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1.
Materials (Basel) ; 17(4)2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38399128

RESUMO

This study aims to establish a correlation between the fragmentation process and the growth mechanisms of a coating deposited on a fluoropolymer. Deposition was carried out using dielectric barrier discharge at atmospheric pressure, employing an oxygen-containing organic precursor in a nitrogen environment. The findings reveal that the impact of precursor concentration on the formation of specific functionalities is more significant than the influence of treatment time. The X-ray photoelectron spectroscopy (XPS) results obtained indicate a reduction in the N/O ratio on the coating's surface as the precursor concentration in the discharge increases. Fourier transform infrared spectroscopy (FTIR) analysis, conducted in the spectral range of 1500 cm-1 to 1800 cm-1, confirmed the connection between the chemical properties of plasma-deposited thin films and the concentration of organic precursors in the discharge. Furthermore, the emergence of nitrile moieties (C≡N) in the FTIR spectrum at 2160 cm-1 was noted under specific experimental conditions.

2.
Polymers (Basel) ; 16(3)2024 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-38337253

RESUMO

Significant progress has been made in recent years in the use of atmospheric pressure plasma techniques for surface modification. This research focused on the beneficial effects of these processes on natural by-products, specifically those involving natural fiber-based materials. The study explored the deposition of hydrophobic organosilicon-like thin films onto flax fibres through plasma-enhanced chemical vapour deposition (PECVD), using tetramethylcyclotetrasiloxane (TMCTS) as the precursor. After the successful deposition of hydrophobic organosilicon-like thin films onto the flax fibres, polylactic acid (PLA) composite materials were fabricated. This fabrication process sets the stage for an in-depth analysis of the modified materials. Subsequently, these flax fabrics were subjected to meticulous characterization through scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and contact angle measurements. The results demonstrated successful TMCTS deposition on the surface which led to a complete hydrophobization of the flax fibers. Mechanical tests of the PLA/flax fibre composites revealed a significant improvement in load transfer and interfacial compatibility following the surface modification of the flax fibres. This improvement was attributed to the enhanced adhesion between the modified fibres and the PLA matrix. The findings highlight the potential of TMCTS-based PECVD as a practical surface modification technique, effectively enhancing the mechanical properties of PLA/flax fibre composites. These developments open exciting possibilities for sustainable and high-performance composite materials in various industries.

3.
Materials (Basel) ; 16(3)2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36769949

RESUMO

Due to their chemical inertness and low friction coefficient, fluoropolymers are today widely employed in sectors of activity as diverse and distinct as the textile industry, architectural sector, and medicine. However, their low surface energy results in poor adhesion, for example, when used for a component in a composite device with multiple other materials. Among the techniques used to enhance their adhesion, atmospheric pressure discharges provide a fast and low-cost method with a reduced environmental impact. Although this approach has proven to be efficient, the different chemical and physical processes in the discharge remain not fully understood. In this study, fluoropolymer surfaces were modified using an atmospheric pressure dielectric barrier discharge in a nitrogen and organic precursor environment. To prevent any damage to fluoropolymer surfaces, the dissipated power in the discharges was tuned by applying a duty cycle. Evidence shows that plasma treatment allows for the incorporation of oxygen and nitrogen in the surface resulting in the formation of hydrophilic functionalities such as carbonyl groups both in ketone and amide form, amine, and hydroxyl groups after 180 s of treatment. Overall, the data reveal that the discharge duty cycle has more effect on the oxygen and carbon content in the coating than the precursor concentration. In addition, increasing the precursor concentration limits the molecular fragmentation and nitrogen incorporation into the coating. These experiments enable the building of a better fundamental understanding of the formation mechanism of such chemical moieties at the fluoropolymer surface.

