Abstract
Chronic inflammation is a common underlying factor in osteoarthritis (OA) and most age-related degenerative diseases. As conventional therapies help only in partial alleviation of symptoms in OA, stem cell-based therapies and herbal supplements are being widely explored. Thymoquinone (TQ), an active ingredient of Nigella sativa is reported to have immunomodulatory, anti-inflammatory and antioxidant properties. We evaluated the effects of TQ on bone marrow MSCs (BM-MSCs) derived from OA patients and its interrelated pathways in inflammation and age-related degenerative diseases using Ingenuity Pathway Analysis (IPA) as well as possible molecular targets using SwissTargetPrediction. BM-MSCs were derived from OA patients and their stemness properties were characterized by studying the MSCs related CD surface marker expression and differentiation into adipocytes, osteoblasts, and chondrocytes. Treatment with TQ (100 nM–5 μM) demonstrated cell death, especially at higher concentrations. MTT assay demonstrated a significant concentration-dependent decrease in cell viability which ranged from 20.04% to 69.76% with higher doses (300 nM, 1 μM, and 5 μM), especially at 48h and 72h. Additional cell viability testing with CellTiter-Blue also demonstrated a significant concentration-dependent decrease in cell viability which ranged from 27.80 to 73.67% with higher doses (300 nM, 1 μM, 3 μM, and 5 μM). Gene expression analysis following treatment of BM-MSCs with TQ (1 and 3 μM) for 48h showed upregulation of the anti-inflammatory genes IL-4 and IL-10. In contrast, the pro-inflammatory genes namely IFN-γ, TNF-α, COX-2, IL-6, IL-8, IL-16, and IL-12A although were upregulated, compared to the lower concentration of TQ (1 μM) they were all decreased at 3 μM. The pro-apoptotic BAX gene was downregulated while the SURVIVIN gene was upregulated. IPA of the molecular interaction of TQ in inflammation and age-related degenerative diseases identified canonical pathways directly related to synaptogenesis, neuroinflammation, TGF-β, and interleukin signaling. Further screening led to the identification of 36 molecules that are involved in apoptosis, cell cycle regulation, cytokines, chemokines, and growth factors. SwissTargetPrediction of TQ identified potential molecular targets with high probability. TQ exerted anti-inflammatory effects and therefore can be a useful adjuvant along with conventional therapies against inflammation in OA and other age-related degenerative diseases.
Highlights
Physiological age-related decline in function of the various tissues in the human body is inevitable
The bone-marrow mesenchymal stem cells (BM-Mesenchymal stem cells (MSCs)) were successfully isolated with all samples obtained from OA patients
The non-adherent cells, dead cells, and RBCs were removed with medium changes allowing expansion of the BM-MSCs
Summary
Physiological age-related decline in function of the various tissues in the human body is inevitable. Age-related degenerative diseases are associated with cellular and biochemical damage contributing to both functional and structural degeneration of the organ systems (da Costa et al, 2016; Cole and Franke, 2017; Ramalingam et al, 2018). Such degenerative processes become commonly manifested as neurological diseases (Alzheimer’s and Parkinson’s) bone and joint diseases (OA, rheumatoid arthritis, and osteoporosis), retinal diseases (macular degeneration, glaucoma, cataract, and diabetic retinopathy) or cardiovascular diseases (atherosclerosis, hypertension, and valvular stenosis). Aging related chronic diseases and degenerative disorders have major economic and social implications. They need serious interventional strategies (Tarailo-Graovac et al, 2016; Ramalingam et al, 2018)
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