title: Carbodiimide cross-linking inactivates soluble and matrix-bound MMPs, in vitro.
abstract: Matrix metalloproteinases (MMPs) cause collagen degradation in hybrid layers created by dentin adhesives. This in vitro study evaluated the feasibility of using a cross-linking agent, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), to inactivate soluble rhMMP-9, as an example of dentin MMPs, and matrix-bound dentin proteases. The inhibitory effects of 5 EDC concentrations (0.01-0.3 M) and 5 incubation times (1-30 min) on soluble rhMMP-9 were screened with an MMP assay kit. The same EDC concentrations were used to evaluate their inhibitory effects on endogenous proteinases from completely demineralized dentin beams that were incubated in simulated body fluid for 30 days. Decreases in modulus of elasticity (E) and dry mass of the beams, and increases in hydroxyproline content of hydrolysates derived from the incubation medium were used as indirect measures of matrix collagen hydrolysis. All EDC concentrations and pre-treatment times inactivated MMP-9 by 98% to 100% (p < 0.05) compared with non-cross-linked controls. Dentin beams incubated in 0.3 M EDC showed only a 9% decrease in E (45% decrease in control), a 3.6% to 5% loss of dry mass (18% loss in control), and significantly less solubilized hydroxyproline when compared with the control without EDC cross-linking (p < 0.05). It is concluded that EDC application for 1 min may be a clinically relevant and effective means for inactivating soluble rhMMP-9 and matrix-bound dentin proteinases if further studies demonstrate that EDC is not toxic to pulpal tissues.
authors: Department of Prosthodontics, University of Turku, Institute of Dentistry, Lemminkaisenkatu 2, FI-20520 Turku, Finland. arztez@utu.fi
authors: A
authors: A
authors: Tezvergil-Mutluay
authors: M M
authors: MM
authors: Mutluay
authors: K A
authors: KA
authors: Agee
authors: R
authors: R
authors: Seseogullari-Dirihan
authors: T
authors: T
authors: Hoshika
authors: M
authors: M
authors: Cadenaro
authors: L
authors: L
authors: Breschi
authors: P
authors: P
authors: Vallittu
authors: F R
authors: FR
authors: Tay
authors: D H
authors: DH
authors: Pashley
mesh_terms: D000293:Adolescent; D002234:Carbodiimides / Q000008:administration & dosage / Q000494:pharmacology*; D003094:Collagen / Q000032:analysis; D003432:Cross-Linking Reagents / Q000008:administration & dosage / Q000494:pharmacology*; D003804:Dentin / Q000187:drug effects / Q000201:enzymology; D003890:Desiccation; D004305:Dose-Response Relationship, Drug; D055119:Elastic Modulus; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000201:enzymology; D005240:Feasibility Studies; D006801:Humans; D006868:Hydrolysis; D006909:Hydroxyproline / Q000032:analysis; D008422:Materials Testing; D061965:Matrix Metalloproteinase Inhibitors; D012995:Solubility; D013314:Stress, Mechanical; D013997:Time Factors; D055815:Young Adult
chemical_list: D002234:Carbodiimides; D003432:Cross-Linking Reagents; D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; C074139:1-ethyl-3-(3-dimethylaminoethyl)carbodiimide; D003094:Collagen; D006909:Hydroxyproline