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Focus Ion Beam/Scanning Electron Microscopy Characterization of Osteoclastic Resorption of Calcium Phosphate Substrates

Authors :
Diez-Escudero, Anna
Espanol, Montserrat
Montufar, Edgar B.
Di Pompo, Gemma
Ciapetti, Gabriela
Baldini, Nicola
Ginebra, Maria-Pau
Source :
Tissue Engineering Part C: Methods; February 2017, Vol. 23 Issue: 2 p118-124, 7p
Publication Year :
2017

Abstract

This article presents the application of dual focused ion beam/scanning electron microscopy (FIB-SEM) imaging for preclinical testing of calcium phosphates with osteoclast precursor cells and how this high-resolution imaging technique is able to reveal microstructural changes at a level of detail previously not possible. Calcium phosphate substrates, having similar compositions but different microstructures, were produced using low- and high-temperature processes (biomimetic calcium-deficient hydroxyapatite [CDHA] and stoichiometric sintered hydroxyapatite, respectively). Human osteoclast precursor cells were cultured for 21 days before evaluating their resorptive potential on varying microstructural features. Alternative to classical morphological evaluation of osteoclasts (OC), FIB-SEM was used to observe the subjacent microstructure by transversally sectioning cells and observing both the cells and the substrates. Resorption pits, indicating OC activity, were visible on the smoother surface of high-temperature sintered hydroxyapatite. FIB-SEM analysis revealed signs of acidic degradation on the grain surface under the cells, as well as intergranular dissolution. No resorption pits were evident on the surface of the rough CDHA substrates. However, whereas no degradation was detected by FIB sections in the material underlying some of the cells, early stages of OC-mediated acidic degradation were observed under cells with more spread morphology. Collectively, these results highlight the potential of FIB to evaluate the resorptive activity of OC, even in rough, irregular, or coarse surfaces where degradation pits are otherwise difficult to visualize.

Details

Language :
English
ISSN :
19373384 and 19373392
Volume :
23
Issue :
2
Database :
Supplemental Index
Journal :
Tissue Engineering Part C: Methods
Publication Type :
Periodical
Accession number :
ejs41331874
Full Text :
https://doi.org/10.1089/ten.tec.2016.0361