Journal: Nanomaterials

Article Title: Surface Characteristics and Biological Evaluation of Si-DLC Coatings Fabricated Using Magnetron Sputtering Method on Ti6Al7Nb Substrate

doi: 10.3390/nano9060812

Figure Lengend Snippet: Endothelial cells’ proliferation on the surface of the deposited Si-DLC coatings (live/dead assay).

Article Snippet: The biocompatibility of the DLC and Si-DLC films was evaluated using human endothelial cell line, EA.hy926, (ATCC - American Type Culture Collection).

Techniques: Live Dead Assay

Journal: Nanomaterials

Article Title: Surface Characteristics and Biological Evaluation of Si-DLC Coatings Fabricated Using Magnetron Sputtering Method on Ti6Al7Nb Substrate

doi: 10.3390/nano9060812

Figure Lengend Snippet: Cytotoxicity of the deposited Si-DLC coatings towards the endothelial cells (live/dead assay).

Article Snippet: The biocompatibility of the DLC and Si-DLC films was evaluated using human endothelial cell line, EA.hy926, (ATCC - American Type Culture Collection).

Techniques: Live Dead Assay

Journal: Nanomaterials

Article Title: Surface Characteristics and Biological Evaluation of Si-DLC Coatings Fabricated Using Magnetron Sputtering Method on Ti6Al7Nb Substrate

doi: 10.3390/nano9060812

Figure Lengend Snippet: Endothelial cells’ viability in direct and indirect contact with the deposited Si-DLC coatings (XTT assay).

Article Snippet: The biocompatibility of the DLC and Si-DLC films was evaluated using human endothelial cell line, EA.hy926, (ATCC - American Type Culture Collection).

Techniques: XTT Assay

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: Diamond nanoparticles are cytotoxic to HUVEC but not to HS-5 and HMEC. HUVEC, HS-5, and HMEC 48-h proliferation ( A ) and 24-h membrane perforation ( B ) after treatment with NDs at concentrations of 5, 10, 20, 50, and 100 μg/mL. Statistical significance is indicated with asterisks: *P < 0.033, **P < 0.002, ***P < 0.001 (multifactor ANOVA; P < 0.05; n = 3 with 4 individual replicates for proliferation and 6 individual replicates for membrane perforation). All values are expressed as mean ± standard deviation. ( C ) HUVEC metabolic activity after treatment with NDs at concentrations of 5, 10, 20, 50, and 100 mg/l for 24 h. Statistical significance is indicated with different superscripts: P<0.001 (a, b, c, d, e) (ANOVA; P < 0.05; n = 3 with 6 individual replicates). ( D ) Live/Dead assay confocal microscopy images of nontreated HUVEC cells (“C”) and treated cells with 50 mg/l for 24 h (“ND”). The nuclei of dead cells are labelled with a green stain (NucGreen Dead 488), whereas all nuclei are stained with a red stain (NucRed Live 647). Images taken using Nomarski interference contrast (“NOM”) showed light-reflecting nanoparticle agglomerates, suggesting strong endocytosis of NDs.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Membrane, Standard Deviation, Activity Assay, Live Dead Assay, Confocal Microscopy, Staining

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: Toxicity of diamond nanoparticles to HUVEC depends on cell density and surface conjugation of nanoparticles. ( A ) HUVEC 48-h proliferation was analysed for untreated cells (“C”) or cells treated with NDs (“ND”, “-RGD -FBS”) and NDs conjugated with a peptide containing an RGD motive (Gly-Arg-Gly-Asp-Ser; “ND”, “+RGD”) and at concentrations of 5 and 50 mg/l in standard HUVEC culture medium and in medium with the addition of 10% FBS (“+FBS”). Statistical significance is indicated with different superscripts: P<0.001 (a, b, c, d, e) (ANOVA; P < 0.05; n = 3 with 4 individual replicates). ( B ) Dependence of ND toxicity on HUVEC cell density was assessed in a 96-well plate at densities of 5 x 10 3 , 1 x 10 4 , 1.5 x 10 4 , and 2×10 4 . After 24-h incubation, cells were treated with NDs at a concentration of 0.4 ng/cell (the final ND concentration was 20, 40, 60, and 80 mg/L in 100 µL medium, respectively), and the metabolic activity was assessed. The relative toxicity of HUVEC was expressed as the ratio of the metabolic activity value of the ND-treated cells to the metabolic activity of the untreated cells. Statistical significance is indicated with different superscripts: P<0.001 (a, b, c, d) (ANOVA; P < 0.05; n = 3 with 6 individual replicates). All values are expressed as mean ± standard deviation. ( C ) HUVEC time-lapse images of a 12-h cell culture with NDs at a final concentration of 20 mg/l. The full video of time-lapse images is available as Supplementary Video . Abbreviations: RV, relative value; FUs, fluorescent units; C, control; NDs, diamond nanoparticles; RGD, Gly-Arg-Gly-Asp-Ser peptide; FBS, fetal bovine serum. ( D ) HUVEC spheroid morphology after 24-h treatment with NDs at a concentration of 10 and 100 mg/l. HUVEC spheroid membrane perforation ( E ) and spheroid size ( F ) for a after 24-h incubation with NDs at a concentration of 5, 10, 20, 50, and 100 mg/l. Statistical significance is indicated with different superscripts:: P<0.001 (a, b, c) (ANOVA; P < 0.05; n = 3 with 6 individual replicates).

