Effects of tissue destructive proteinases on conective tissue attachment

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The tissue destructive neutral proteinases in the gingival crevice of 18 patients were determined by ELISA assay. The average concentrations of neutrophil elastase and cathepsin G in sites of minimum probing depth (1-2mm) were least. In crevices with moderate (3-5mm) and severe (6mm of more) depth there was increase of their concentrations in proportion to the depth. Those enzymes may play a significant role in tissue breakdown in gingival crevicular area.As the following experiment, were examined the factors which influenced connective tissue attachment. Root slices were prepared from surgically extracted human normal and periodontally involved teeth. Each treated root slice was incubated in a well of 24-well plate containing 1x10^4 cells in 1ml of a-MEM. Morphological changes, the numbers of attached cells to the root slices, DNA synthesis and Alkaline phosphatase activity of the attached cells were determined. Fibroblasts on normal (unaffected) roots appeared flat and spindle-shaped. The cells on diseased root surfaces were rounded or slightly elongated. The number of cells attached on normal root slices was higher than on periodontally involved root slices on day 1 and 4 culture. The quantity of DNA of the cells on diseased root surfaces was inhibited to 50% of normal roots on day 1, 4 and 7 culture. Significant differences in the alkaline phosphatase activities of cells were found between cells attached on normal root slices and on diseased root surfaces through the 56 days of culture period. The amount of endotoxin eluted from periodontally involved root surfaces into media was significantly higher than from normal root surfaces.It is suggested that our experimental system should be useful to determine the effects of tissue destructive proteinases on connective tissue attachment.Furthermore, traumatic forces affected connective tissue attachment are continued to examine by means of the study of bruxism.