Previous studies have demonstrated that the mechanical compressive loading affects the biosynthesis of chondrocytes seeded in three dimensional scaffolds. In this study, the level of type II collagen mRNA expression was increased by a continuous dynamic compression at 10% compressive strain and 0.1 Hz in chondrocytes seeded in a biodegradable, elastomeric scaffold, poly(L-lactide-co-epsilon-caprolactone) (PLCL). To further examine this molecular mechanism, the promoter region of COL2A1 gene, which is encoding type II collagen, was analyzed using rabbit chondrocytes transfected with luciferase reporter vectors containing the 5'-flanking regions of human COL2A1 gene. A deletion mutant analysis revealed that the most active short promoter in response to continuous dynamic compression is in the region between -509 and -109 base pairs, where the transcription factor Sp1 is located. Additionally, an mRNA decay experiment using transcription inhibitor actinomycin D demonstrated that dynamic compression do not stabilize type II collagen mRNA. Our results indicate that mechanical compression increases the level of type II mRNA expression by transcriptional activation possibly through the Sp1 binding sites residing in the proximal region of the COL2A1 gene promoter. (c) 2006 Elsevier Inc. All rights reserved.