Effects of TNFα and prostaglandin E₂ on the expression of MMPs in human periodontal ligament fibroblasts

Abstract

Accumulating evidence indicates that TNFalpha plays an important role in the pathogenesis of periodontitis, but the effect of TNFalpha on the degradation of the periodontal ligament is not well understood. This study used reverse transcriptase-PCR to investigate the effects of TNFalpha on matrix metalloproteinase (MMP) mRNA expression in human periodontal ligament fibroblasts. TNFalpha increased MMP-1, MMP-3 and MMP-13 mRNA levels in both a time-dependent (0-24 h) and a dose-dependent (0.1-10 ng/ml) manner. TNFalpha also increased COX-2 mRNA levels. Because elevation of COX-2 mRNA levels enhances the production of prostaglandins, we therefore investigated whether endogenous prostaglandins are involved in the MMP mRNA expression that is enhanced by TNFalpha. Pretreatment with the selective COX-2 inhibitor, NS-398, increased MMP-13 mRNA levels, while prostaglandin E2 and dibutyryl cyclic AMP decreased MMP-13 mRNA levels. Neither MMP-1 nor MMP-3 mRNA levels were affected by these chemicals. These findings indicate that prostaglandin E2 has a lowering effect on TNFalpha-enhanced MMP-13 mRNA levels, and that this effect is dependent on cAMP. Our results suggest that TNFalpha participates in periodontal ligament destruction by stimulating the production of MMPs (MMP-1, MMP-3 and MMP-13), while endogenous prostaglandin E2 has a negative feedback role in TNFalpha-enhanced MMP-13 production.

Effect of Porphyromonas gingivalis and Lactobacillus acidophilus on Secretion of IL1B, IL6, and IL8 by Gingival Epithelial Cells

“…Functionally, IL1B can promote the formation of osteoclasts, leading to destruction of bone and periodontal connective tissue and loss of periodontal tissue repair capacity. IL6 is capable of promoting the absorption of alveolar bone as well as inhibiting osteoblast alkaline phosphatase activity and collagen synthesis [23][24][25][26]. It is also known to weaken fibroblast adhesion, induce periodontal tissue expression of vascular endothelial growth factor, and promote angiogenesis, resulting in an increase in the number of inflammatory cells within the tissue leading to increased energy consumption [27].…”

Matrix metalloproteinases in the brain and blood–brain barrier: Versatile breakers and makers

“…107 To detect other MMPs, gelatin is replaced with another substrate such as collagen, carboxymethylated transferrin, or casein. 104,[108][109][110][111] Another method to detect MMP activity in vitro is by using fluorogenic MMP substrates. These artificial MMP substrates are composed of a fluorescent dye that is connected via a peptide to a quencher.…”

Dexamethasone-induced up-regulation of two-pore domain K+ channel genes, TASK-1 and TWIK-2, in cultured human periodontal ligament fibroblasts

“…After obtaining an approval of the present experiment by the Ethical Committee of the Shiga University of Medical Science, human PDL fibroblasts were prepared from healthy erupted third molars and cultured in Dulbecco's modified Eagle's Medium (DMEM) containing 10% fetal calf serum (FCS), 100 units/ml penicillin, and 100 μg/ml streptomycin under a humidified atmosphere of 95% air and 5% CO 2 at 37°C, as previously reported (Nishikawa et al 2002). The PDL fibroblasts at passages two through seven were used for the present experiments.…”

“…The PDL fibroblasts at passages two through seven were used for the present experiments. In addition, the PDL fibroblasts thus prepared were stained positive with monoclonal anti-human fibroblast surface protein antibody (Nishikawa et al 2002).…”