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Mar. Drugs 2020, 18, 217 4 of 11 Mar. Drugs 2019, 17, x FOR PEER REVIEW 4 of 11 IIntthiissssttudy, ,aasssshhoownnininTaabblele11, ,ththeepprreesseenncceeooffcchhitiotossaannaalslsoorreedduucceedttheemeembrraneflfluiidiitty off nnaanolliipossomes,, as tthiis new llaayeerr aarroound tthee lilpipoossoomee waass pprroobbaabbllyy ininccoorrporraatteed wiitthiin tthee membrranebiillayer,,tthussiinccrreeaassiingttheerriigiidiittyoffttheebbiillaayeerrssaanddeeccrreeaassiingttheemooveementtofftthee FAcchhaaininss. .Consequenttlly,,ttheemeembbrraaneebbiillaayeerrflfluiidiittydeeccrreeaasseedaandttheemottiionallffrreeedomofftthee phoossphhaatteeggrroouuppwaassrreedduucceedd[4[411].]. Taabblele11.. Membranefflluiidiittyooffththeecchhitiotosasann-c-ocoaatetdedlilpiposoosmomeses(e(aecahchvavlauleuererperpersesnetnsttshtehme emaenaonf of triplicates). triplicates). 2.4. Morphology of the Liposomes 2.4. Morphology of the Liposomes Sampllee Soya CEL MembraneFlluuididitiyty 3.21 ± 0.10 Soya CEL 3.21 ± 0.10 2.56± 0..100 SoyaCLLL Salmon CEL 2.70 ± 0.10 2.70 ± 0.10 Salmon CEL 2.62 ± 0.20 Salmon CLL 2.62 ± 0.20 Salmon CLL 3.21 ± 0.10 Rapeseed CEL 3.21 ± 0.10 Rapeseed CEL 2.71 ± 0.10 2.71 ± 0.10 Rapeseed CLL Rapeseed CLL CEL: chitosan-coated empty liposomes; CLL: chitosan-coated curcumin-loaded liposomes. CEL: chitosan-coated empty liposomes; CLL: chitosan-coated curcumin-loaded liposomes. Small unilamellar vesicles (SUV) can be observed in the TEM images of nanoliposomes that were Small unilamellar vesicles (SUV) can be observed in the TEM images of nanoliposomes that were prepared via sonication followed by high-pressure homogenization (Figure 2a). A small quantity (10%) prepared via sonication followed by high-pressure homogenization (Figure 2a). A small quantity of oil droplets in each formulation, in the form of nanoemulsions, can be observed. Figure 2b shows a (10%) of oil droplets in each formulation, in the form of nanoemulsions, can be observed. Figure 2b chitosan contrasting band surrounding the nanoliposomes. The TEM images confirm the size results shows a chitosan contrasting band surrounding the nanoliposomes. The TEM images confirm the measured via the dynamic light scattering (DLS) technique. size results measured via the dynamic light scattering (DLS) technique. Figure 2. Transmission Electron Microscopic (TEM) images of a curcumin-loaded nanoliposome before Figure 2. Transmission Electron Microscopic (TEM) images of a curcumin-loaded nanoliposome (a) and after (b) coating with chitosan. before (a) and after (b) coating with chitosan. 2.5. Growth-Inhibition by Real-time Cell Analysis 2.5. Growth-Inhibition by Real-time Cell Analysis The growth-inhibition of uncoated and chitosan-coated nanoliposomes, that were empty or loaded The growth-inhibition of uncoated and chitosan-coated nanoliposomes, that were empty or with curcumin, was measured via an impedance-based analysis method. Their composition and loaded with curcumin, was measured via an impedance-based analysis method. Their composition concentration effects on MCF-7 cells’ growth was investigated. Curcumin concentrations of 12 μM or and concentration effects on MCF-7 cells’ growth was investigated. Curcumin concentrations of 12 higher showed a significant effect on cell index (CI), whereas the 5 μM curcumin concentration showed μM or higher showed a significant effect on cell index (CI), whereas the 5 μM curcumin concentration no significant difference (Figure 3). Reports on the anti-tumor activity of curcumin were based on two showed no significant difference (Figure 3). Reports on the anti-tumor activity of curcumin were opposite mechanisms: inhibiting anti-apoptosis proteins and activating the pro-apoptosis proteins [42]. based on two opposite mechanisms: inhibiting anti-apoptosis proteins and activating the pro- Some studies suggest that curcumin acts as an antioxidant and prevents reactive oxygen species apoptosis proteins [42]. (ROS) production [43,44]. Other studies suggest that curcumin induces ROS production at high Some studies suggest that curcumin acts as an antioxidant and prevents reactive oxygen species concentrations and quenches ROS production at low concentrations [45,46]. The antioxidant mechanism (ROS) production [43,44]. Other studies suggest that curcumin induces ROS production at high mediates NF-κB-suppressive effects and the pro-oxidant mechanism mediates apoptotic effects [45]. concentrations and quenches ROS production at low concentrations [45,46]. The antioxidant Other studies report that tumor cells show the preferential uptake of curcumin compared to normal mechanism mediates NF-κB-suppressive effects and the pro-oxidant mechanism mediates apoptotic cells and that the toxicity clearly increases with increasing curcumin uptake; therefore, this advantage effects [45]. Other studies report that tumor cells show the preferential uptake of curcumin compared to normal cells and that the toxicity clearly increases with increasing curcumin uptake; therefore, thisPDF Image | Growth-Inhibitory Effect of Chitosan-Coated Liposomes Encapsulating Curcumin
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