Down Regulation of MMP-2 Expression and Invasion Potential of Human Gingival Fibroblasts

M.W. Roomi, V. Ivanov, A. Niedzwiecki, M. Rath
Matthias Rath Research., Cancer Research Division, Santa Clara, CA 95050

Presented at: XXVIII Scientific Meeting of the South African Division of the International Association for Dental Research, Pretoria, South Africa, September 2-3, 2004

Published in: Conference proceedings, Abstract #37, pg 42

Abstract

Background:
Gingivitis, a condition in which gums swell, turn red, and bleed easily, is the earliest stage of gum disease, affecting three out of four adults over the age of 35. Left untreated, it progresses to periodontitis, an advanced stage of gum disease characterized by additional destruction of gum tissue and progression to the bone. MMPs play a critical role in this process as, in concert, they degrade the majority of the extracellular matrix components.

Objective:
We investigated the effect of a unique formulation (NM) containing ascorbic acid, lysine, proline, arginine, and epigallocatechin gallate on gingival fibroblast growth, MMP-2 secretion, and invasion potential.

Materials and Methods:
Human gingival fibroblasts CRL-2014 (ATCC) were grown in DME medium supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 mg/ml) in 24-well tissue culture plates. At near confluence, the cells were treated with NM dissolved in media and tested at 0, 10, 100, 500, and 1000 ?g/ml in triplicate at each dose. Cell proliferation/ cytotoxicity was evaluated by MTT assay, MMP expression by gelatinase zymography, and invasion through Matrigel.

Results:
The nutrient mixture demonstrated slight, dose-dependent inhibition of gingival fibroblast growth with maximum inhibition of 30% at 1000 µg/ml (p=0.001). NM inhibited gingival fibroblast MMP-2 expression in a dose-dependent fashion with total virtual inhibition at 500 µg/ml. The synergistically acting nutrient mixture significantly reduced the invasion of human gingival cells through Matrigel in a dose-dependent manner, with 76% inhibition at 50 µg/ml and 100% at 500 µg/ml NM (p<0.0001).

Conclusion:
Our results suggest that NM has potential as a therapeutic tool in treatment of periodontal inflammation, through inhibition of gingival fibroblast MMP-2 expression and matrix invasion.