[The effect of matrix metalloproteinase-1 on root surface dentin matrix: a scanning electron microscope observation].

OBJECTIVE: To observe the effect of matrix metalloproteinase-1 (MMP-1) from human host on degradation of dentin organic matrix of root dentin. METHODS: The freshly extracted caries-free impacted teeth were selected. Teeth were cut transversely under the enamel-cementum junction into dentin sections with a thickness of about 5 mm. Then all sections with removal of cementum, pulp and predentin were randomly divided into four groups. In the first group, dentin sections were demineralized with acid solution for 21 days, and then incubated with MMP-1 solution for 7 days; the second group were only treated with acid solution for 21 days; the third group were only attacked by MMP-1 solution for 7 days; and the fourth group were untreated as a control. Then all sections were dehydrated in ascending strength of alcohol, critically dried, coated with platinum, and then observed under scanning electron microscope(SEM). RESULTS: The dentin sections of root surface attacked by acid and MMP-1 showed that demineralization of dentin mineral and degradation of dentin matrix fibrae synchronously happened. The dentin matrix fibrae wasn't degradated in the groups treated with acid or MMP-1. CONCLUSION: The proteinases from human host may play an important role in the development of root surface caries. MMP-1 may distinctly degradate the organic matrix of demineralized dentin.

Host-derived proteinases and degradation of dentine collagen in situ.

Dentine root caries is a process of demineralization and degradation of the organic matrix by proteinases. In this in situ study, the presence and activity of the matrix metalloproteinases 1, 2 and 9 (MMP-1, MMP-2, MMP-9) in saliva and in completely demineralized dentine specimens were investigated. Furthermore, the activity of cathepsin B was determined in saliva. A correlation between these enzymes and the level of degraded collagen was investigated. Demineralized dentine specimens were mounted in the partial prosthesis of 17 volunteers. Saliva samples were taken at 0, 2 and 4 weeks. After 4 weeks, the enzymes were extracted from the dentine specimens and the collagen loss was assessed. The collagen loss varied between 0 and 40.3%. Zymography of the saliva and the dentine extract samples showed that (pro-)MMP-2 and (pro-)MMP-9 were present. The levels of active MMPs were assessed, using fluorogenic MMP-specific substrates. All but 3 of the 51 saliva samples showed MMP-1 activity ranging from 1.5 to 101.1 relative fluorescence units (RFU)/s. Forty-eight saliva samples showed gelatinolytic MMP-2/MMP-9 activity (1.7-141.1 RFU/s). MMP-1 activity was shown in all dentine extracts varying between 3.5 and 295.0 RFU/s. From the dentine extracts, 15 showed MMP-2/MMP-9 activity (0.2-13.7 RFU/s). The MMP activity from both saliva and dentine extracts did not correlate with the collagen loss. The activity of salivary cathepsin B varied from 4.8 to 42.2 arbitrary units/min. A positive correlation was found between salivary MMP activity and cathepsin B activity. This study revealed that gelatinolytic enzyme activity was present both in saliva and dentine collagen. No correlation could be observed, however, between the level of enzyme activity and the collagen loss of the dentine specimens.

Effects of collagenase on root demineralization.

The role of proteolytic enzymes in the root caries process remains unclear. The aim of this study was to investigate collagenase activity during tooth root demineralization and remineralization in an in vitro demineralization/remineralization pH-cycling model, Human tooth roots were subjected to pH cycling (alternating demineralization and remineralization) in one of two different time cycles for five days. Collagenase at 90, 180, or 360 micrograms per root was placed into either the demineralizing solution or the remineralizing solution in the pH-cycling system. The effects of additional exposure to collagenase before or after pH cycling were also studied. After the exposure, thin sections of the roots were examined histologically by polarized light microscopy. Changes of calcium and phosphate in the solutions were analyzed chemically. Surface erosion occurred only in the groups where collagenase was contained in the remineralizing solution and in which the root samples were exposed to severe demineralization. However, no differences among the control and experimental groups were found in calcium and phosphate changes in the pH-cycling solutions. These findings suggest that collagenase works during the remineralizing phase and predominantly attacks the organic matrix of the root after demineralization. Additional exposure to collagenase before or after pH cycling did not increase surface erosion except for exposure to collagenase in the absence of phosphate following pH cycling.