*4.5. Statistical Analysis*

All experimental data were expressed as mean ± standard deviation (mean ± SD). Data analysis was performed using Excel and GraphPad Prism software version 6.0 (Graph-Pad Software, La Jolla, CA, USA). The statistical analyses were T-test and one-way or

two-way ANOVA with the component of Dunnett's multiple comparisons test. A *p*-value less than 0.05 was considered a statistically significant difference (SSD) between the experimental and control groups.

#### **5. Conclusions**

To improve the low oral bioavailability of catechins, EGCG was loaded in liposomes in this study. It is found that PS-containing liposomes were smaller and more stable. It has higher encapsulation efficiency, and the addition of vitamin E can protect EGCG from oxidation and improve encapsulation efficiency. In the in vitro study, expressions of TNF-α and NO production from BV-2 cells were all reduced after pretreatment of EGCG-loaded liposomes on LPS-induced BV-2 cells. Therefore, the EGCG-loaded liposomes have played an essential role in the neuroinflammatory response as an inhibitor. The beneficial effects have been to prevent cells from apoptosis in the neuroinflammatory reactions.

In in vivo study, the Parkinsonian syndrome in rats induced by LPS to the unilateral midbrain substantia nigra region has been improved. The neuro-inflammation mechanism, including the TNF-α secretion, has been inhibited by post-treatment of EGCG-loaded liposomes. We demonstrated that EGCG would be a more valuable candidate for treating neurodegenerative diseases by alleviating the inflammation in the brain. The subsequent investigation should focus on the inflammatory pathways by using a lysosome tracker to track the liposome entrance into the cell and the release of EGCG from the liposome.

**Supplementary Materials:** The following are available online at https://www.mdpi.com/1422-006 7/22/6/3037/s1.

**Author Contributions:** C.-Y.C.: Investigation; Methodology; Validation. L.B.: Validation; Data curation; Formal analysis; Writing—review and editing. S.-T.T.: Data curation; Visualization; Roles/Writing—original draft; Writing—review and editing. T.-W.F.: Investigation; Methodology; Validation. X.-Y.W.: Data curation; Formal analysis; Investigation; Methodology; Software; Validation; Visualization. C.-W.C.: Data curation; Formal analysis; Investigation; Methodology; Software; Validation; Visualization. R.-S.Y.: Investigation; Resources. T.-Y.C.: Funding acquisition; Methodology; Project administration; Resources; Writing—review and editing. M.F.H.: Conceptualization; Funding acquisition; Methodology; Project administration; Resources; Supervision; Roles/Writing—original draft; Writing—review and editing. All authors have read and agreed to the published version of the manuscript.

**Funding:** This work was funded by the Ministry of Science and Technology of Taiwan, grant number MOST-108-2119-M-033-002 and the Changhua Christian Hospital, grant number 107CYCU-CCH. The APC was funded by MOST-109-2124-M-033-001.

**Data Availability Statement:** The data presented in this study are available in this article *Int. J. Mol. Sci.* **2021**, *22*, 3037 or Supplementary Materials here.

**Conflicts of Interest:** The authors declare no competing interests. The funding bodies played no role in the design of the study, collection, analysis, and interpretation of the data, or writing of the manuscript.
