الفهرس 
AbstractOnly 14 pages are availabe for public view
 |  | from | 138 |  |  |
| | | from | 138 | | |
Abstract
Rheumatoid arthritis (RA) is a syndrome of ongoing inflammation that is categorized with joint rubefaction, edema, and impairment of synovial joints. It is considered a chief cause of permanent disability, augmented mortality, and socioeconomic costs. Its prevalence is around 1% of the global population which is in continuous increase with time and propagates in females 3 times more than males and usually develops in the fourth and fifth decades of life. Former research and studies suggested that the imbalanced immunological responses in addition to genetic factors play a fundamental role in RA development. However, the exact mechanism of RA pathogenesis and its etiology remains generally indefinite. Complete Freund’s Adjuvant (CFA)-induced arthritis is a model of chronic polyarthri¬tis with features that resemble human RA in many aspects and is one of the best experi¬mental models for detection and evaluation of the compounds with anti-inflammatory or anti-rheumatic activity.
Currently, there is no optimal therapy for RA except for systemic immune suppressants. The most widely used medications such as non-steroidal anti-inflammatory drugs (NSAIDS) besides the biological agents (e.g., antitumor necrosis-α antibody) help to relieve the symptoms, but serious adverse reactions are associated with extended use of these medications. Furthermore, these drugs are costly, and not all patients are reacting well to them. Consequently, it became so critical to explore promise mechanisms and seek potential, safer alternative therapies to improve the inflammatory pathological progress in RA patients.
The present work aims to evaluate the convenience and bioavailability of BM-MSCs, IMC, and BM-MSCs plus IMC on CFA-induced arthritis in experimental rats.
To perform the experiment, all groups except the healthy group all animals were inoculated by subcutaneous injection of 0.1 mL/rat CFA, (Sigma Chemical Co., St Louis, Mo., USA) into the footpad of the right hind paw at the dose of 1 mg/ kg b.w. For more extensive severity of arthritis, a booster dose (0.1 ml) was injected on the second day.
The experimental design contained five groups each consists of ten rats. These groups are listed as follow:
group 1 (Normal): It consists of healthy rats that was given the equivalent volumes carboxy methylcellulose (CMC) daily and orally for 3 weeks and Dulbecco’s Modified Eagles Medium (DMEM) intravenously at the 1st, 6th, 12th and 18th days.
group 2 (CFA): It is composed of CFA-induced arthritic rats and was orally given the equivalent volumes CMC daily and orally for 3 weeks and DMEM intravenously at the 1st, 6th, 12th and 18th days.
group 3 (CFA+BM-MSCs): This group consists of CFA-induced arthritic rats that received four doses of BM-MSCs (1 × 106 cells /rat/dose) by intravenous injection through lateral tail vein per rat. Each dose suspended in 0.2 ml DMEM (Dulbecco’s modified Eagles medium). Doses were given at the 1st, 6th, 12th and 18th days after CFA injection.
group 4 (CFA+IMC): This group is composed of CFA-induced arthritic rats supplemented orally with IMC daily in a dose of 2 mg/kg body weight (b.w.). IMC was freshly prepared immediately before administration by dissolving in 5 mL of 1% CMC for three weeks. IMC was acquired from Sigma Chemical Company (Sigma Chemical Co., St Louis, Mo., USA).
group 5 (AIA+BM-MSCs+IMC): This group consists of CFA-induced arthritic rats that were concurrently supplemented with BM-MSCs and IMC as described in groups 3 and 4.
The CFA-administered group was compared with the normal one while the arthritic rats treated with BM-MSCs, IMC, and BM-MSCs plus IMC were compared with the CFA-arthritic group.
Results showed that CFA injection caused a marked elevation in paw diameter, circumference, and volume in the arthritic rats in comparison with the normal rats. These parameters were improved significantly as a result of BM-MSCs and/or IMC treatments which suggest the effectiveness of these treatments on treating RA.
The TLC and DLC hematological alterations were evidenced by an obvious leukocytosis with higher lymphocyte and neutrophil counts in the arthritic control animals. It was found that the three treatments BM-MSCs, IMC, and BM-MSCs plus IMC, decreased the TLC as well as the lymphocyte, monocyte and neutrophils counts near to their normal levels, indicating the anti-inflammatory effect of the treatments on the arthritis course.
Concerning oxidative stress and antioxidant defense system, the LPO level was significantly elevated in the serum of CFA-induced arthritic rats while the GSH content and the activities of antioxidant enzymes including SOD, GPx, and GST were decreased. Conversely, the treatment with BM-MSCs and/or IMC caused marked decline of LPO and increase in GSH content as well the GPx, GST, and SOD activities. These results refer that the three therapies play an important role in treating the RA course through inhibition of the oxidative stress and enhancement of the antioxidant defense system.
Similarly, the transforming growth factor-β1 (TGF-β1) was enhanced while the antioxidant, nuclear factor erythroid 2-related factor, (Nrf-2) expression was remarkably decreased in ankle articular joint tissue of the CFA-induced arthritic rats. Conversely, the supplementation with BM-MSCs and/or IMC succeeded in inhibiting the TGF-β1 and improving the Nrf-2 levels in the samples of the treated animals.
Moreover, in the CFA-arthritic group, the levels of serum RF and Anti-CCP autoantibodies were increased. Also, the levels of the inflammatory markers involving IL-1β, TNF-α, iNOS and MMP-9 mRNA expression level were significantly elevated. Besides, the anti-inflammatory IL-10 and IL-4 mRNA expression levels were extremely decreased. However, all these previously altered mediators were effectively ameliorated by the three therapies and BM-MSCs plus IMC therapy was the most potent in improving the disease.
Histopathologically, the former results of the immunological indices are supported by the histopathological examination of the ankle joint. The BM-MSCs and/or IMC treated arthritic rats showed an obvious reduction in the extent of inflammatory cell infiltration and pannus formation in the synovial membrane of the ankle joint sections when compared with the CFA-arthritic group. This is consistent with the reduction of the pro-inflammatory and inflammatory cytokines and enhancement of the anti-inflammatory status as a result of the treatments.
In conclusion, BM-MSCs and/or IMC treatments has a powerful therapeutic action against CFA-induced arthritis in Wistar rats. Thus, BM-MSCs plus IMC might have the potential to be developed as a safer alternative to NSAIDs and DMARDs for the treatment of RA. Furthermore, the anti-arthritic effects of BM-MSCs and/or IMC treatments may be mediated via their abilities to suppress inflammation, reduce oxidative stress and enhance the antioxidant defense system.