4.
J Biomed Mater Res A ; 111(5): 688-700, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36680491

RESUMO

The long-term success of intraosseous transcutaneous amputation prostheses (ITAPs) mainly relies on dermal attachment of skin cells to the implant. Otherwise, bacteria can easily penetrate through the interface between the implant and the skin. Therefore, infection at this implant/skin interface remains a significant complication in orthopedic surgeries in which these prostheses are required. Two main strategies were investigated to prevent these potential infection problems which consist in either establishing a strong sealing at the skin/implant interface or on eradicating infections by killing bacteria. In this work, two adhesion peptides, either KRGDS or KYIGSR and one antimicrobial peptide, Magainin 2 (Mag 2), were covalently grafted via phosphonate anchor arms to the surface of the Ti6Al4V ELI (extra low interstitials) material, commonly used to manufacture ITAPs. X-ray photoelectron spectroscopy, contact angle, and confocal microscopy analyses enabled to validate the covalent and stable grafting of these three peptides. Dermal fibroblasts cultures on bare Ti6Al4V ELI surfaces and functionalized ones displayed a good cell adhesion and proliferation on all samples. However, cell spreading and viability appeared to be improved on grafted surfaces, especially for those conjugated with KRGDS and Mag 2. Moreover, the dermal sheet attachment, was significantly higher on surfaces functionalized with Mag 2 as compared to the other surfaces. Therefore, the surface functionalization with the antimicrobial peptide Mag 2 seems to be the best approach for the targeted application, as it could play a dual role, inducing a strong skin adhesion while limiting infections on Ti6Al4V ELI materials.


Assuntos
Próteses e Implantes , Titânio , Titânio/química , Adesão Celular , Peptídeos , Amputação Cirúrgica , Peptídeos Antimicrobianos , Propriedades de Superfície
5.
PLoS One ; 17(8): e0269316, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36040884

RESUMO

Maximizing the re-endothelialization of vascular implants such as prostheses or stents has the potential to significantly improve their long-term performance. Endothelial progenitor cell capture stents with surface-immobilized antibodies show significantly improved endothelialization in the clinic. However, most current antibody-based stent surface modification strategies rely on antibody adsorption or direct conjugation via amino or carboxyl groups which leads to poor control over antibody surface concentration and/or molecular orientation, and ultimately bioavailability for cell capture. Here, we assess the utility of a bioaffinity-based surface modification strategy to immobilize antibodies targeting endothelial cell surface antigens. A cysteine-tagged truncated protein G polypeptide containing three Fc-binding domains was conjugated onto aminated polystyrene substrates via a bi-functional linking arm, followed by antibody immobilization. Different IgG antibodies were successfully immobilized on the protein G-modified surfaces. Covalent grafting of the protein G polypeptide was more effective than surface adsorption in immobilizing antibodies at high density based on fluorophore-labeled secondary antibody detection, as well as endothelial colony-forming cell capture through anti-CD144 antibodies. This work presents a potential avenue for enhancing the performance of cell capture strategies by using covalent grafting of protein G polypeptides to immobilize IgG antibodies.


Assuntos
Células Progenitoras Endoteliais , Anticorpos Imobilizados , Imunoglobulina G , Peptídeos , Stents
6.
Biomater Sci ; 10(17): 4978-4996, 2022 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-35801706

RESUMO

The aim of this study is to investigate the impact of the stiffness and stress relaxation of poly(acrylamide-co-acrylic acid) hydrogels on the osteogenic differentiation of human mesenchymal stem cells (hMSCs). Varying the amount of the crosslinker and the ratio between the monomers enabled the obtainment of hydrogels with controlled mechanical properties, as characterized using unconfined compression and atomic force microscopy (AFM). Subsequently, the surface of the hydrogels was functionalized with a mimetic peptide of the BMP-2 protein, in order to favor the osteogenic differentiation of hMSCs. Finally, hMSCs were cultured on the hydrogels with different stiffness and stress relaxation: 15 kPa - 15%, 60 kPa - 15%, 140 kPa - 15%, 100 kPa - 30%, and 140 kPa - 70%. The cells on hydrogels with stiffnesses from 60 kPa to 140 kPa presented a star-like shape, typical of osteocytes, which has only been reported by our group for two-dimensional substrates. Then, the extent of hMSC differentiation was evaluated by using immunofluorescence and by quantifying the expression of both osteoblast markers (Runx-2 and osteopontin) and osteocyte markers (E11, DMP1, and sclerostin). It was found that a stiffness of 60 kPa led to a higher expression of osteocyte markers as compared to stiffnesses of 15 and 140 kPa. Finally, the strongest expression of osteoblast and osteocyte differentiation markers was observed for the hydrogel with a high relaxation of 70% and a stiffness of 140 kPa.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Diferenciação Celular , Células Cultivadas , Humanos , Hidrogéis/química , Hidrogéis/farmacologia , Osteoblastos
7.
Biomed Tech (Berl) ; 67(4): 317-330, 2022 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-35611716

