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1.
Anal Chem ; 96(17): 6764-6773, 2024 04 30.
Artículo en Inglés | MEDLINE | ID: mdl-38619911

RESUMEN

Tremendous efforts have been made to develop practical and efficient microfluidic cell and particle sorting systems; however, there are technological limitations in terms of system complexity and low operability. Here, we propose a sheath flow generator that can dramatically simplify operational procedures and enhance the usability of microfluidic cell sorters. The device utilizes an embedded polydimethylsiloxane (PDMS) sponge with interconnected micropores, which is in direct contact with microchannels and seamlessly integrated into the microfluidic platform. The high-density micropores on the sponge surface facilitated fluid drainage, and the drained fluid was used as the sheath flow for downstream cell sorting processes. To fabricate the integrated device, a new process for sponge-embedded substrates was developed through the accumulation, incorporation, and dissolution of PMMA microparticles as sacrificial porogens. The effects of the microchannel geometry and flow velocity on the sheath flow generation were investigated. Furthermore, an asymmetric lattice-shaped microchannel network for cell/particle sorting was connected to the sheath flow generator in series, and the sorting performances of model particles, blood cells, and spiked tumor cells were investigated. The sheath flow generation technique developed in this study is expected to streamline conventional microfluidic cell-sorting systems as it dramatically improves versatility and operability.


Asunto(s)
Separación Celular , Técnicas Analíticas Microfluídicas , Humanos , Separación Celular/instrumentación , Separación Celular/métodos , Técnicas Analíticas Microfluídicas/instrumentación , Porosidad , Dimetilpolisiloxanos/química , Dispositivos Laboratorio en un Chip , Polimetil Metacrilato/química
2.
Analyst ; 147(8): 1622-1630, 2022 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-35297918

RESUMEN

Numerous attempts have been made to develop efficient systems to purify trace amounts of circulating tumor cells (CTCs) from blood samples. However, current technologies are limited by complexities in device fabrication, system design, and process operability. Here we describe a facile, scalable, and highly efficient approach to physically capturing CTCs using a rationally designed microfluidic isolator with an array of microslit channels. The wide but thin microslit channels with a depth of several micrometers selectively capture CTCs, which are larger and less deformable than other blood cells, while allowing other blood cells to just flow through. We investigated in detail the effects of the microchannel geometry and operating parameters on the capture efficiency and selectivity of several types of cultured tumor cells spiked in blood samples as the CTC model. Additionally, in situ post-capture staining of the captured cells was demonstrated to investigate the system's applicability to clinical cancer diagnosis. The presented approach is simple in operation but significantly effective in capturing specific cells and hence it may have great potential in implementating cell physics-based CTC isolation techniques for cancer liquid biopsy.


Asunto(s)
Técnicas Analíticas Microfluídicas , Células Neoplásicas Circulantes , Línea Celular Tumoral , Separación Celular/métodos , Humanos , Técnicas Analíticas Microfluídicas/métodos , Microfluídica , Células Neoplásicas Circulantes/patología
3.
Anal Chem ; 92(3): 2580-2588, 2020 02 04.
Artículo en Inglés | MEDLINE | ID: mdl-31822057

RESUMEN

Visualization and quantification of intracellular molecules of mammalian cells are crucial steps in clinical diagnosis, drug development, and basic biological research. However, conventional methods rely mostly on labor-intensive, centrifugation-based manual operations for exchanging the cell carrier medium and have limited reproducibility and recovery efficiency. Here we present a microfluidic cell processor that can perform four-step exchange of carrier medium, simply by introducing a cell suspension and fluid reagents into the device. The reaction time period for each reaction step, including fixation, membrane permeabilization, and staining, was tunable in the range of 2 to 15 min by adjusting the volume of the reaction tube connecting the neighboring exchanger modules. We double-stained the cell nucleus and cytoskeleton (F-actin) using the presented device with an overall reaction period of ∼30 min, achieving a high recovery ratio and high staining efficiency. Additionally, intracellular cytokine (IL-2) was visualized for T cells to demonstrate the feasibility of the device as a pretreatment system for downstream flow-cytometric analysis. The presented approach would facilitate the development of laborless, automated microfluidic systems that integrate cell processing and analysis operations and would pave a new path to high-throughput biological experiments.


