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Three 5-μm-thick paraffin-embedded horizontal bone sections were cut from the proximal end of the diaphysis, stained with haematoxylin eosin and examined by light microscopy.
Additionally, four to five sequential frozen bone sections were cut at 10 μm thick and were stained with safranin-O/fast green to visualize gross pathologic changes in cartilage and bone induced by CFA.
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About 5 millimeter thick blocks containing a sagittal vertebral bone section were cut using a low speed diamond saw (Buehler Isomed, Lake Pluff, USA).
All specimen preparation was done with blinding as to treatment group. 2 transverse bone-implant sections were cut perpendicular to the implant axis, using a water-cooled diamond-band saw and implant-based alignment post (Exact Apperatebau, Nordstedt, Germany).
Sections were cut in the middle of the bone defect and perpendicular to the original drill hole revealing the round original bone defect for further histological assessment with light microscopy.
Ankle joints were embedded in paraffin and sections were cut and stained with hematoxylin and eosin for evaluation of inflammation and bone erosion, as previously described [ 21].
Sections were cut and stained with periodic acid and Schiff's reagent for evaluation of new bone formation.
Sections were cut and stained with anti-VEGF antibodies to evaluate VEGF expression, and with periodic acid and Schiff's reagent to evaluate new bone formation.
Sections were cut and stained with hematoxylin and eosin.
Paraffin sections were cut at 5 µm.
Sections were cut at 5 µm thickness.
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