THP-1 cell adhesion to HUVEC exposed to the supernatants of oxLDL-treated macrophages and mast cells.

<p>(A) Fluorescent images of adherent THP-1 cells (green) on resting (control) and activated HUVEC. (B) Number of adherent THP-1 cells (mean ± SD, n = 4) on HUVEC per image field (904 μm × 675 μm). The HUVEC activation groups are: 1) “Control”—RPMI-1640 medium alone; 2) “S_L1“—the supernatant of 8 μg/ml oxLDL-treated mast cells for two hours; 3) “S_L1 + H^-“—S_L1 combined with the antagonists of histamine receptors for two hours; 4) “S_M“—the supernatant of 8 μg/ml oxLDL-treated macrophages for five hours; 5) “S_M + H^-“—S_M combined with the antagonists of histamine receptors for five hours; 6) “S_M + S_L1“—S_M for five hours with S_L1 added two hours before the end of the incubation time; 7) “S_M + S_L1 + H^-“—S_M combined with the antagonists of histamine receptors for five hours and with S_L1 added two hours before the end of the incubation time; 8) “S_M + S_L2“—S_M for five hours with the supernatant of 25 μg/ml oxLDL-treated mast cells (S_L2) added two hours before the end of the incubation time; and 9) S_M + S_L2 + H^-—S_M combined with the antagonists of histamine receptors for five hours and with S_L2 added two hours before the end of the incubation time.</p

Number of firmly adherent THP-1 cells on resting or activated HUVEC under shear flow conditions, according to micrfluidic channel-based detachment assays.

<p>Here, “Control”—resting endothelium, “S_L1“—the supernatant of 8 μg/ml oxLDL-treated mast cells, “S_M“—the supernatant of 8 μg/ml oxLDL-treated macrophages, and “S_M + S_L1“—the combination of the supernatants of the oxLDL-treated macrophages and mast cells. Mean μ SD of three independent tests. More details about the HUVEC activation groups are given in Figs. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123088#pone.0123088.g001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123088#pone.0123088.g007" target="_blank">7</a>.</p

Surface expression of adhesion molecules on resting HUVEC (“Control”) and on HUVEC exposed to the supernatant of 8 μg/ml oxLDL-treated mast cells (“S_L1”), the supernatant of 8 μg/ml oxLDL-treated macrophages (“S_M”), or a combination of these supernatants (S_M + S_L1).

<p>(A) Histogram of the expression of ICAM-1, VCAM-1 and E-selectin on the HUVEC surface; (B) Percentage of HUVEC with positive expression of these adhesion molecules (mean ± SD, n = 3). * <i>p</i> < 0.05, ** <i>p</i> < 0.01, *** <i>p</i> < 0.001. Details about the HUVEC activation groups are given in Figs. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123088#pone.0123088.g001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123088#pone.0123088.g007" target="_blank">7</a>.</p

Concentration of histamine in the supernatants of untreated mast cells (“Ctrl”), 8 μg/ml oxLDL-treated mast cells (“OxLDL(8 μg/ml)”), 25 μg/ml oxLDL-treated mast cells (“OxLDL(25 μg/ml)”), and mast cells exposed to the supernatant of 8 μg/ml oxLDL-treated monocytes, according to ELISA.

<p>Mean ± SD of three independent tests.</p

THP-1 cell adhesion to HUVEC exposed to oxLDL, histamine, or the macrophage supernatant.

<p>(A) Fluorescent images of adherent THP-1 cells (green) on resting (control) or activated HUVEC. (B) Number of adherent THP-1 cells (mean ± SD, n = 4) on resting or activated HUVEC per image field (904 μm × 675 μm). The HUVEC activation groups are: 1) “Control”—RPMI-1640 medium alone; 2) “Hist”—RPMI-1640 medium with 10^-6 mol/l histamine for four hours; 3) “OxLDL”- 80 μg/ml of oxLDL for 20 hours; 4) “OxLDL + Hist”- 80 μg/ml of oxLDL for 20 hours with 10^-6 mol/l histamine added 4 hours before the end of the incubation time; 5) “S_T“—the supernatant of 8 μg/ml oxLDL-treated THP-1 cells for five hours; 6) “S_T + Hist”—S_T for five hours with 10^-6 mol/l histamine added four hours before the end of the incubation time; 7) “S_M“—the supernatant of 8 μg/ml oxLDL-treated THP-1 macrophages for five hours; and 8) “S_M + Hist”—S_M for five hours with 10^-6 mol/l histamine added four hours before the end of the incubation time.</p