Asunto(s)
Automatización , Citocinas/análisis , Técnicas Analíticas Microfluídicas , Animales , Línea Celular , Núcleo Celular/química , Núcleo Celular/metabolismo , Citocinas/biosíntesis , Citoesqueleto/química , Citoesqueleto/metabolismo , Diseño de Equipo , Citometría de Flujo/instrumentación , Ratones , Técnicas Analíticas Microfluídicas/instrumentación , Células 3T3 NIH
4.
Anal Chem ; 91(21): 13377-13382, 2019 11 05.
Artículo en Inglés | MEDLINE | ID: mdl-31585029

RESUMEN

Lateral flow immunoassay devices have revolutionized the style of on-site disease detection and point-of-care testing in the past few decades. The surface nanotopography of a solid substrate is a dominant parameter in the efficiency of antibody immobilization, but precise control over surface roughness has not been fully investigated. Here we presented lateral flow immunoassay platforms with nanometer-scale surface roughness, reproducibly engineered using thermal nanoimprinting lithography, and investigated the effects of surface nanotopography on immunoadsorption and immunoassay performance. We fabricated three types of imprinted polycarbonate sheets with microcone array structures having different degrees of surface roughness using three types of molds fabricated by micromachining or laser ablation. The structures fabricated by laser-ablated nickel mold exhibited numerous bumps measuring several tens of nanometers, which enhanced antibody adsorption. We performed sandwich immunoassays of C-reactive protein in serum samples and achieved highly sensitive detection with a detection limit of ∼0.01 µg mL-1 and a broad dynamic range. The present results provide useful information on the remarkable effect of nanoengineered surfaces on biomolecule adsorption, and the platforms presented here will widen the applicability and versatility of lateral flow immunoassay devices.


Asunto(s)
Proteína C-Reactiva/química , Inmunoensayo/instrumentación , Nanoestructuras , Polímeros/química , Adsorción , Bioensayo/métodos , Inmunoensayo/métodos , Límite de Detección , Propiedades de Superficie
5.
Analyst ; 144(5): 1519-1526, 2019 Feb 25.
Artículo en Inglés | MEDLINE | ID: mdl-30539188

RESUMEN

Although various types of on-site immunoassay platforms have been developed, facile and reliable sample-to-answer immunoassay systems are still under development. In this study, we proposed a lateral-flow immunoassay system utilizing a polymer sheet with microcone array structures fabricated by thermal imprinting. By adding a surfactant to the running/washing buffer, we were able to dispense with complicated chemical modification protocols, which are usually necessary to enhance antibody adsorption. We investigated three types of polymeric materials and confirmed that polycarbonate is most suitable as an imprinted polymer substrate. Experiments using microcone arrays with different distances revealed that the increased surface area with nanometer-scale surface roughness was key to achieving stable immobilization of antibodies. To assess the applicability of this assay to clinical diagnosis, C-reactive protein in a pure buffer and in a serum sample was analyzed as a model, with a ∼0.1 µg mL-1 lower limit of detection. The presented approach would open a new path to the development of immunoassay-based on-site point-of-care testing by virtue of its simplicity of operation, high sensitivity, and versatility.


Asunto(s)
Proteína C-Reactiva/análisis , Inmunoensayo/métodos , Adsorción , Anticuerpos Inmovilizados/química , Anticuerpos Inmovilizados/inmunología , Biomarcadores/análisis , Proteína C-Reactiva/inmunología , Humanos , Inmunoensayo/instrumentación , Límite de Detección , Cemento de Policarboxilato/química , Polimetil Metacrilato/química , Propiedades de Superficie , Tensoactivos/química , Temperatura
6.
J Hepatol ; 68(4): 744-753, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29288124

