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1.
Int J Biol Macromol ; 265(Pt 2): 130938, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38493814

RESUMO

The rapid manufacturing of biocomposite scaffold made of saturated-Poly(ε-caprolactone) (PCL) and unsaturated Polyester (PE) blends with gelatin and modified gelatin (NCO-Gel) is demonstrated. Polyester blend-based scaffold are fabricated with and without applying potential in the melt electrowriting system. Notably, the applied potential induces phase separation between PCL and PE and drives the formation of PE rich spots at the interface of electrowritten fibers. The objective of the current study is to control the phase separation between saturated and unsaturated polyesters occurring in the melt electro-writing process and utilization of this phenomenon to improve efficiency of biofunctionalization at the interface of scaffold via Aza-Michael addition reaction. Electron-deficient triple bonds of PE spots on the fibers exhibit good potential for the biofunctionalization via the aza-Michael addition reaction. PE spots are found to be pronounced in which blend compositions are PCL-PE as 90:10 and 75:25 %. The biofunctionalization of scaffold is monitored through CN bond formation appeared at 400 eV via X-ray photoelectron spectroscopy (XPS) and XPS chemical mapping. The described biofunctionalization methodology suggest avoiding use of multi-step chemical modification on additive manufacturing products and thereby rapid prototyping of functional polymer blend based scaffolds with enhanced biocompatibility and preserved mechanical properties. Additionally one-step additive manufacturing method eliminates side effects of toxic solvents and long modification steps during scaffold fabrication.


Assuntos
Poliésteres , Polímeros , Poliésteres/química , Alicerces Teciduais/química , Gelatina/química , Engenharia Tecidual/métodos
2.
Environ Geochem Health ; 46(2): 58, 2024 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-38277072

RESUMO

This research aimed to determine and evaluate the concentrations of As, Cu, Hg, Ni, and Pb, and the physicochemical properties of 48 agricultural soil samples, to identify potential ecological risks and their sources associated with heavy metals contamination in Usak, western Türkiye. Various methods were used to assess ecological risks, including geoaccumulation index (Igeo), enrichment factor (EF), degree of contamination (Cdeg), potential ecological risk (RI), and pollution load index (PLI). The heavy metals concentrations ranged from 4 to 61 mg/kg for As, 8-48 mg/kg for Cu, 0.01-0.06 mg/kg for Hg, 30-813 mg/kg for Ni, and 4-30 mg/kg for Pb. The mean As and Ni concentrations were much greater than Earth's crustal average, the world's mean values, and mean values from many other emerging countries. Igeo and EF values for As, Ni, and Pb indicate various degrees of contamination. Cdeg values show that 96% of the study area is affected to some degree by contamination. For RI values, 38% indicate ecological risks ranging from moderate to considerable degrees. PLI values show that 75% of the agricultural soils are moderately polluted. Spatial distribution maps of Cdeg, RI, and PLI show that the northeastern and southwestern parts of the study area have been polluted to different levels by As, Ni, and Pb. Industrial activities and excessive use of fertilizers, pesticides, fungicides, and herbicides were identified as major sources of heavy metals contamination in the agricultural soils of Usak.


Assuntos
Mercúrio , Metais Pesados , Poluentes do Solo , Solo/química , Chumbo/análise , Turquia , Monitoramento Ambiental/métodos , Poluentes do Solo/análise , Medição de Risco , Metais Pesados/análise , Mercúrio/análise , Análise Multivariada , China
3.
Macromol Biosci ; 24(4): e2300402, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38102867

