Journal: Experimental and Therapeutic Medicine

Article Title: Effects of ozone stimulation of bronchial epithelial cells on proliferation and collagen synthesis of co-cultured lung fibroblasts

doi: 10.3892/etm.2018.6122

Figure Lengend Snippet: Representative microscopic images of HBECs and HLFs cultured alone or co-cultured with or without O 3 stimulation. (A) HBECs cultured alone without O 3 stimulation, (B) HBECs cultured alone with O 3 stimulation, (C) HLFs cultured alone, (D) HLFs co-cultured with HBECs without O 3 stimulation and (E) HLFs co-cultured with O 3 -stimulated HBECs (magnification, ×50). O 3 stimulation was given at a dosage of 2 ppm, 30 min for each treatment. Scale bar=100 µm. HBECs, human bronchial epithelial cells; HLFs, human lung fibroblasts; O 3 , ozone.

Article Snippet: Following this, cells were treated with TGF-β1 antibodies (1:1,000; cat. no. BA0290) at 37°C for 1–2 h and biotinylated goat anti-rabbit IgG antibodies (1:500; cat. no. BA1003; both Wuhan Boster Biological Technology Ltd.) at room temperature for 20 min. Reagent SABC (Wuhan Boster Biological Technology Ltd.) was added at room temperature for 20 min.

Techniques: Cell Culture

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of ozone stimulation of bronchial epithelial cells on proliferation and collagen synthesis of co-cultured lung fibroblasts

doi: 10.3892/etm.2018.6122

Figure Lengend Snippet: Time course of TGF-β1, TNF-α and PGE2 cytokine secretion in the supernatant of the co-culture system after human bronchial epithelial cells were stimulated ozone. TGF-β1 and TNF-α secretion was measured using ELISA and PGE2 secretion was measured using a radioimmunoassay. The levels of TGF-β1 remained high and continued to gradually increase for up to 24 h. TNF-α levels were relatively low and did not vary significantly over the 24 h. However, the secretion of PGE2 increased significantly between 6 and 12 h and thereafter plateaued until 24 h. Data are presented as the mean ± standard deviation (n=4). **P<0.01, *P<0.05 vs. 6 h. TGF-β1, transforming growth factor-β1; TNF-α, tumor necrosis factor-α; PGE2, prostaglandin E2.

Article Snippet: Following this, cells were treated with TGF-β1 antibodies (1:1,000; cat. no. BA0290) at 37°C for 1–2 h and biotinylated goat anti-rabbit IgG antibodies (1:500; cat. no. BA1003; both Wuhan Boster Biological Technology Ltd.) at room temperature for 20 min. Reagent SABC (Wuhan Boster Biological Technology Ltd.) was added at room temperature for 20 min.

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay, RIA Assay, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of ozone stimulation of bronchial epithelial cells on proliferation and collagen synthesis of co-cultured lung fibroblasts

doi: 10.3892/etm.2018.6122

Figure Lengend Snippet: Effect of O 3 stimulation on the concentration of TGF-β1, TNF-α and PGE2 cytokines in HBECs cultured alone. The concentration levels of the cytokines TGF-β1 and TNF-α were measured using ELISA, and PGE2 was measured using a radioimmunoassay. The white bars represent HBECs cultured alone without O 3 -stimulation (control) and the filled bars represent HBECs cultured alone with O 3 -stimulation, respectively. Data are presented as the mean ± standard deviation (n=4). *P<0.05 vs. HBECS. TGF-β1, transforming growth factor-β1; TNF-α, tumor necrosis factor-α; PGE2, prostaglandin E2; O 3 , ozone.

Article Snippet: Following this, cells were treated with TGF-β1 antibodies (1:1,000; cat. no. BA0290) at 37°C for 1–2 h and biotinylated goat anti-rabbit IgG antibodies (1:500; cat. no. BA1003; both Wuhan Boster Biological Technology Ltd.) at room temperature for 20 min. Reagent SABC (Wuhan Boster Biological Technology Ltd.) was added at room temperature for 20 min.

Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, RIA Assay, Control, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of ozone stimulation of bronchial epithelial cells on proliferation and collagen synthesis of co-cultured lung fibroblasts

doi: 10.3892/etm.2018.6122

Figure Lengend Snippet: Correlation between cytokine concentration and cell proliferation and collagen synthesis in HLFs co-cultured with O 3 -stressed HBECs. (A and B) A positive linear correlation between proliferation of HLFs and the concentration of TGF-β1 in the co-culture supernatant. Collagen synthesis capacity in the culture supernatant was also positively correlated with TGF-β1 concentration (r=0.758, P=0.018). (C and D) There was no correlation between the proliferation and TNF-α concentration (r=0.209, P=0.589), nor between collagen synthesis and TNF-α concentration (r=0.311, P=0.415). (E and F) A negative linear correlation between the proliferation and PGE2 concentration of HLFs (r=0.783, P=0.013) and between collagen synthesis and PGE2 concentration (r=0.817, P=0.007) was indicated. TGF-β1, transforming growth factor-β1; TNF-α, tumor necrosis factor-α; PGE2, prostaglandin E2; O 3 , ozone; HBECs, human bronchial epithelial cells; HLFs, human lung fibroblasts; OD, optical density.

Article Snippet: Following this, cells were treated with TGF-β1 antibodies (1:1,000; cat. no. BA0290) at 37°C for 1–2 h and biotinylated goat anti-rabbit IgG antibodies (1:500; cat. no. BA1003; both Wuhan Boster Biological Technology Ltd.) at room temperature for 20 min. Reagent SABC (Wuhan Boster Biological Technology Ltd.) was added at room temperature for 20 min.

Techniques: Concentration Assay, Cell Culture, Co-Culture Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of ozone stimulation of bronchial epithelial cells on proliferation and collagen synthesis of co-cultured lung fibroblasts

doi: 10.3892/etm.2018.6122

Figure Lengend Snippet: Representative images and corresponding quantitative expression level of TGF-β1 in HBECs with or without O 3 stimulation. HBECs were cultured alone without O 3 stimulation (control) or cultured alone with O 3 stimulation. (A) TGF-β1 in HBECs was labeled by immunocytochemistry and imaged by photomicroscope. (B) Expression level of TGF-β1 in HBECs was quantified according to the averaged photodensity of the images. Data are presented as the mean ± standard deviation (n=4). *P<0.05 vs. HBECs. HBECs, human bronchial epithelial cells; HLFs, human lung fibroblasts; O 3 , ozone; TGF-β1, transforming growth factor-β1.

Article Snippet: Following this, cells were treated with TGF-β1 antibodies (1:1,000; cat. no. BA0290) at 37°C for 1–2 h and biotinylated goat anti-rabbit IgG antibodies (1:500; cat. no. BA1003; both Wuhan Boster Biological Technology Ltd.) at room temperature for 20 min. Reagent SABC (Wuhan Boster Biological Technology Ltd.) was added at room temperature for 20 min.

Techniques: Expressing, Cell Culture, Control, Labeling, Immunocytochemistry, Standard Deviation