RESUMEN

BACKGROUND & AIMS: Since the first account of the myth of Prometheus, the amazing regenerative capacity of the liver has fascinated researchers because of its enormous medical potential. Liver regeneration is promoted by multiple types of liver cells, including hepatocytes and liver non-parenchymal cells (NPCs), through complex intercellular signaling. However, the mechanism of liver organogenesis, especially the role of adult hepatocytes at ectopic sites, remains unknown. In this study, we demonstrate that hepatocytes alone spurred liver organogenesis to form an organ-sized complex 3D liver that exhibited native liver architecture and functions in the kidneys of mice. METHODS: Isolated hepatocytes were transplanted under the kidney capsule of monocrotaline (MCT) and partial hepatectomy (PHx)-treated mice. To determine the origin of NPCs in neo-livers, hepatocytes were transplanted into MCT/PHx-treated green fluorescent protein transgenic mice or wild-type mice transplanted with bone marrow cells isolated from green fluorescent protein-mice. RESULTS: Hepatocytes engrafted at the subrenal space of mice underwent continuous growth in response to a chronic hepatic injury in the native liver. More than 1.5 years later, whole organ-sized liver tissues with greater mass than those of the injured native liver had formed. Most remarkably, we revealed that at least three types of NPCs with similar phenotypic features to the liver NPCs were recruited from the host tissues including bone marrow. The neo-livers in the kidney exhibited liver-specific functions and architectures, including sinusoidal vascular systems, zonal heterogeneity, and emergence of bile duct cells. Furthermore, the neo-livers successfully rescued the mice with lethal liver injury. CONCLUSION: Our data clearly show that adult hepatocytes play a leading role as organizer cells in liver organogenesis at ectopic sites via NPC recruitment. LAY SUMMARY: The role of adult hepatocytes at ectopic locations has not been clarified. In this study, we demonstrated that engrafted hepatocytes in the kidney proliferated, recruited non-parenchymal cells from host tissues including bone marrow, and finally created an organ-sized, complex liver system that exhibited liver-specific architectures and functions. Our results revealed previously undescribed functions of hepatocytes to direct liver organogenesis through non-parenchymal cell recruitment and organize multiple cell types into a complex 3D liver at ectopic sites. Transcript profiling: Microarray data are deposited in GEO (GEO accession: GSE99141).


Asunto(s)
Hepatocitos/fisiología , Riñón/citología , Hígado/embriología , Organogénesis , Animales , Movimiento Celular , Proliferación Celular , Hepatocitos/trasplante , Regeneración Hepática , Ratones , Ratones Endogámicos C57BL
7.
Langmuir ; 33(49): 14087-14092, 2017 12 12.
Artículo en Inglés | MEDLINE | ID: mdl-29140704

RESUMEN

Direct observation of double emulsion droplet permeation through a microchannel that mimicked 100 µm membrane pores with a porosity of 66.7% provided insights regarding splitting mechanisms in porous membranes. The microchannel was fabricated by standard soft lithography, and the oil-in-water-in-oil double emulsion droplets were prepared with a glass capillary device. By changing the flow rate from 0.5 to 5.0 × 10-2 m s-1, three characteristic behaviors were observed: (a) passage into one channel without splitting; (b) division into two smaller components; and (c) stripping of the middle water phase of the double emulsion droplets into a smaller double emulsion droplet and a smaller water-in-oil single emulsion droplet. The mechanisms are discussed with respect to the balance of viscous forces and interfacial tension, the contact point with the tip of the channel, and the relative position of the innermost droplet within the middle droplet.

8.
Anal Chem ; 88(11): 5637-43, 2016 06 07.
Artículo en Inglés | MEDLINE | ID: mdl-27152997

RESUMEN

When analyzing the kinetics of liquid-liquid extraction (LLE), the change in the concentration of extracted target molecules over time should be monitored for a known interfacial area. Herein, we developed a microfluidic system for precisely analyzing the kinetics of LLE using droplets of a constant size even in the presence of additives. Extraction is initiated by exchanging the carrier fluid for the droplets with a target solution and then terminated by phase separation, based on the principle of hydrodynamic filtration. By using one out of several pairs of outlet/buffer inlet at a given time, the extraction time period is tuned stepwise without changing the flow rate condition. We successfully demonstrated droplet-based LLE by controlling the extraction period from ∼0.03 to ∼1.2 s and evaluated the extraction kinetics of rhodamine B from the continuous aqueous phase to droplets of 1-octanol with a diameter of ∼40 µm. In addition, the effect of additives on extraction efficiency was evaluated. The system presented in this study would be useful for determining rate constants for interfacial mass transfer in general LLE kinetic studies as well as for developing new extraction chemistries and optimizing microfluidic chemical/biochemical analysis systems that involve an LLE process.

