Effects of tissue destructive proteinases on conective tissue attachment

采用ELISA法对18例患者龈缝组织破坏中性蛋白酶进行检测。中性粒细胞弹性蛋白酶和组织蛋白酶G在最小探测深度（1 ~ 2mm）部位的平均浓度最低。在中等深度（3-5mm）和严重深度（大于6mm）的裂缝中，它们的浓度随深度的增加而增加。这些酶可能在龈沟区组织破裂中起重要作用。在接下来的实验中，我们考察了影响结缔组织附着的因素。根片由手术拔除的人类正常牙和牙周受累牙制备。每个处理过的根片在含有1ml a- mem的1 × 10^4个细胞的24孔板中孵育。测定其形态变化、贴壁细胞数、DNA合成及碱性磷酸酶活性。正常（未受影响）根上的成纤维细胞呈扁平和纺锤状。病根表面的细胞呈圆形或微伸长。在培养第1天和第4天，正常根片上的细胞数量明显高于患牙周根片。在培养第1、4和7天，病根表面细胞的DNA含量被抑制到正常根的50%。在56 d的培养过程中，正常根片上的细胞与病根片上的细胞碱性磷酸酶活性有显著差异。从牙周受累根表面洗脱到介质中的内毒素量明显高于正常根表面。提示本实验系统可用于确定组织破坏蛋白酶对结缔组织附着的影响。此外，创伤力影响结缔组织附着继续检查的手段研究磨牙症。

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.