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1.
Langmuir ; 37(48): 14043-14049, 2021 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-34818018

RESUMO

Hierarchically structured films comprise a growing section of the field of surface-enhanced Raman spectroscopy (SERS). Here, we report a novel, powerfully enhancing hierarchical plasmonic substrate featuring patterned multilayers of magnetic iron oxide nanospheres using an external magnetic field to create sets of radial ridges. This new substrate allows for effective analyte adsorption and significant Raman signal enhancement, thanks to the contribution of both the magnetic and plasmonic components to the electromagnetic hotspots. We demonstrate significant and reliable Raman enhancement for polycyclic aromatic hydrocarbons (PAHs), dilute but persistent environmental pollutants, in a complex and real-world matrix of produced water (PW). The substrate activity for PAHs is validated by gas chromatography-mass spectrometry analysis. An impressive signal-to-noise ratio (SNR) of several dB enables detection of the analyte below 1 ppm. This multilayer magnetic film sensor substrate shows remarkable stability and robustness suitable for real-world applications while boasting simple methods and strong potential to scale up fabrication.

2.
BMC Biotechnol ; 15: 112, 2015 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-26666739

RESUMO

BACKGROUND: Triple helical collagens are the most abundant structural protein in vertebrates and are widely used as biomaterials for a variety of applications including drug delivery and cellular and tissue engineering. In these applications, the mechanics of this hierarchically structured protein play a key role, as does its chemical composition. To facilitate investigation into how gene mutations of collagen lead to disease as well as the rational development of tunable mechanical and chemical properties of this full-length protein, production of recombinant expressed protein is required. RESULTS: Here, we present a human type II procollagen expression system that produces full-length procollagen utilizing a previously characterized human fibrosarcoma cell line for production. The system exploits a non-covalently linked fluorescence readout for gene expression to facilitate screening of cell lines. Biochemical and biophysical characterization of the secreted, purified protein are used to demonstrate the proper formation and function of the protein. Assays to demonstrate fidelity include proteolytic digestion, mass spectrometric sequence and posttranslational composition analysis, circular dichroism spectroscopy, single-molecule stretching with optical tweezers, atomic-force microscopy imaging of fibril assembly, and transmission electron microscopy imaging of self-assembled fibrils. CONCLUSIONS: Using a mammalian expression system, we produced full-length recombinant human type II procollagen. The integrity of the collagen preparation was verified by various structural and degradation assays. This system provides a platform from which to explore new directions in collagen manipulation.


Assuntos
Colágeno Tipo II/biossíntese , Colágeno Tipo II/genética , Eucariotos/genética , Eucariotos/metabolismo , Catepsina K/química , Catepsina K/metabolismo , Linhagem Celular Tumoral , Dicroísmo Circular , Células Clonais , Matriz Extracelular/metabolismo , Fibrossarcoma/genética , Fibrossarcoma/metabolismo , Fibrossarcoma/patologia , Humanos , Microscopia de Força Atômica , Pinças Ópticas , Pró-Colágeno/biossíntese , Pró-Colágeno/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transfecção
3.
Sensors (Basel) ; 13(6): 7021-32, 2013 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-23760086

RESUMO

We present and analyze a method to improve the morphology and mechanical properties of gold thin films for use in optical sensors or other settings where good adhesion of gold to a substrate is of importance and where controlled topography/roughness is key. To improve the adhesion of thermally evaporated gold thin films, we introduce a gold deposition step on SU-8 photoresist prior to UV exposure but after the pre-bake step of SU-8 processing. Shrinkage and distribution of residual stresses, which occur during cross-linking of the SU-8 polymer layer in the post-exposure baking step, are responsible for the higher adhesion of the top gold film to the post-deposition cured SU-8 sublayer. The SU-8 underlayer can also be used to tune the resulting gold film morphology. Our promoter-free protocol is easily integrated with existing sensor microfabrication processes.


