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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 tekniska universitet, Institutionen för teknikvetenskap och matematik, Strömningslära och experimentell mekanik.
Luleå tekniska universitet, Institutionen för teknikvetenskap och matematik, Strömningslära och experimentell mekanik.ORCID-id: 0000-0001-9789-6293
CMS, Imperial College, London, UK.
Vise andre og tillknytning
2018 (engelsk)Inngår i: Sensors, ISSN 1424-8220, E-ISSN 1424-8220, Vol. 10, artikkel-id 3238Artikkel i tidsskrift (Fagfellevurdert) 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.

sted, utgiver, år, opplag, sider
Basel, Switzerland: MDPI, 2018. Vol. 10, artikkel-id 3238
Emneord [en]
Raman spectroscopy, hypoxia, microfluidic system, redox reactions on single cell level
HSV kategori
Forskningsprogram
Experimentell mekanik; Strömningslära
Identifikatorer
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
Merknad

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

Tilgjengelig fra: 2018-10-03 Laget: 2018-10-03 Sist oppdatert: 2019-04-24bibliografisk kontrollert

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