Histological Evaluation of Articular Cartilage

The fresh samples were fixed in 4% paraformaldehyde for 48 h followed with decalcification in 10% EDTA-2Na for 3 weeks, and then paraffin-embedded for further routine histological preparation. Three micrometer-thick sections were deparaffnized in xylene and rehydrated in graded alcohols and distilled water prior to H.E. and Safranin O –Fast green stainings as previously described [18 (link), 19 (link)]. The articular cartilage thickness of each femur condyle (from superficial zone to tidemark) was measured using Image-Pro Plus 6.0 software. According to OARSI scoring system established for grading OA changes [25 (link)], and semi-quantitative histopathological grading was performed using VS120-S6 (Olympus, Japan) by two different blinded pathologists, for a maximum possible score of 6. Grade 0 represents normal articular cartilage and increasing grade indicates a more biologically cartilage degeneration.

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Cai H., Qu N., Chen X., Zhou Y., Zheng X., Zhang B, & Xia C. (2017). The inhibition of PLCγ1 protects chondrocytes against osteoarthritis, implicating its binding to Akt. Oncotarget, 9(4), 4461-4474.

Publication 2017

VS120-S6

Alcohols Articular cartilage Cartilage Condyle Edta Embedded paraffin Fast green Femur Paraformaldehyde Pathologists Safranin o Stainings Xylene

Corresponding Organization :

Other organizations : Zhongshan Hospital of Xiamen University, Xiamen University

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Variable analysis

independent variables

Decalcification in 10% EDTA-2Na for 3 weeks

dependent variables

Articular cartilage thickness of each femur condyle (from superficial zone to tidemark)
Histopathological grading using OARSI scoring system (maximum possible score of 6)

control variables

Fixation in 4% paraformaldehyde for 48 h
Paraffin-embedding for routine histological preparation
H.E. and Safranin O–Fast green stainings

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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