Effects of oxygen tension and dextran-shelled/2H,3H-decafluoropentane-cored oxygen-loaded nanodroplets on secretion of gelatinases and their inhibitors in term human placenta

Figures & data

Fig. 1. OLND morphology and size distribution.

Notes: OLNDs were checked for morphology by optical microscopy and for size distribution by light scattering. Results are shown as representative images from three different preparations. Panel A. OLND image by optical microscopy. Magnification: 60×. Panel B. OLND size distribution.

Fig. 2. Oxygen tension and OLND effects on the secretion of latent forms of gelatinases (proMMP-2 and proMMP-9) by human placental explants.

Notes: Small portions of human placental chorionic villous explants (35 mg) from normal term pregnancies were cultured for 8 h at 37 °C either in 20% O₂ (panel A: lanes 1–2; panels B-C: black columns) or 3% O₂ (panel A: lanes 3–4; panels B-C: white columns) in serum-free Ham’s F12 medium with or without 10% v/v OLNDs. After collection, explant supernatants were analyzed by gelatin zymography (panel A) and subsequent densitometry (panels B–C). Recombinant human proMMP-9 (92 kDa) and MMP-9 (83 kDa) were employed as standard markers (st). Results are shown as representative gel (panel A) or means + SEM (panels B–C) from three independent experiments. For every experiment, each condition was performed in quadruplicate. Densitometric data were also evaluated for significance by ANOVA. Panel B: *vs. 20% O₂ ctrl, p < 0.0001; ° vs. 3% O₂ ctrl, p < 0.0001. Panel C: * vs. 20% O₂ ctrl, p < 0.0001; ° vs. 3% O₂ ctrl, p < 0.001; °° vs. 3% O₂ ctrl, p < 0.0001.

Fig. 3. Oxygen tension and OLND effects on the levels of secreted gelatinases (MMP-2 and -9) in human placental explants.

Notes: Small portions of human placental chorionic villous explants (35 mg) from normal term pregnancies were cultured for 8 h at 37 °C either in 20% O₂ (black columns, all panels) or 3% O₂ (white columns, all panels) in serum-free Ham’s F12 medium with or without 10% v/v OLNDs. After supernatant collection, the protein levels of MMP-9 (panel A) and MMP-2 (panel B) were evaluated by ELISA. Results are shown as means + SEM from three independent experiments. For every experiment, each condition was performed in quadruplicate. All data were evaluated for significance by ANOVA. Panel A: *vs. 20% O₂ ctrl, p < 0.0001; ° vs. 3% O₂ ctrl, p < 0.0001. Panel B: *vs. 20% O₂ ctrl, p < 0.003; ° vs. 3% O₂ ctrl, p < 0.004; °° vs. 3% O₂ ctrl, p < 0.003.

Fig. 4. Oxygen tension and OLND effects on the levels of secreted MMP tissue inhibitors (TIMP-1 and -2) in human placental explants.

Notes: Small portions of human placental chorionic villous explants (35 mg) from normal term pregnancies were cultured for 8 h at 37 °C either in 20% O₂ (black columns, all panels) or 3% O₂ (white columns, all panels) in serum-free Ham’s F12 medium with or without 10% v/v OLNDs. After supernatant collection, the protein levels of TIMP-1 (panel A) and TIMP-2 (panel B) were evaluated by ELISA. Results are shown as means + SEM from three independent experiments. For every experiment, each condition was performed in quadruplicate. All data were evaluated for significance by ANOVA. Panel A: * vs. 20% O₂ ctrl, p < 0.001; ° vs. 3% O₂ ctrl, p < 0.003; °° vs. 3% O₂ ctrl, p < 0.002. Panel B: * vs. 20% O₂ ctrl, p < 0.003; ° vs. 3% O₂ ctrl, p < 0.002; °° vs. 3% O₂ ctrl, p < 0.001.

Fig. 5. Oxygen tension and OLND effects on secreted MMP/TIMP balances in human placental explants.

Notes: Small portions of human placental chorionic villous explants (35 mg) from normal term pregnancies were cultured for 8 h at 37 °C either in 20% O₂ (black columns, all panels) or 3% O₂ (white columns, all panels) in serum-free Ham’s F12 medium with or without 10% v/v OLNDs. After supernatant collection and measurement of the protein levels of MMP-9, MMP-2, TIMP-1, and TIMP-2 (see Figs. 2 and 3), MMP-9/TIMP-1 (panel A) and MMP-2/TIMP-2 (panel B) stoichiometric ratios were calculated. Results are shown as means + SEM from three independent experiments. For every experiment, each condition was performed in quadruplicate. All data were evaluated for significance by ANOVA. Panel A: * vs. 20% O₂ ctrl, p < 0.005; ° vs. 3% O₂ ctrl, p < 0.005. Panel B: * vs. 20% O₂ ctrl, p < 0.0001; ° vs. 3% O₂ ctrl, p < 0.003; °° vs. 3% O₂ ctrl, p < 0.002.