Concentration of TNF-α in the supernatants of untreated monocytes (“THP-1 sup”), untreated macrophages (“Macro. Sup”), 8 μg/ml oxLDL-treated monocytes (“OxLDL-THP-1 sup”), and 8 μg/ml oxLDL-treated macrophages (“OxLDL-Macro. Sup”), according to ELISA.

<p>Mean ± SD of three independent tests.</p

Surface expression of adhesion molecules on resting HUVEC (“Control”) and on HUVEC exposed to 10^-6 mol/l histamine (“Hist”), the supernatant of 8 μg/ml oxLDL-treated macrophages (“S_M”), or a combination of the oxLDL-treated macrophage supernatant and histamine (“S_M+Hist”).

<p>(A) Histogram of the expression and (B) fluorescence intensity relative to the isotype control (mean ± SD, n = 3) of ICAM-1, VCAM-1 and E-selectin. (C) Percentage of HUVEC with positive expression of these adhesion molecules (mean ± SD, n = 3). Details about the HUVEC activation groups are given in Figs. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123088#pone.0123088.g001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123088#pone.0123088.g004" target="_blank">4</a>.</p

Number of adherent THP-1 cells on HUVEC activated by TNF-α with a concentration of 250, 500, 1000, 2000, 4000, or 10000 pg/ml.

<p>HUVEC were exposed to TNF-α for five hours. Mean ± SD per image field (904 μm × 675 μm) of three independent tests.</p

Oxidized Low-Density Lipoprotein Contributes to Atherogenesis via Co-activation of Macrophages and Mast Cells - Fig 1

<p>(A) Flow chart of static adhesion experiments in which HUVECs were exposed to oxLDL, histamine, and/or the supernatant of oxLDL-treated monocytes or macrophages. (B) Flow chart of static and flow adhesion experiments in which HUVECs were exposed to the supernatant of oxLDL-treated macrophages and/or the supernatant of oxLDL-treated mast cells.</p

Ablative Focused Ultrasound Synergistically Enhances Thermally Triggered Chemotherapy for Prostate Cancer <i>in Vitro</i>

High-intensity focused ultrasound (HIFU) can locally ablate biological tissues such as tumors, i.e., induce their rapid heating and coagulative necrosis without causing damage to surrounding healthy structures. It is widely used in clinical practice for minimally invasive treatment of prostate cancer. Nonablative, low-power HIFU was established as a promising tool for triggering the release of chemotherapeutic drugs from temperature-sensitive liposomes (TSLs). In this study, we combine ablative HIFU and thermally triggered chemotherapy to address the lack of safe and effective treatment options for elderly patients with high-risk localized prostate cancer. DU145 prostate cancer cells were exposed to chemotherapy (free and liposomal Sorafenib) and ablative HIFU, alone or in combination. Prior to cell viability assessment by trypan blue exclusion and flow cytometry, the uptake of TSLs by DU145 cells was verified by confocal microscopy and cryogenic scanning electron microscopy (cryo-SEM). The combination of TSLs encapsulating 10 μM Sorafenib and 8.7W HIFU resulted in a viability of less than 10% at 72 h post-treatment, which was significant less than the viability of the cells treated with free Sorafenib (76%), Sorafenib-loaded TSLs (63%), or HIFU alone (44%). This synergy was not observed on cells treated with Sorafenib-loaded nontemperature sensitive liposomes and HIFU. According to cryo-SEM analysis, cells exposed to ablative HIFU exhibited significant mechanical disruption. Water bath immersion experiments also showed an important role of mechanical effects in the synergistic enhancement of TSL-mediated chemotherapy by ablative HIFU. This combination therapy can be an effective strategy for treatment of geriatric prostate cancer patients