RESUMO

This study describes the formation, size control, and penetration behavior of polymer nanodots (Pdots) consisting of single or few chain polythiophene-based conjugated polyelectrolytes (CPEs) via nanophase separation between good solvent and poor solvent of CPE. Though the chain singularity may be associated with dilution nanophase separation suggests that molecules of a good solvent create a thermodynamically driven solvation layer surrounding the CPEs and thereby separating the single chains even in their poor solvents. This statement is therefore corroborated with emission intensity/lifetime, particle size, and scattering intensity of polyelectrolyte in good and poor solvents. Regarding the augmented features, Pdots are implemented into cell imaging studies to understand the nuclear penetration and to differentiate the invasive characteristics of breast cancer cells. The python based red, green, blue (RGB) color analysis   depicts that Pdots have more nuclear penetration ability in triple negative breast cancer cells due to the different nuclear morphology in shape and composition and Pdots have penetrated cell membrane as well as extracellular matrix in spheroid models. The current Pdot protocol and its utilization in cancer cell imaging are holding great promise for gene/drug delivery to target cancer cells by explicitly achieving the very first priority of nuclear intake.


Assuntos
Corantes Fluorescentes , Pontos Quânticos , Semicondutores , Polímeros , Polieletrólitos , Solventes
4.
ACS Omega ; 8(40): 37413-37420, 2023 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-37841123

RESUMO

The study describes a simple yet robust methodology for forming gradients in polymer coatings with nanometer-thickness precision. The thickness gradients of 0-20 nm in the coating are obtained by a reactive layer-by-layer assembly of polyester and polyethylenimine on gold substrates. Three parameters are important in forming thickness gradients: (i) the incubation time, (ii) the incubation concentration of the polymer solutions, and (iii) the tilt angle of the gold substrate during the dipping process. After examining these parameters, the characterization of the anisotropic surface obtained under the best conditions is presented in the manuscript. The thickness profile and nanomechanical characterization of the polymer gradients are characterized by atomic force microscopy. The roughness analysis has demonstrated that the coating exhibited decreasing roughness with increasing thickness. On the other hand, Young's moduli of the thin and thick coatings are 0.50 and 1.4 MPa, respectively, which assured an increase in mechanical stability with increasing coating thickness. Angle-dependent infrared spectroscopy reveals that the C-O-C ester groups of the polyesters exhibit a perpendicular orientation to the surface, while the C≡C groups are parallel to the surface. The surface properties of the polymer gradients are explored by fluorescence microscopy, proving that the dye's fluorescence intensity increases as the coating thickness increases. The significant benefit of the suggested methodology is that it promises thickness control of gradients in the coating as a consequence of the fast reaction kinetics between layers and the reaction time.

5.
Molecules ; 28(4)2023 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-36838895

RESUMO

Exosomes released into the extracellular matrix have been reported to contain metabolic biomarkers of various diseases. These intraluminal vesicles are typically found in blood, urine, saliva, breast milk, cerebrospinal fluid, semen, amniotic fluid, and ascites. Analysis of exosomal content with specific profiles of DNA, microRNA, proteins, and lipids can mirror their cellular origin and physiological state. Therefore, exosomal cargos may reflect the physiological processes at cellular level and can potentially be used as biomarkers. Herein, we report an optical detection method for assaying exosomal biomarkers that supersedes the state-of-the-art time consuming and laborious assays such as ELISA and NTA. The proposed assay monitors the changes in optical properties of poly(3-(4-methyl-3'-thienyloxy) propyltriethylammonium bromide) upon interacting with aptamers/peptide nucleic acids in the presence or absence of target biomarkers. As a proof of concept, this study demonstrates facile assaying of microRNA, DNA, and advanced glycation end products in exosomes isolated from human plasma with detection levels of ~1.2, 0.04, and 0.35 fM/exosome, respectively. Thus, the obtained results illustrate that the proposed methodology is applicable for rapid and facile detection of generic exosomal biomarkers for facilitating diseases diagnosis.


