## 3 Results
### Effects of NEM on Inflammation, Apoptosis, and Cell Proliferation In Vivo  
Figures 9 and 10 show the immunostained images of the inflammatory markers TNF\(\alpha\) and COX-2, the apoptosis marker PARP, and the cell proliferation marker BrdU in the femoral and tibial AC and SM. In the OA group, the number of immunosuppressive cells for TNF\(\alpha\), COX-2, and PARP was significantly increased in the femoral and tibial AC and SM. However, this increment was attenuated in the DS, NEM100, and NEM50 groups. Figure 6: Gene expression levels of Col-2, aggrecan, and Sox9 in the femoral and tibial articular cartilage (AC) and the synovial membrane (SM). We assessed the gene expression related to cartilage composition from the femoral and tibial articular cartilage (AC) and synovial membrane (SM). Total RNA was isolated and the mRNA levels were measured using real-time PCR. Gene expression levels were quantified relative to a vehicle control and normalized to \(\beta\)-actin mRNA. Values are expressed as means \(\pm\) SD in 10 rats. a \(p<0.01\) as compared with sham by DT3; b \(p<0.01\) as compared with OA by DT3. \begin{table}
\begin{tabular}{l c c c c c} \hline \hline \multicolumn{1}{c}{**Items**} & \multicolumn{4}{c}{**Rankin Scores**} \\ \cline{2-6}
**Groups** & **Surface** & **Cellularity** & **Clone** & **Stain intensity** & **Total \({}^{*}\)** \\ \hline
**Controls** & & & & & \\ \hline Sham & 0.20 \(\pm\) 0.42 & 0.40 \(\pm\) 0.52 & 0.00 \(\pm\) 0.00 & 0.50 \(\pm\) 0.52 & 1.10 \(\pm\) 1.10 \\ OA & 2.80 \(\pm\) 0.42\({}^{\text{a}}\) & 2.70 \(\pm\) 0.48\({}^{\text{c}}\) & 2.80 \(\pm\) 0.42\({}^{\text{a}}\) & 11.00 \(\pm\) 0.94\({}^{\text{a}}\) \\ Diclofenac sodium & & & & \\ \hline
2 mg/kg & 1.60 \(\pm\) 0.52\({}^{\text{ab}}\) & 1.50 \(\pm\) 0.53\({}^{\text{cd}}\) & 1.70 \(\pm\) 0.48\({}^{\text{cd}}\) & 1.80 \(\pm\) 0.42\({}^{\text{ab}}\) & 6.60 \(\pm\) 1.07\({}^{\text{ab}}\) \\ NEM & & & & \\ \hline
100 mg/kg & 1.30 \(\pm\) 0.48\({}^{\text{ab}}\) & 1.00 \(\pm\) 0.00\({}^{\text{d}}\) & 1.10 \(\pm\) 0.32\({}^{\text{cd}}\) & 1.00 \(\pm\) 0.67\({}^{\text{b}}\) & 4.40 \(\pm\) 0.84\({}^{\text{ab}}\) \\
50 mg/kg & 1.50 \(\pm\) 0.53\({}^{\text{ab}}\) & 1.60 \(\pm\) 0.52\({}^{\text{cd}}\) & 1.70 \(\pm\) 0.48\({}^{\text{cd}}\) & 1.70 \(\pm\) 0.48\({}^{\text{ab}}\) & 6.50 \(\pm\) 0.97\({}^{\text{ab}}\) \\
25 mg/kg & 1.90 \(\pm\) 0.74\({}^{\text{ab}}\) & 2.10 \(\pm\) 0.57\({}^{\text{c}}\) & 2.10 \(\pm\) 0.57\({}^{\text{c}}\) & 1.90 \(\pm\) 0.74\({}^{\text{ab}}\) & 8.00 \(\pm\) 1.25\({}^{\text{ab}}\) \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Femoral AC Mankin scores in sham-operated or OA rats. \begin{table}
\begin{tabular}{l c c c c c} \hline \hline \multicolumn{1}{c}{**Items**} & \multicolumn{4}{c}{**Rankin Scores**} \\ \cline{2-6}
**Groups** & **Surface** & **Cellularity** & **Clone** & **Stain intensity** & **Total \({}^{*}\)** \\ \hline Controls & & & & \\ \hline Sham & 0.30 \(\pm\) 0.48 & 0.40 \(\pm\) 0.52 & 0.00 \(\pm\) 0.00 & 0.50 \(\pm\) 0.53 & 1.20 \(\pm\) 1.03 \\ OA & 2.60 \(\pm\) 0.52\({}^{\text{a}}\) & 2.60 \(\pm\) 0.52\({}^{\text{a}}\) & 2.80 \(\pm\) 0.42\({}^{\text{a}}\) & 2.70 \(\pm\) 0.48\({}^{\text{a}}\) & 10.70 \(\pm\) 0.82\({}^{\text{a}}\) \\ \hline \hline \end{tabular}
\end{table} 
Table 3: Tibial AC Mankin scores in sham-operated or OA rats. Figure 7: Histopathological analysis in femoral and tibial articular cartilage (AC) and synovial membrane (SM). (**A**) Representative histological images of the femoral and tibial AC with SM stained with hematoxylin and eosin (H&E). Black and white arrows indicate the clone formation in the articular surfaces and SM-lining epithelium, respectively. GP = Growth plate; MN = Meniscus; CL = Crucial ligament. Scale bars = 160 \(\mu\)m. (**B**) Thickness of both AC and the SM-lining epithelium and the number of infiltrated inflammatory (IF) cells in the SM. Values are expressed as means \(\pm\) SD in 10 rats.