Assuntos
Ouro/química , Sistemas Microeletromecânicos/métodos , Módulo de Elasticidade , Sistemas Microeletromecânicos/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Microscopia de Força Atômica , Polímeros/química , Propriedades de Superfície
4.
Chem Commun (Camb) ; 53(88): 12024-12027, 2017 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-29058738

RESUMO

We demonstrate the power of Kelvin probe force microscopy (KPFM) in enabling a comprehensive study of enhancement mechanisms of surface enhanced Raman scattering (SERS) through the correlation of surface electrical and topographical effects. Local electric fields generated on Au/ZnO nanohybrid films impact analyte adsorption, while roughness is linked to hotspot generation. Optimizing the interplay between these two effects yields SERS enhancement factors (EFs) of 106, enabling ppb detection of polycyclic aromatic hydrocarbons (PAHs) in water.

5.
Macromol Biosci ; 13(3): 348-55, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23335515

RESUMO

The in vitro viability, osteogenic differentiation, and mineralization of four different equine mesenchymal stem cells (MSCs) from bone marrow, periosteum, muscle, and adipose tissue are compared, when they are cultured with different collagen-based scaffolds or with fibrin glue. The results indicate that bone marrow cells are the best source of MSCs for osteogenic differentiation, and that an electrochemically aggregated collagen gives the highest cell viability and best osteogenic differentiation among the four kinds of scaffolds studied.


Assuntos
Colágeno/farmacologia , Adesivo Tecidual de Fibrina/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteócitos/efeitos dos fármacos , Engenharia Tecidual/métodos , Tecido Adiposo/citologia , Animais , Células da Medula Óssea/citologia , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/química , Adesivo Tecidual de Fibrina/química , Cavalos , Células-Tronco Mesenquimais/citologia , Músculo Esquelético/citologia , Osteócitos/citologia , Periósteo/citologia , Isoformas de Proteínas/química , Isoformas de Proteínas/farmacologia , Alicerces Teciduais
6.
Macromol Biosci ; 12(3): 360-6, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22213568

RESUMO

Therapies for corneal disease and injury often rely on artificial implants, but integrating cells into synthetic corneal materials remains a significant challenge. The electrochemically formed collagen-based matrix presented here is non-toxic to cells and controls the proliferation in the corneal fibroblasts seeded onto it. Histology and biomolecular studies show a behavior similar to corneal stromal cells in a native corneal environment. Not only is this result an important first step toward developing a more realistic, multi-component artificial cornea, but it also opens possibilities for using this matrix to control and contain the growth of cells in engineered tissues.


Assuntos
Colágeno/química , Córnea/citologia , Fibroblastos/citologia , Engenharia Tecidual/métodos , Biomarcadores/metabolismo , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Técnicas Eletroquímicas , Fibroblastos/fisiologia , Humanos , Imuno-Histoquímica , Poliestirenos/química , Alicerces Teciduais
7.
Langmuir ; 24(10): 5418-21, 2008 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-18407679

RESUMO

Ordered arrays of collagen microfibrils form rapidly and spontaneously from a solution of monomers deposited onto a mica substrate. These arrays are well-ordered and apparently continuous over the entire substrate. Correlated atomic force microscope images and Laué diffraction patterns indicate that the protein alignment and microfibril formation is controlled by the crystal orientation of the mica substrate rather than fluid flow or drying effects. This surface-induced mechanism allows for immediate, robust, and reproducible pattern formation.


Assuntos
Silicatos de Alumínio/química , Colágeno/química , Microfibrilas/química , Proteínas/química , Adsorção , Biofísica/métodos , Cátions , Cristalização , Concentração de Íons de Hidrogênio , Microscopia de Força Atômica , Nanopartículas/química , Nanotecnologia/métodos , Análise Serial de Proteínas , Propriedades de Superfície , Difração de Raios X
8.
J Am Chem Soc ; 127(41): 14154-5, 2005 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-16218588

RESUMO

A 1,3,5,7-octatetrayne-linked bis(fullerene) compound has been synthesized. Through a thermally induced solid-state polymerization reaction on a surface, the solid thin film of this compound was transformed into a highly uniform and well organized polymer nanosphere array. This finding suggests a simple and useful method for the preparation of carbon-rich, fullerene-containing nanoparticles.


Assuntos
Alcinos/síntese química , Fulerenos/química , Nanoestruturas/química , Temperatura , Alcinos/química , Estrutura Molecular , Tamanho da Partícula , Propriedades de Superfície
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