Populations of cells in liver isolates (light scatters).

<p>Exemplary flow cytometric dot plots showing populations of cells isolated from human liver tissue in light scatters: forward—FSC and side—SSC. Depending on specimen, there are minimum 2 and maximum 4 distinct cells populations (labeled P2 –P5) identified in the P1 gate (cellular debris and doublets excluded). Population P2 –blue dots; P3 –yellow dots; P4 –green dots; P5 –purple dots. The percentage of cells in the individual populations of the P1 gate—specimen H27-14: P2 = 35.9%, P3 = 62.8%; H26-14: P2 = 32.3%, P3 = 52.8%, P4 = 6.1%; H16-13: P2 = 47.6%, P3 = 46.4%, P4 = 1.7%, P5 = 1.3%; H22-13: P2 = 40.6%, P3 = 41.6%, P4 = 11%, P5 = 1.1%.</p

Double immunostaining of the isolated cells.

<p>Exemplary double immunostainings of the cells isolated from liver specimen (H31-14, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0182846#pone.0182846.t001" target="_blank">Table 1</a>). (<b>A</b>) In the P2 population cells labeled with anti-Alb antibody expressed neither CD45 (83.5% vs.1.3%, left panel) nor CD14 (81.3% vs. 0.5%, right panel) markers. (<b>B</b>) Nonparenchymal cells in the P3 population (left panel) expressed CD45 marker but not CD14 (46.7% vs. 0.1%). Some of the cells from the P4 population (right panel) expressed both CD45 and CD14 markers (CD45 = 23.9%, CD14 = 16%, and CD45/CD14 = 15.1%) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0182846#pone.0182846.t002" target="_blank">Table 2</a> for the list of antibodies). Alb-positive cells–green dots; CD45-positive cells–pink dots; CD14-positive cells–blue dots; CD54/CD14-positive cells–grey dots; unstained cells–yellow dots. Abbr.: Alb, albumin; FITC, PerCP, APC—fluorophores.</p

The impact of sex, chemotherapy, and age on the percentage of A1AT-positive cells in P2 population.

<p>Comparison of the percentage of A1AT-positive cells in the P2 population between groups of patients. F (n = 15), M (n = 14), Ch- (n = 7), Ch+ (n = 22), A60- (n = 16), A60+ (n = 13). The differences between two independent groups were analyzed using the Welch's two-sample t-test. The results are presented as the median values with the range for each variable. Abbr.: A1AT, α-1-antitrypsin; F, female; M, male; Ch-, non-treated patients; Ch+, patients after chemotherapy; A60-, patients under 60 years old; A60+, patients 60 years old and older.</p

Antibodies used in this study.

<p>Antibodies used in this study.</p

The characteristic of liver tissue used and the main results of liver cells isolations.

<p>The characteristic of liver tissue used and the main results of liver cells isolations.</p

The <i>p</i>-values for selected interactions from two-way Analysis of Variance (ANOVA) confirmed using Tukey HSD test.

<p>The <i>p</i>-values for selected interactions from two-way Analysis of Variance (ANOVA) confirmed using Tukey HSD test.</p

Transmission electron microscopy images of nanoparticles.

<p>Images of (<b>A</b>) diamond (ND) and (<b>B</b>) graphite (NG) nanoparticles. Scale bar: 100 nm.</p

Glioblastoma and hepatoma cells status (CI).

<p>U87, U87-EGFP, C3A and C3A-EGFP CI monitored by a real-time cell analyzer (RTCA) after 1 and 2 h of exposure to the culture medium containing graphite and diamond nanoparticles at different concentrations: diamond 20 μg/mL (ND 20), 50 μg/mL (ND 50) and 100 μg/mL (ND 100), and graphite 20 μg/mL (NG 20), 50 μg/mL (NG 50) and 100 μg/mL (NG 100); nontreated cells were used as a control (NT). (<b>A</b>) U87 (blue lines) and C3A cells (red lines) treated with graphite (NG) after 1 h (vertical blue marker line) and 2 h (vertical red marker line) and control (vertical green marker line); (<b>B</b>) U87-EGFP cells (violet lines) and C3A-EGFP cells (green lines) treated with graphite after 1 h (vertical blue marker line) and 2 h (vertical red marker line) and control (vertical green marker line); (<b>C</b>) U87 (blue lines) and C3A cells (red lines) treated with diamond (ND) after 1 h (vertical blue marker line) and 2 h (vertical red marker line) and control (vertical green marker line); (<b>D</b>) U87-EGFP cells (violet lines) and C3A-EGFP cells (green lines) treated with diamond (ND) after 1 h (vertical blue marker line) and 2 h (vertical red marker line) and control (vertical green marker line). Data analysis was carried out by two-way ANOVA and the differences between the groups were tested by Duncan’s test. Data were averaged from three replicates (n = 3); <i>P</i>< 0.05. No significant interaction (nanoparticles with transduction) was observed.</p

Glioblastoma and hepatoma cells morphology.

<p>U87, U87-EGFP, C3A and C3A-EGFP cells morphology after 2 h exposure to medium containing nanoparticles at the highest concentration: graphite 100 μg/mL (NG 100) and diamond 100 μg/mL (ND 100), nontreated cells were used as a control (Control). Analysis of the cells morphology was performed using an inverted fluorescence microscope. Images of the labeled cells, U87-EGFP and C3A-EGFP, were captured using a green fluorescence filter. Scale bar: 200 μm.</p

Human serum albumin concentration in culture medium.

<p>Albumin secreted by the C3A and C3A-EGFP cells treated for 24 h with the culture medium containing nanoparticles at different concentrations measured by ELISA test: diamond 20 μg/mL (ND 20), 50 μg/mL (ND 50) and 100 μg/mL (ND 100), and graphite 20 μg/mL (NG 20), 50 μg/mL (NG 50) and 100 μg/mL (NG 100). Albumin secretion by cells after 24 h exposure to the medium with different concentrations of diamond and graphite nanoparticles: (<b>A</b>) C3A cells and (<b>B</b>) C3A-EGFP cells. Nontreated cells were used as a control (Control). Data were analyzed using a two-way ANOVA with the Duncan’s test. Data were averaged from three replicates (n = 3); <i>P</i> < 0.05. No significant interaction (nanoparticles with transduction) was observed.</p