9.
Langmuir ; 31(8): 2334-41, 2015 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-25669326

RESUMEN

A new microfluidic process was used to generate unique micrometer-sized hierarchical lipid particles having spherical lipid-core and multilamellar-shell structures. The process includes three steps: (1) formation of monodisperse droplets in a nonequilibrium state at a microchannel confluence, using a phospholipid-containing water-soluble organic solvent as the dispersed phase and water as the continuous phase; (2) dissolution of the organic solvent of the droplet into the continuous phase and concentration of the lipid molecules; and (3) reconstitution of multilamellar lipid membranes and simultaneous formation of a lipid core. We demonstrated control of the lipid particle size by the process conditions and characterized the obtained particles by transmission electron microscopy and microbeam small-angle X-ray scattering analysis. In addition, we prepared various types of core-shell and core-core-shell particles incorporating hydrophobic/hydrophilic compounds, showing the applicability of the presented process to the production of drug-encapsulating lipid particles.


Asunto(s)
Técnicas Analíticas Microfluídicas , Interacciones Hidrofóbicas e Hidrofílicas , Tamaño de la Partícula , Propiedades de Superficie
10.
Lab Chip ; 24(2): 171-181, 2024 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-38050757

RESUMEN

Although droplet microfluidics has been studied for the past two decades, its applications are still limited due to the low productivity of microdroplets resulting from the low integration of planar microchannel structures. In this study, a microfluidic system implementing inverse colloidal crystals (ICCs), a spongious matrix with regularly and densely formed three-dimensional (3D) interconnected micropores, was developed to significantly increase the throughput of microdroplet generation. A new bottom-up microfabrication technique was developed to seamlessly integrate the ICCs into planar microchannels by accumulating non-crosslinked spherical PMMA microparticles as sacrificial porogens in a selective area of a mold and later dissolving them. We have demonstrated that the densely arranged micropores on the spongious ICC of the microchannel function as massively parallel micronozzles, enabling droplet formation on the order of >10 kHz. Droplet size could be adjusted by flow conditions, fluid properties, and micropore size, and biopolymer particles composed of polysaccharides and proteins were produced. By further parallelization of the unit structures, droplet formation on the order of >100 kHz was achieved. The presented approach is an upgrade of the existing droplet microfluidics concept, not only in terms of its high throughput, but also in terms of ease of fabrication and operation.

11.
J Endourol ; 38(6): 564-572, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38545752

RESUMEN

Purpose: We herein describe the superficial preprostatic vein (SPV) anatomy and determine its relationship with the accessory pudendal artery (APA). Materials and Methods: We reviewed 500 patients with localized prostate cancer who underwent conventional robot-assisted radical prostatectomy between April 2019 and March 2023 at our institution. SPV was defined as "any vein coming from the space between the puboprostatic ligaments and running within the retropubic adipose tissue anterior to the prostate toward the vesical venous plexus or pelvic side wall." While APA was defined as "any artery located in the periprostatic region running parallel to the dorsal vascular complex and extending caudal toward the anterior perineum." The intraoperative anatomy of each SPV and APA was described. Results: SPVs had a prevalence rate of 88%. They were preserved in 252 men (58%) and classified as I-, reversed-Y (rY)-, Y-, or H-shaped (64%, 22%, 12%, and 2%, respectively) based on their intraoperative appearance. Overall, 214 APAs were found in 142 of the 252 men with preserved SPV (56%; 165 lateral and 50 apical APAs in 111 and 41 men, respectively). SPVs were pulsatile in 39% men perhaps due to an accompanying tiny artery functioning as a median APA. Pulsations seemed to be initially absent in most SPVs but become apparent late during surgery possibly due to increased arterial and venous blood flow after prostate removal. Pulsations were common in men with ≥1 APA. Conclusions: This study, which described the anatomical variations in arteries and veins around the prostrate and their preservation techniques, revealed that preserving this vasculature may help preserve postprostatectomy erection. ClinicalTrials: The Clinical Research Registration Number is 230523D.


