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. 2022 Jun 21;13:912933. doi: 10.3389/fimmu.2022.912933

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Copyright © 2022 Li, Wang, Liu, Wu, He, Li, Wang, Su, Yan, Su, Zhang and Lin

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Therapeutic effects of BHGZD, two-BAC combination, and MTX on the severity of arthritis in AIA-M rats (Control, AIA-M, BHGZD, MG+CA, and MTX). (A) Representative images of arthritis. (B) Infrared thermography. (C) Arthritis incidence (n = 3 per group), the diameter of the limb (n = 5 per group), and arthritis score on the 27th day after immunization (n ≥ 4 per group, all experiments were performed in triplicate). (D) The pain thresholds (mechanical-, acetone-, and thermal-induced hyperalgesia, n ≥ 4 per group, all experiments were performed in triplicate). (E) Quantitative micro-computed tomography (micro-CT) analysis of bone mineral density (BMD), tissue mineral density (TMD), bone volume/tissue volume (BV/TV), bone surface/bone volume (BS/BV), trabecular separation (Tb.Sp), and trabecular thickness (Tb.Th) on the 31st day after immunization (n ≥ 5 per group, all experiments were performed in triplicate). (F) Representative micro-CT images of knee joints showing bone erosion levels (the red arrow indicates the position of the bone destruction). (G) Pathological changes in the knee joints using hematoxylin and eosin (H&E, scale bar represents 1 mm), safranin O fast green (scale bar represents 200 μm), and Masson trichrome staining (scale bar represents 200 μm) in different groups (n = 5 per group). (H) Quantitative analysis of H&E staining in inflammatory cell infiltration, bone destruction, and synovial hyperplasia. Cartilage destruction of safranin O fast, and Masson trichrome staining (n = 5 per group). Data are expressed as the mean ± SD. ***, p < 0.001, comparison with the normal control group; #, ##, ###, p < 0.05, p < 0.01, p < 0.001, respectively, comparison with the AIA-M model group.