Assuntos
Exossomos , MicroRNAs , Feminino , Humanos , Colorimetria , MicroRNAs/metabolismo , Biomarcadores/metabolismo , Proteínas/metabolismo , Exossomos/metabolismo , Biomarcadores Tumorais/metabolismo
6.
J Pharm Biomed Anal ; 219: 114914, 2022 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-35843186

RESUMO

Lab-on-a-chip devices have now-a-days become an important aspect of analytical/bioanalytical chemistry having wide range of applications including clinical diagnosis, drug screening, cell biology, environmental monitoring, food safety analysis etc. Conventional lab-on-a-chip devices generally employ chemicals that are not environmentally friendly and were commonly fabricated on hard plastic platform which are non-degradable and hence ignore the importance of green analytical chemistry. In today's scenario, it is highly imperative to protect our environment by using less toxic and environmentally friendly chemicals/solvents and biocompatible platforms. Accordingly, the present article comprehensively reviews on the various green aspects of lab-on-a-chip devices for analytical processes which aim at fabricating environmentally friendly and cost-effective downsized devices so that the risk factor at the user's end upon longer exposure as well as to the environment can be reduced. The decisive factors for the accomplishment of green aspects of lab-on-a-chip devices including sample preparation using lab-on-a-chip systems to minimize the amount of sample/solvents to few microliters only, substitution of harmful solvents with green alternatives, minimal waste generation or proper treatment of waste and biodegradable and biocompatible platforms for fabricating lab-on-a-chip devices have been discussed in details. Additionally, the challenges that may hinder their commercialization are also critically discussed.


Assuntos
Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas , Técnicas Analíticas Microfluídicas/métodos , Solventes
8.
Nanomaterials (Basel) ; 12(4)2022 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-35214969

RESUMO

A rapid and one-pot synthesis of poly 3-thiopheneacetic acid (PTAA) functionalized polyurea polymer dots (Pdots) using polyethyleneimine and isophorone diisocyanate is reported. The one-pot mini-emulsion polymerization technique yielded Pdots with an average diameter of ~20 nm. The size, shape, and concentration of the surface functional groups could be controlled by altering the synthesis parameters such as ultrasonication time, concentration of the surfactant, and crosslinking agent, and the types of isocyanates utilized for the synthesis. Colloidal properties of Pdots were characterized using dynamic light scattering and zeta potential measurements. The spherical geometry of Pdots was confirmed by scanning electron microscopy. The Pdots were post-functionalized by 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid for chelating gadolinium nanoparticles (Gd3+) that provide magnetic properties to the Pdots. Thus, the synthesized Pdots possess fluorescent and magnetic properties, imparted by PTAA and Gd3+, respectively. Fluorescence spectroscopy and microscopy revealed that the synthesized dual-functional Gd3+-Pdots exhibited detectable fluorescent signals even at lower concentrations. Magnetic levitation experiments indicated that the Gd3+-Pdots could be easily manipulated via an external magnetic field. These findings illustrate that the dua- functional Gd3+-Pdots could be potentially utilized as fluorescent reporters that can be magnetically manipulated for bioimaging applications.

9.
ACS Sens ; 7(3): 686-703, 2022 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-35226461

RESUMO

Poly(3-alkylthiophene) (PT)-based conjugated polyelectrolytes (CPEs) constitute an important class of responsive polymers with excellent optical properties. The electrostatic interactions between PTs and target analytes trigger complexation and concomitant conformational changes of the PT backbones that produce distinct optical responses. These conformation-induced optical responses of the PTs enable them to be utilized as reporters for detection of various analytes by employing simple UV-vis spectrophotometry or the naked eye. Numerous PTs with unique pendant groups have been synthesized to tailor their interactions with analytes such as nucleotides, ions, surfactants, proteins, and bacterial and viral pathogens. In this perspective, we discuss PT-target analyte complexation for bioanalytical applications and highlight recent advancements in point-of-care and field deployable assays. Subsequently, we highlight a few areas of critical importance for future applications of PTs as reporters, including (i) design and synthesis of specific PTs to advance the understanding of the mechanisms of interaction with target analytes, (ii) using arrays of PTs and linear discriminant analysis for selective and specific detection of target analytes, (iii) translation of conventional homogeneous solution-based assays into heterogeneous membrane-based assay formats, and finally (iv) the potential of using PT as an alternative to conjugated polymer nanoparticles and dots in bioimaging.


