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Flow Cytometry Analysis of Immune Cell Subsets within the Murine Spleen, Bone Marrow, Lymph Nodes and Synovial Tissue in an Osteoarthritis Model
Published on: April 24, 2020
Commiphora wightii and its formulations extenuate macrophage-mediated inflammatory pathology in osteoarthritis
Vanshika Srivastava1, Ritik Raj1, Ankit Ganeshpurkar2
1Department of Pharmaceutical Biotechnology, Poona College of Pharmacy, Bharati Vidyapeeth Deemed to Be University, Erandwane, Pune, 411038, India.
Background:
Osteoarthritis (OA) treatment often focuses on symptom management rather than addressing underlying inflammation and cartilage degeneration. In search of safer, long-term options, many patients turn to Ayurvedic remedies like Guggul (Commiphora wightii) and its formulations-Amritadi Guggul (AG) and Rasnadi Guggul (RG)-though biochemical validation remains limited.
Objective:
This study evaluates the immunomodulatory effects of standard guggulsterone extract (SGE), AG, and RG on synovial inflammation, mitochondrial stress, and complement activation, using the U937 monocyte cell line.
Materials And Methods:
Cells were stimulated with Phorbol 12-myristate 13 acetate (PMA) and treated with varying concentrations of the extracts. Anti-inflammatory effects were measured via mRNA expression of iNOS, MMP-1, MMP-13, and VEGF-1. Macrophage polarization markers (CD68, CD86, CD163), mitochondrial membrane potential (JC-1 assay), collagenase activity (gelatinase spot assay), and molecular docking with complement factor B (CFB) were also assessed.
Results:
Results showed that SGE, AG, and RG significantly reduced nitric oxide and pro-inflammatory gene expression. Treatments suppressed M1 macrophage markers without promoting M2 differentiation. 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimi- dazolylcarbocyanine iodide (JC) - 1 assays indicated improved mitochondrial stability, while all formulations inhibited collagenase activity. Docking studies revealed strong interactions between guggulsterone and CFB, suggesting complement inhibition.
Conclusions:
These findings highlight the potential of Guggul and its formulations to modulate macrophage activity, reduce inflammation, and support joint preservation in OA. Though limited by the use of a monocyte-derived cell line, the study lays the groundwork for future validation in primary cells and in vivo models.
