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Diamond Nanoparticles Modify Curcumin Activity: In Vitro Studies on Cancer and Normal Cells and In Ovo Studies on Chicken Embryo Model

Strojny, Barbara, Grodzik, Marta, Sawosz, Ewa, Winnicka, Anna, Kurantowicz, Natalia, Jaworski, Sławomir, Kutwin, Marta, Urbańska, Kaja, Hotowy, Anna, Wierzbicki, Mateusz, Chwalibog, André·2016·PLOS ONE

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Fig 1 TEM examination of Cur (A), DN (B) and a bio-complex of Cur:DN in a ratio of 1:3 (C).
Fig 1 TEM examination of Cur (A), DN (B) and a bio-complex of Cur:DN in a ratio of 1:3 (C).CC BY · PubMed Central
Fig 2 HepG2 cell viability measured by mitochondrial activity depletion after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complexes the DN and Cur concentrations in each of the bio-complexes were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.4813 for Cur x DN effect); B–Cur25:DN75 (P < 0.0001 for Cur effect, P = 0.0001 for DN effect, P = 0.3422 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = 0.0043 for DN effect, P = 0.1610 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P = 0.1207 for DN effect, P = 0.0722 for Cur x DN effect). Graphs show mean ± SD, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors.
Fig 2 HepG2 cell viability measured by mitochondrial activity depletion after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complexes the DN and Cur concentrations in each of the bio-complexes were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.4813 for Cur x DN effect); B–Cur25:DN75 (P < 0.0001 for Cur effect, P = 0.0001 for DN effect, P = 0.3422 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = 0.0043 for DN effect, P = 0.1610 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P = 0.1207 for DN effect, P = 0.0722 for Cur x DN effect). Graphs show mean ± SD, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors.CC BY · PubMed Central
Fig 3 Fibroblast cell viability measured by mitochondrial activity depletion after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complexes the DN and Cur concentrations in each of the bio-complex were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.1568 for Cur x DN effect); B–Cur25:DN75 (P < 0.0001 for Cur effect, P < 0.0001 for DN effect, P < 0.0001 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.0615 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.0495 for Cur x DN effect). Graphs show mean ± SD, ** P <0.01, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors and #P <0.05, ###P <0.001 for the Cur x DN interaction.
Fig 3 Fibroblast cell viability measured by mitochondrial activity depletion after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complexes the DN and Cur concentrations in each of the bio-complex were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.1568 for Cur x DN effect); B–Cur25:DN75 (P < 0.0001 for Cur effect, P < 0.0001 for DN effect, P < 0.0001 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.0615 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P = < 0.0001 for DN effect, P = 0.0495 for Cur x DN effect). Graphs show mean ± SD, ** P <0.01, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors and #P <0.05, ###P <0.001 for the Cur x DN interaction.CC BY · PubMed Central
Fig 4 HepG2 membrane integrity measured by LDH leakage after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complexs the DN and Cur concentrations in each of the bio-complexes were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P < 0.0001 for Cur effect, P = 0.0667 for DN effect, P = 0.4338 for Cur x DN effect); B–Cur25:DN75 (P < 0.0001 for Cur effect, P = 0.0004 for DN effect, P = 0.0024 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = 0.1844 for DN effect, P = 0.8649 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P = 0.4403 for DN effect, P = 0.3246 for Cur x DN effect). Graphs show mean ± SD, ** P <0.01, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors and ###P <0.001 for the Cur x DN interaction.
Fig 4 HepG2 membrane integrity measured by LDH leakage after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complexs the DN and Cur concentrations in each of the bio-complexes were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P < 0.0001 for Cur effect, P = 0.0667 for DN effect, P = 0.4338 for Cur x DN effect); B–Cur25:DN75 (P < 0.0001 for Cur effect, P = 0.0004 for DN effect, P = 0.0024 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = 0.1844 for DN effect, P = 0.8649 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P = 0.4403 for DN effect, P = 0.3246 for Cur x DN effect). Graphs show mean ± SD, ** P <0.01, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors and ###P <0.001 for the Cur x DN interaction.CC BY · PubMed Central
