scholarly journals 3D Printed Lab-on-a-Chip Platform for Chemical Stimulation and Parallel Analysis of Ion Channel Function

Micromachines ◽  
2019 ◽  
Vol 10 (8) ◽  
pp. 548 ◽  
Author(s):  
Daniel Aschenbrenner ◽  
Oliver Friedrich ◽  
Daniel F. Gilbert
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
Low Cost ◽  
Lab On A Chip ◽  
Chemical Stimulation ◽  
Hek293 Cells ◽  
Solution Exchange ◽  
3D Printed

Functional imaging has been a widely established method for the assessment of ion channel function in vitro. Conventional infrastructure used for in vitro functional analysis of ion channels is typically proprietary, non-customizable, expensive, and requires a high level of skill to use and maintain. 3D desktop printing, which is employed in the rapid prototyping field, allows for quick engineering of alternatives to conventional imaging infrastructure that are customizable, low cost, and user friendly. Here, we describe an ultra-low-cost microfluidic lab-on-a-chip (LOC) device manufactured using acrylonitrile butadiene styrene (ABS) for in vitro functional imaging of ion channels that can quickly and easily be reconstructed using three-dimensional (3D) desktop printing. The device is light weight (<5 g), small (20 mm × 49 mm), and extremely low cost (<EUR 1). We simulate fluidics within the printed channels and assess the suitability of the engineered chamber to generate homogeneous mixtures during solution exchange. We demonstrate the usability of the 3D printed microfluidic device in a case study using Fluo-4-loaded human embryonal kidney-derived (HEK293) cells, recombinantly expressing the capsaicin receptor, transient receptor potential vanilloid receptor type 1 (TRPV1), as a model system. In the case study, we confirm its applicability to solution exchange for chemical stimulation and parallel functional time-lapse fluorescence microscopy-based calcium imaging. We assess the suitability of ABS for culturing HEK293 cells inside the microfluidic LOC, based on qualitative analysis of microscopic transmission light images of ABS-exposed HEK293 cells and confirm the previously reported biocompatibility of ABS. To highlight the versatility of the 3D printed microfluidic device, we provide an example for multiplication of the shown concept within a 3D printed multichannel microfluidic LOC to be used, for example, in a higher throughput format for parallelized functional analysis of ion channels. While this work focusses on Ca2+ imaging with TRPV1 channels, the device may also be useful for application with other ion channel types and in vitro models.

The Sparthan Three-Dimensional Printed Exo-Glove: A Preliminary Evaluation of Performance Via Case Study

2019 ◽  
Vol 13 (3) ◽  
Author(s):  
Tomás A. Georgiou ◽  
Davide Asnaghi ◽  
Alva Liang ◽  
Alice M. Agogino
Keyword(s):  
Light Intensity ◽  
Daily Living ◽  
Low Cost ◽  
Three Dimensional ◽  
Preliminary Evaluation ◽  
Evaluation Of Performance ◽  
3D Printed ◽  
Hand Exoskeleton ◽  
And Performance

This paper describes the development and testing of a low-cost three-dimensional (3D) printed wearable hand exoskeleton to assist people with limited finger mobility and grip strength. The function of the presented orthosis is to support and enable light intensity activities of daily living and improve the ability to grasp and hold objects. The Sparthan Exoskeleton prototype utilizes a cable-driven design applied to individual digits with motors. The initial prototype is presented in this paper along with a preliminary evaluation of durability and performance efficacy.

A Case Study in 3D Printed Porous Ceramics: Infant Incubator Humidification System

2012 ◽  
Author(s):  
Olaf Diegel ◽  
Andrew Withell ◽  
Deon Debeer ◽  
Mark Wu
Keyword(s):  
Material Properties ◽  
Low Cost ◽  
Ceramic Materials ◽  
Porous Ceramics ◽  
Burn Out ◽  
Infant Incubator ◽  
3D Printers ◽  
3D Printed ◽  
Factorial Design Experiment

This paper describes research in adapting 3D printers to operate with low-cost ceramic materials. The components produced with these clay-based ceramic powders can be fired to produce strong, complex and lightweight ceramic parts. The final material properties, including the porosity of the parts, can be controlled through the part design and, potentially, through additives to the material that burn out during firing. The paper begins with a brief description of the 3D printing process and how it can be used with clay powders. It then introduces a factorial design experiment initiated to explore the effect of ingredient and parameter variations on the dimensional stability and material properties of green and fired ceramic parts. It then presents a case study in which 3D printed ceramic parts are used in the humidification system for an infant incubator for developing countries.

scholarly journals Fluorescence Microscopy of Piezo1 in Droplet Hydrogel Bilayers

2018 ◽  
Author(s):  
Oskar B. Jaggers ◽  
Pietro Ridone ◽  
Boris Martinac ◽  
Matthew A. B. Baker
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
Simultaneous Recording ◽  
Mechanical Stimuli ◽  
Channel Activity ◽  
Channel Proteins ◽  
Mechanosensitive Ion Channel ◽  
Lymphatic Dysplasia ◽  
Hydrogel Bilayer