RESUMO

Non-woven textile has been largely used as medical implant material over the last decades, especially for scaffold manufacturing purpose. This material presents a large surface area-to-volume ratio, which promotes adequate interaction with biological tissues. However, its strength is limited due to the lack of cohesion between the fibers. The goal of the present work was to investigate if a non-woven substrate can be reinforced by embroidery stitching towards strength increase. Non-woven samples were produced from both melt-blowing and electro-spinning techniques, reinforced with a stitching yarn and tested regarding several performances: ultimate tensile strength, burst strength and strength loss after fatigue stress. Several stitching parameters were considered: distance between stitches, number of stitch lines (1, 2 or 3) and line geometry (horizontal H, vertical L, cross X). The performance values obtained after reinforcement were compared with values obtained for control samples. Results bring out that reinforcement can increase the strength by up to 50% for a melt-blown mat and by up to 100% for an electro-spun mat with an X reinforcement pattern. However, after cyclic loading, the reinforcement yarn tends to degrade the ES mat in particular. Moreover, increasing the number of stitches tends to fragilize the mats.


Assuntos
Próteses e Implantes , Têxteis , Resistência à Tração
8.
J Biomed Mater Res B Appl Biomater ; 110(9): 2110-2120, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35420261

RESUMO

Transcatheter aortic valve replacement (TAVR) is an alternative technique to surgical valve replacement for over 300,000 patients worldwide. The valve material used in the TAVR is made of biological tissues, whose durability remains unknown. The success of the TAVR favors the research toward synthetic valve leaflet materials as an alternative to biological tissues. In particular, polyethylene terephthalate (PET) textile valves have recently proven durability over a 6-month period in animal sheep models. Excessive fibrotic tissue formation remains, however, a critical issue to be addressed. The aim of this work was therefore to investigate the potential of PET textiles covalently conjugated with polyethylene glycol (PEG), known for its antifouling properties, to modulate the fibrosis formation both in vitro and in vivo. For this purpose, the surfaces of heart valves made of PET textiles were functionalized with an atmospheric pressure plasma, leading to the formation of carboxylic acid (COOH) groups, further used for PEG-NH2 conjugation. Surface modification efficiency was assessed by X-ray photoelectron spectroscopy and water contact angle measurements. The biological behavior of the as-modified surfaces was evaluated by in vitro assays, using rat cardiac fibroblast cells. The results show that PEG treated substrates restrained the fibroblasts adhesion and proliferation. The PEG treated valve, implanted in a juvenile sheep model, showed a significant fibrosis reduction. The explant also revealed calcification issues that need to be addressed.


Assuntos
Próteses Valvulares Cardíacas , Substituição da Valva Aórtica Transcateter , Animais , Valva Aórtica/cirurgia , Fibrose , Polietilenoglicóis/farmacologia , Polietilenotereftalatos , Desenho de Prótese , Ratos , Ovinos , Têxteis
9.
J Biomed Mater Res A ; 109(11): 2187-2198, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-33931940

RESUMO

Intraosseous transcutaneous amputation prosthesis is a new approach in orthopedic implants that overcomes socket prosthesis problems. Its long-term performance requires a tight skin-implant seal to prevent infections. In this study, fibronectin (Fn), a widely used adhesion protein, was adsorbed or grafted onto titanium alloy. Fn grafting was performed using two different linking arms, dopamine/glutaric anhydride or phosphonate. The characterization of Fn-modified surfaces showed that Fn grating via phosphonate has led to the highest amount of Fn cell-binding site (RGD, arginine, glycine, and aspartate) available on the surface. Interestingly, cell culture studies revealed a strong correlation between the amount of available RGD ligands and cellular behavior, since enhanced proliferation and spreading of fibroblasts were noticed on Fn-grafted surfaces via phosphonate. In addition, an original in vitro mechanical test, inspired from the real situation, to better predict clinical outcomes after implant insertion, has been developed. Tensile test data showed that the adhesion strength of a bio-engineered dermal tissue was significantly higher around Fn-grafted surfaces via phosphonate, as compared to untreated surfaces. This study sheds light on the importance of an appropriate selection of the linking arm to tightly control the spatial conformation of biomolecules on the material surface, and consequently cell interactions at the interface tissue/implant.


