A1 Refereed original research article in a scientific journal
In vitro model of bone to facilitate measurement of adhesion forces and super-resolution imaging of osteoclasts
Authors: Deguchi T, Alanne MH, Fazeli E, Fagerlund KM, Pennanen P, Lehenkari P, Hanninen PE, Peltonen J, Nareoja T
Publisher: NATURE PUBLISHING GROUP
Publication year: 2016
Journal: Scientific Reports
Journal name in source: SCIENTIFIC REPORTS
Journal acronym: SCI REP-UK
Article number: ARTN 22585
Volume: 6
Number of pages: 13
ISSN: 2045-2322
eISSN: 2045-2322
DOI: https://doi.org/10.1038/srep22585(external)
To elucidate processes in the osteoclastic bone resorption, visualise resorption and related actin reorganisation, a combination of imaging technologies and an applicable in vitro model is needed. Nanosized bone powder from matching species is deposited on any biocompatible surface in order to form a thin, translucent, smooth and elastic representation of injured bone. Osteoclasts cultured on the layer expressed matching morphology to ones cultured on sawed cortical bone slices. Resorption pits were easily identified by reflectance microscopy. The coating allowed actin structures on the bone interface to be visualised with super-resolution microscopy along with a detailed interlinked actin networks and actin branching in conjunction with V-ATPase, dynamin and Arp2/3 at actin patches. Furthermore, we measured the timescale of an adaptive osteoclast adhesion to bone by force spectroscopy experiments on live osteoclasts with bone-coated AFM cantilevers. Utilising the in vitro model and the advanced imaging technologies we localised immunofluorescence signals in respect to bone with high precision and detected resorption at its early stages. Put together, our data supports a cyclic model for resorption in human osteoclasts.
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