Asunto(s)
Arterias , Próstata , Prostatectomía , Procedimientos Quirúrgicos Robotizados , Venas , Humanos , Masculino , Prostatectomía/métodos , Procedimientos Quirúrgicos Robotizados/métodos , Próstata/irrigación sanguínea , Próstata/cirugía , Persona de Mediana Edad , Anciano , Arterias/anatomía & histología , Venas/anatomía & histología , Venas/cirugía , Neoplasias de la Próstata/cirugía , Neoplasias de la Próstata/patología
12.
J Robot Surg ; 18(1): 25, 2024 Jan 13.
Artículo en Inglés | MEDLINE | ID: mdl-38217741

RESUMEN

OBJECTIVES: Patients with inguinal hernia (IH) may have voiding dysfunction and weak urethra-stabilizing periurethral fascial tissues, contributing to urinary incontinence. This study aimed to review the association between IH and urinary continence after robotic-assisted radical prostatectomy (RARP). METHODS: This single-institution retrospective study included 251 consecutive cases of RARP between April 2019 and June 2022. Patients with concurrent IH or a history of adult IH repair were examined. The urine loss rate (ULR), defined as 24-h urine loss volume divided by the total urine volume immediately after urinary catheter removal (i.e., 6 or 7 postoperative days), was compared between the groups with (n = 33) and without IH (n = 214). Possible contributing factors for ULR were assessed, including age, body mass index (BMI), after benign prostatic hyperplasia surgery, prostate weight, and nerve-sparing. ULR was compared intergroup after propensity score matching countering selection biases. RESULTS: Patients with IH were older (71.3 versus. 66.8 years, p < 0.01), had lower BMI (22.8 versus. 24.3, p < 0.01), and had higher ULR (14.5% versus. 5.1%, p < 0.01). In a multiple linear regression analysis (adjusted R2 = 0.084), IH (p < 0.01) was an independent contributing factor for ULR besides advancing age (p < 0.03). After propensity score matching adjusted for patient's age and nerve-sparing, patients with IH had higher ULR (14.1% versus. 5.7%, p < 0.03) as well. CONCLUSIONS: This study first reported that IH may be one of the risk factors of urinary incontinence after RARP.


Asunto(s)
Hernia Inguinal , Neoplasias de la Próstata , Procedimientos Quirúrgicos Robotizados , Robótica , Incontinencia Urinaria , Masculino , Adulto , Humanos , Próstata , Estudios Retrospectivos , Procedimientos Quirúrgicos Robotizados/métodos , Hernia Inguinal/cirugía , Incontinencia Urinaria/etiología , Incontinencia Urinaria/cirugía , Prostatectomía/efectos adversos , Neoplasias de la Próstata/cirugía
13.
Anal Chem ; 85(16): 7666-73, 2013 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-23875607

RESUMEN

A simple microfluidic system has been presented to perform continuous two-parameter cell sorting based on size and surface markers. Immunomagnetic bead-conjugated cells are initially sorted based on size by utilizing the hydrodynamic filtration (HDF) scheme, introduced into individual separation lanes, and simultaneously focused onto one sidewall by the hydrodynamic effect. Cells are then subjected to magnetophoretic separation in the lateral direction, and finally they are individually recovered through multiple outlet branches. We successfully demonstrated the continuous sorting of JM (human lymphocyte cell line) cells using anti-CD4 immunomagnetic beads and confirmed that accurate size- and surface marker-based sorting was achieved. In addition, the sorting of cell mixtures was performed at purification ratios higher than 90%. The proposed system enables two-dimensional cell sorting without necessitating complicated setups and operations, and thus, it can be a useful tool for general biological experiments including cell-based disease diagnosis, stem cell engineering, and cellular physiological studies.


Asunto(s)
Separación Celular/métodos , Filtración/instrumentación , Hidrodinámica , Magnetismo , Microfluídica/instrumentación , Antígenos CD4/inmunología , Línea Celular , Humanos , Separación Inmunomagnética
14.
Lab Chip ; 23(9): 2257-2267, 2023 05 02.
Artículo en Inglés | MEDLINE | ID: mdl-37038847

RESUMEN

Spheroid formation assisted by microengineered chambers is a versatile approach for morphology-controlled three-dimensional (3D) cell cultivation with physiological relevance to human tissues. However, the limitation in diffusion-based oxygen/nutrient transport has been a critical issue for the densely packed cells in spheroids, preventing maximization of cellular functions and thus limiting their biomedical applications. Here, we have developed a multiscale microfluidic system for the perfusion culture of spheroids, in which porous microchambers, connected with microfluidic channels, were engineered. A newly developed process of centrifugation-assisted replica molding and salt-leaching enabled the formation of single micrometer-sized pores on the chamber surface and in the substrate. The porous configuration generates a vertical flow to directly supply the medium to the spheroids, while avoiding the formation of stagnant flow regions. We created seamlessly integrated, all PDMS/silicone-based microfluidic devices with an array of microchambers. Spheroids of human liver cells (HepG2 cells) were formed and cultured under vertical-flow perfusion, and the proliferation ability and liver cell-specific functions were compared with those of cells cultured in non-porous chambers with a horizontal flow. The presented system realizes both size-controlled formation of spheroids and direct medium supply, making it suitable as a precision cell culture platform for drug development, disease modelling, and regenerative medicine.