Assuntos
Nanopartículas , Polímeros , Conformação Molecular , Polieletrólitos , Eletricidade Estática
10.
ACS Appl Mater Interfaces ; 14(1): 94-103, 2022 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-34964349

RESUMO

Profiling of advanced glycation end products (AGEs) is an emerging area of clinical significance for disease diagnosis and prognosis. Typically, concentrations of AGEs are estimated in laboratories by trained personnel using sophisticated equipment. Herein, a facile approach for colorimetric and fluorometric profiling of AGEs is reported for rapid and on-site analysis. The concentrations of AGE levels in plasma are estimated via changes in optical properties of polythiophenes (PTs) upon interaction with aptamers (Apts) in the presence and in the absence of AGEs. To validate the proposed approach, glyceraldehyde-derived AGEs (AGE class 1 [AGE1]), the biomarker associated with cardiovascular diseases and diabetes, are used as a model system. Colorimetric analysis yielded linear responses for AGE1 for clinically relevant concentration ranges between 1.5 and 300 µg/mL with a limit of detection (LOD) of ∼1.3 µg/mL. Subsequently, an approach utilizing PTs with four different pendant groups in conjunction with four different Apts is demonstrated for qualitative colorimetric profiling and for quantitative fluorometric profiling of up to four AGEs in clinical matrices. Principal component analysis (PCA) of fluorometric responses of AGE-spiked samples yielded distinct responses for the different AGEs tested. Thus, the proposed approach ascertains rapid profiling of spiked AGEs in plasma samples without the requirement of preanalytical processing and advanced instrumentation, thereby facilitating on-site diagnosis.


Assuntos
Colorimetria/métodos , Produtos Finais de Glicação Avançada/sangue , Espectrometria de Fluorescência/métodos , Aptâmeros de Nucleotídeos/química , Biomarcadores/sangue , Biomarcadores/química , Corantes Fluorescentes/química , Produtos Finais de Glicação Avançada/química , Humanos , Limite de Detecção , Polímeros/química , Tiofenos/química
11.
Langmuir ; 37(37): 10902-10913, 2021 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-34477388

RESUMO

We here demonstrate the utilization of reactive layer-by-layer (rLBL) assembly to form a nanogel coating made of branched polyethylenimine (BPEI) and alkyne containing polyester (PE) on a gold surface. The rLBL is generated by the rapid aza-Michael addition reaction of the alkyne group of PE and the -NH2 groups of BPEI by yielding a homogeneous gel coating on the gold substrate. The thickness profile of the nanogel revealed that a 400 nm thick coating is formed by six multilayers of rLBL, and it exhibits 50 nm roughness over 8 µm distance. The LBL characteristics were determined via depth profiling analysis by X-ray photoelectron spectroscopy, and it has been shown that a 70-100 nm periodic increase in gel thickness is a consequence of consecutive cycles of rLBL. A detailed XPS analysis was performed to determine the yield of the rLBL reaction: the average yield was deduced as 86.4% by the ratio of the binding energies at 286.26 eV, (C═CN-C bond) and 283.33 eV, (C≡C triple bond). The electrochemical characterization of the nanogels ascertains that up to the six-multilayered rLBL of BPEI-PE is electroactive, and the nanogel permeability had led to drive mass and charge transfer effectively. These results promise that nanogel formation by rLBL films may be a straightforward modification of electrodes approach, and it exhibits potential for the application of soft biointerfaces.