Assuntos
Ligas/química , Materiais Revestidos Biocompatíveis/química , Derme/metabolismo , Fibroblastos/metabolismo , Fibronectinas/química , Implantes Experimentais , Receptores Imunológicos/química , Receptores de Peptídeos/química , Titânio/química , Humanos
10.
Macromol Biosci ; 21(6): e2100069, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33870650

RESUMO

The aim of this study is to investigate polyacrylamide-based hydrogels stress relaxation and the subsequent impact on the osteogenic differentiation of human mesenchymal stem cells (hMSCs). Different hydrogels are synthesized by varying the amount of cross-linker and the ratio between the monomers (acrylamide and acrylic acid), and characterized by compression tests. It has been found that hydrogels containing 18% of acrylic acid exhibit an average relaxation of 70%, while pure polyacrylamide gels show an average relaxation of 15%. Subsequently, hMSCs are cultured on two different hydrogels functionalized with a mimetic peptide of the bone morphogenetic protein-2 to enable cell adhesion and favor their osteogenic differentiation. Phalloidin staining shows that for a constant stiffness of 55 kPa, a hydrogel with a low relaxation (15%) leads to star-shaped cells, which is typical of osteocytes, while a hydrogel with a high relaxation (70%) presents cells with a polygonal shape characteristic of osteoblasts. Immunofluorescence labeling of E11, strongly expressed in early osteocytes, also shows a dramatically higher expression for cells cultured on the hydrogel with low relaxation (15%). These results clearly demonstrate that, by fine-tuning hydrogels stress relaxation, hMSCs differentiation can be directed toward osteoblasts, and even osteocytes, which is particularly rare in vitro.


Assuntos
Acrilamidas/farmacologia , Hidrogéis/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteoblastos/efeitos dos fármacos , Osteócitos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Alicerces Teciduais , Acrilamidas/síntese química , Proteína Morfogenética Óssea 2/química , Proteína Morfogenética Óssea 2/farmacologia , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Reagentes de Ligações Cruzadas/química , Humanos , Hidrogéis/síntese química , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteócitos/citologia , Osteócitos/metabolismo , Peptidomiméticos/química , Peptidomiméticos/farmacologia , Estresse Mecânico , Relação Estrutura-Atividade
11.
Nanotechnology ; 32(17): 175601, 2021 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-33470988

RESUMO

The proof of the concept of a new, onestep and safe by design process to synthesize metal-polymer nanocomposites thin films on a large surface is presented. It is based on the injection of an aerosol of a solution of metal (gold) salts dissolved in a polymerizable solvent (isopropanol) into an argon atmospheric pressure dielectric barrier discharge. The main novelty of this method resides in the fact that the nanoparticles are formed in situ, inside the plasma reactor, in the gas phase. Consequently, the nanoparticle synthesis and deposition are concomitant with the solvent polymerization used to produce the matrix, which makes it possible to obtain homogeneous layers of non-agglomerated nanoparticles (NPs) with high NPs density. By toggling between low and high-frequency discharges, gold/polymer nanocomposites with different morphologies and optical properties are synthesized. The effect of the concentration of gold in the aerosol and the gas residence time in the plasma as well as the ratio of high and low-frequency discharge and their repetition rate are presented. The thin films are systematically characterized by AFM and UV-visible spectroscopy to analyze their morphologies along with their plasmonic resonances.

12.
Biomater Sci ; 8(19): 5465-5475, 2020 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-32902522

RESUMO

Antibody surface immobilization is a promising strategy to capture cells of interest from circulating fluids in vitro and in vivo. An application of particular interest in vascular interventions is to capture endothelial progenitor cells (EPCs) on the surface of stents to accelerate endothelialization. The clinical impact of EPC capture stents has been limited by the lack of efficient selective cell capture. Here, we describe a simple method to immobilize a variety of immunoglobulin G antibodies through their fragment crystallizable (Fc) regions via surface-conjugated RRGW peptides for cell capture applications. As an EPC capture model, peripheral blood endothelial colony-forming cells suspended in cell culture medium with up to 70% serum were captured by immobilized anti-CD144, anti-CD34 or anti-CD309 antibodies under laminar flow. The endothelial colony-forming cells were successfully enriched from a mixture with peripheral blood mononuclear cells using surfaces with anti-CD309 but not anti-CD45. This antibody immobilization approach holds great promise to engineer vascular biomaterials with improved EPC capture potential. The ease of immobilizing different antibodies using the same Fc-binding peptide surface grafting chemistry renders this platform suitable to screen antibodies that maximize cell capture efficiency and selectivity.


