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Molecules 2018, 23, 1436 6 of 13 2.4. Photodynamic Activity Against Cells In Vitro Molecules 2018, 23, x FOR PEER REVIEW 6 of 13 Finally, in vitro photodynamic activities of PS1 to PS5 against two cell lines, including murine (PDT) with 25.8 J/cm2 of light (λ 660 nm) or not (dark) by MTT bioassay [33]. As shown in Figure 4, breast adenocarcinoma cell 4T1 and normal murine fibroblast cell NIH-3T3, were tested by irradiated all the tested compou2 nds did not display significant toxicity towards both NIH-3T3 and 4T1 cells in (PDT)with25.8J/cm oflight(λ660nm)ornot(dark)byMTTbioassay[33].AsshowninFigure4, the dark with inhibition ratio less than 50% within the concentration range of 2.5 to 40 μM. The all the tested compounds did not display significant toxicity towards both NIH-3T3 and 4T1 cells in the absence of toxicity in the dark is a requirement for a desirable PS, as it avoids that non-irradiated dark with inhibition ratio less than 50% within the concentration range of 2.5 to 40 μM. The absence of tissues become affected during a PDT protocol. Regarding the photodynamic activity, it is remarkable toxicity in the dark is a requirement for a desirable PS, as it avoids that non-irradiated tissues become that both PS1 and PS4 were robustly phototoxic against 4T1 cancer cell in a concentration-dependent affected during a PDT protocol. Regarding the photodynamic activity, it is remarkable that both manner (p < 0.05). Compound PS3 presented no evident photodynamic activity against 4T1 cells. PS1 and PS4 were robustly phototoxic against 4T1 cancer cell in a concentration-dependent manner Interesting results were also obtained with PS5, which showed a significant phototoxicity against 4T1 (p < 0.05). Compound PS3 presented no evident photodynamic activity against 4T1 cells. Interesting cells but no activity against NIH-3T3 in PDT experiment, indicating that this compound might have results were also obtained with PS5, which showed a significant phototoxicity against 4T1 cells but a better selectivity towards the cancerous 4T1 cells tested in this study. However, further no activity against NIH-3T3 in PDT experiment, indicating that this compound might have a better investigation on the selectivity of PS5 towards cancerous cells and the mechanism of action is selectivity towards the cancerous 4T1 cells tested in this study. However, further investigation on the Figure 4. Toxicity of the compounds PS1 to PS5 at different concentrations against 4T1 and NIH-3T3 Figure 4. Toxicity of the compounds PS1 to PS5 at different concentrations against 4T1 and NIH-3T3 cells treated in the dark or irradiated with red light (λ 660 nm, 25.8 J/cm2 2). cells treated in the dark or irradiated with red light (λ 660 nm, 25.8 J/cm ). warranted. selectivity of PS5 towards cancerous cells and the mechanism of action is warranted. 3. Materials and Methods 3. Materials and Methods 3.1. General Information 3.1. General Information Dimethyl sulfoxide (DMSO), 1,3-diphenylisobenzofuran (DPBF) and Kholipho®r® HS were Dimethyl sulfoxide (DMSO), 1,3-diphenylisobenzofuran (DPBF) and Kholiphor HS were obtained from Sigma (St. Louis, Missouri, USA). Roswell Park Memorial Institute (RPMI) medium obtained from Sigma (St. Louis, Missouri, USA). Roswell Park Memorial Institute (RPMI) medium and Dulbecco’s modified Eagle medium (DMEM) were obtained from Gibco (Waltham, MA, USA). and Dulbecco’s modified Eagle medium (DMEM) were obtained from Gibco (Waltham, MA, The 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from USA). The 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Invitrogen (Carlsbad, CA, USA). The phosphate buffered saline (PBS) was supplied by Laborclin (Rio Invitrogen (Carlsbad, CA, USA). The phosphate buffered saline (PBS) was supplied by Laborclin de Janeiro, Brazil). Commercially available reagents were used without further purification. Organic (Rio de Janeiro, Brazil). Commercially available reagents were used without further purification. solvents were evaporated with reduced pressure using Büchi evaporators. Reactions were monitored by TLC using Yantai JingYou (Yantai, China) GF254 silica gel plates. Silica gel column chromatography was performed on an Isolera One system (Biotage, Uppsala, Sweden), and silica gel (200–300 mesh) from Qingdao Hailang Inc. (Qingdao, China). NMR spectra were measured on an Avance III 600 MHz spectrometer (Bruker, Fällanden, Switzerland). Chemical shifts were expressedPDF Image | Photosensitizers for Anticancer Photodynamic Therapy
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