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2018 (English)In: Sensors, 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
Keywords
Raman spectroscopy, hypoxia, microfluidic system, redox reactions on single cell level
National Category
Applied Mechanics Fluid Mechanics
Research subject
Experimental Mechanics; Fluid Mechanics
Identifiers
urn:nbn:se:ltu:diva-71090 (URN)10.3390/s18103238 (DOI)000448661500066 ()30261634 (PubMedID)2-s2.0-85054841733 (Scopus ID)
Note
Validerad;2018;Nivå 2;2018-10-03 (svasva)
2018-10-032018-10-032025-10-22Bibliographically approved