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Matrix Metalloproteinase-3 Differences in Oral and Skin Fibroblasts

S.T.W. McKeown^1,, J.J. Barnes^1,, P.L. Hyland², F.T. Lundy¹, M.J. Fray³, and C.R. Irwin^1,*

¹ Oral Science Research Centre, School of Dentistry, Queen’s University Belfast, Northern Ireland;
² Centre for Cancer Research and Cell Biology, Queen’s University, Belfast; and
³ Department of Discovery Chemistry, Pfizer Global Research and Development, Sandwich, Kent, England

View larger version (8K): [in this window] [in a new window]   Figure 1. Kinetics of fibroblast-induced gel contraction. (a) Collagen gel contraction by paired oral and skin fibroblasts under control conditions. Oral fibroblasts induced significantly increased contraction compared with skin cells at all concentrations studied (p < 0.05). (b,c) Effect of total MMP (GM1489) and MMP-3 inhibition on collagen gel contraction kinetics by (b) oral and (c) skin fibroblasts over a seven-day study period. Both inhibitors significantly reduced gel contraction by oral fibroblasts at all time-points (p < 0.05); GM1489 significantly inhibited gel contraction by skin fibroblasts at all time-points (p < 0.05). Studies were carried out in triplicate on 3 paired skin and oral fibroblast populations. Data are expressed as mean ± SE.

View larger version (34K): [in this window] [in a new window]   Figure 2. Effect of total MMP (B-MMP-I) and MMP-3 (MMP-3-I) inhibition on SMA expression by oral and skin fibroblasts cultured (a) in monolayer, (b) in attached collagen gels, and (c) in floating collagen gels (representative of duplicate experiments for 3 paired oral and skin populations).

View larger version (55K): [in this window] [in a new window]   Figure 3. MMP-3 mRNA expression. (a) MMP-3 RT-PCR products in oral and skin fibroblasts relative to ß-actin products for control and TGF-ß-treated conditions. Representative of duplicate experiments for 3 paired oral and skin populations. Appropriate PCR-negative blanks and RT controls were used during all electrophoresis runs (data not shown). (b) Effects of TGF-ß1 and TGF-ß3 on MMP-3 mRNA expression indices. Data are expressed compared with control cultures following normalization of control values for each cell population to 100%. Treatment with both factors, at all concentrations, induced a statistically significant increase in MMP-3 expression (p < 0.05).

View larger version (24K): [in this window] [in a new window]   Figure 4. Effects of TGF-ß1 and TGF-ß3 on MMP-3 protein and activity levels in oral and skin fibroblast cultures. Each plot represents the mean ± SE of 3 paired fibroblast populations. Data are expressed as percentage change from control, where control values have been normalized to 100%. All culture conditions resulted in significantly increased protein and activity levels compared with controls (P < 0.05).