## 3 Results and Discussions
### Cell Culture and Treatment  
#### 3.4.1 Cell Viability and Proliferation Assay (MTS) of Samples  
Determination of cell viability represents a key stage in the formation of cell cultures [72]. A high cell viability and a degree of confluence over 80% allow the application of the scratch method in favorable conditions [72]. The human dermal fibroblasts viability (%) according to the samples used based on trypan blue exclusion test is shown in Table 4.  
The dye exclusion test is used to determine the number of viable cells. The principle of test is that live cells possess intact cell membranes that exclude trypan blue dye whereas dead cells do not [73]. Based on our results, all the samples shown no significant difference compared with CTRL. Cellular proliferation was performed by the MTS test, using different doses of mulberry extract (between 25-200 \(\upmu\)g/mL), as well as liposome samples with or without extract (MnL and L respectively). \begin{table}
\begin{tabular}{c c c c} \hline
**Liposomal Formula \({}^{*}\)** & **Size up to 500 nm (\%)** & **Zeta Potential (mV)** & **EE\%** \\ \hline L & 87.12 & \(-\)2.80 & - \\ \hline MnL & 78.68 & \(-\)2.5 & 88.25 \\ \hline \end{tabular} \({}^{*}\)L—empty liposomes, MnL—mulberry-loaded liposomes.  
\end{table} 
Table 3: Characterisation of liposomes. The samples tested did not exhibit proliferation inhibition on NHDF cells. Significant differences were obtained between all samples compared to the control. It should be noted that a significant difference was obtained in the case of the MnL sample whose cell viability was increased by 54.15% compared to CTRL. No statistical significances were obtained between the mulberry extracts, regardless of the concentration used (Figure 3). The authors [PERSON] et al., 2022 [74] shown that the high content of compounds such as anthocyanins, respectively antioxidant compounds and vitamins, including vitamin C, have beneficial effects on the stimulation and proliferation of dermal fibroblasts. [PERSON] et al., 2021 [75] also confirm the beneficial and non-toxic effects of mulberry extracts on the process of growth, development and proliferation of fibroblasts and other dermal and hair follicle cells, demonstrating the beneficial effects on hair growth and thus combating alopecia [75]. Also, liposomal formulas composed of phospholipids and cholesterol are highly biocompatible with the human body, results that are confirmed by data from the literature [20, 62, 76]. \begin{table}
\begin{tabular}{c c} \hline \hline
**Samples** & **\% of Cell Viability** \\ \hline CTRL & \(95.25\pm 7.31\) \\ ALA\_50 & \(94.77\pm 5.51\) \\ MnE\_25 & \(98.55\pm 5.90\) \\ MnE\_50 & \(97.21\pm 7.11\) \\ MnE\_100 & \(95.48\pm 8.24\) \\ MnE\_200 & \(96.78\pm 7.48\) \\ L & \(95.15\pm 6.20\) \\ MnL & \(95.30\pm 6.50\) \\ \hline CTRL—untreated cells; ALA\_50—alanine 50 ug/mL; L—empty liposomes; MnE\_25—mulberry extract 25 ug/mL; MnE\_50—mulberry extract 50 ug/mL; MnE\_100—mulberry extract 100 ug/mL; MnE\_200—mulberry extract 200 ug/mL; MnL— liposomes with mulberry extract. \\ \end{tabular}
\end{table} 
Table 4: The effect of human dermal fibroblasts viability (%) depending on the samples used. Figure 3: The effect of mulberry extract concentrations (25, 50, 100 and 200 ug/mL) and liposomes on cell viability of NHDF after 24 h using MTS assay. CTRL—untreated cells; L—empty liposomes; ALA\_50—alanine 50 ug/mL; MnE\_25—mulberry extract 25 ug/mL; MnE\_50—mulberry extract 50 ug/mL; MnE\_100—mulberry extract 100 ug/mL; MnE\_200—mulberry extract 200 ug/mL; MnL—liposomes with mulberry extract. Different lowercase letters indicate significant differences between samples (\(p<0.05\)). 4.2 In Vitro Testing of the Healing Effect of Mulberry Extracts and Its Liposome Using the Scratch Method  
The scratch method an in vitro technique is widely used to evaluate the wound healing capacity of different plant extracts [19; 40].