【作者】 毛钊；

【导师】 吴织芬；

【作者基本信息】 第四军医大学 ， 口腔临床医学， 2002， 博士

【摘要】 老年人牙周炎的发病率及轻重程度随年龄递增，且远高于其它年龄组。一般认为这与老年人组织抗感染能力、免疫功能、组织修复能力、抗氧化能力、一氧化氮水平和细胞凋亡等多种局部和全身因素变化有关。但目前国内外对老年人牙周炎病因病理学研究较少。据此，我们进行了老年人牙周膜细胞生物学特性及老年牙周炎病理过程研究，采用人牙周膜细胞原代培养法进行了老年人牙周膜细胞增殖（MTT法）和碱性磷酸酶活性的测定；采用免疫组织化学染色法观察了老年人牙周炎牙龈组织中T和B淋巴细胞、NK细胞、凋亡细胞、Fas/Apo-1(CD₅₉)抗原和诱导型一氧化氮合酶（iNOS）的分布和阳性表达情况；测定了外周血中性粒细胞功能和NK细胞活性，牙龈组织中超氧化物歧化酶（SOD）和丙二醛（MDA）的含量及一氧化氮的水平。得出如下结果和结论： 1 老年组牙周膜细胞生长极其缓慢，细胞显衰老状态，较青年组牙周膜细胞表现明显不同。老年组牙周膜细胞在1、3、5、7、9天时增殖的光密度值(A₅₇₀)均明显低于青年组，比较有显著性差异（P＜0.01）。结论：由于增龄的影响，老年人牙周膜细胞生长增殖功能明显降低。 2 老年组牙周膜细胞碱性磷酸酶活性明显低于青年组（P＜0．01）；且对小牛血清刺激增殖的反应也明显低于青年组（P＜0.01）。结论：老年人牙周膜细胞成骨活性可能降低，对外源性促细胞生长因素的敏感性降低。 3 老年组牙周膜细胞Fas/Apo-1(CD₉₅)抗原细胞核周胞浆强阳性表达，且强于青年组；老年组牙周膜细胞iNOS阳性表达，而青年组牙周膜细胞iNOS阴性表达。结论：增龄造成的老年人牙周膜细胞生长增殖能力的减弱与老年人牙周膜细胞对细胞凋亡的敏感性增强有关。 训)‘博厂大学博十学位论义《老年人牙J刷Q圳胞生物学特忡及老年牙川炎刑理过程研究》4 老年慢性牙周炎组中性粒细胞移动指数明显低于成人慢性牙周炎组和 青少年牙周炎组o刃．05，o．01卜吞噬能力和吞噬活性均较成人慢性牙周炎组。 和青少年牙周炎组明显减低爬吻．0八。结论：老年牙周炎患者中性粒细胞功 能明显降低5 老年慢性牙周炎组外周血NK细胞活性明显低于成年慢性牙周炎组 o吻．01卜 也低于青少年牙周炎组o吻．05卜结论：老年慢性牙周炎患者外 周血NK细胞活性低下，功能降低。6 成人慢性牙周炎组NK阳性细胞计数明显高于老年慢性牙周炎组 （＜0．01卜成人慢性牙周炎组＊K阳性细胞表达率也高于老年慢性牙周炎组。 结论：增龄使老年慢性牙周炎患者局部牙龈组织中出现兔疫调节功能的紊 乱，NK细胞功能降低。7 老年慢性牙周炎组IgM阳性表达例数0 例）多于IgA阳性表达例数o 例)，而成人慢性牙周炎组则相反。成人慢性牙周炎组CD4”和CDS”细胞 数明显多于老年慢性牙周炎组o功．05卜 结论：老年馒性牙周炎组患者牙 龈组织局部兔疫功能降低，免疫调节功能紊乱，有其独特的兔疫组织化学 表现。吕 老年慢性牙周炎组SOD水平明显低于健康老年人组、成人慢性牙周炎 组和青少年牙周炎组o吻．05)；其M＊A水平明显高于健康老年人组、成人 慢性牙周炎组和青少年牙周炎组（P＜o．05）。结论：增龄和牙周炎症双重 作用造成老年慢性牙周炎患者牙周组织局部抗氧化能力降低，氧化物质堆 积。9 老年慢性牙周炎组NO水平明显低于健康老年人组、成人慢性牙周炎 组和青少年牙周炎组o刃．05卜 结论：由于增龄和牙周炎症的双重作用， ．老年慢性牙周炎患者牙龈组织生成和分泌NO明显减少，导致抗感染和免 疫调节功能的改变，非特异性免疫功能降低。 6 第叫军医人学博D：学位冰义《老年人牙协w驯胞生物学什叮及老年牙川炎灿I坐过汉盯1卜状10 老年慢性牙周炎组阳性凋亡细胞总例数明显高于健康老年人组和成人 慢性牙周炎组（P＜0．05）；老年慢性牙周炎组阳性凋亡细胞计数明显高于成 人慢性牙周炎组、青少年牙周炎组及健康老年人组(P＜0．05卜结论：牙周 炎症和增龄均能促进老年人牙周炎时牙龈组织细胞凋亡。11 在老年慢性牙周炎组完整的牙龈鳞状上皮间桥、核周胞浆 Fas／Apo－l(CD，，航原阳性表达和结缔组织中淋巴细胞FaslApo－l（CD，s）抗 原阳性表达为4组中最强；老年慢性牙周炎组上皮细胞和上皮细胞的细胞 间桥FaslAPo＊(CD帖航原阳性表达例数明显高于成人慢性牙周炎组和青 少年牙周炎组o叩．05、结论：由于牙周炎症和增龄的影响，在老年慢性牙 周炎患者牙龈组织中上皮细胞和炎性细胞对细胞凋亡易感性明显增强。12 老年慢性牙周炎组血管壁内皮细胞、结缔组织内炎症细胞、上皮乳头 诱导型一氧化氮合酶阳性表达例数明显低于青少年牙周炎组和成人慢性牙 周炎组（P＜o．05），血管壁内皮细胞和胶原纤维阳性表达例数低于健康老年 人组(P＜0．05卜结论：由于增龄和炎症的双重影响，老年慢性牙周炎患者 牙龈组织中诱导型一氧化氮合酶的表达明显降低，造成了局部一氧化氮更多还原