Assuntos
Poliésteres , Polietilenoimina , Nanogéis , Polietilenoglicóis
12.
Int J Biol Macromol ; 180: 729-738, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33757854

RESUMO

Natural gums and mucilages from plant-derived polysaccharides are potential candidates for a tissue-engineering scaffold by their ability of gelation and biocompatibility. Herein, we utilized Glucuronoxylan-based quince seed hydrogel (QSH) as a scaffold for tissue engineering applications. Optimization of QSH gelation was conducted by varying QSH and crosslinker glutaraldehyde (GTA) concentrations. Structural characterization of QSH was done by Fourier Transform Infrared Spectroscopy (FTIR). Furthermore, morphological and mechanical investigation of QSH was performed by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). The protein adsorption test revealed the suitability of QSH for cell attachment. Biocompatibility of QSH was confirmed by culturing NIH-3T3 mouse fibroblast cells on it. Cell viability and proliferation results revealed that optimum parameters for cell viability were 2 mg mL-1 of QSH and 0.03 M GTA. SEM and DAPI staining results indicated the formation of spheroids with a diameter of approximately 300 µm. Furthermore, formation of extracellular matrix (ECM) microenvironment was confirmed with the Collagen Type-I staining. Here, it was demonstrated that the fabricated QSH is a promising scaffold for 3D cell culture and tissue engineering applications provided by its highly porous structure, remarkable swelling capacity and high biocompatibility.


Assuntos
Hidrogéis/química , Rosaceae/química , Sementes/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Xilanos/química , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Matriz Extracelular/química , Matriz Extracelular/metabolismo , Matriz Extracelular/ultraestrutura , Hidrogéis/metabolismo , Camundongos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Células NIH 3T3 , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier , Xilanos/metabolismo
13.
ACS Sens ; 6(3): 950-957, 2021 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-33621051

RESUMO

This study presents a nonamplification-based nucleic acid assay for the detection of single-nucleotide polymorphism (SNP) associated with familial Mediterranean fever (FMF) besides polymerase chain reaction (PCR)-based methodologies. The major objective is to show the potential of the proposed assay for rapid screening of FMF in a Mediterranean region of 400 million population. The assay relies on binding difference of specially designed wild and mutant primers to the target genomic DNA, followed by determination of unbound primers by quick titration of a cationic polythiophene reporter. The fluorescent reporter exhibits signal transition from 525 to 580 nm in the presence of unbound primers, and it correlates the binding affinity of label-free primers to the homozygous wild and mutant genomes. As a proof of concept, 26 real samples are studied relying on the ON and OFF fluorescence signals of the cationic polythiophene reporter. The results are analyzed by principal component analysis (PCA), which provides clear separation of healthy and patient individuals. The further analysis by support vector machine (SVM) classification has revealed that our assay converges to 96% overall accuracy. These results support that the PCR-free nucleic acid assay has a significant potential for rapid and cost-effective screening of familial Mediterranean fever.


Assuntos
Polímeros , Tiofenos , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único
14.
ACS Appl Mater Interfaces ; 12(28): 31270-31281, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32551533

RESUMO

Over the past few decades, colorimetric assays have been developed for cost-effective and rapid on-site urinalysis. Most of these assays were employed for detection of biomarkers such as glucose, uric acid, ions, and albumin that are abundant in urine at micromolar to millimolar levels. In contrast, direct assaying of urinary biomarkers such as glycated proteins, low-molecular-weight reactive oxygen species, and nucleic acids that are present at significantly lower levels (nanomolar to picomolar) remain challenging due to the interferences from the urine sample matrix. State-of-the-art assays for detection of trace amounts of urinary biomarkers typically utilize time-consuming and equipment-dependent sample pretreatment or clean-up protocols prior to assaying, which limits their applicability for on-site analysis. Herein, we report a colorimetric assay for on-site detection of trace amount of generic biomarkers in urine without involving tedious sample pretreatment protocols. The detection strategy is based on monitoring the changes in optical properties of poly(3-(4-methyl-3'-thienyloxy)propyltriethylammonium bromide) upon interacting with an aptamer or a peptide nucleic acid in the presence and absence of target biomarkers of relevance for the diagnosis of metabolic complications and diabetes. As a proof of concept, this study demonstrates facile assaying of advanced glycation end products, 8-hydroxy-2'-deoxyguanosine and hepatitis B virus DNA in urine samples at clinically relevant concentrations, with limits of detection of ∼850 pM, ∼650 pM, and ∼ 1 nM, respectively. These analytes represent three distinct classes of biomarkers: (i) glycated proteins, (ii) low-molecular-weight reactive oxygen species, and (iii) nucleic acids. Hence, the proposed methodology is applicable for rapid detection of generic biomarkers in urine, without involving sophisticated equipment and skilled personnel, thereby enabling on-site urinalysis. At the end of the contribution, we discuss the opportunity to translate the homogeneous assay into a paper-based format.