Assuntos
Células Progenitoras Endoteliais , Anticorpos , Endotélio , Leucócitos Mononucleares , Peptídeos
13.
J Biomed Mater Res A ; 108(7): 1479-1492, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32170899

RESUMO

Synthetic grafts do not provide an appealing surface for endothelial cells to adhere and colonize the inner surface. To promote in situ endothelialization the following aspect has to be taken into account, endothelial progenitor cells (EPCs) needs to be mobilized on the surface of the graft. The surface of the graft has to be sufficiently biocompatible to create a prone environment for the EPCs to adhere, proliferate and, differentiate to form a layer and subsequently improve graft patency. In this work, two active molecules GRGDS and sitagliptin, were chosen for their abilities to recruit, enhance adhesion and induce differentiation of endothelial progenitor cells. They were grafted on PET surfaces in order to provide restrained cues triggering cell alignment and evaluate the influence of such structuration on EPCs fate. We then analyze cell behavior onto functionalized biomaterials. Their abilities to control EPCs fate were demonstrated via RT-qPCR, immunofluorescence, and enzymatic tests. The GRGDS/sitagliptin 100 × 10 surface enables to reduce the stemness phenotype on EPCs and induce the expression of endothelial lineage markers. These results highlight the importance of spatial patterning cues in guiding EPCs organization and function, which may have clinical relevance in the development of vascular grafts that promote patency.


Assuntos
Materiais Biocompatíveis/farmacologia , Diferenciação Celular , Células Progenitoras Endoteliais/citologia , Oligopeptídeos/farmacologia , Fosfato de Sitagliptina/farmacologia , Materiais Biocompatíveis/química , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Células Progenitoras Endoteliais/efeitos dos fármacos , Humanos , Oligopeptídeos/química , Fosfato de Sitagliptina/química , Propriedades de Superfície
14.
Colloids Surf B Biointerfaces ; 189: 110847, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32086024

RESUMO

The use of polylactic acid (PLA) has attracted growing interest, particularly in recent years, for biomedical applications because of its mechanical properties, biocompatibility, and biodegradability. Despite this, features such as surface hydrophobicity and the absence of suitable functional groups for covalent immobilization of bioactive molecules, make it challenging to endow PLA-based medical devices with additional features and thus broaden their range of applicability. In the present study, we demonstrate the suitability of atmospheric pressure dielectric barrier discharges operating in the Townsend regime as a promising alternative to other surface treatments, such as diazonium and alkali hydrolytic treatments, for carboxyl functionalization of PLA. Chemical changes in PLA surfaces are evaluated by contact angle measurements and by X-ray photoelectron spectroscopy while physical changes are investigated by scanning electron microscopy and atomic force microscopy. The amount of carboxyl groups generated on PLA surfaces is assessed by toluidine blue O assay and substantiated by grafting, through carboxyl groups, a fluorescent probe containing amino functionalities. All of the surface treatments have proven to be very effective in generating carboxylic groups on the PLA surface. Nevertheless, plasma treatment is shown to not degrade the PLA surface, in sharp contrast with diazonium and alkali hydrolytic treatments.


Assuntos
Gases em Plasma/química , Poliésteres/química , Pressão Atmosférica , Interações Hidrofóbicas e Hidrofílicas , Estrutura Molecular , Tamanho da Partícula , Propriedades de Superfície , Molhabilidade
15.
J Biomed Mater Res A ; 108(2): 201-211, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31595677

RESUMO

The native microenvironment of mesenchymal stem cells (hMSCs)-the extracellular matrix (ECM), is a complex and heterogenous environment structured at different scales. The present study aims at mimicking the hierarchical microorganization of proteins or growth factors within the ECM using the photolithography technique. Polyethylene terephthalate substrates were used as a model material to geometrically defined regions of RGD + BMP-2 or RDG + OGP mimetic peptides. These ECM-derived ligands are under research for regulation of mesenchymal stem cells osteogenic differentiation in a synergic manner. The hMSCs osteogenic differentiation was significantly affected by the spatial distribution of dually grafted peptides on surfaces, and hMSCs cells reacted differently according to the shape and size of peptide micropatterns. Our study demonstrates the presence of a strong interplay between peptide geometric cues and stem cell differentiation toward the osteoblastic lineage. These tethered surfaces provide valuable tools to investigate stem cell fate mechanisms regulated by multiple ECM cues, thereby contributing to the design of new biomaterials and improving hMSCs differentiation cues.