AbstractMechanosensitive ion channels are membrane gated pores which are activated by mechanical stimuli. The focus of this study is on Piezo1, a newly discovered, large, mammalian, mechanosensitive ion channel, which has been linked to diseases such as dehydrated hereditary stomatocytosis (Xerocytosis) and lymphatic dysplasia. Here we utilize an established in-vitro artificial bilayer system to interrogate single Piezo1 channel activity. The droplet-hydrogel bilayer (DHB) system uniquely allows the simultaneous recording of electrical activity and fluorescence imaging of labelled protein. We successfully reconstituted fluorescently labelled Piezo1 ion channels in DHBs and verified activity using electrophysiology in the same system. We demonstrate successful insertion and activation of hPiezo1-GFP in bilayers of varying composition. Furthermore, we compare the Piezo1 bilayer reconstitution with measurements of insertion and activation of KcsA channels to reproduce the channel conductances reported in the literature. Together, our results showcase the use of DHBs for future experiments allowing simultaneous measurements of ion channel gating while visualising the channel proteins using fluorescence.

scholarly journals Kv1.3 channel inhibition limits uremia-induced calcification in mouse and human vascular smooth muscle

Function ◽  
2020 ◽  
Author(s):  
Violeta Cazaña-Pérez ◽  
Pilar Cidad ◽  
Juan F Navarro-González ◽  
Jorge Rojo-Mencía ◽  
Frederic Jaisser ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Ion Channels ◽  
Vascular Smooth Muscle ◽  
Ion Channel ◽  
Mineral Metabolism ◽  
K Channel ◽  
Human Aorta ◽  
Genetic Ablation ◽  
Channel Inhibition

Abstract Chronic kidney disease (CKD) significantly increases cardiovascular risk. In advanced CKD stages, accumulation of toxic circulating metabolites and mineral metabolism alterations triggers vascular calcification, characterized by vascular smooth muscle cell (VSMC) transdifferentiation and loss of the contractile phenotype. Phenotypic modulation of VSMC occurs with significant changes in gene expression. Even though ion channels are an integral component of VSMC function, the effects of uremia on ion channel remodeling has not been explored. We used an in vitro model of uremia-induced calcification of human aorta smooth muscle cells (HASMC) to study the expression of 92 ion channel subunit genes. Uremic serum induced extensive remodeling of ion channel expression consistent with loss of excitability but different from the one previously associated to transition from contractile to proliferative phenotypes. Among the ion channels tested, we found increased abundance and activity of voltage-dependent K+ channel Kv1.3. Enhanced Kv1.3 expression was also detected in aorta from a mouse model of CKD. Pharmacological inhibition or genetic ablation of Kv1.3 decreased the amount of calcium phosphate deposition induced by uremia, supporting an important role for this channel on uremia-induced VSMC calcification.

scholarly journals Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets

Cancers ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 446 ◽  
Author(s):  
Daniela D’Arcangelo ◽  
Francesca Scatozza ◽  
Claudia Giampietri ◽  
Paolo Marchetti ◽  
Francesco Facchiano ◽  
...  
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
In Silico ◽  
Molecular Targets ◽  
In Silico Analysis ◽  
Human Melanoma ◽  
Antifungal Drug ◽  
Characteristic Analysis ◽  
Discrimination Ability

Expression of 328 ion channel genes was investigated, by in silico analysis, in 170 human melanoma samples and controls. Ninety-one members of this gene-family (i.e., about 28%) show a significant (p < 0.05) differential expression in melanoma- vs. nevi-biopsies, taken from the GEO database. ROC (receiver operating characteristic) analysis selected 20 genes as potential markers showing the highest discrimination ability of melanoma vs. nevi (AUC > 0.90 and p < 0.0001). These 20 genes underwent a first in silico-validation round in an independent patients-dataset from GEO. A second-in silico-validation step was then carried out on a third human dataset in Oncomine. Finally, five genes were validated, showing extremely high sensitivity and specificity in melanoma detection (>90% in most cases). Such five genes (namely, SCNN1A, GJB3, KCNK7, GJB1, KCNN2) are novel potential melanoma markers or molecular targets, never previously related to melanoma. The “druggable genome” analysis was then carried out. Miconazole, an antifungal drug commonly used in clinics, is known to target KCNN2, the best candidate among the five identified genes. Miconazole was then tested in vitro in proliferation assays; it dose-dependently inhibited proliferation up to 90% and potently induced cell-death in A-375 and SKMEL-28 melanoma cells, while it showed no effect in control cells. Moreover, specific silencing of KCNN2 ion channel was achieved by siRNA transfection; under such condition miconazole strongly increases its anti-proliferative effect. In conclusion, the present study identified five ion channels that can potentially serve as sensitive and specific markers in human melanoma specimens and demonstrates that the antifungal drug miconazole, known to target one of the five identified ion channels, exerts strong and specific anti-melanoma effects in vitro.

scholarly journals Development of a Low-Cost, Wireless Smart Thermostat for Isothermal DNA Amplification in Lab-On-A-Chip Devices