Assuntos
Biomarcadores/urina , Técnicas Biossensoriais/métodos , Colorimetria/métodos , Humanos , Limite de Detecção , Polímeros , Tiofenos/urina , Urinálise
15.
Langmuir ; 36(24): 6828-6836, 2020 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-32459493

RESUMO

This study describes surface-assisted (SurfAst) urethane polymerization, providing a modular/postfunctionalizable, biorepellent, electroactive ∼10 to 100 nm-thick polyurethane (PU) interface on a gold surface. SurfAst is a functionalization methodology based on sequential incubation steps of alkane diisocyanates and alkanediol monomers. The gold surface is functionalized by alkane diisocyanates in the first incubation step, and our theoretical calculations reveal that while the isocyanate group atoms (N, C, and O) at one end of the molecule exhibits strong interactions (∼900 meV) with surface atoms, the other end group remains unreacted. After the first incubation step, sequential alkanediol and alkane diisocyanate incubations provide formation of the PU interface. The extensive analysis of the PU interface has been conducted via X-ray photoelectron spectroscopy, and the chemical mapping verifies that the interface is made of PU moieties. The topographical analysis of the surface conducted by the atomic force microscopy shows that the PU interface consists of mostly a nanoporous texture with 150 nm total roughness. The adherence force mapping of the PU interface reveals that the nanoporous matrix exhibits an adhesion force of about 14 nN. The electrostatic force microscopy characterizing long-range electrostatic interactions (40 nm) shows that the PU interface has been attracted by positively charged species as compared to negative objects. Finally, it is demonstrated that the PU interface is readily postfunctionalizable by polyethylene glycol (PEG 1000), serving as a biorepellent interface and preserving electroactivity. We foresee that SurfAst polymerization will have potential for the facile fabrication of a postfunctionalizable and modular biointerface which might be utilized for biosensing and bioelectronic applications.

16.
Talanta ; 209: 120581, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31892020

RESUMO

Conjugated polyelectrolytes (CPEs) have been widely used as reporters in colorimetric assays targeting nucleic acids. CPEs provide naked eye detection possibility by their superior optical properties however, as concentration of target analytes decrease, trace amounts of nucleic acid typically yield colorimetric responses that are not readily perceivable by naked eye. Herein, we report a pixelated analysis approach for correlating colorimetric responses of CPE with nucleic acid concentrations down to 1 nM, in plasma samples, utilizing a smart phone with an algorithm that can perform analytical testing and data processing. The detection strategy employed relies on conformational transitions between single stranded nucleic acid-cationic CPE duplexes and double stranded nucleic acid-CPE triplexes that yield distinct colorimetric responses for enabling naked eye detection of nucleic acids. Cationic poly[N,N,N-triethyl-3-((4-methylthiophen-3-yl)oxy)propan-1-aminium bromide] is utilized as the CPE reporter deposited on a polyvinylidene fluoride (PVDF) membrane for nucleic acid assay. A smart phone application is developed to capture and digitize the colorimetric response of the individual pixels of the digital images of CPE on the PVDF membrane, followed by an analysis using the algorithm. The proposed pixelated approach enables precise quantification of nucleic acid assay concentrations, thereby eliminating the margin of error involved in conventional methodologies adopted for interpretation of colorimetric responses, for instance, RGB analysis. The obtained results illustrate that a ubiquitous smart phone could be utilized for point of care colorimetric nucleic acids assays in complex matrices without requiring sophisticated software or instrumentation.


