acid)-poly(methyl methacrylate) (PAA-PMMA) gel) synthesized from two immiscible monomers-acrylic acid (AA) and methyl methacrylate (MMA)-through a simple free radical polymerization in an aqueous medium. The developed hydrogel, with a specific molar ratio of MMA and AA, is self-healable, which is attributed to the hydrophobic interaction arising from methyl groups of PMMA, as well as the breakdown and reformation of sacrificial noncovalent cross-linking through the weak hydrogen bonds between the carboxylic acid groups of PAA and methoxy groups of PMMA. The energy dissipation values in the hysteresis test signify the excellent self-recoverability of the hydrogel. The developed hydrogel showed adhesive behavior to the surfaces of polystyrene, glass, wood, metal, stone, ceramics, pork skin, and human skin. The physical and mechanical properties of the PAA-PMMA gel were fine-tuned through changes in the MMA/AA ratio and pH. Moreover, the PAA-PMMA hydrogel can serve as a template for calcium phosphate mineralization to yield a hydrogel composite, which improved MC3T3 cell adhesion and proliferation. Overall, we propose that depending on synthesis parameters and other scenarios, the synthesized PAA-PMMA hydrogel could potentially be employed in varying biomedical and industrial applications.


Subject(s)
Adhesives/chemistry , Cell Proliferation/physiology , Drug Development/methods , Hydrogels/chemistry , Polymers/chemistry , Surface-Active Agents/chemistry , Acrylic Resins/chemistry , Acrylic Resins/metabolism , Adhesives/metabolism , Animals , Calcium Chloride/chemistry , Calcium Chloride/metabolism , Cell Line , Hydrogels/metabolism , Mice , Phosphates/chemistry , Phosphates/metabolism , Polymerization , Polymers/metabolism , Polymethyl Methacrylate/chemistry , Polymethyl Methacrylate/metabolism , Potassium Compounds/chemistry , Potassium Compounds/metabolism , Skin/metabolism , Spectroscopy, Fourier Transform Infrared/methods , Surface-Active Agents/metabolism , Swine
12.
Colloids Surf B Biointerfaces ; 188: 110787, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31954269

ABSTRACT

Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer.


Subject(s)
Acrylic Resins/chemistry , alpha-Amylases/chemistry , Acrylic Resins/metabolism , Molecular Structure , Molecular Weight , alpha-Amylases/metabolism
13.
J Mater Chem B ; 8(34): 7685-7695, 2020 09 14.
Article in English | MEDLINE | ID: mdl-33456778

ABSTRACT

Synthetic hydrogels with the ability to recognize and bind target proteins are useful for a number of applications, including biosensing and therapeutic agent delivery. One popular method for fabricating recognitive hydrogels is molecular imprinting. A long-standing hypothesis of the field is that these molecularly imprinted polymers (MIPs) retain the chemical and geometric profile of their protein template, resulting in subsequent ability to recognize the template in solution. Here, we systematically determined the influence of network composition, as well as the identity, amount, and extraction of imprinting templates, on the protein binding of MIPs. Network composition (i.e. the relative number of ionizable and hydrophobic groups) explained the extent of protein adsorption in all cases. The identity and amount of imprinting template, albeit a protein or synthetic polymer (PEG) of similar molecular weight, did not significantly influence the amount of protein bound. While the purification method influenced the extent of template adsorption, it did so by chemically modifying the network (acrylamide hydrolysis, increasing the acid content by up to 21%) and not by voiding occupied MIP pores. Therefore, our results indicate that material composition determines the extent to which MIPs bind template and non-template proteins.


Subject(s)
Acrylic Resins/metabolism , Hydrogels/metabolism , Molecular Imprinting/methods , Proteins/metabolism , Acrylic Resins/chemistry , Adsorption , Animals , Cattle , Chickens , Hydrogels/chemistry , Hydrophobic and Hydrophilic Interactions , Muramidase/chemistry , Polyethylene Glycols/chemistry , Polyethylene Glycols/metabolism , Protein Binding , Proteins/chemistry , Serum Albumin, Bovine/chemistry , Static Electricity , gamma-Globulins/chemistry
14.
ACS Synth Biol ; 9(1): 76-83, 2020 01 17.
Article in English | MEDLINE | ID: mdl-31880928

ABSTRACT

Herein we report a new type of artificial cells capable of long-term protein expression and regulation. We constructed the artificial cells by grafting anti-His-tag aptamer into the polymer backbone of the hydrogel particles, and then immobilizing the His-tagged proteinaceous factors of the transcription and translation system into the hydrogel particles. Long-term protein expression for at least 16 days was achieved by continuously flowing feeding buffer through the artificial cells. The effect of various metal ions on the protein expression in the artificial cells was investigated. Utilizing the lac operator-repressor system, we could regulate the expression level of eGFP in the artificial cells by controlling the ß-D-1-thiogalatopyranoside (IPTG) concentration in the feeding buffer. The artificial cells based on the aptamer grafted hydrogel provide a useful platform for gene circuit engineering, metabolic engineering, drug delivery, and biosensors.


