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Mar. Drugs 2020, 18, 217 3 of 11 Mar. Drugs 2019, 17, x FOR PEER REVIEW 3 of 11 Uncoated nanoliposomes showed negative ζ-potential values, which became positive after Uncoated nanoliposomes showed negative ζ-potential values, which became positive after coating with chitosan. The ζ-potential increased from -43.3, -40.9 and -43.5 mV for soya, salmon and coating with chitosan. The ζ-potential increased from -43.3, -40.9 and -43.5 mV for soya, salmon and rapeseed liposomes, respectively, to 60.9, 66.2 and 66.8 mV after coating with chitosan, respectively. rapeseed liposomes, respectively, to 60.9, 66.2 and 66.8 mV after coating with chitosan, respectively. The ζ-potential of uncoated nanoliposomes was negative, probably due to the anionic fractions of The ζ-potential of uncoated nanoliposomes was negative, probably due to the anionic fractions of lecithin [20]. The increase in the surface charge of chitosan-coated nanoliposomes was attributed to the lecithin [20]. The increase in the surface charge of chitosan-coated nanoliposomes was attributed to increase in the positively-charged amino groups of chitosan molecules, proving that the nanoliposome the increase in the positively-charged amino groups of chitosan molecules, proving that the coating was successfully achieved [39]. Regarding the stability of the formulations, no significant nanoliposome coating was successfully achieved [39]. Regarding the stability of the formulations, no variation in particle size or charge was observed during storage periods of 30 days at 4 ◦C and 37 ◦C, significant variation in particle size or charge was observed during storage periods of 30 days at 4 °C which suggests that the liposomes can be stored without lyophilization for a minimum of 1 month and 37 °C, which suggests that the liposomes can be stored without lyophilization for a minimum of without showing any changes in their properties. 1 month without showing any changes in their properties. Figure 1. Schematic and physicochemical characterization of salmon, rapeseed and soya Figure 1. Schematic and physicochemical characterization of salmon, rapeseed and soya nanoliposomes, nanoliposomes, curcumin-loaded nanoliposomes and curcumin-loaded nanoliposomes coated curcumin-loaded nanoliposomes and curcumin-loaded nanoliposomes coated with chitosan. with chitosan. 2.2. Encapsulation Efficiency of Curcumin 2.2. Encapsulation Efficiency of Curcumin The encapsulation efficiency of curcumin was 87.15, 88.61 and 88.72% in rapeseed, soya and The encapsulation efficiency of curcumin was 87.15, 88.61 and 88.72% in rapeseed, soya and salmon chitosan-coated nanoliposomes, respectively. Compared to our previous study [23], these salmon chitosan-coated nanoliposomes, respectively. Compared to our previous study [23], these results indicate that the encapsulation efficiency of curcumin significantly increases when liposomes results indicate that the encapsulation efficiency of curcumin significantly increases when liposomes are coated with chitosan. are coated with chitosan. 2.3. Membrane Fluidity 2.3. Membrane Fluidity The fluidity of the liposomes reflects the order and dynamics of the phospholipids’ alkyl chains in The fluidity of the liposomes reflects the order and dynamics of the phospholipids’ alkyl chains the vesicle’s bilayer. The fatty acid (FA) composition tunes the membrane’s fluidity level. Membrane in the vesicle’s bilayer. The fatty acid (FA) composition tunes the membrane’s fluidity level. fluidity decreases when saturated FAs are present, due to an increase in the packing between the Membrane fluidity decreases when saturated FAs are present, due to an increase in the packing phospholipids, whilst unsaturated FAs increase membrane fluidity by reducing the packing between between the phospholipids, whilst unsaturated FAs increase membrane fluidity by reducing the the phospholipids [40]. packing between the phospholipids [40]. To recognize the action of curcumin and chitosan on membrane fluidity, it is necessary to To recognize the action of curcumin and chitosan on membrane fluidity, it is necessary to understand the behavior of curcumin and chitosan with respect to solution composition variation. understand the behavior of curcumin and chitosan with respect to solution composition variation. According to our previous study [23], membrane fluidity depends on the lipid composition of According to our previous study [23], membrane fluidity depends on the lipid composition of nanoliposomes, as a lower membrane fluidity was found for rapeseed and salmon liposomes tocsoomypaalirpedostomsoesy.aTlihpiosscoamnebse. TexhpislacianedbebeyxtphleaihniegdhberypthroephoigrthioernporfoPpUorFtAiosnwofitPhUshFoArst wchiathinsshforut nd nanoliposomes, as a lower membrane fluidity was found for rapeseed and salmon liposomes compared incshoayinaslifpoousnodminesocyoamlpipaorseodmtoesrcaopmespeaerdedantodrsaaplemseoendliapnodsosamlmeso.n liposomes. AAs sshsohwownnininoouurrpprreeviious results [23],, currccumininddecerceraesaesdedthtehemmemebmrabnraenfelufliduiitdyitoyf oalfl all nanaonloipliopsoosmomese,sa,sasitistspprreesseenncceeccanweakenthehydrrophoobbicicininteteraractcitoinosnsamamonogngacayclyclhcahinasin.s.PDF Image | Growth-Inhibitory Effect of Chitosan-Coated Liposomes Encapsulating Curcumin
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