RESUMEN
Polydimethylsiloxane (PDMS) is extensively used to fabricate biocompatible microfluidic systems due to its favorable properties for cell culture. Recently, ultraviolet-curable PDMS (UV-PDMS) has shown potential for enhancing manufacturing processes and final optical quality while retaining the benefits of traditional thermally cured PDMS. This study investigates the biocompatibility of UV-PDMS under static and flow conditions using human umbilical vein endothelial cells (HUVECs). UV-PDMS samples were treated with oxygen plasma and boiling deionized water to assess potential improvements in cell behavior compared with untreated samples. We evaluated HUVECs adhesion, growth, morphology, and viability in static cultures and microchannels fabricated with UV-PDMS to test their resistance to flow conditions. Our results confirmed the biocompatibility of UV-PDMS for HUVECs culture. Moreover, plasma-oxygen-treated UV-PDMS substrates exhibited superior cell growth and adhesion compared to untreated UV-PDMS. This enhancement enabled HUVECs to maintain their morphology and viability under flow conditions in UV-PDMS microchannels. Additionally, UV-PDMS demonstrated improved optical quality and more efficient handling and processing, characterized by shorter curing times and simplified procedures utilizing UV light compared to traditional PDMS.
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There is a need to develop new and versatile fabrication methods to achieve efficient mixing of fluids in microfluidic channels using microstructures. This work presents a new technique that combines stereolithography (SLA) and pulsed laser ablation (PLA) to manufacture a straight micromixer for uniform mixing of two samples. Computational fluid dynamics (CFD) simulation is performed to deeply understand the physical mechanisms of the process. The results suggest that this new optical technique holds the potential to become a versatile hybrid technique for manufacturing remarkable mixing microfluidic devices.
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Subaquatic indirect Laser-Induced Plasma-Assisted Ablation (SLIPAA) is proposed as a laser-based technique for glass processing. In this configuration, a water layer is added between a metallic target and a soda-lime glass substrate, so the processing of the glass is due to a combination of the ablation mechanism, the shock waves, and the cavitation bubbles. Thus, this method makes it possible to produce higher depth structures than those performed up to now by other standard laser techniques based on ablation, achieving structures in glass with rectangular cross-sectional profiles. Channels of 1 mm width are fabricated, reaching an average maximal depth value of almost 1400 µm at 30 passes of the laser beam while keeping the focal position fixed. Furthermore, the difference between processing the material with and without the addition of the water layer is presented. The influence of the processing parameters on the shape and quality of the fabricated structures is studied by optical and confocal microscopy, microcomputed tomography, and scanning electron microscopy. Compositional analysis of the glass is performed by energy dispersive X-ray technique to assess the transference of material from the metallic target to the fabricated channels. Deeper and more complex structures are obtained by refocusing the laser beam on the target and adding a pulsed flowing water film.
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Diabetes is associated with a high mortality rate due to vascular complications. Chronic hyperglycemia in diabetes leads to enhanced oxidative stress and glycation. Here, we explored the impact of glycation on human erythrocyte characteristics and capacity to affect endothelial cell function following erythrophagocytosis. Native and glucose-mediated glycated erythrocytes were prepared and characterized in terms of structural and deformability modifications. Erythrocyte preparations were tested for their binding and phagocytosis capacity as well as the potential functional consequences on human endothelial cell lines and primary cultures. Oxidative modifications were found to be enhanced in glycated erythrocytes after determination of their deformability, advanced glycation end-product content and eryptosis. Erythrophagocytosis by endothelial cells was significantly increased when incubated in the presence of glycated erythrocytes. In addition, higher iron accumulation, oxidative stress and impaired endothelial cell permeability were evidenced in cells previously incubated with glycated erythrocytes. When cultured under flow conditions, cellular integrity was disrupted by glycated erythrocytes at microvessel bifurcations, areas particularly prone to vascular complications. This study provides important new data on the impact of glycation on the structure of erythrocytes and their ability to alter endothelial cell function. Increased erythrophagocytosis may have a deleterious impact on endothelial cell function with adverse consequences on diabetic vascular complications.
Asunto(s)
Diabetes Mellitus , Células Endoteliales , Diabetes Mellitus/metabolismo , Células Endoteliales/metabolismo , Eritrocitos/metabolismo , Productos Finales de Glicación Avanzada/metabolismo , Humanos , Fagocitosis/fisiologíaRESUMEN
Homing of circulating tumour cells (CTC) at distant sites represents a critical event in metastasis dissemination. In addition to physical entrapment, probably responsible of the majority of the homing events, the vascular system provides with geometrical factors that govern the flow biomechanics and impact on the fate of the CTC. Here we mathematically explored the distribution of velocities and the corresponding streamlines at the bifurcations of large blood vessel and characterized an area of low-velocity at the carina of bifurcation that favours the residence of CTC. In addition to this fluid physics effect, the adhesive capabilities of the CTC provide with a biological competitive advantage resulting in a marginal but systematic arrest as evidenced by dynamic in vitro recirculation in Y-microchannels and by perfusion in in vivo mice models. Our results also demonstrate that viscosity, as a main determinant of the Reynolds number that define flow biomechanics, may be modulated to limit or impair CTC accumulation at the bifurcation of blood vessels, in agreement with the apparent positive effect observed in the clinical setting by anticoagulants in advanced oncology disease.
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Velocidad del Flujo Sanguíneo , Hemodinámica , Células Neoplásicas Circulantes , Animales , Adhesión Celular , Línea Celular Tumoral , Células Endoteliales de la Vena Umbilical Humana , Humanos , Leucocitos Mononucleares , Ratones , Modelos Cardiovasculares , Modelos TeóricosRESUMEN
Stereolithographic printers have revolutionized many manufacturing processes with their capacity to easily produce highly detailed structures. In the field of microfluidics, this technique avoids the use of complex steps and equipment of the conventional technologies. The potential of low force stereolithography technology is analysed for the first time using a Form 3B printer and seven printing resins through the fabrication of microchannels and pillars. Manufacturing performance of internal and superficial channels and pillars is studied for the seven printing resins in different configurations. A complete characterization of printed structures is carried out by optical, confocal and SEM microscopy, and EDX analysis. Internal channels with unobstructed lumen are obtained for diameters and angles greater than 500 µm and 60°, respectively. Outward and inward superficial channels in the range of hundreds of microns can be fabricated with an accurate profile, printing them with a perpendicular orientation respect to the base, allowing a proper uncured resin evacuation. Outward channels are replicated by soft lithography using polydimethylsiloxane. Clear, Model and Tough resins show a good behaviour to be used as master, but Amber and Dental resins present a poor topology transference from the master to the replica. According to the needs of devices used for biological and biomedical research, transparency as well as superficial biocompatibility of some resins is evaluated. Human umbilical vein endothelial cells (HUVEC) adhesion is confirmed on Amber, Dental and Clear resins, but these cells were only able to grow and progress as a cell culture over the Amber resin. Therefore, Amber showed an adequate biocompatibility, in terms of cell adhesion and growth for HUVEC.