Novel Use of a Dektak 150 Surface Profiler Unmasks Differences in Resorption Pit Profiles Between Control and Charcot Patient Osteoclasts

We hypothesized that newly formed osteoclasts from patients with acute Charcot osteoarthropathy can resorb surfaces of bone more extensively compared with controls. Peripheral blood monocytes, isolated from eight Charcot patients and nine controls, were cultured in vitro on 24-well plates and bovine...

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Bibliographic Details
Published in:Calcified tissue international 2014-04, Vol.94 (4), p.403-411
Main Authors: Petrova, Nina L., Petrov, Peter K., Edmonds, Michael E., Shanahan, Catherine M.
Format: Article
Language:English
Subjects:
Acid phosphatase (tartrate-resistant)
Adult
Amyotrophic Lateral Sclerosis - pathology
Animals
Biochemistry
Biomedical and Life Sciences
Bone and Bones
Bone density
Bone Resorption
Case-Control Studies
Cattle
Cell Biology
Cellular biology
Culture Media
Diabetes
Endocrinology
Feet
Female
Humans
Leukocytes, Mononuclear - cytology
Life Sciences
Macrophage Colony-Stimulating Factor - pharmacology
Male
Middle Aged
Monocytes - cytology
Original Research
Orthopedics
Osteoclasts - cytology
RANK Ligand - pharmacology
Surface Properties
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Summary:We hypothesized that newly formed osteoclasts from patients with acute Charcot osteoarthropathy can resorb surfaces of bone more extensively compared with controls. Peripheral blood monocytes, isolated from eight Charcot patients and nine controls, were cultured in vitro on 24-well plates and bovine bone discs in duplicate with macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor κβ ligand (RANKL). Osteoclast formation was assessed by tartrate-resistant acid phosphatase staining (TRAcP) at day 17. Resorption was measured at day 21 after toluidine blue staining by two methods: (1) area of resorption at the surface by image analysis (%) and (2) area of resorption under the surface (μm 2 ) measured by a Dektak 150 Surface Profiler. Ten 1,000 μm-long scans were performed per disc. Pits were classified as unidented, bidented, and multidented according to their shape. Although the number of newly formed TRAcP positive multinucleated cells (>3 nuclei) was similar in M-CSF + RANKL-treated cultures between controls and Charcot patients, the latter exhibited increased resorbing activity. The area of resorption on the surface by image analysis was significantly greater in Charcot patients compared with controls (21.1 % [14.5–26.2] vs. 40.8 % [35.4–46.0], median [25–75th percentile], p  
ISSN:0171-967X
1432-0827
DOI:10.1007/s00223-013-9820-9