## 4 Discussion  
This study demonstrated that NEM exhibits dose-dependent anti-inflammatory and cytoprotective effects on extracellular matrix preservation in primary cultured rat articular chondrocytes, without cytotoxicity up to a concentration of 10 mg/mL in vitro. These findings provide direct evidence of NEM's beneficial effects in mitigating chondrocyte damage induced by inflammatory stress under experimental conditions. Historically, surgical techniques have been utilized in animal models to induce OA, facilitating the preclinical evaluation of potential disease-modifying drugs. Such models, developed in rodents since the late 1970s, enable researchers to assess drug efficacy within specific joint tissues and determine associated side effects [32]. Surgical methods, including anterior cruciate ligament transection and partial medial meniscectomy in rats, effectively mimic post-traumatic OA, offering critical insights into disease progression and the potential of therapeutic interventions. These surgical models are instrumental in reproducing the conditions of post-traumatic OA, thereby enhancing the translational relevance of preclinical findings for therapeutic development [33]. Incorporating surgical techniques in OA models aids in the practical reproduction of post-traumatic OA conditions, enhancing the relevance of preclinical trials for potential therapeutic interventions [29; 30]. In our study, NEM demonstrated a dose-dependent mitigation of surgically induced OA in Sprague-Dawley rats, suggesting its potential as an alternative therapy or functional food component for OA management. Comparing NEM's effects to those of diclofenac sodium (DS), a well-established COX inhibitor and NSAID, further underscores NEM's therapeutic promise. Ten animals per group, exhibiting consistent weight changes 13 days post-OA surgery, were selected for analysis. At the study's conclusion, no significant differences were observed in body weight and weight gain among the test substances, including the highest NEM concentration tested (NEM100), indicating the absence of adverse effects at therapeutic doses. The metabolism of arachidonic acid is pivotal in maintaining bodily homeostasis and orchestrating inflammatory responses. This metabolic pathway is responsible for the synthesis of prostaglandamins, which serve as chemical mediators of inflammation, and leukotrienes, which modulate immune responses, including those linked to inflammation, asthma, and allergies [34]. The enzyme 5-LPO aids in leukotriene biosynthesis [35], while PGE\({}_{2}\) is produced by the enzyme COX-2, in coordination with IL-1 and TNF-\(\alpha\). PGE\({}_{2}\) synthesis is implicated in the etiopathogenesis of OA, contributing to AC degradation [36; 37]. Additionally, PGE\({}_{2}\) is implicated in abnormal bone growth and osteophyte formation, hallmark features of OA, leading to joint pain and discomfort [28; 38]. These inflammatory processes induce edema in surrounding tissues, significantly increasing joint thickness--a symptom commonly observed in OA patients. Such insights underscore the complex interplay between arachidonic acid metabolism and OA development, highlighting potential therapeutic targets for mitigating disease progression and associated symptoms [28; 31; 38; 39]. In some instances, NSAIDs may adversely affect cartilage health by hindering the synthesis of PGs, which are essential for maintaining cartilage integrity and function [19]. Considering this, it is important to discuss the implications of NEM treatment on either increasing or inhibiting PG synthesis. To better understand the impact of NEM on PG synthesis and cartilage health, future studies should investigate the long-term effects of NEM treatment on cartilage function and structure. Additionally, exploring the specific pathways through which NEM modulates PG synthesis will provide deeper insights into its mechanisms of action.