## 2 Results
### Cell Viability Assay
#### 2.4.2 ANOVA Statistics  
Based on the image analysis, the following parameters were taken into account, which were statistically processed: relative wound width, wound area evolution, and normalized  
Figure 4: The evolution over time (0, 12, 24, 36, and 48 h) of cell wound coverage at different concentrations of _Stellaria media_ (L) Vill extract (range 200 \(\mathrm{\SIUnitSymbolMicro g}\)/mL–12.5 \(\mathrm{\SIUnitSymbolMicro g}\)/mL) compared to positive control (CTRL)—treated with allantoin 50 \(\mathrm{\SIUnitSymbolMicro g}\)/mL and untreated wound control (CTRL0). Fibroblasts marked in pink are those that have mobilized into the wound. The scale of the processed images is 100 \(\mathrm{\SIUnitSymbolMicro m}\).  
cell density inside the wounds, depending on the time and the concentration of the SM extract (Supplementary Materials Tables S1-S3).  
The values of relative wound width and area depended on the concentrations and the time of exposure to the SM extract. After 12 h, cell numbers in the wounded width were significantly different compared to the control, while at 48 h, no difference was recorded between samples (Tables S1 and S2).  
The density of NHDF is a defining parameter in the wound-healing process, being the one that highlights the most effective treatment [18; 19; 20]. Figure 5 shows the cell density inside the wound depending on both the treatment applied and the time.  
The normalized cell density factor is the one that best differentiates the effects of in vitro treatments. Figure 5 and Table S3 show the evolution of the cell density (%) compared with the control (untreated samples) for each treatment applied. Only the SM extract with a concentration of 100 \(\upmu\)g/mL shows a significant percentage increase (\(p=0.05\)) in cell density compared to the initial density (after 24 h). On the other hand, in the case of the SM extract of the highest concentration (200 \(\upmu\)g/mL) and 50 \(\upmu\)g/mL, an increase in cell density compared to the initial one is only observed after 36 h.  
Figure 5: The evolution of the cell density parameter inside the wound depending on the applied treatment and time. The horizontal red line represents the initial density of the cells before the application of the “scratch” test, and the dotted red line represents the evolution in time of the cell density (%) depending on the applied treatment. _Stellaria media_ (L.) Vill extract was applied in concentrations of: 200 \(\upmu\)g/mL (**A**), 100 \(\upmu\)g/mL (**B**), 50 \(\upmu\)g/mL (**C**), 25 \(\upmu\)g/mL (**D**), 12.5 \(\upmu\)g/mL (**E**), Positive Control (CTRL), 50 \(\upmu\)g/mL Allantoin (**F**), and control (CTRL0) (**G**), without any treatment.  
The value of the LP norm--a quantitative parameter that allows the classification of samples taking into account the three normalized variables: coverage of the relative width of the wound (%), coverage of the wound area (%), and the normalized density of the cells inside the wound (%) for the removal of errors that may occur in univariate statistical analysis--was calculated with the data presented in Figure 6.  
Thus, all the results obtained by image and statistical analyses were represented in Figure 6 to highlight the most effective and fastest treatment that led to wound healing.