Assuntos
Colorimetria/métodos , Ácidos Nucleicos/sangue , Polieletrólitos/química , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Cátions/química , Colorimetria/instrumentação , Desenho de Equipamento , Humanos , Ácidos Nucleicos/análise , Papel , Sistemas Automatizados de Assistência Junto ao Leito , Polímeros/química , Polivinil/química , Smartphone , Tiofenos/química
17.
Int J Biol Macromol ; 139: 1054-1062, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31404597

RESUMO

Electrospun collagen is commonly used as a scaffold in tissue engineering applications since it mimics the content and morphology of native extracellular matrix (ECM) well. This report describes "toxic solvent free" fabrication of electrospun hybrid scaffold consisting of Collagen (Col) and Poly(l-lactide-co-ε-caprolactone) (PLLCL) for three-dimensional (3D) cell culture. Biomimetic hybrid scaffold was fabricated via co-spinning approach where simultaneous electrospinning of PLLCL and Collagen was mediated by polymer sacrificing agent Polyvinylpyrrolidone (PVP). Acidified aqueous solution of PVP was used to solubilize collagen without using toxic solvents for electrospinning, and then PVP was readily removed by rinsing in water. Mechanical characterizations, protein adsorption, as well as biodegradation analysis have been conducted to investigate feasibility of biomimetic hybrid scaffold for 3D cell culture applications. Electrospun biomimetic hybrid scaffold, which has 3D-network structure with 300-450 nm fiber diameters, was found to be maximizing cell adhesion through assisting NIH 3T3 mouse fibroblast cells. 3D cell culture studies confirmed that presence of collagen in biomimetic hybrid scaffold have created a major impact on cell proliferation compared to conventional 2D systems on long-term, also cell viability increased with the increasing amount of collagen.


Assuntos
Materiais Biomiméticos/química , Colágeno/química , Eletricidade , Poliésteres/química , Alicerces Teciduais/química , Adsorção , Animais , Materiais Biomiméticos/metabolismo , Materiais Biomiméticos/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Fenômenos Mecânicos , Camundongos , Células NIH 3T3 , Poliésteres/metabolismo , Poliésteres/toxicidade , Soroalbumina Bovina/química
18.
Anal Chem ; 91(16): 10357-10360, 2019 08 20.
Artigo em Inglês | MEDLINE | ID: mdl-31334629

RESUMO

This letter describes formation of single chain cationic polymer dots (Pdots) made of poly[1,4-dimethyl-1-(3-((2,4,5-trimethylthiophen-3-yl)oxy)propyl)piperazin-1-ium bromide] conjugated polyelectrolyte (CPE). The single chain Pdot formation relies on a simple process which is a rapid nanophase separation between CPE solution of ethylene glycol and water. Pdots show narrow monodisperse size distribution with a 3.6 nm in diameter exhibiting high brightness and excellent colloidal and optical stability. It has been demonstrated that photoluminescent Pdots provide selective nuclear translocation to hepatocellular carcinoma cells as compared to healthy liver cells. The Pdot labeling effectively discriminates cancer cells in the coculture media. Pdots hold great promise as a luminescent probe to diagnose cancer cells in histology and may guide surgeons during operations to precisely separate out cancerous tissue due to augmented fluorescence brightness.