Assuntos
Materiais Biocompatíveis/química , Células-Tronco Mesenquimais/citologia , Peptidomiméticos/química , Proteína Morfogenética Óssea 2/química , Linhagem Celular , Matriz Extracelular/química , Humanos , Oligopeptídeos/química , Propriedades de Superfície
16.
Mater Sci Eng C Mater Biol Appl ; 107: 110335, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31761211

RESUMO

In order to stimulate the cellular response to implant materials, extracellular matrix (ECM) proteins, such as collagen and fibronectin (FN), are immobilized on the implant surface. Amongst all ECM proteins used for biomimetic materials for medical applications, FN is one of the most investigated proteins thanks to its ability to promote cell adhesion and its contribution to important physiological processes. However, its conformation and hence its bioactivity strongly depend on the hydrophilic/hydrophobic nature of the surface as well as on immobilization strategies. This work investigates the effect of these two parameters, as well as the effect of the crosslinker length. FN was grafted onto silicon wafers using eights different linking arms presenting different lengths, hydrophilic/hydrophobic characters and binding sites. The protein was linked through either its amino groups (lysine amino acids) or sulfhydryl functionalities (cysteine amino acids). The grafting of each crosslinker and subsequent FN conjugation onto the surfaces was evidenced by X-ray photoelectron spectroscopy, while the surface hydrophilicity was determined by contact angle measurements. Moreover, atomic force microscopy images revealed that the conformation of surface conjugated FN only depends on the hydrophilicity of the linking arm. The FN conformation was also probed by enzyme-linked immunosorbent assays (ELISA). ELISA data demonstrated that all of the three investigated parameters linking arm parameter (length, hydrophobic/hydrophilic character, and terminal end-group) somewhat influence the RGD accessibility.


Assuntos
Fibronectinas/química , Proteínas Imobilizadas/química , Oligopeptídeos/química , Sítios de Ligação , Reagentes de Ligações Cruzadas/química , Ensaio de Imunoadsorção Enzimática , Fibronectinas/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Proteínas Imobilizadas/metabolismo , Microscopia de Força Atômica , Espectroscopia Fotoeletrônica , Propriedades de Superfície
17.
Biotechnol Bioeng ; 116(10): 2598-2609, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31286475

RESUMO

The expansion of human peripheral blood endothelial progenitor cells to obtain therapeutically relevant endothelial colony-forming cells (ECFCs) has been commonly performed on xeno-derived extracellular matrix proteins. For cellular therapy applications, xeno-free culture conditions are desirable to improve product safety and reduce process variability. We have previously described a novel fluorophore-tagged RGD peptide (RGD-TAMRA) that enhanced the adhesion of mature endothelial cells in vitro. To investigate whether this peptide can replace animal-derived extracellular matrix proteins in the isolation and expansion of ECFCs, peripheral blood mononuclear cells from 22 healthy adult donors were seeded on RGD-TAMRA-modified polystyrene culture surfaces. Endothelial colony formation was significantly enhanced on RGD-TAMRA-modified surfaces compared to the unmodified control. No phenotypic differences were detected between ECFCs obtained on RGD-TAMRA compared to ECFCs obtained on rat-tail collagen-coated surfaces. Compared with collagen-coated surfaces and unmodified surfaces, RGD-TAMRA surfaces promoted ECFC adhesion, cell spreading, and clonal expansion. This study presents a platform that allows for a comprehensive in vitro evaluation of peptide-based biofunctionalization as a promising avenue for ex vivo ECFC expansion.


Assuntos
Células Sanguíneas/citologia , Separação Celular , Células Progenitoras Endoteliais/citologia , Oligopeptídeos/química , Poliestirenos/química , Células Sanguíneas/metabolismo , Células Progenitoras Endoteliais/metabolismo , Feminino , Humanos , Masculino , Propriedades de Superfície
18.
Adv Colloid Interface Sci ; 263: 68-94, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30521982