Micromachines ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 437
Author(s):  
Pardy ◽  
Sink ◽  
Koel ◽  
Rang
Keyword(s):  
Dna Analysis ◽  
Low Cost ◽  
Lab On A Chip ◽  
Dna Amplification ◽  
Nucleic Acid Amplification ◽  
Battery Life ◽  
Worst Case ◽  
3D Printed ◽  
Living Organisms ◽  
Smart Thermostat

Nucleic acid amplification tests (NAAT) are widely used for the detection of living organisms, recently applied in Lab-on-a-Chip (LoC) devices to make portable DNA analysis platforms. While portable LoC-NAAT can provide definitive test results on the spot, it requires specialized temperature control equipment. This work focuses on delivering a generalized low-cost, wireless smart thermostat for isothermal NAAT protocols in 2 cm × 3 cm LoC cartridges. We report on the design, prototyping, and evaluation results of our smart thermostat. The thermostat was evaluated by experimental and simulated thermal analysis using 3D printed LoC cartridges, in order to verify its applicability to various isothermal NAAT protocols. Furthermore, it was tested at the boundaries of its operating ambient temperature range as well as its battery life was evaluated. The prototype thermostat was proven functional in 20–30 °C ambient range, capable of maintaining the required reaction temperature of 12 isothermal NAAT protocols with 0.7 °C steady-state error in the worst case.

scholarly journals Manufacturing of 3D-Printed Microfluidic Devices for the Synthesis of Drug-Loaded Liposomal Formulations

2021 ◽  
Vol 22 (15) ◽  
pp. 8064
Author(s):  
Giulia Ballacchino ◽  
Edward Weaver ◽  
Essyrose Mathew ◽  
Rossella Dorati ◽  
Ida Genta ◽  
...  
Keyword(s):  
Encapsulation Efficiency ◽  
Liquid Crystal Display ◽  
Low Cost ◽  
In Vitro Release ◽  
Microfluidic Devices ◽  
Fused Deposition Modelling ◽  
Ζ Potential ◽  
Fused Deposition ◽  
3D Printed

Microfluidic technique has emerged as a promising tool for the production of stable and monodispersed nanoparticles (NPs). In particular, this work focuses on liposome production by microfluidics and on factors involved in determining liposome characteristics. Traditional fabrication techniques for microfluidic devices suffer from several disadvantages, such as multistep processing and expensive facilities. Three-dimensional printing (3DP) has been revolutionary for microfluidic device production, boasting facile and low-cost fabrication. In this study, microfluidic devices with innovative micromixing patterns were developed using fused deposition modelling (FDM) and liquid crystal display (LCD) printers. To date, this work is the first to study liposome production using LCD-printed microfluidic devices. The current study deals with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) liposomes with cholesterol (2:1) prepared using commercial and 3D-printed microfluidic devices. We evaluated the effect of microfluidic parameters, chip manufacturing, material, and channel design on liposomal formulation by analysing the size, PDI, and ζ-potential. Curcumin exhibits potent anticancer activity and it has been reported that curcumin-loaded liposomes formulated by microfluidics show enhanced encapsulation efficiency when compared with other reported systems. In this work, curcumal liposomes were produced using the developed microfluidic devices and particle sizing, ζ-potential, encapsulation efficiency, and in vitro release studies were performed at 37 °C.

scholarly journals Microfluidic Lab-on-a-Chip Based on UHF-Dielectrophoresis for Stemness Phenotype Characterization and Discrimination among Glioblastoma Cells

Biosensors ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 388
Author(s):  
Elisa Lambert ◽  
Rémi Manczak ◽  
Elodie Barthout ◽  
Sofiane Saada ◽  
Elena Porcù ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Ex Vivo ◽  
Low Cost ◽  
Low Frequency ◽  
Lab On A Chip ◽  
Microfluidic System ◽  
Frequency Range ◽  
Phenotype Transformation

Glioblastoma (GBM) is one of the most aggressive solid tumors, particularly due to the presence of cancer stem cells (CSCs). Nowadays, the characterization of this cell type with an efficient, fast and low-cost method remains an issue. Hence, we have developed a microfluidic lab-on-a-chip based on dielectrophoresis (DEP) single cell electro-manipulation to measure the two crossover frequencies: fx01 in the low-frequency range (below 500 kHz) and fx02 in the ultra-high-frequency range (UHF, above 50 MHz). First, in vitro conditions were investigated. An U87-MG cell line was cultured in different conditions in order to induce an undifferentiated phenotype. Then, ex vivo GBM cells from patients’ primary cell culture were passed through the developed microfluidic system and characterized in order to reflect clinical conditions. This article demonstrates that the usual exploitation of low-frequency range DEP does not allow the discrimination of the undifferentiated GBM cells from the differentiated one. However, the presented study highlights the use of UHF-DEP as a relevant discriminant parameter. The proposed microfluidic lab-on-a-chip is able to follow the kinetics of U87-MG phenotype transformation in a CSC enrichment medium and the cancer stem cells phenotype acquirement.

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