Assuntos
Núcleo Celular/ultraestrutura , Corantes Fluorescentes/química , Hepatócitos/ultraestrutura , Piperazinas/química , Pontos Quânticos/química , Tiofenos/química , Cátions , Linhagem Celular Transformada , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Técnicas de Cocultura , Corantes Fluorescentes/análise , Hepatócitos/metabolismo , Humanos , Imagem Óptica/métodos , Tamanho da Partícula , Piperazinas/análise , Polieletrólitos/química , Pontos Quânticos/análise , Coloração e Rotulagem/métodos , Tiofenos/análise
19.
Anal Chim Acta ; 1066: 102-111, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31027524

RESUMO

A flow-through colorimetric assay for detection of nucleic acids in plasma is reported. The proposed assay features an array of four polyvinylidene fluoride (PVDF) membranes impregnated with cationic poly (3-alkoxy-4-methylthiophene) (PT) as an optical reporter. The sensing strategy is based on monitoring the changes in optical properties of PT, upon complexation with target nucleic acids in the presence and in the absence of their corresponding complementary peptide nucleic acids (PNAs). As a proof of concept, the proposed methodology is validated using two biomarkers; lung cancer associated microRNA (mir21) and hepatitis B virus DNA (HBV-DNA). The flow-through colorimetric assay enabled detection of mir21 and HBV-DNA in plasma without requiring tedious sample pre-treatment and clean up protocols. Colorimetric responses for mir21 and HBV-DNA were obtained at nanomolar concentrations over five orders of magnitudes (from 1 nM to 10 µM), with a limit of detection of ∼0.6 nM and ∼2 nM in DI water and plasma, respectively. A logic gate system was developed to utilize the colorimetric assay responses as inputs for discrimination of mir21 and HBV-DNA and subsequently to obtain a profile of nucleic acids in samples that exceed respective clinical threshold limits, thereby enabling rapid and point of care (POC) disease diagnosis. Furthermore, the proposed methodology can be utilized for detection of a large number of nucleic acids in plasma by extending the array of PT impregnated membranes incorporated with their corresponding complementary PNAs.


Assuntos
Colorimetria , DNA Viral/sangue , MicroRNAs/sangue , Vírus da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Humanos , Sistemas Automatizados de Assistência Junto ao Leito
20.
Anal Chem ; 90(8): 5122-5129, 2018 04 17.
Artigo em Inglês | MEDLINE | ID: mdl-29557164

RESUMO

This study reports on a hand-held volatilome analyzer for selective determination of clinically relevant biomarkers in exhaled breath. The sensing platform is based on electrospun polymer nanofiber-multiwalled carbon nanotube (MWCNT) sensing microchannels. Polymer nanofibers of poly(vinylidene fluoride) (PVDF), polystyrene (PS), and poly(methyl methacrylate) (PMMA) incorporated with MWCNT exhibits a stable response to interferences of humidity and CO2 and provides selective deformations upon exposure of exhaled breath target volatilomes acetone and toluene, exhibiting correlation to diabetes and lung cancer, respectively. The sensing microchannels "P1" (PVDF-MWCNT), "P2" (PS-MWCNT), and "P3" (PMMA-MWCNT) are integrated with a microfluidic cartridge (µ-card) that facilitates collection and concentration of exhaled breath. The volatilome analyzer consists of a conductivity monitoring unit, signal conditioning circuitries and a low energy display module. A combinatorial operation algorithm was developed for analyzing normalized resistivity changes of the sensing microchannels upon exposure to breath in the concentration ranges between 35 ppb and 3.0 ppm for acetone and 1 ppb and 10 ppm for toluene. Subsequently, responses of volatilomes from individuals in the different risk groups of diabetes were evaluated for validation of the proposed methodology. We foresee that proposed methodology provides an avenue for rapid detection of volatilomes thereby enabling point of care diagnosis in high-risk group individuals.


Assuntos
Testes Respiratórios/métodos , Nanofibras/análise , Compostos Orgânicos Voláteis/análise , Acetona/análise , Testes Respiratórios/instrumentação , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/metabolismo , Humanos , Pneumopatias/diagnóstico , Pneumopatias/metabolismo , Técnicas Analíticas Microfluídicas , Nanotubos de Carbono/química , Sistemas Automatizados de Assistência Junto ao Leito , Polimetil Metacrilato/química , Poliestirenos/química , Tolueno/análise , Compostos Orgânicos Voláteis/metabolismo
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