Subject(s)
Acrylic Resins/metabolism , Aptamers, Nucleotide/metabolism , Artificial Cells/metabolism , Hydrogels/chemistry , Protein Biosynthesis , Cell-Free System/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Regulatory Networks , Green Fluorescent Proteins/metabolism , Ions/metabolism , Isopropyl Thiogalactoside/metabolism , Lac Operon , Lac Repressors/metabolism , Luminescent Proteins/metabolism , Metals/metabolism , Synthetic Biology/methods , Transcription, Genetic , Red Fluorescent Protein
16.
Chem Commun (Camb) ; 56(4): 647-650, 2020 Jan 14.
Article in English | MEDLINE | ID: mdl-31840153

ABSTRACT

Here, we demonstrate use of a Mg2+-dependent, site-specific DNA enzyme (DNAzyme) to cleave oligos from polyacrylamide gel beads, which is suitable for use in drop-based assays. We show that cleavage efficiency is improved by use of a tandem-repeat cleavage site. We further demonstrate that DNAzyme-released oligos function as primers in reverse transcription of cell-released mRNA.


Subject(s)
DNA, Catalytic/metabolism , Nucleic Acids/metabolism , Acrylic Resins/chemistry , Acrylic Resins/metabolism , Gels/chemistry , Gels/metabolism , Immobilized Nucleic Acids/chemistry , Immobilized Nucleic Acids/metabolism , Magnesium/chemistry , Magnesium/metabolism , Nucleic Acid Amplification Techniques , Nucleic Acids/chemistry , Particle Size , Surface Properties
17.
J Mater Chem B ; 8(1): 136-145, 2020 01 07.
Article in English | MEDLINE | ID: mdl-31778137

ABSTRACT

Glycan-lectin interactions drive a diverse range of biological signaling and recognition processes. The display of glycans in multivalent format enables their intrinsically weak binding affinity to lectins to be overcome by the cluster glycoside effect, which results in a non-linear increase in binding affinity. As many lectins have multiple binding sites, upon interaction with glycosylated nanomaterials either aggregation or surface binding without aggregation can occur. Depending on the application area, either one of these responses are desirable (or undesirable) but methods to tune the aggregation state, independently from the overall extent/affinity of binding are currently missing. Herein, we use gold nanoparticles decorated with galactose-terminated polymer ligands, obtained by photo-initiated RAFT polymerization to ensure high end-group fidelity, to show the dramatic impact on agglutination behaviour due to the chemistry of the polymer linker. Poly(N-hydroxyethyl acrylamide) (PHEA)-coated gold nanoparticles, a polymer widely used as a non-ionic stabilizer, showed preference for aggregation with lectins compared to poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA)-coated nanoparticles which retained colloidal stability, across a wide range of polymer lengths and particle core sizes. Using biolayer interferometry, it was observed that both coatings gave rise to similar binding affinity and hence provided conclusive evidence that aggregation rate alone cannot be used to measure affinity between nanoparticle systems with different stabilizing linkers. This is significant, as turbidimetry is widely used to demonstrate glycomaterial activity, although this work shows the most aggregating may not be the most avid, when comparing different polymer backbones/coating. Overall, our findings underline the potential of PHPMA as the coating of choice for applications where aggregation upon lectin binding would be problematic, such as in vivo imaging or drug delivery.


Subject(s)
Acrylic Resins/metabolism , Plant Lectins/metabolism , Polymethacrylic Acids/metabolism , Soybean Proteins/metabolism , Binding Sites , Glycosylation , Metal Nanoparticles , Protein Binding
18.
Lab Chip ; 19(17): 2874-2884, 2019 09 07.
Article in English | MEDLINE | ID: mdl-31343025

ABSTRACT

Neutrophils combat infections and promote healing of damaged tissues while protecting the surrounding healthy tissue through a process called swarming. Swarming neutrophils release soluble factors that recruit additional neutrophils and shape the inflammation response. Additionally, neutrophils release extracellular vesicles (EVs), which are gaining attention as important intercellular mediators. We developed a large-scale array of bioparticles on a glass substrate that triggers neutrophil swarming in vitro in a spatially and temporally controlled manner that facilitates the analysis of neutrophil migration. Our platform can generate 30 000 neutrophil swarms on a glass slide in a highly reproducible manner (98% patterning efficiency), which produces an EV-rich supernatant that enables quantitative characterization of inflammation-specific EVs. Healthy neutrophils were able to form uniform swarms across the bioparticle array, which demonstrates a high degree of intercellular coordination. However, neutrophils swarming on the bioparticle array tended to have a lower radial velocity than neutrophils swarming toward a single target. After collecting and isolating EVs released by swarming and non-swarming neutrophils, we found that neutrophils constitutively release exosomes and microvesicles. Furthermore, EVs released by swarming neutrophils cause neutrophil activation and contain the proinflammatory mediator galectin-3, suggesting that EVs have an active role during neutrophil swarming. Ultimately, understanding EVs' role in intercellular communication during swarming will improve understanding of the complex signaling pathways involved in the regulation of inflammation.


