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1.
Methods Mol Biol ; 2125: 1-13, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-30539347

RESUMEN

Endothelial cell culture under flow, to mimic physiological conditions within blood vessels, has gained particular attention for the formation of a homogeneous endothelium in vitro. Here, we report on the design of a setup for simultaneous culture of up to nine electrospun membranes or thin polymer films in custom-made holders under flow on an orbital shaker. The versatile design of the device allows for the use of electrospun membranes/polymer films of choice and subsequent analysis with commonly used methods such as immunofluorescence or scanning electron microscopy.


Asunto(s)
Técnicas de Cultivo de Célula/instrumentación , Técnicas de Cultivo de Célula/métodos , Membranas Artificiales , Polímeros/química , Reología , Células Cultivadas , Técnica del Anticuerpo Fluorescente , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/ultraestructura , Humanos , Esterilización , Fijación del Tejido
2.
Nanoscale ; 11(15): 7176-7187, 2019 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-30919869

RESUMEN

A dedicated nanofiber design for applications in the biomedical domain is based on the understanding of nanofiber structures. The structure of electrospun nanofibers strongly influences their properties and functionalities. In polymeric nanofibers X-ray scattering and diffraction methods, i.e. SAXS and WAXD, are capable of decoding their structural insights from about 100 nm down to the Angström scale. Here, we present a comprehensive X-ray scattering and diffraction based study and introduce new data analysis approaches to unveil detailed structural features in electrospun poly(vinylidene fluoride-co-hexafluoropropylene) (PVDFhfp) nanofiber membranes. Particular emphasis was placed on anisotropic morphologies being developed during the nanofiber fabrication process. Global analysis was performed on SAXS data to derive the nanofibrillar structure of repeating lamella crystalline domains with average dimensions of 12.5 nm thickness and 7.8 nm spacing along with associated tie-molecules. The varying surface roughness of the nanofiber was evaluated by extracting the Porod exponent in parallel and perpendicular direction to the nanofiber axis, which was further validated by Atomic Force Microscopy. Additionally, the presence of a mixture of the monoclinic alpha and the orthorhombic beta PVDFhfp phases both exhibiting about 6% larger unit cells compared to the corresponding pure PVDF phases was derived from WAXD. The current study shows a generic approach in detailed understanding of internal structures and surface morphology for nanofibers. This forms the basis for targeted structure and morphology steering and the respective controlling during the fabrication process with the aim to engineer nanofibers for different biomedical applications with specific requirements.

3.
ACS Appl Mater Interfaces ; 11(6): 5740-5751, 2019 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-30668107

RESUMEN

Despite major technological advances within the field of cardiovascular engineering, the risk of thromboembolic events on artificial surfaces in contact with blood remains a major challenge and limits the functionality of ventricular assist devices (VADs) during mid- or long-term therapy. Here, a biomimetic blood-material interface is created via a nanofiber-based approach that promotes the endothelialization capability of elastic silicone surfaces for next-generation VADs under elevated hemodynamic loads. A blend fiber membrane made of elastic polyurethane and low-thrombogenic poly(vinylidene fluoride- co-hexafluoropropylene) was partially embedded into the surface of silicone films. These blend membranes resist fundamental irreversible deformation of the internal structure and are stably attached to the surface, while also exhibiting enhanced antithrombotic properties when compared to bare silicone. The composite material supports the formation of a stable monolayer of endothelial cells within a pulsatile flow bioreactor, resembling the physiological in vivo situation in a VAD. The nanofiber surface modification concept thus presents a promising approach for the future design of advanced elastic composite materials that are particularly interesting for applications in contact with blood.


Asunto(s)
Materiales Biomiméticos/química , Nanofibras/química , Adsorción , Materiales Biomiméticos/farmacología , Reactores Biológicos , Coagulación Sanguínea/efectos de los fármacos , Células Endoteliales/citología , Células Endoteliales/metabolismo , Fibrinógeno/química , Humanos , Membranas Artificiales , Microscopía Confocal , Polivinilos/química , Resistencia al Corte , Silicio/química , Propiedades de Superficie
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