In-vitro and ex-vivo evaluation of plant sterol in osteoarthritis by CAM Assay

Abstract

The aim of the study was to evaluate the pharmacological effect of stigmasterol and β-sitosterol using anti-inflammatory and anti-arthritic in-vitro and ex-vivo methods. Stigmasterol and β-sitosterol was obtained commercially and the CAM assay was done to determine the anti-arthritic and anti-inflammatory effects. The DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2, 2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid) assay was performed for anti-oxidant by in-vitro method. An evaluation of toxicity was determined by the lethality study assessment of Brine Shrimp larvae in which the administration of test (Phytosterols) and standard (NaCl, NaOH) drugs was estimated, by estimating the toxicity of the phytosterols the Chick chorioallantoic membrane (CAM) assay was studied for chondrogenesis in correlation to infiltration of inflammatory mediators. The CAM assay shows the anti-arthritic activity and anti-inflammatory activity by mTORC1 signalling pathway and deactivates certain inflammatory mediators thereby increasing cell recovery and joint health. The CAM assay in chick embryo done by using 10 μg and 20μg of each phytosterol respectively. The toxicity LC50 measured at 87.7 μg/ml of phytosterols and 1.99 μg/ml for potassium dichromate (Std). At higher dose of 20 μg/ml the CAM membrane was observed in reducing the inflammation caused in cartilages of synovial joint. From the study, Stigmasterol and β-sitosterol managed the inflammation and an oxidative property plays a significant role in human health care system. Keywords: Osteoarthritis, Stigmasterol, ?-Sitosterol, Inflammation, Anti-Oxidant.