• Title/Summary/Keyword: Spectrophotometric collagenase assay

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Collagenolytic Activity Of Gingival Crevicular Fluid In Progressive Periodontitis (진행성 치주염에서의 치은열구액내 교원질분해효소 활성)

  • Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.161-175
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    • 1996
  • There were many reports that elevations in the levels of active and latent collagenase in gingival crevicular fluid(GCF) have been correlated positively with periodontal disease activity. To provide a simple diagnostic approach for testing GCF collagenolytic activity, the detection limit of enzyme activity was compared using radiofibril assay(Sodek et.al.1981) and spectrophotometric collagenolytic assay(Nethery et al. 1986). The detection limits of both assay for standard bacterial enzyme were similar and the radiofibril assay showed a little (1/2) lower detection limit for tad pole collagenase. To evaluate the relationship between periodontal tissue destruction and the collagenolytic activity, GCF was collected, and latent and active enzyme activities were measured by a spectrophotometric collagenolytic assay. Twelve subjects showing progressive lesions were selected according to the presence of immediate tissue destruction, frequent abscess formation, and increasing need for tooth extraction, and the absence of underlying systemic disease and previous antibiotic medication history within 6 months. Comparisons were made between sites with either: 1) inflammation with a previous history of progressive loss of periodontal tissue and bone support(2l progressive sites): 2) previous history of bone loss and periodontal destruction but now clinically stable(12 comparably stable sites); or 3) no loss of periodontal tissue and bone support(11 control sites including 5 gingivitis sites and 6 healthy sites). Active collagenase activity was the highest in the progressive sites and decreased in the order of the gingivitis sites, the stable sites, and the healthy sites. The total enzyme activity was $2{\sim}3$ fold higher in the progressive sites and the gingivitis sites, compared to the stable and the healthy sites. The ratio of active to total collagenolytic activity was twice in the progressive sites. Analysis of active collagenase level(5mU) and the ratio of active to total collagenolytic activity(0.8) as a diagnositic test indicates that these measurements have the sensitivity of 0.81 and 0.86, the specificity of 0.70 and 0.65, and the overall agreement of 0.75 and 0.73, respectively. Thus, this method has significant merits as a diagnostic tool to determine wherher the site is in a state of remission or progression.

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In Vitro Effects of Female Sex Hormones on Collagenase Activity of Gingival Fibroblast and Periodontal Ligament Fibroblast (여성 호르몬의 변화가 치은 섬유아세포와 치주인대세포의 교원질 분해 효소의 활성에 미치는 영향)

  • Sin, Ji-Yearn;Lee, Chul-Woo;Han, Soo-Boo
    • Journal of Periodontal and Implant Science
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    • v.29 no.1
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    • pp.31-40
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    • 1999
  • Many factors may affect periodontal changes during the physiologic conditions of woman(e.g. puberty, menstrual cycle, pregnancy, menopause). Recently many research has focused on the immunological changes of host, but the exact mechanism is not clear. Collagen is a major constituent of periodontium, and collagenase specifically digests the collagen and plays a role in destruction of periodontal tissue. So, I suppose that it participates with the cytokines in the inflammation of gingiva and vascular response during the changes of female sex hormones. Because there are some evidences of the existence of the receptors of estrogen and progesterone in the gingiva, it may be a target tissue of female sex hormones. In this experiment, gingival fibroblast and periodontal ligament cell were cultured in the presence of various concentrations of estrogen or progesterone corresponding to the menstrual cycle and pregnancy. Collagenase activity of the supernatant of culture media was determined by Spectrophotometric collagenase assay. The enzyme activity was calculated by the % decrease of the coated collagen. 1. The estrogen at both concentrations had no effect on the activity of collagenase of the gingival fibroblast. 2. The progesterone had some effect on the collagenase activity of the gingival fibroblast at low and high concentration of menstrual cycle, and elevated the enzyme activity at all range of pregnancy concentrations. 3. In periodontal ligament cells, estrogen elevated the enzyme activity at the early pregnancy concentration and progesterone elevated at the concentration just before menstruation. In this experiment, pregesterone elevated the collagenase activity of gingival fibroblast and periodontal ligament cells. But the mechanism of the up-regulation of the enzyme activity was not confirmed. The more experiments of direct effect of progesterone on gingival at the molecular level(e.g. northern blot analysis) can reveal the exact mechanism.

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