【Abstract】 The periodontitis in elderly peoples and injure is increasing with aging , moreover it is far higher in elderly people than in other peoples.Generally it is thought these are related to the changes of ability of anti-infection and repairment of tissues , function immunology , ability of anti-oxidation , level of NO and apoptsis. According to those theories , we design a series of pexperiments to study on biological character of periodontal ligament cells in elderly people and pathologic changes of periodontitis of elderly peoples with methods of immunohistochenical , MTT and spectrphotometry. We can get conclusions as:1 Growth of elderly healthy donors periodontal ligament cells were much slower than that of young healthy donors ; proliferation(As7o) of elderly healthy donors periodontal ligament cells were considerably lower than that of young healthy donors on 1 ,3 , 5, 7, 9 day (P<0.01) . Conclusions: Under the influence of ageing , proliferation ability of elderly peoples periodontal ligament cells was significantly decreased and was significantly different from that of young peoples. These showed that ageing might be one of important reasons which result in decreasing in repairing and proliferation ability of elderly peoples periodontal tissues.2 Elderly peoples PDL cells exhibited significantly lower alkalinephosphatase activity than the young peoples PDL cells ( P<0.01). Their ability to respond to fetal calf serum ( PCS ) was also lower than that of young peoples PDL cells ( P<0.01 ). Conclusion: These results suggest that ability to form mineralized tissue in elderly peoples PDL cells may be altered , functions affecting the maintenance and regeneration of the periodontium .3 The expression of Fas-antigen in periodontal ligament cells of elderly peoples was obviously heavier than that in periodontal ligament cells of young peoples . The expression of iNOS in periodontal ligament cells of elderly peoples was positive, but the expression of iNOS in periodontal ligament cells of young peoples was negative. Conclusions: The decrease of proliferation of periodontal ligament cells of elderly peoples was relative to increasing of sensitivity to apoptosis in periodontal ligament cells of elderly peoples.4 Chemotaxis and phagocytosis of neutrophil leukocytes in elderly patients with periodontitis were much lower than those in adult and juvenile patients with periodontitis(P < 0.05 , P < 0.01). Conclusion : Defect of host defense may be one of the major factors causing periodontitis in elderly patients.5 The activity of NK cells of venous blood in elderly patients with periodontitis was significantly lower than that in adult patients (P < 0.01) and juvenile patients (P < 0.05 ) with perodontitis. Conclusion: These data suggested that the low activity of NK cells of venous blood in elderly patients with periodontitis may be one of the major factors causing serious periodontitis in elderly patients with periodontitis.6 The incidence ol NK ceils positive cases in gingiva in elderly patients with periodontitis was less then the incidence of adult patients.The number of positive NK cells was less than the number of positive in adult patients. Conclusion: These results suggest that the functions NK cells in gingiva may be low in elderly patients with periodontitis.7 In elderly patients with periodontitis, the cases of IgM positive cells in gingiva (15 cases) were more than those of IgA positive cells(4cases), which was contrary to the adult patients with periodontitis.the number of CD4+ and CDs" lymphocytes in gingiva in adult patients with periodontitis was much greater than that in elderly patients (P <0.05). Conclusion: Elderly patients with periodontitis have certain specific characteristics in immunohistochemitry , such as low immunological function in local gingival and poor function of immunological regulation.8 Level of SOD in eldly patients group was significantly lower than those in health elder group, adult patients group and j更多还原