Lingzhi and San-Miao-San with hyaluronic acid gel mitigate cartilage degeneration in anterior cruciate ligament transection induced osteoarthritis

Man Chu; Ping Wu; Ming Hong; Huasong Zeng; Chun Kwok Wong; Yu Feng; Zhe Cai; William Weijia Lu

doi:10.1016/j.jot.2020.07.008

Lingzhi and San-Miao-San with hyaluronic acid gel mitigate cartilage degeneration in anterior cruciate ligament transection induced osteoarthritis

J Orthop Translat. 2020 Oct 21:26:132-140. doi: 10.1016/j.jot.2020.07.008. eCollection 2021 Jan.

Authors

Man Chu¹, Ping Wu², Ming Hong^{3

4}, Huasong Zeng², Chun Kwok Wong⁵, Yu Feng⁶, Zhe Cai^{2

5

7

8}, William Weijia Lu^{9

10}

Affiliations

¹ Faulty of Medical Technology, Shaanxi University of Chinese Medicine, Xianyang, China.
² Department of Allergy, Immunology and Rheumatology, Guangzhou Women and Children's Medical Center, Guangzhou, Guangdong, China.
³ Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou, China.
⁴ Institute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou, China.
⁵ Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong, China.
⁶ Department of Traumatology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China.
⁷ The Joint Center for Infection and Immunity, Guangzhou Institute of Pediatrics, Guangzhou Women and Children's Medical Center, Guangzhou, China.
⁸ The Joint Center for Infection and Immunity, Institute Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
⁹ Department of Orthopaedics and Traumatology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong.
¹⁰ Shenzhen Institutes of Advanced Technology, Chinese Academy of Science, Shenzhen, China.

Abstract

Objective: To investigate the mitigate efficacy of Chinese medicine Lingzhi (LZ) and San-Miao-San (SMS) combined with hyaluronic acid (HA)-gel in attenuating cartilage degeneration in traumatic osteoarthritis (OA).

Methods: The standardized surgery of anterior cruciate ligament transection (ACLT) was made from the medial compartment of right hind limbs of 8-week-old female SD rats and resulted in a traumatic OA. Rats (n = 5/group) were treated once intra-articular injection of 50 μl HA-gel, 50 μl HA-gel+50 μg LZ-SMS, 50 μl of saline+50 μg LZ-SMS and null (ACLT group) respectively, except sham group. Limbs were harvested for μCT scan and histopathological staining 3-month post-treatment. Inflammatory cytokines from plasma and synovial fluid were detected using Immunology Multiplex Assay kit. The putative targets of active compounds in LZ-SMS and known therapeutic targets for OA were combined to construct protein-protein interaction network. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was adopted to predict the potential targets and signaling pathway of LZ-SMS in OA through the tool of DAVID Bioinformatics.

Results: In vivo, HA-gel + LZ-SMS treatment resulted in a higher volume ratio of hyaline cartilage (HC)/calcified cartilage (CC) and HC/Sum (total volume of cartilage), compared to ACLT and HA-gel groups. In addition, histological results showed the elevated cartilage matrix, chondrogenic and osteoblastic signals in HA-gel + LZ-SMS treatment. Treatment also significantly altered subchondral bone (SCB) structure including an increase in BV/TV, Tb.Th, BMD, Conn.Dn, Tb.N, and DA, as well as a significant decrease in Tb.Sp and Po(tot), which implied a protective effect on maintaining the stabilization of tibial SCB microstructure. Furthermore, there was also a down-regulated inflammatory cytokines and upregulated anti-inflammatory cytokine IL-10 in HA+LZ-SMS group. Finally, 64 shared targets from 37 active compounds in LZ-SMS related to the core genes for the development of OA. LZ-SMS has a putative role in regulating inflammatory circumstance through influencing the MAPK signaling pathway.

Conclusion: Our study elucidated a protective effect of HA-gel + LZ-SMS in mitigating cartilage degradation and putative interaction with targets and signaling pathway for the development of traumatic OA.

The translational potential of this article: Our results provide a biological rationale for the use of LZ-SMS as a potential candidate for OA treatment.

Keywords: 3D, Three-dimensional; AC, Articular cartilage; ACLT, Anterior cruciate ligament transection; Acan, Aggrecan; Articular cartilage; BMD, Bone mineral density; BV/TV, Bone volume fraction; CC, Calcified cartilage; Conn.Dn, Connectivity density; DA, Degree of anisotropy; DL, Drug-likeness; ECM, Extracellular matrix; FDR, False discovery rate; GO, Gene ontology; HA, Hyaluronic acid; HC, Hyaline cartilage; Hyaluronic acid gel; KEGG, Kyoto Encyclopedia of Genes and Genomes; LZ-SMS, Lingzhi-San-Miao-San; Lingzhi and San-Miao-San; MZ, Middle zone area of articular cartilage; NC, Negative control; OA, Osteoarthritis; OB, Oral bioavailability; OMIM, Online Mendelian Inheritance in Man; Osteoarthritis; PPI, Protein–protein interaction; Po(tot), Total porosity; ROI, Region of Interest; SC, Superficial cartilage; SCB, Subchondral bone; SZ, Superficial zone of articular cartilage; Subchondral trabecular bone; Sum, Whole cartilage; TCM, Traditional Chinese medicine; TCMSP, Traditional Chinese Medicine Systems Pharmacology Database; Tb.N, Trabecular number; Tb.Pf, Trabecular bone pattern factor; Tb.Sp, Trabecular separation; Tb.Th, Trabecular thickness.