## 4 Conclusions  
The conventional treatments for _Staphylococcus_ spp. infections mainly involve the use of antibiotics. Some bacterial strains have become resistant to almost all commonly available antibiotics and some kinds of disinfectants [93]. Chronic infections, particularly those involving biofilms, pose a significant challenge in clinical settings. Nowadays, nanomaterial approaches, able to interfere with bacterial adherence, colonization, and biofilm formation, represent emerging alternatives to address these challenges [94]. The mass spectrometry approach, involving the combined application of ESI FT-ICR-MS and GC-MS, led to a quali-quantitative assessment of the (semi)polar and volatile fractions of LO. Phytochemical profiling has revealed the presence of numerous metabolites recognized for their antimicrobial, antibacterial and antibiofilm activities. In particular, palmitic and oleic acids, along with stearic and linoleic acids, were found to be the most abundant fatty acids in LO. Moreover, a wide range of secondary metabolites have been identified, including flavonoids, which are largely studied for their synergism with antibiotics; monoterpenes, primarily represented by alpha-pinene and beta-myrorene; and sesquiterpenes, mainly depicted by beta-elemene, alpha-humulene and alpha-cubebene. This rich composition in bioactive compounds makes LO a potential antimicrobial/antibiofilm agent useful as an adjuvant in topical formulations against bacterial strains. The use of an oil-based nanocarrier, combined with antibiofilm agents, represents a promising strategy to enhance the efficacy of topical treatments for these persistent infections. In this context, our study using an antibiotic with high susceptibility rates and anti-staphylococcal activity, loaded into NEs, can serve as a starting point for developing new strategies that are currently under investigation. NEsLs have been thoroughly characterized, and it should be emphasized that they exhibited suitable physical-chemical characteristics for the proposed application, and interesting antibiofilm effects against clinical strains at low sub-inhibiting concentrations. Thus, a reduction in the antibiotic dose required for treatment could be achieved, leading to a decrease in side effects. In this context, further studies are required to determine the following: (i) the cytotoxicity of these preparations and (ii) the optimal semisolid topical formulation capable of incorporating loaded nanoemulsions for in vivo skin disease treatment. Furthermore, although the microtiter plate is a versatile and easy-to-use biofilm reactor that exhibits good reproducibility [95], and the crystal violet staining assay is a method extensively applied in biofilm studies, the present results can be influenced by minor experiment variation [96]. Further studies are planned to establish the origin of the high variability observed in our results and to explore the molecular mechanisms involved in the eradication process. However, despite the extensive ongoing research, only a few of the promising preclinical studies have translated into clinical trials [97]. Our preliminary study could represent a starting point for future treatment opportunities against _S. aureus_ infections using the innovative tools of nanomedicine. **Supplementary Materials:** The following supporting information can be downloaded at [[https://www.mdpi.com/article/10.3390/pharmaceutics16070927/s1](https://www.mdpi.com/article/10.3390/pharmaceutics16070927/s1)]([https://www.mdpi.com/article/10.3390/pharmaceutics16070927/s1](https://www.mdpi.com/article/10.3390/pharmaceutics16070927/s1)), Table S1. Untargeted metabolic profiling of Lentisk oil by ESI(+) FT-ICR MS; Table S2. Untargeted metabolic profiling of Lentisk oil by ESI(-) FT-ICR MS; Table S3. Relative abundance of Lentisk seed oil components revealed in static-HS/GC-MS analysis; Table S4. Relative abundance of Lentisk seed oil components revealed in HS-SPME/GC-MS analysis; Figure S1. LVX release profile from NEsL by cellulose dialysis tubing.