Subject(s)
Cell-Derived Microparticles/metabolism , Extracellular Vesicles/metabolism , High-Throughput Screening Assays , Microfluidic Analytical Techniques , Neutrophils/metabolism , Acrylic Resins/chemistry , Acrylic Resins/metabolism , Blood Proteins , Cell-Derived Microparticles/chemistry , Extracellular Vesicles/chemistry , Galectin 3/chemistry , Galectin 3/metabolism , Galectins , Healthy Volunteers , High-Throughput Screening Assays/instrumentation , Humans , Inflammation/metabolism , Microfluidic Analytical Techniques/instrumentation , Neutrophils/chemistry
19.
Colloids Surf B Biointerfaces ; 181: 539-548, 2019 Sep 01.
Article in English | MEDLINE | ID: mdl-31185446

ABSTRACT

The biological synthesis of reduced graphene oxide (rGO) from graphene oxide (GO) is an emerging phenomenon for developing biocompatible nanomaterials for its potential applications in nanomedicine. In this study, we demonstrated a simple, green, and non-toxic method for graphene synthesis using the live biomass of Lysinibacillus sphaericus as the reducing and stabilizing agent under ambient conditions. Ultraviolet-visible spectroscopic analysis confirmed the formation of graphene from GO suspension. X-ray diffraction studies showed the disappearance of the GO peak and the appearance of characteristic graphene broad peak at 2θ = 22.8°. Infrared analysis showed the decrease/disappearance of peaks corresponding to the oxygen-containing functionalities, and appearance of a peak at 1620 cm-1 from unoxidized graphitic domains. Scanning electron microscopic images showed that L. sphaericus-reduced graphene oxide (L-rGO) contains aggregated graphene nanoflakes. Evaluation of the in vitro cytotoxicity of L-rGO nanosheets on human skin fibroblasts using the WST-1 assay did not show any significant effects after 24 h of exposure, which is indicative of biocompatibility. Polyacrylamide hydrogels with L-rGO were synthesized and used as scaffolds to support the growth and proliferation of skin fibroblasts. Cell viability assays and DAPI staining showed proliferation of fibroblasts and exhibited 83% of cell viability even after 28 days. Biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus was enhanced in nanocomposite hydrogels in the presence of 0.25 mg/mL GO and L-rGO in 48 h. Overall, this study showed that microbially-synthesized L-rGO can be used as a dopant in polymeric scaffolds for tissue engineering and highlighted their role in biofilm formation.


Subject(s)
Acrylic Resins/metabolism , Bacillaceae/metabolism , Carbon/metabolism , Graphite/metabolism , Hydrogels/metabolism , Nanocomposites/chemistry , Skin/metabolism , Tissue Engineering , Acrylic Resins/chemistry , Bacillaceae/chemistry , Bacillaceae/growth & development , Carbon/chemistry , Cell Survival/drug effects , Graphite/chemistry , Graphite/pharmacology , Humans , Hydrogels/chemistry , Particle Size , Skin/chemistry , Surface Properties
20.
Chemosphere ; 231: 184-193, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31129399

ABSTRACT

Hydrolyzed polyacrylamide (HPAM) is the most widely used water-soluble linear polymer with high molecular weight in polymer flooding. Microbiological degradation is an environment-friendly and effective method of treating HPAM-containing oilfield produced water. In this study, a strain SZK-5 that could degrade HPAM was isolated from soil contaminated by oilfield produced water. Based on morphological, biochemical characteristics and 16S rDNA sequence homology analysis, the strain was identified as Bacillus megaterium. The biodegradation capability of strain SZK-5 was determined by incubation in a mineral salt medium (MSM) containing HPAM under different environmental conditions, showing 55.93% of the HPAM removed after 7 d of incubation under the optimum conditions ((NH4)2SO4 = 1667.9 mg L-1, temperature = 24.05 °C and pH = 8.19). Cytochrome P450 (CYP) and urease (URE) played significant roles in biological carbon and nitrogen removal, respectively. The strain SZK-5 could resist the damages caused by oxidative stress given by crude oil and HPAM. To our knowledge, this is the first report about the biodegradation of HPAM by B. megaterium. These results suggest that strain SZK-5 might be a new auxiliary microbiological resource for the biodegradation of HPAM residue in wastewater and soil.


Subject(s)
Acrylic Resins/metabolism , Bacillus megaterium/metabolism , Biodegradation, Environmental , Acrylic Resins/analysis , Antioxidants/metabolism , Bacillus megaterium/enzymology , Carbon , Hydrolysis , Nitrogen , Oil and Gas Fields , Petroleum , Wastewater
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