## 4 Discussion
Supporting the results of quantitative uptake and MTS assay, confocal images displayed an increase in proliferation of cells as seen with R123 labeled liposomes (Figure 5A,B), thus confirming the elimination of cytotoxicity of EFA at a concentration of 400 uM and a further improvement in the presence of GSH (100 uM). Finally, flow cytometry data reveal that fluorescence intensity was higher when the cells were treated with pure EFA. Loading of EFA into liposomes was effective in reducing the intracellular accumulation of ROS with a synergistic effect by additional GSH (2.5 fold). This finding suggests that the presence of glutathione moieties within liposomes could lead to an increase in the cellular uptake of liposomes. The results of measurement of the reduction in oxidative stress demonstrated that the anti-oxidative effect of GSH was not impaired by the liposome carrier system and its encapsulation along with EFA within the lipid layers. Furthermore, EFA loaded into liposomes along with GSH exhibited generally a small but significant increased activity in comparison with the same concentration of free forms. Thusencapsulation of glutathione and efavirenz in liposomes apparently facilitate intracellular delivery and prolong the retention time of entrapped agents inside the cell. Nonetheless, co-administration of GSH favoured the reduction of ROS fluorescence when delivered via liposomes form to a greater extent than in the pure form of EFA + GSH (Figures 4 and 5). Studies using three different humanized mouse models demonstrated provided important insights confirming the roles of macrophages, and other myeloid cells can sustain HIV replication and produce HIV in vivo in the absence of CD4 T cells [58; 59]. These study findings indicate that macrophages can serve as a genuine target for HIV infection in vivo by sustaining and transmitting HIV infection [59]. [PERSON] et al. also determined the specific roles of macrophages in HIV persistence during long-term suppressive ART [60]. Their study findings reveal that ART induced the suppression of HIV infection in tissue macrophages, as evident from a significant decrease in the plasma viral load and a reduction in the levels of cell-associated virus. There was no viral rebound in the plasma of the majority of the ART-treated animals' several weeks after ART interruption. However, in a small subset of animals, a delayed viral rebound was observed that is consistent with the establishment of persistent infection in tissue macrophages [60]. These observations further signify the persistence of HIV in tissue macrophages in vivo. The data presented in this manuscript are findings from our initial studies exclusively using differentiated human macrophage cell lines. While the uptake of formulations, toxicity, oxidative stress, and GSH levels in macrophages were thoroughly investigated, our compelling results have propelled us to continue future efficacy studies in HIV-infected cells such as monocytes, dendritic cells and CD4 T cells. Both in vitro studies using isolated CD4 T cells, dendritic cells and monocytes from the peripheral blood and in vivo studies using humanized mice will be performed to further characterize the effects of these formulations against HIV.