Quantitative Evaluation of Arthritis Severity

After euthanasia, knee joints of mice were collected and stained with haematoxylin-eosin (H&E). To quantitatively evaluate the severity of arthritis, a scoring system was employed according to the reported protocol.4 (link) Immunohistochemical analysis was performed as previously described with some modifications.4 (link) Knee joint sections on slides were incubated with antibodies against PIAS3, p-STAT3, STAT3 or VEGF (Wanleibio, Shenyang, China). Subsequently, the sections were stained using a polymer HRP detection system (PV9001, ZSGB-BIO, Beijing, China) and visualized with a DAB peroxidase substrate kit (ZLI-9017, ZSGB-BIO, Beijing, China). Following immunostaining, the synovial area in the joint of each section was evaluated under a microscope (LEICA DMi8, Germany) in three randomly selected areas at a magnification of 100×. Image-Pro Plus 6 (Media Cybernetics, Inc.) was used to analyse the average integrated optical density (IOD) according to a previously described protocol.2 (link)

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Zhang J., Zhang Y., Ma Y., Luo L., Chu M, & Zhang Z. (2021). Therapeutic Potential of Exosomal circRNA Derived from Synovial Mesenchymal Cells via Targeting circEDIL3/miR-485-3p/PIAS3/STAT3/VEGF Functional Module in Rheumatoid Arthritis. International Journal of Nanomedicine, 16, 7977-7994.

Publication 2021

p-STAT3 STAT3 VEGF PV9001 ZLI-9017 Image-Pro Plus 6

Antibodies Arthritis Eosin Euthanasia Joint Knee joint Mice Microscope Optical Peroxidase Pias3 Polymer Stat3 Vegf

Corresponding Organization :

Other organizations : Harbin Medical University, First Affiliated Hospital of Harbin Medical University, Second Affiliated Hospital of Harbin Medical University

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Variable analysis

independent variables

Type of staining (haematoxylin-eosin (H&E))
Antibodies used for immunohistochemical analysis (PIAS3, p-STAT3, STAT3, VEGF)

dependent variables

Severity of arthritis (evaluated using a scoring system)
Average integrated optical density (IOD) of the synovial area in the joint

control variables

Knee joint sections on slides
Magnification (100×) used for microscope evaluation

controls

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

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