Fig 5 Fibroblast membrane integrity measured by LDH leakage after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complex the DN and Cur concentrations in each of the bio-complexes were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P = 0.0336 for Cur effect, P = 0.0171 for DN effect, P = 0.0952 for Cur x DN effect); B–Cur25:DN75 (P = 0.0136 for Cur effect, P = 0.0003 for DN effect, P = 0.7073 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = 0.0001 for DN effect, P = 0.0019 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P < 0.0001 for DN effect, P = 0.0001 for Cur x DN effect). Graphs show mean ± SD, * P <0.05, ** P <0.01, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors and ##P <0.01, ###P <0.001 for the Cur x DN interaction.
Fig 5 Fibroblast membrane integrity measured by LDH leakage after DN, Cur and DN:Cur bio-complexes treatment, separately for each of the used bio-complex the DN and Cur concentrations in each of the bio-complexes were 15, 25, 45 and 75, defining the concentrations in mg/L: A–Cur15:DN45 (P = 0.0336 for Cur effect, P = 0.0171 for DN effect, P = 0.0952 for Cur x DN effect); B–Cur25:DN75 (P = 0.0136 for Cur effect, P = 0.0003 for DN effect, P = 0.7073 for Cur x DN effect); C–Cur45:DN15 (P < 0.0001 for Cur effect, P = 0.0001 for DN effect, P = 0.0019 for Cur x DN effect); D–Cur75:DN25 (P < 0.0001 for Cur effect, P < 0.0001 for DN effect, P = 0.0001 for Cur x DN effect). Graphs show mean ± SD, * P <0.05, ** P <0.01, *** P <0.001, for Cur effect by Bonferroni post-tests following two-way ANOVA with Cur and DN factors and ##P <0.01, ###P <0.001 for the Cur x DN interaction.CC BY · PubMed Central
Fig 6 Dot blot/scatter diagrams of PI/AnnV staining of HepG2 cells treated with curcumin (Cur), diamond nanoparticles (DN) and their bio-complexes (Cur:DN) at concentrations of 15 and 45 mg/L. The transition of cells to AnnV quadrants visible for Cur and bio-complex-treated cells.
Fig 6 Dot blot/scatter diagrams of PI/AnnV staining of HepG2 cells treated with curcumin (Cur), diamond nanoparticles (DN) and their bio-complexes (Cur:DN) at concentrations of 15 and 45 mg/L. The transition of cells to AnnV quadrants visible for Cur and bio-complex-treated cells.CC BY · PubMed Central
Fig 7 Dot blot/scatter diagrams of PI/AnnV staining of fibroblasts treated with curcumin (Cur), diamond nanoparticles (DN) and their bio-complexes (Cur:DN) at concentrations of 15 and 45 mg/L. The transition of cells to AnnV quadrants visible for bio-complex-treated cells.
Fig 7 Dot blot/scatter diagrams of PI/AnnV staining of fibroblasts treated with curcumin (Cur), diamond nanoparticles (DN) and their bio-complexes (Cur:DN) at concentrations of 15 and 45 mg/L. The transition of cells to AnnV quadrants visible for bio-complex-treated cells.CC BY · PubMed Central
Fig 8 HepG2 cell morphology. Control (A), Cur45 (B), DN45 (C), Cur75:DN25 (D), Cur25:DN75 (E). DN visible on the surface of wells (C–E). Scale bar indicates 200 μm.
Fig 8 HepG2 cell morphology. Control (A), Cur45 (B), DN45 (C), Cur75:DN25 (D), Cur25:DN75 (E). DN visible on the surface of wells (C–E). Scale bar indicates 200 μm.CC BY · PubMed Central
Fig 9 Fibroblast cell morphology. Control (A), Cur45 (B), DN45 (C), Cur75:DN25 (D), Cur25:DN75 (E). Scale bar indicates 500 μm.
Fig 9 Fibroblast cell morphology. Control (A), Cur45 (B), DN45 (C), Cur75:DN25 (D), Cur25:DN75 (E). Scale bar indicates 500 μm.CC BY · PubMed Central
Fig 10 Images of stained chicken blood smears. Groups: Control (A), Placebo (B), Cur (C), DN (D), 3Cur:1DN (E), 1Cur:3DN (F). Heterophils (black arrows) and thrombocytes (white arrows). Scale bar indicates 20 μm.
Fig 10 Images of stained chicken blood smears. Groups: Control (A), Placebo (B), Cur (C), DN (D), 3Cur:1DN (E), 1Cur:3DN (F). Heterophils (black arrows) and thrombocytes (white arrows). Scale bar indicates 20 μm.CC BY · PubMed Central
Fig 11 Brain histology of chicken embryo. Groups: Control (A), Placebo (B), Cur (C), DN (D), 3Cur:1DN (E), 1Cur:3DN (F). Silver-grey particles were observed in the DN group (black arrows) and single necrotic-like neurons (white arrows). Scale bar indicates 100 μm.
Fig 11 Brain histology of chicken embryo. Groups: Control (A), Placebo (B), Cur (C), DN (D), 3Cur:1DN (E), 1Cur:3DN (F). Silver-grey particles were observed in the DN group (black arrows) and single necrotic-like neurons (white arrows). Scale bar indicates 100 μm.CC BY · PubMed Central
Fig 12 Liver histology of chicken embryos. Groups: Control (A), Placebo (B), Cur (C), DN (D), 3Cur:1DN (E), 1Cur:3DN (F). Heterophils and primary hematopoietic cells were present in the proximity of wider blood vesicles (black arrows). Scale bar indicates 100 μm.
Fig 12 Liver histology of chicken embryos. Groups: Control (A), Placebo (B), Cur (C), DN (D), 3Cur:1DN (E), 1Cur:3DN (F). Heterophils and primary hematopoietic cells were present in the proximity of wider blood vesicles (black arrows). Scale bar indicates 100 μm.CC BY · PubMed Central

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Authors
Strojny, B., Grodzik, M., Sawosz, E., Winnicka, A., Kurantowicz, N., Jaworski, S., Kutwin, M., Urbańska, K., Hotowy, A., Wierzbicki, M., & Chwalibog, A.
Venue
PLOS ONE
Published
2016-10-13
Volume
11
Issue
10
Pages
e0164637
ISSN
1932-6203
Cited by
33
References
0
Language
en
Funders
Narodowe Centrum Nauki (NN311540840)

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