Characteristics of fibroblasts donors.

<p>Lung function data presented as % of predicted values for donor's gender, age and height. F: Female, M: Male, NSCLC: non small cell lung carcinoma, COPD: chronic obstructive pulmonary disease, Ca: carcinoma, IPF: idiopathic pulmonary fibrosis, R: resection, T: Transplant. FEV₁ forced expiratory volume in 1 second. FVC: forced vital capacity. N/A: data not available.</p

Chemical inhibition of the ERK signalling pathway attenuates BME induced IL-8 release.

<p>Fibroblasts were pretreated for 1(1, 10 µM) or UO126 (0.5, 5 µM) in DMSO (vehicle control) before stimulation with 10% (A) and 20% (B) BME in the presence of inhibitors for 72 hours, prior to analysis of IL-8 release by ELISA. Data expressed as pg/ml. Bars represent mean ± SEM. Data analysed by one-way ANOVA with Bonferroni's post-test.*p<0.05 vs vehicle control, n = 5.</p

Biomass smoke exposure upregulates release of IL-8 from fibroblasts.

<p>Release of interleukin (IL)-8 from human lung fibroblasts (n = 10) in response to 72 hours stimulation with 1, 5, 10 or 20% biomass smoke extract (BME) in 0.1% FBS/DMEM as measured by IL-8 ELISA. Data expressed as pg/ml. Bars represent mean ± SEM. Data analysed by one-way ANOVA with Dunnet's post-test. *p<0.05 vs unstimulated, n = 10.</p

Chemical inhibition of the ERK signalling pathway attenuates biomass smoke induced fibronectin deposition.

<p>Fibroblasts were pretreated for 1(10 µM) (A,B) or UO126 (5 µM) (C,D) in DMSO (vehicle control) before stimulation with 10% (A,C) and 20% BME (B,D) in the presence of inhibitors for 72 hours, prior to analysis of fibronectin deposition by ECM ELISA. Data expressed as % of unstimulated. Bars represent mean ± SEM. Data analysed by T-Test. *p<0.05 vs vehicle control, n = 5.</p

BME activates ERK1/2 signalling molecules.

<p>Fibroblasts were stimulated for 2% CSE or 1, 5, 10 or 20% BME in 0.1% FBS/DMEM, before whole cell lysates were collected and ERK1 (A) or ERK/2 (B) phosphorylation was assessed by western blotting. Data expressed as the ratio of pERK over GAPDH (housekeeping protein). Bars represent mean ± SEM. Data analysed by one-way ANOVA with Dunnet's post-test *p<0.05 vs unstimulated, n = 5. Image at top of graph is a representative composite western blot.</p

Biomass smoke extract enhances deposition of fibronectin from fibroblasts.

<p>Deposition of fibronectin (n = 16) (A) or perlecan (n = 6) (B) from human lung fibroblasts as measured by ECM ELISA following 72 hours stimulation with 5% cigarette smoke extract (CSE) or 1, 5, 10 or 20% biomass smoke extract (BME) in 0.1% FBS/DMEM. Data expressed as absorbance at 405 nm with media alone baseline removed (A,B). Bars represent mean ± SEM. Data analysed by one-way ANOVA with Dunnet's post-test.*p<0.05 vs unstimulated, n = 6–16.</p

Biomass smoke and cigarette smoke have similar particle consistency.

<p>Quantity and distribution of particles from the smoke of one cigarette (A) or one unit of biomass (B) as measured by laser particle counter. Data expressed as particles per cubic metre or as % of total particle counts (C–D) for particles in the size ranges of 0.3–0.5, 0.5–1, 1–5, 5–10, 10–25 or <25 uM diameter. Bars represent mean ± SEM. Each experiment was performed in triplicate.</p

LPS does not induce fibronectin deposition (A) or IL-8 release (B).

<p>Human lung fibroblasts were stimulated for 72 µg/ml lipopolysaccharide (LPS) prior to analysis of fibronectin deposition by ECM ELISA (A) or IL-8 release by ELISA (B). The profibrotic cytokine TGF-β₁ (1 ng/ml) was used as a positive control for fibronectin deposition. Data expressed as % of unstimulated (A) or pg/ml (B). Bars represent mean ± SEM. Data analysed by one-way ANOVA with Dunnet's post-test.*p<0.05 vs unstimulated, n = 5.</p

High concentrations of biomass smoke are cytotoxic.

<p>Assessment of cytotoxicity of biomass smoke extract (BME) to human lung fibroblasts (n = 3) as measured by LDH assay (A), MTT assay (B) or manual cell counts (C) following stimulation with 5% cigarette smoke extract (CSE) or 1, 5, or 20% BME in 0.1% FBS/DMEM. Data expressed as % of unstimulated. Bars represent mean ± SEM. Data analysed by one-way ANOVA with Dunnet's post test.*p<0.05 vs unstimulated, n = 6.</p