## 3 Results
While both compounds exhibit inhibitory effects on _Acanthamoeba_, chlorogenic acid demonstrates a more stable and prolonged inhibitory action, whereas rosmarinic acid's effect diminishes over time, with higher doses even stimulating growth (Table 4). Both chlorogenic acid and rosmarinic acid (Figure 2D,E), tested individually in Table 4, are also present in the callus extract of _E. planum_ analyzed in Table 5. These comparisons suggest that the presence of chlorogenic and rosmarinic acids alone is insufficient to inhibit _Acanthamoeba_ growth and that other compounds in the _E. planum_ extract may either counteract or mask any weak inhibitory effect, potentially even stimulating growth. The callus extract, which contains both of these acids, caused a pronounced stimulation of \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline
**Extracts** & \multicolumn{2}{c}{**1 st Day**} & \multicolumn{2}{c}{**2 nd Day**} & \multicolumn{2}{c}{**3 rd Day**} \\ \cline{2-7}
**Concentration** & **MN \(\pm\) SD** & **GI [\%]** & **MN \(\pm\) SD** & **GI [\%]** & **MN \(\pm\) SD** & **GI [\%]** \\ \hline control & 1.61 \(\pm\) 1.41 a & 0 & 3.61 \(\pm\) 1.38 a & 0 & 6.00 \(\pm\) 1.33 a & 0 \\
0.01 mg/mL & 2.44 \(\pm\) 1.44 a & 6.52 & 2.75 \(\pm\) 1.19 b & 23.83 & 4.00 \(\pm\) 2.47 b & 43.33 \\
0.1 mg/mL & 2.42 \(\pm\) 1.54 a & 7.28 & 2.61 \(\pm\) 0.95 b & 27.71 & 3.76 \(\pm\) 1.70 b & 37.34 \\
0.2 mg/mL & 2.17 \(\pm\) 1.57 a & 16.86 & 2.17 \(\pm\) 0.96 b & 39.89 & 2.95 \(\pm\) 0.91 b & 50.84 \\ \hline \hline \end{tabular}
\end{table} 
Table 8: Effect of acteoside on inhibition of _Acanthamoeba_ trophozoites during three days of treatment. trophozoite growth, especially at higher doses. The fact that both the individual compounds (Table 4) and the complex extract containing them (Table 5) show trophozoite proliferation suggests that rosmarinic and chlorogenic acids may contribute to, or synergize with, other compounds in the extract to enhance growth rather than inhibit it. The effect of _Eryngium planum_ callus culture extract on _Acanthamoeba_ growth is positive for all the tested concentrations (Table 4). The stimulation of _Acanthamoeba_ growth is dose- and time-dependent, increasing with both concentration and duration of treatment, reaching 59.35% stimulation at 10 mg/mL on day 3. Given its growth-promoting effects rather than inhibition, _E. planum_ extract does not appear to be a viable candidate for combating _Acanthamoeba_ infections. Further studies would be needed to understand the underlying mechanism of this stimulation and assess its potential implications. Table 6 evaluates how two hydroxycinamic acids--ferulic acid and _p_-coumaric acid (Figure 2B,C)--influence the growth of _Acanthamoeba_ trophozoites over a 3-day period (Table 6). The results demonstrate that both compounds exhibit moderate to strong inhibitory effects, which are dose-dependent and generally increase over time. Ferulic acid displays moderate inhibition, with effectiveness improving with dose and time. Interestingly, the strongest inhibition for _p_-coumaric acid occurs early (day 1) and slightly decreases over time, although it remains significant. _p_-Coumaric acid is more potent than ferulic acid, particularly at lower doses, suggesting rapid initial action but with less cumulative effect over three days. The extract from _Lychnis flos-cuculi_ callus culture shows strong growth inhibition (Table 7). Growth inhibition increases over time for all concentrations. The highest inhibition (87.52%) occurs on day 3 at 10 mg/mL, suggesting a cumulative effect of the extract. Even at lower concentrations (1 mg/mL, 5 mg/mL), inhibition trends upward each day, indicating sustained effectiveness. This extract appears to be a promising candidate for anti-_Acanthamoeba_ applications, as it effectively suppresses growth with increasing efficacy over time. Both the individual compounds (Table 6) and the whole callus extract (Table 7) of _L. flos-cuculi_ demonstrate significant inhibition of _Acanthamoeba_ trophozoite growth over a 3-day period.