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Conjugation Assay, Incubation, Concentration Assay, Activity Assay, Standard Deviation, Cell Culture, Control, Membrane

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: Diamond nanoparticles led to an NADPH-dependent induction of ROS in HUVEC. ( A ) Intercellular level of NADPH analysed in lysates of control HUVEC cells (“C”) and HUVEC cells incubated for 4 h with NDs at a concentration of 20 and 50 mg/l (“ND”). Statistical significance is indicated with different superscripts: P<0.001 (a, b, c) (ANOVA; P < 0.05; n = 2 with 4 individual replicates). ( B ) Merged images of mitochondrial superoxide staining (red channel; MitoSox Red, Thermo Fisher Scientific) and transmitted light with Nomarski interference contrast of control HUVEC cells (“C”) and HUVEC cells incubated for 4 h with NDs at a concentration of 50 mg/l (“ND 50”). ( C ) Confocal images of ROS and NO in control HUVEC cells (“C”) and HUVEC cells incubated for 4 h with NDs at a concentration of 20 and 50 mg/l (“ND 20”, “ND 50”) detected using the general oxidative stress indicator CM-H2CFDA (green channel; Thermo Fisher Scientific) and the NO synthesis level indicator DAF-FM (green channel; Thermo Fisher Scientific). Cell nuclei were stained with NucRed Live 647 (red channel; Thermo Fisher Scientific). Graphs showing ROS ( D ) and NO ( E ) levels expressed as a sum of the pixel values per cell. Statistical significance is indicated with different superscripts: P<0.001 (a, b) (ANOVA; P < 0.05; n = 2 with 7 individual replicates, and each well was imaged in 4 fields of view). All values are expressed as mean ± standard deviation.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Control, Incubation, Concentration Assay, Staining, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: NDs increase the synthesis level of stress-related proteins. ( A ) Antibody array analysis of cytokine synthesis in control HUVEC cells (“C”) incubated with NDs at a concentration of 20 mg/l for 24 h (“ND”). One assay consists of two membranes, which are labelled “I” and “II”, and the proteins are analysed in duplicate. The localization of the positive controls: I_A1,2; I_B1,2; I_H7,8; II_A1,2; II_B1,2; II_H7,8. The localization of the selected proteins in the order of their fold (log(2)) change shown in the graph ( B ): IL-6, I_D3,4; EGF, I_F1,2; VEGF, I_G5,6; TIMP2, I_E5,6; IL-8, I_E3,4; endostatin II_H1,2; IGF-1, I_C3,4; CCL-5, I_B5,6; PECAM-1, II_F5,6; VEGF-D, I_H5,6; uPAR, II_A7,8; VEGFR3, II_C7,8; PLG, II_G1,2; MCP-1, I_G3,4; CXCL11, II_A5,6; ANGPT1, II_E1,2; TIMP1, I_D5,6; GRO A, I_A1,2; MMP-1, II_D5,6; VEGFR2, II_B7,8; TIE-2, II_G5,6; bFGF, I_H1,2; MMP-9, II_E5,6; ANGPT2 II_F1,2. The full array map is available in Tables S1 and S2 . ( C ) Images of control HUVEC tube formation (“C”) and tube formation during treatment with NDs at a final concentration of 20 mg/l (“ND”). Graphs show the mean number of junctions of HUVEC tubes in the field of view ( D ), mean tube length in the field of view ( E ), and mean number of meshes in the field of view ( F ). Data were obtained by analysing the images using ImageJ software and the Angiogenesis Analyzer macro. Statistical significance is indicated with asterisks: *P = 0.003, P = 0.002 and P < 0.001, respectively ( t -test; P < 0.05; n = 2 with 4 individual replicates). All values are expressed as mean ± standard deviation.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Ab Array, Control, Incubation, Concentration Assay, Software, Standard Deviation