152 Adhesives activate MMP-2 and MMP-9: ELISA and zymographic assay

Thursday, March 22, 2012: 10:45 a.m. - 12:15 p.m.
Presentation Type: Oral Session
A. MAZZONI1, V. PAPA2, F. NATO3, M. CARRILHO4, A. TEZVERGIL-MUTLUAY5, K. AGEE6, L. TJADERHANE7, F. TAY8, D. PASHLEY6, and L. BRESCHI9, 1University of Bologna, Bologna, Italy, 2Department of Health Sciences, University of Cassino, Cassino, Italy, 3University of Bologna, University of Urbino, Bologna, Italy, 4Bandeirante University of Sao Paulo, Sao Paulo, Brazil, 5University of Turku, Turku, Finland, 6Dept. Oral Biology, Georgia Health Sciences University, Augusta, GA, 7Institute of Dentistry, University of Oulu, Oulu, Finland, 8Oral Biology & Maxillofacial Pathology, Georgia Health Sciences University, College of Dental Medicine, Augusta, GA, 9Department of Medical Sciences, University of Trieste, Trieste, Italy
Objectives: MMP-2 and -9 were claimed to participate in the degradation of collagen fibrils that compromises the integrity of the hybrid layer. This study tested the null hypothesis that there are no differences in MMP-2 and -9 activity after treatment with etch-and-rinse or self-etch adhesives.

Methods : Powdered dentin prepared from extracted human teeth was treated as follow: 1) untreated mineralized powder; 2) demineralized with 10% H3PO4 or demineralized as for group 2 then treated with the following etch-and-rinse adhesives: 3) Adper Scotchbond 1XT (3M ESPE); 4) PQ1 (Ultradent); 5) Peak LC (Ultradent); 6) Prime&Bond NT (Dentsply); or left mineralized and treated with self-etch adhesives: 7) Adper Easy Bond (3M ESPE) or 8) TRIS (Kuraray). MMP-2 and -9 activity in adhesive-treated dentin powder was determined using gelatin zymography and ELISA test (Biotrak™ activity assay system; GE Healthcare, Buckinghamshire, UK). Assays were performed in triplicate.

Results : For etch-and-rinse adhesives (G3-6), gelatin zympography showed increased activity of MMP-2, while MMP-9 was adhesive dependent. For self-etch adhesives (G7-G8), zymography showed activation for both MMP-2 and -9. Elisa test revealed increased MMP-2 and -9 activity after all adhesive treatments. Increased MMP-2 and -9 activity was found after treatment either with etch-and-rinse and self-etch adhesives and the extent of activation was adhesive-dependent.

Conclusions : The hypothesis tested was rejected since differences in MMP-2 and -9 activities were recorded. It is concluded that all two-step etch-and-rinse and the one-step self-etch adhesives tested can activate endogenous MMP-2 and MMP-9 in dentin. These results support the role of endogenous MMPs in the degradation of hybrid layers created by these adhesives.

Supported, in part, by grants FIRB RBAP1095CR, PRIN 2009SAN9K5 and 2009FXT3WL from MIUR, Italy, and R01DE015306 from the NIDCR to DP (PI) and grant #8126472 from the Academy of Finland to AT-M (PI).

This abstract is based on research that was funded entirely or partially by an outside source: Supported, in part, by grants FIRB RBAP1095CR, PRIN 2009SAN9K5 and 2009FXT3WL from MIUR, Italy, and R01DE015306 from the NIDCR to DP (PI) and grant #8126472 from the Academy of Finland to AT-M (PI)

Keywords: Collagen, Dentin, Dentin bonding agents and Enzymes