Asunto(s)
Microfluídica , Esferoides Celulares , Humanos , Hepatocitos , Perfusión , Hígado
15.
J Biosci Bioeng ; 135(5): 417-422, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-36931921

RESUMEN

Technologies for efficiently expanding Chinese hamster ovary (CHO) cells, the primary host cells for antibody production, are of growing industrial importance. Various processes for the use of microcarriers in CHO suspension cultures have been developed, but there have been very few studies on cell-adhesive microcarriers that are similar in size to cells. In this study, we proposed a new approach to suspension cultures of CHO cells using cell-sized condensed and crosslinked gelatin microparticles (GMPs) as carriers. Unlike commercially available carriers with sizes typically greater than 100 µm, each cell can adhere to the surface of multiple particles and form loose clusters with voids. We prepared GMPs of different average diameters (27 and 48 µm) and investigated their effects on cell adhesion and cluster formation. In particular, small GMPs promoted cell proliferation and increased IgG4 production by the antibody-producing CHO cell line. The data obtained in this study suggest that cell-sized particles, rather than larger ones, enhance cell proliferation and function, providing useful insights for improving suspension-culture-based cell expansion and cell-based biologics production for a wide range of applications.


Asunto(s)
Técnicas de Cultivo de Célula , Gelatina , Cricetinae , Animales , Cricetulus , Células CHO , Proliferación Celular
16.
Biomed Microdevices ; 14(4): 751-7, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22544390

RESUMEN

Separation/purification of nuclei from cells is a critical process required for medical and biochemical research applications. Here, we report a flow-through microfluidic device for isolating cell nuclei by selectively digesting the cell membrane by using the concept of hydrodynamic filtration (HDF). When a cell suspension is continuously introduced into a microchannel (main channel) possessing multiple side channels, cells flow through the main channel, whereas the carrier medium of the cells is drained through the side channels. Introductions of a cell treatment solution containing a surfactant and a washing buffer enable the two-step exchange of the carrier-medium and the cell treatment by the surfactant for a short span of time. The precise control of the treatment time by changing the flow rate and/or the size of the microchannel enables the selective digestion of cell membranes, resulting in the isolation of cell nuclei after separation from membrane debris and cytoplasmic components according to size. We examined several surfactant molecules and demonstrated that Triton X-100 exhibited high efficiency regarding nucleus isolation for both adherent (HeLa) and nonadherent (JM) cells, with a recovery ratio of ~80 %. In addition, the isolation efficiency was evaluated by western blotting. The presented flow-through microfluidic cell-nucleus separator may be a useful tool for general biological applications, because of its simplicity in operation, high reproducibility, and accuracy.


Asunto(s)
Fraccionamiento Celular/instrumentación , Núcleo Celular/efectos de los fármacos , Técnicas Analíticas Microfluídicas/instrumentación , Células HeLa , Humanos , Octoxinol/farmacología , Dodecil Sulfato de Sodio/farmacología , Tensoactivos/farmacología , Factores de Tiempo
17.
Langmuir ; 28(39): 14073-80, 2012 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-22991929

RESUMEN

This study presents a simple but highly versatile method of fabricating picoliter-volume hydrogel patterns on poly(dimethylsiloxane) (PDMS) substrates. Hydrophilic regions were prepared on hydrophobic PDMS plates by trapping and melting functional polymer particles and performing subsequent reactions with partially oxidized dextran. Small aliquots of a gelation solution were selectively trapped on the hydrophilic areas by a simple dipping process that was utilized to make thin hydrogel patterns by the in situ gelation of a sol solution. Using this process, we successfully formed calcium alginate, collagen I, and chitosan hydrogels with a thickness of several micrometers and shapes that followed the hydrophilized regions. In addition, alginate and collagen gel patterns were used to capture cells with different adhesion properties selectively on or off the hydrogel structures. The presented strategy could be applicable to the preparation of a variety of hydrogels for the development of functional biosensors, bioreactors, and cell cultivation platforms.


