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Pharmaceutics 2020, 12, 1102 6 of 17 Pharmaceutics 2020, 12, x FOR PEER REVIEW 6 of 16 μL) of HeLa cells (3 × 104 cells/mL) in 4DMEM (containing 1% FBS, 1% P/S) were seeded in V-shaped Aliquots(100μL)ofHeLacells(3×10 cells/mL)inDMEM(containing1%FBS,1%P/S)wereseeded 96-well plates designed to induce the formation of 3D multicellular spheroids (Shimadzu Scientific in V-shaped 96-well plates designed to induce the formation of 3D multicellular spheroids (Shimadzu Instruments, Kyoto, Japan) and incubated for 36 h. The cell spheroids were treated with Scientific Instruments, Kyoto, Japan) and incubated for 36 h. The cell spheroids were treated with UCNPs@SiO2-NH2@FA/PEG/TCPP (30 μg/mL of TCPP). After a 24-h incubation, the spheroids were UCNPs@SiO2-NH2@FA/PEG/TCPP (30 μg/mL of TCPP). After a 24-h incubation, the spheroids were transferred to an 8-well plate with PBS, and cells were observed using a CLSM. transferred to an 8-well plate with PBS, and cells were observed using a CLSM. 2.11. Data Analysis 2.11. Data Analysis Data are presented as the mean ± standard deviation (SD). The significance of differences was Data are presented as the mean ± standard deviation (SD). The significance of differences was determined by a Student’s t-test. p-values < 0.05 were considered statistically significant. determined by a Student’s t-test. p-values < 0.05 were considered statistically significant. 3. Results and Discussion 3.1. Synthesis and Characterization of Core@shell UCNPs 3.1. Synthesis and Characterization of Core@shell UCNPs The conventional UCNPs doped with the core or shell element of yttrium (Y), ytterbium (Yb), The conventional UCNPs doped with the core or shell element of yttrium (Y), ytterbium (Yb), and neodymium (Nd) have split fluorescence lights of ~545 nm and 660 nm, which is considered to be and neodymium (Nd) have split fluorescence lights of ~545 nm and 660 nm, which is considered to a main reason for a reduced PDT efficiency [37]. To this end, the first specific aim of this study was to be a main reason for a reduced PDT efficiency [37]. To this end, the first specific aim of this study was developaprototypeofUCNPs@SiO thatemitted~660nmredlightandwassurface-modifiedwith to develop a prototype of UCNPs@S2iO2 that emitted ~660 nm red light and was surface-modified folic acid for an efficient PDT treatment. As a first step, we aimed to produce a hexagonal core of with folic acid for an efficient PDT treatment. As a first step, we aimed to produce a hexagonal core UCNPs to achieve optimal fluorescence efficacy values [50]. As shown in Figure 2A, changing the of UCNPs to achieve optimal fluorescence efficacy values [50]. As shown in Figure 2A, changing the molar ratio of OA and ODE resulted in a significant morphological change of core UCNPs: the core molar ratio of OA and ODE resulted in a significant morphological change of core UCNPs: the core UCNPs were spherical at a ratio of 3:7 but had sharpened edges and exhibited an optimal hexagonal UCNPs were spherical at a ratio of 3:7 but had sharpened edges and exhibited an optimal hexagonal shape at a ratio of 4:15. Therefore, this ratio was chosen for the core UCNPs. shape at a ratio of 4:15. Therefore, this ratio was chosen for the core UCNPs. Figure22..(A(A))TETMEMimiamgaegseosfUoCfNUPCsNdPespednedpienngduipnognuthpeornattihoebertawtieoenbeotlweieceancidol(eOicAa)acindd(1O-oAct)adanecden1e- (oOctDadEe)c(3e:n7efo(rOlDefEt,)6(:135:7foforrmliedfdt,le6,:1a5ndfo4r:1m5fiodrdrleig,hatn).d(B4):1T5EfMorimriagghet)s.o(fBc)orTeE@MsheimllaUgCeNsPofsdcoerpe@nsdhinelgl UupCoNnPtshedempoelnadrirnagtioupboetnwtehenmthoelacroraeti(oerbeiutwme)eanntdheshceolrle(l(uetrebtium)aconmdpshoenleln(ltust(e1t:i0u.5mf)ocrolmefpt,o1n:1enfotsr (m1:i0d.d5lfeo,ralnedft,11:1:1.5ffoorrmriigdhdtl)e.,and1:1.5forright). The surface characteristics of UCNPs are of prime importance because lanthanide-doped core-only The surface characteristics of UCNPs are of prime importance because lanthanide-doped core- UCNPs have a relatively weak upconverting efficiency. Among several methods, the utilization of only UCNPs have a relatively weak upconverting efficiency. Among several methods, the utilization a core/shell structure is considered the most effective for enhancing the efficiency [14]. The shells of a core/shell structure is considered the most effective for enhancing the efficiency [14]. The shells of UCNPs were made of lutetium using Lu(CH CO ) to produce UCNPs with a red fluorescence. of UCNPs were made of lutetium using Lu(CH3C3O2)32to3 produce UCNPs with a red fluorescence. At molar ratios (core to shell = Er(CH3CO2)3 to Lu(CH3CO2)3) of 1:0.5, 1:1, and 1:1.5, the morphologies of core@shell UCNPs were observed by TEM. Increasing the lutetium amount led to a size increase ofPDF Image | Red LED Erbium Upconverting Nanoparticles v Cervical Cancer
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