Increased pro-inflammatory mediators and ECM deposition, are key features of the airways in asthma. Matrix metalloproteinases (MMPs) are produced by airway smooth muscle (ASM) cells and have multiple roles in inflammation and tissue remodelling. We hypothesised that components of the asthmatic airway would stimulate MMP production and activation by ASM and contribute to airway remodelling. We measured human ASM derived MMP mRNA, protein and activity by real-time RT-PCR, zymography, western blotting and MMP activity assay. Collagen I and thrombin caused a synergistic increase in MMP-2 protein and total MMP activity but paradoxically decreased MMP-2 mRNA. Additionally collagen I activated MMP-2 in culture supernatants independent of the cell surface. Together collagen I and thrombin strongly enhanced MMP-14 mRNA and protein but had no effect individually suggesting increased MMP-14, the activating protease for MMP-2, may be partially responsible for MMP-2 activation. Further, collagen I reduced TIMP-2 protein, an MMP-2 inhibitor. We examined the role of MMPs in functions of ASM related to airway remodelling and found migration and proliferation were MMP dependent, whereas adhesion and apoptosis were not. Ilomastat inhibited migration by 25% which was also inhibited by TIMPs 1-4 and increased by the MMP-2 activator thrombin. These in vitro findings suggest the environment within the airways of patients with asthma enhances MMP-2 and -14 protein and activity by a complex interaction of transcriptional and post transcriptional mechanisms which may contribute to ASM migration.