RESUMO

Glass- and polymer-based materials have become essential in the fabrication of a multitude of elements, including eyeglasses, automobile windshields, bathroom mirrors, greenhouses, and food packages, which unfortunately mist up under typical operating conditions. Far from being an innocuous phenomenon, the formation of minute water drops on the surface is detrimental to their optical properties (e.g., light-transmitting capability) and, in many cases, results in esthetical, hygienic, and safety concerns. In this context, it is therefore not surprising that research in the field of fog-resistant surfaces is gaining in popularity, particularly in recent years, in view of the growing number of studies focusing on this topic. This review addresses the most relevant advances released thus far on anti-fogging surfaces, with a particular focus on coating deposition, surface micro/nanostructuring, and surface functionalization. A brief explanation of how surfaces fog up and the main issues of interest linked to fogging phenomenon, including common problems, anti-fogging strategies, and wetting states are first presented. Anti-fogging mechanisms are then discussed in terms of the morphology of water drops, continuing with a description of the main fabrication techniques toward anti-fogging property. This review concludes with the current and the future perspectives on the utility of anti-fogging surfaces for several applications and some remaining challenges in this field.

19.
Exp Cell Res ; 370(2): 389-398, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-30146063

RESUMO

In the last ten years, endothelial progenitor cells (EPCs) have gained interest as an attractive cell population in regenerative medicine for vascular applications. This population is defined as the precursor of endothelial mature cells (ECs) through a process of differentiation. To our knowledge, no single marker can be used to discriminate them from mature ECs. To effectively study their differentiation kinetics, gene expression must be assessed. Quantitative real-time PCR (RT-qPCR) is widely used to analyze gene expression. To minimize the impact of variances from RT-qPCR, a rigorous selection of reference genes must be performed prior to any experiments due to variations in experimental conditions. In this study, CD34+ mononuclear cells were extracted from human cord blood and differentiated into EPCs after seeding for a maximum period of 21 days. To choose the best combinations of reference genes, we compared the results of EPCs, CD34+ mononuclear cells, and mature endothelial cells to ensure that the differentiation kinetics did not affect the expression of our selected reference genes. The expression levels of seven genes, namely, YWHAZ, GAPDH, HPRT1, RPLP0, UBC, B2M, and TBP were thus compared. The algorithms geNorm, NormFinder, BestKeeper, and the Comparative ΔCt method were employed to assess the expression of each candidate gene. Overall results reveal that the expression stability of reference genes may differ depending on the statistical program used. YWHAZ, GAPDH, and UBC composed the optimal set of reference genes for the gene expression studies performed by RT-qPCR in our experimental conditions. This work can thus serve as a starting point for the selection of candidate reference genes to normalize the levels of gene expression in endothelial progenitor cell populations.


Assuntos
Células Progenitoras Endoteliais/metabolismo , Perfilação da Expressão Gênica , Reação em Cadeia da Polimerase em Tempo Real , Padrões de Referência , Algoritmos , Diferenciação Celular/genética , Células Cultivadas , Sangue Fetal/metabolismo , Expressão Gênica/genética , Perfilação da Expressão Gênica/métodos , Humanos , Leucócitos Mononucleares/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos
20.
Biomed Tech (Berl) ; 63(3): 221-230, 2018 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-29664732

RESUMO

With over 150,000 implantations performed over the world, transcatheter aortic valve replacement (TAVR) has become a surgical technique, which largely competes with open surgery valve replacement for an increasing number of patients. The success of the procedure favors the research toward synthetic valve leaflet materials as an alternative to biological tissues, whose durability remains unknown. In particular, fibrous constructions have recently proven to be durable in vivo over a 6-month period of time in animal sheep models. Exaggerated fibrotic tissue formation remains, however, a critical issue to be addressed. This work investigates the design of a composite fibrous construction combining a woven polyethylene terephthalate (PET) layer and a non-woven PET mat, which are expected to provide, respectively, strength and appropriate topography toward limited fibrotic tissue ingrowth. For this purpose, a specific equipment has been developed to produce non-woven PET mats made from fibers with small diameter. These mats were assembled with woven PET substrates using various assembling techniques in order to obtain hybrid fibrous constructions. The physical and mechanical properties of the obtained materials were assessed and valve samples were manufactured to be tested in vitro for hydrodynamic performances. The results show that the composite fibrous construction is characterized by properties suitable for the valve leaflet function, but the durability of the assembling is however limited under accelerated cyclic loading.


Assuntos
Valva Aórtica/fisiologia , Próteses Valvulares Cardíacas , Animais , Humanos , Desenho de Prótese , Ovinos , Têxteis , Resultado do Tratamento
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