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Molecules 2019, 24, x Molecules 2019, 24, 1388 9 of 14 Molecules 2019, 24, x 9 of 14 2.5. In Vitro Cellular Uptake 2.5. In Vitro Cellular Uptake The cellular uptake of curcumin and Cur-CS/Alg NPs was investigated in HaCaT cells. The cellular uptake of curcumin and Cur-CS/Alg NPs was investigated in HaCaT cells. The cellular uptake of curcumin and Cur-CS/Alg NPs was investigated in HaCaT cells. Fluorescence microscopy images of each treatment are shown in Figure 4. Curcumin and Cur-CS/Alg Fluorescence microscopy images of each treatment are shown in Figure 4. Curcumin and Cur-CS/Alg Fluorescence microscopy images of each treatment are shown in Figure 4. Curcumin and Cur-CS/Alg 9 of 14 Figure 3. Physical stability of Cur-CS/Alg NPs in cell culture medium (pH 7.4) (n = 3). Figure 3. Physiicallssttaabbiliiltiytyoof fCCuur-rC-CS/SA/AlglgNPNsPisnicnelcleclul lctulrteumreemdieudmiu(mpH(p7H.4)7(.n4)=(3n).= 3). 2.5. In Vitro Cellular Uptake NPs emitted fluorescent light under a fluorescence microscope, whereas the control cells treated with NPs emitted fluorescent light under a fluorescence microscope, whereas the control cells treated with NPs emitted fluorescent light under a fluorescence microscope, whereas the control cells treated with DMSO or empty nanoparticles had no fluorescent signal. Interestingly, the Cur-CS/Alg NPs-treated DMSO or empty nanoparticles had no fluorescent signal. Interestingly, the Cur-CS/Alg NPs-treated DMSO or empty nanoparticles had no fluorescent signal. Interestingly, the Cur-CS/Alg NPs-treated groups had higher fluorescent intensity than curcumin at the equivalent concentration of curcumin. groups had higher fluorescent intensity than curcumin at the equivalent concentration of curcumin. groups had higher fluorescent intensity than curcumin at the equivalent concentration of curcumin. These results indicated that the Cur-CS/Alg NPs improved the cellular uptake of curcumin. These results indicated that the Cur-CS/Alg NPs improved the cellular uptake of curcumin. These results indicated that the Cur-CS/Alg NPs improved the cellular uptake of curcumin. Figure4.CelllulalarruupptatakkeeooffcucurcrucmumininanadndCuCru-Cr-SC/AS/lgANlgPNsvPisewviedwbeydfbluyoflreusocerenscceemncicermosiccorpoyscionpyin HaCaT celllss.. Figure 4. Cellular uptake of curcumin and Cur-CS/Alg NPs viewed by fluorescence microscopy in HaCaT cells. 3. Materials and Methods 3. Materials and Methods 3. Materials and Methods 3.1. Materials 3.1. Materials CS, with a molecular weight of 22,000 Da and degree of deacetylation of 90%, was donated by 3.1. MCaSte,rwiailtsh a molecular weight of 22,000 Da and degree of deacetylation of 90%, was donated by Marine Bio Resources Co., Ltd., Samut Sakorn, Thailand. Sodium alginate with a medium viscosity Marine Bio Resources Co., Ltd., Samut Sakorn, Thailand. Sodium alginate with a medium viscosity CS, with a molecular weight of 22,000 Da and degree of deacetylation of 90%, was donated by waspurchasedffromSSigigmaa,,SSttLLouoiusi,sM,MOO,U,SUAS.AC.uCrcuurmcuimnwinawsparsepraerpedaraenddacnhdarcahcaterraicztedrifzreodmfrthoemthe Marine Bio Resources Co., Ltd., Samut Sakorn, Thailand. Sodium alginate with a medium viscosity ®® previously publliissheeddmeeththoodd[1[61]6.]N. Nono-nio-nioicniscusrufarcftacnttanat mnaemd eTdwTeewnee8n0 (o80r P(orlyPsolrybsaoterb8a0t)ew8a0s) was was purchased from Sigma, St Louis, MO, USA. Curcumin was prepared and characterized from the suppliedfromAccrrossOrrggaannicisc,sT,ThheremrmooFiFshisehreSrcSiecnietinfitci,fiMc,oMrroisrrPilsaPinlas,inNsJ,,NUJS,AU.SAll.oAthlleortchermcihceamlsicals previously published method [16]. Non-ionic surfactant named Tween® 80 (or Polysorbate 80) was were of analytical grade. were of analytical grade. supplied from Acros Organics, Thermo Fisher Scientific, Morris Plains, NJ, USA. All other chemicals w3e.2r.ePorefpaanratliyotnicoaflthgeraCduer.-CS/Alg NPs 3.2. Preparation of the Cur-CS/Alg NPs Na-Alg and CaCl2 solutions were dissolved in ultrapure water, while CS was dissolved in 1% Na-Alg and CaCl2 solutions were dissolved in ultrapure water, while CS was dissolved in 1% 3.2. Preparation of the Cur-CS/Alg NPs (v/v) acetic solution. The pH of the Alg and CS solutions were adjusted to 4.9 and 4.6, respectively, (v/v) acetic solution. The pH of the Alg and CS solutions were adjusted to 4.9 and 4.6, respectively, ® usinNg a1-%Al(gv/avn) dacCetaicCal2cisdolauntdion4%s w(wer/ve) dNisasOoHlv,erdesinpeuctlitvraeplyu. rTeweaenter,8w0®whiales CusSedwtaos edmisusolslivfyedthien 1% using 1% (v/v) acetic acid and 4% (w/v) NaOH, respectively. Tween 80 was used to emulsify the (v/v) acetic solution. The pH of the Alg and CS solutions were adjusted to 4.9 and 4.6, respectively, ethanolic curcumin and Alg solution through micelle formation. The Cur-CS/Alg NP formulations using 1% (v/v) acetic acid and 4% (w/v) NaOH, respectively. Tween® 80 was used to emulsify thePDF Image | Synergistic Effects of Photo-Irradiation and Curcumin-Chitosan
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