Asunto(s)
Dimetilpolisiloxanos/química , Hidrogeles/química , Alginatos/química , Quitosano/química , Colágeno Tipo I/química , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Hidrogeles/síntesis química , Interacciones Hidrofóbicas e Hidrofílicas , Soluciones
18.
J Biosci Bioeng ; 133(3): 265-272, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34903469

RESUMEN

Numerous attempts have been made to organize isolated primary hepatocytes into functional three-dimensional (3D) constructs, but technologies to introduce extracellular matrix (ECM) components into such assemblies have not been fully developed. Here we report a new approach to forming hepatocyte-based 3D tissues using fibrillized collagen microparticles (F-CMPs) as intercellular binders. We created thick tissues with a thickness of ∼200 µm simply by mixing F-CMPs with isolated primary rat hepatocytes and culturing them in cell culture inserts. Owing to the incorporated F-CMPs, the circular morphology of the formed tissues was stabilized, which was strong enough to be manually manipulated and retrieved from the chamber of the insert. We confirmed that the F-CMPs dramatically improved the cell viability and hepatocyte-specific functions such as albumin production and urea synthesis in the formed tissues. The presented approach provides a versatile strategy for hepatocyte-based tissue engineering, and will have a significant impact on biomedical applications and pharmaceutical research.


Asunto(s)
Colágeno , Hepatocitos , Animales , Técnicas de Cultivo de Célula , Supervivencia Celular , Células Cultivadas , Ratas , Ingeniería de Tejidos/métodos
19.
Sci Rep ; 12(1): 22354, 2022 Dec 26.
Artículo en Inglés | MEDLINE | ID: mdl-36572667

RESUMEN

Tsunamis are commonly generated by earthquakes beneath the ocean floor, volcanic eruptions, and landslides. The tsunami following the Tonga eruption of 2022 is believed to have been excited by atmospheric pressure fluctuations generated by the explosion of the volcano. The first, fast-traveling tsunami was excited by Lamb waves; however, it has not been clarified observationally or theoretically which type of atmospheric fluctuations excited more prominent tsunami which followd. In this study, we investigate atmospheric gravity waves that possibly excited the aforementioned subsequent tsunami based on observations and atmosphere-ocean coupling simulations. The atmospheric fluctuations are classified as Lamb waves, acoustic waves, or gravity waves. The arrival time of the gravity wave and the simulation shows that the gravity wave propagated at a phase speed of 215 m/s, coinciding with the tsunami velocity in the Pacific Ocean, and suggesting that the gravity wave resonantly excited the tsunami (Proudman resonance). These observations and theoretical calculations provide an essential basis for investigations of volcano-induced meteotsunamis, including the Tonga event.

20.
Mater Sci Eng C Mater Biol Appl ; 129: 112417, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34579926

RESUMEN

Artificial biological scaffolds made of extracellular matrix (ECM) components, such as type I collagen, provide ideal physicochemical cues to various cell culture platforms. However, it remains a challenge to fabricate micrometer-sized ECM materials with precisely controlled morphologies that could reconstitute the 3-dimensional (3D) microenvironments surrounding cells. In the present study, we proposed a unique process to fabricate fragmented collagen microfibers using a microfluidic laminar-flow system. The continuous flow of an acidic collagen solution was neutralized to generate solid fibers, which were subsequently fragmented by applying a gentle shear stress in a polyanion-containing phosphate buffer. The morphology of the fiber fragment was controllable in a wide range by changing the type and/or concentration of the polyanion and by tuning the applied shear stress. The biological benefits of the fragmented fibers were investigated through the formation of multicellular spheroids composed of primary rat hepatocytes and microfibers on non-cell-adhesive micro-vessels. The microfibers enhanced the survival and functions of the hepatocytes and reproduced proper cell polarity, because the fibers facilitated the formation of cell-cell and cell-matrix interactions while modulating the close packing of cells. These results clearly indicated that the microengineered fragmented collagen fibers have great potential to reconstitute extracellular microenvironments for hepatocytes in 3D culture, which will be of significant benefit for cell-based drug testing and bottom-up tissue engineering.


Asunto(s)
Colágeno , Microfluídica , Animales , Matriz Extracelular , Hepatocitos , Polielectrolitos , Ratas , Ingeniería de Tejidos
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