TY - JOUR
T1 - Real-time quantification of osteoclastic resorptive activity by electric cell-substrate impedance sensing
AU - Jansen, Ineke D.C.
AU - van Velzen, Thijs
AU - de Vries, Teun J.
AU - Szulcek, Robert
AU - van Loon, Jack J.W.A.
N1 - Funding Information: This work was partially supported by the Dutch CardioVascular Alliance (DCVA) (2012-08, 2014-11) awarded to the Phaedra and the RECONNECT consortium as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium. These Grants include collective funding from the Dutch Heart Foundation, the Dutch Federation of University Medical Centers, Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences. Publisher Copyright: Copyright © 2022 Jansen, van Velzen, de Vries, Szulcek and van Loon.
PY - 2022/8/19
Y1 - 2022/8/19
N2 - In several diseases, bone resorption by osteoclasts is dysregulated. Thus far, no simple technique for real-time measurement of resorption is available. Here, we introduce an impedimetric bioassay for real-time monitoring of resorption by making use of the electrical insulating properties of the resorbable substrate calcium phosphate. Different chemical stimuli were applied to (pre)osteoclasts cultured on a layer of calcium phosphate in multi-well plates containing electrodes. By this, osteoclast activity can be measured continuously over days, and the effects of stimulating or inhibiting factors can be quantified. When cells were cultured in the presence of an inflammatory factor such as IL-1β, the resorptive activity started earlier. The measured decline in resistance was higher at culture day 5 than at cultures with M-CSF or M-CSF + RANKL (M-CSF norm. Resistance = 1, M-CSF + RANKL = 0.7, M-CSF + RANKL + IL-1β = 0.5). However, at day 11, this difference had nearly disappeared. Likewise, bisphosphonates were shown to inhibit osteoclastic activity. Our findings illustrate the importance of real-time monitoring; wherefore, this method has high potential not only for the study of osteoclast resorptive activity in the context of osteoclast function and diseases but also could find application in high-throughput drug-testing studies.
AB - In several diseases, bone resorption by osteoclasts is dysregulated. Thus far, no simple technique for real-time measurement of resorption is available. Here, we introduce an impedimetric bioassay for real-time monitoring of resorption by making use of the electrical insulating properties of the resorbable substrate calcium phosphate. Different chemical stimuli were applied to (pre)osteoclasts cultured on a layer of calcium phosphate in multi-well plates containing electrodes. By this, osteoclast activity can be measured continuously over days, and the effects of stimulating or inhibiting factors can be quantified. When cells were cultured in the presence of an inflammatory factor such as IL-1β, the resorptive activity started earlier. The measured decline in resistance was higher at culture day 5 than at cultures with M-CSF or M-CSF + RANKL (M-CSF norm. Resistance = 1, M-CSF + RANKL = 0.7, M-CSF + RANKL + IL-1β = 0.5). However, at day 11, this difference had nearly disappeared. Likewise, bisphosphonates were shown to inhibit osteoclastic activity. Our findings illustrate the importance of real-time monitoring; wherefore, this method has high potential not only for the study of osteoclast resorptive activity in the context of osteoclast function and diseases but also could find application in high-throughput drug-testing studies.
KW - bone
KW - ECIS
KW - osteoclast
KW - real-time quantification
KW - resorption
KW - sensor
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U2 - https://doi.org/10.3389/fcell.2022.921066
DO - https://doi.org/10.3389/fcell.2022.921066
M3 - Article
C2 - 36060806
SN - 2296-634X
VL - 10
SP - 1
EP - 9
JO - Frontiers in cell and developmental biology
JF - Frontiers in cell and developmental biology
M1 - 921066
ER -