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Development of a Gas-Tight Microfluidic System for Raman Sensing of Single Pulmonary Arterial Smooth Muscle Cells Under Normoxic/Hypoxic Conditions
Excellence-Cluster Cardio-Pulmonary System (ECCPS), Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Center for Lung Research (DZL), Justus-Liebig University Giessen, Giessen, Germany.
Luleå University of Technology, Department of Engineering Sciences and Mathematics, Fluid and Experimental Mechanics.
Luleå University of Technology, Department of Engineering Sciences and Mathematics, Fluid and Experimental Mechanics.ORCID iD: 0000-0001-9789-6293
CMS, Imperial College, London, UK.
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2018 (English)In: Sensors, ISSN 1424-8220, E-ISSN 1424-8220, Vol. 10, article id 3238Article in journal (Refereed) Published
Abstract [en]

Acute hypoxia changes the redox-state of pulmonary arterial smooth muscle cells (PASMCs). This might influence the activity of redox-sensitive voltage-gated K⁺-channels (Kv-channels) whose inhibition initiates hypoxic pulmonary vasoconstriction (HPV). However, the molecular mechanism of how hypoxia-or the subsequent change in the cellular redox-state-inhibits Kv-channels remains elusive. For this purpose, a new multifunctional gas-tight microfluidic system was developed enabling simultaneous single-cell Raman spectroscopic studies (to sense the redox-state under normoxic/hypoxic conditions) and patch-clamp experiments (to study the Kv-channel activity). The performance of the system was tested by optically recording the O₂-content and taking Raman spectra on murine PASMCs under normoxic/hypoxic conditions or in the presence of H₂O₂. Oxygen sensing showed that hypoxic levels in the gas-tight microfluidic system were achieved faster, more stable and significantly lower compared to a conventional open system (1.6 ± 0.2%, respectively 6.7 ± 0.7%, n = 6, p < 0.001). Raman spectra revealed that the redistribution of biomarkers (cytochromes, FeS, myoglobin and NADH) under hypoxic/normoxic conditions were improved in the gas-tight microfluidic system (p-values from 0.00% to 16.30%) compared to the open system (p-value from 0.01% to 98.42%). In conclusion, the new redox sensor holds promise for future experiments that may elucidate the role of Kv-channels during HPV.

Place, publisher, year, edition, pages
Basel, Switzerland: MDPI, 2018. Vol. 10, article id 3238
Keywords [en]
Raman spectroscopy, hypoxia, microfluidic system, redox reactions on single cell level
National Category
Applied Mechanics Fluid Mechanics and Acoustics
Research subject
Experimental Mechanics; Fluid Mechanics
Identifiers
URN: urn:nbn:se:ltu:diva-71090DOI: 10.3390/s18103238ISI: 000448661500066PubMedID: 30261634Scopus ID: 2-s2.0-85054841733OAI: oai:DiVA.org:ltu-71090DiVA, id: diva2:1253043
Note

Validerad;2018;Nivå 2;2018-10-03 (svasva)

Available from: 2018-10-03 Created: 2018-10-03 Last updated: 2019-04-24Bibliographically approved

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Wahl, JoelAndersson, Anders G.Ramser, Kerstin

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