Lee W, Aitken S, Sodek J, McCulloch C A
Faculty of Dentistry, University of Toronto, Ontario, Canada.
J Periodontal Res. 1995 Jan;30(1):23-33. doi: 10.1111/j.1600-0765.1995.tb01249.x.
To assess the temporal relationship between periodontal tissue destruction and the activity of collagenase, exudate from inflamed periodontal tissues was collected and latent and active collagenase activities were measured by a functional assay in a longitudinal cohort study. Comparisons were made between human subjects with either: 1) inflammation with a previous history of progressive loss of connective tissue and bone support (n = 14); 2) inflammation and previous history of bone loss but now clinically stable (n = 27); or 3) inflammation and no loss of bone support (n = 17). Experiments using specific enzyme inhibitors, blocking antibodies and SDS-PAGE fluorograph to identify the pattern of collagen substrate degradation demonstrated that the collagenase activity was derived from neutrophils and not from bacteria or other host cells. Active collagenase activity pooled from 6 sites per subject was respectively 5 and 6-fold higher in the group with progressive loss of connective tissue compared to the groups with either inflamed tissues alone or with inflammation and previous bone loss. In contrast, latent collagenase was increased up to 2 fold higher in the group with inflammation but no bone loss compared to the group with progressive lesions. Moreover, the ratio of active to total collagenase activity was 50% higher in the group with progressive lesions. Although in all subjects successive measurements of site-specific active collagenase 1 month apart demonstrated wide variation (r < 0.50), only in sites with progressive periodontal destruction was there significant increase of active collagenase with time (1.28 x 10(-4) collagenase units per day). There were also sharp elevations in active enzyme level at the time of detection of loss of connective tissue attachment in specific sites of 8 subjects. At the time of detection of connective tissue attachment loss, there was an overall 40% increase of pooled active collagenase activity in all subjects with progressive loss of connective tissue compared to pre-breakdown sampling times. These data provide strong in vivo evidence for a direct role of active neutrophil collagenase in the pathological destruction of periodontal connective tissue.
为了评估牙周组织破坏与胶原酶活性之间的时间关系，在一项纵向队列研究中，收集了炎症牙周组织的渗出液，并通过功能测定法测量了潜在和活性胶原酶活性。对以下人类受试者进行了比较：1）有结缔组织和骨支持逐渐丧失既往史的炎症患者（n = 14）；2）有骨丧失既往史但目前临床稳定的炎症患者（n = 27）；或3）有炎症且无骨支持丧失的患者（n = 17）。使用特异性酶抑制剂、阻断抗体和SDS-PAGE荧光成像来鉴定胶原底物降解模式的实验表明，胶原酶活性源自中性粒细胞，而非细菌或其他宿主细胞。与仅患有炎症组织或患有炎症和既往骨丧失的组相比，结缔组织逐渐丧失组中每个受试者6个部位汇集的活性胶原酶活性分别高5倍和6倍。相比之下，与进行性病变组相比，有炎症但无骨丧失组的潜在胶原酶增加了2倍。此外，进行性病变组中活性胶原酶与总胶原酶活性的比率高50%。尽管在所有受试者中，相隔1个月对位点特异性活性胶原酶的连续测量显示出很大差异（r < 0.50），但只有在牙周进行性破坏的位点，活性胶原酶才随时间显著增加（每天1.28×10⁻⁴胶原酶单位）。在8名受试者的特定部位检测到结缔组织附着丧失时，活性酶水平也急剧升高。在检测到结缔组织附着丧失时，与结缔组织丧失前的采样时间相比，所有结缔组织进行性丧失的受试者中汇集的活性胶原酶活性总体增加了40%。这些数据为活性中性粒细胞胶原酶在牙周结缔组织病理破坏中的直接作用提供了有力的体内证据。
