老年牙周炎牙龈组织Fas/Apo-1(CD95)抗原表达的免疫组织化学研究

目的 :观察慢性老年牙周炎牙龈组织中Fas/Apo - 1(CD95 )抗原表达和分布情况。方法 :采用免疫组织化学染色方法对 10例慢性老年牙周炎患者、10例慢性成人牙周炎患者、10例青少年牙周炎患者和 10例健康老年人牙龈组织中Fas/Apo - 1(CD95 )抗原阳性表达和分布情况进行了观察和比较。结果 :在慢性老年牙周炎组完整的牙龈鳞状上皮间桥、核周胞浆Fas/Apo - 1(CD95 )抗原阳性表达和结缔组织中淋巴细胞Fas/Apo - 1(CD95 )抗原阳性表达为 4组中最强 ;健康老年人组上皮角化层Fas/Apo - 1(CD95 )抗原阳性表达为 4组中最强 ;慢性成人牙周炎组和青少年牙周炎组结缔组织中淋巴细胞和吞噬细胞Fas/Apo - 1(CD95 )抗原阳性表达强于健康老年人组 ;慢性老年牙周炎组上皮细胞和上皮细胞的细胞间桥Fas/Apo - 1(CD95 )抗原阳性表达例数明显高于慢性成人牙周炎组和青少年牙周炎组 (P <0 .0 5 ) ;健康老年人组结缔组织中淋巴细胞和吞噬细胞Fas/Apo - 1(CD95 )抗原阳性表达例数明显低于其它 3组 (P <0 .0 5 )。结论 :由于感染性炎症和衰老的影响 ,①牙周炎患者和健康老年人牙龈组织中鳞状上皮凋亡细胞发生部位与身体其他器官鳞状上皮细胞凋亡发生部位不同 ;②在慢性老年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     

老年牙周炎牙龈组织诱导型一氧化氮合酶分布研究

目的 :观察慢性老年牙周炎患者牙龈组织中诱导型一氧化氮合酶 (iNOS)分布。方法 :采用免疫组织化学方法对 10例慢性老年牙周炎患者、10例慢性成人牙周炎患者、10例青少年牙周炎患者和 10例健康老年人牙龈组织中诱导型一氧化氮合酶分布进行了检测并比较研究。结果 :(1)牙周炎时牙龈组织中诱导型一氧化氮合酶主要在鳞状上皮细胞胞浆核周区颗粒状阳性表达 ,毛细血管壁内皮细胞、老化的胶原纤维及上皮下基底膜共同形成了一种乳头状轮廓样阳性表达形态 ,结缔组织和肉芽组织中各类炎症细胞也显阳性表达 ;(2 )慢性老年牙周炎组血管壁内皮细胞、结缔组织内炎症细胞、上皮乳头阳性表达例数明显低于青少年牙周炎组和慢性成人牙周炎组 (P <0 .0 5 )。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 .0 5 )。结论 :慢性老年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(NO)合成减少 ,引起了局部牙龈组织免疫功能降低和免疫调节功能紊乱     

年龄对NK细胞在牙周炎龈组织中分布影响的免疫组化研究

目的：观察老年牙周炎牙龈组织中自然杀伤（NK）细胞的分布。方法：采用免疫组织化学方法对15例老年牙周炎患者10例慢性成人牙周炎患者牙龈组织中NK细胞、T淋巴细胞和B淋巴细胞的分布及阳性计数进行比较研究。结果：慢性成人牙周炎组NK阳性细胞计数明显高于老年牙周炎组（P＜0.01)，慢性成人牙周炎组NK阳性细胞表达率也高于老年牙周炎组。结论：由于年龄增加的影响，老年牙周炎患者局部牙龈组织中出向免疫调节功能的紊乱，NK细胞功能降低。     

老年牙周炎患者牙龈组织NO水平研究

目的：观察老年牙周炎患者牙龈组织中NO水平变化并与健康老年人、慢性成人牙周炎患者、青少年牙周炎患者比较。方法：采用硝酸还原酶法测量老年牙周炎患者、健康老年人、慢性成人牙周炎患者和青少年牙周炎患者牙龈组织中NO含量并比较。结果：老年牙周炎组NO水平明显低于健康老年人组、慢性成人牙周炎组和青少年牙周炎组（P＜0．05）。结论：由于衰老和牙周炎症的双重作用,老年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。     

慢性牙周炎牙龈组织中细胞凋亡的研究

目的：观察慢性牙周炎牙龈组织中的凋亡细胞,并探讨其发生凋亡的机制。方法：采用透射电镜法观察牙龈组织凋亡细胞的超微结构、利用免疫组化方法检测20例慢性牙周炎患者及20例牙周健康者牙龈组织中凋亡蛋白Casepase-9、Cyt-C的表达并进行统计学分析。结果：慢性牙周炎患者牙龈组织中可观察到凋亡细胞,Case-pase-9、Cyt-C的阳性表达显著高于牙周健康组（P〈0.05）。结论：慢性牙周炎较牙周健康者龈组织发生明显的细胞凋亡现象,并依赖于线粒体途径。     

慢性牙周炎牙龈组织细胞凋亡及凋亡相关蛋白Caspase-3的表达

目的:研究慢性牙周炎牙龈组织中细胞凋亡的发生情况和Caspase-3蛋白的表达,探讨其在慢性牙周炎病变发生中的意义。方法:应用脱氧核糖核苷酸末端转移酶介导的原位缺口末端标记法(TUNEL法)、免疫组织化学方法检测21例慢性牙周炎牙龈组织和21例健康牙龈组织中的细胞凋亡指数(apoptosis index,AI)及Caspase-3蛋白的表达。结果:慢性牙周炎组牙龈组织中细胞凋亡指数明显高于正常对照组(P<0.05)。与正常组相比,慢性牙周炎组牙龈组织中细胞caspase-3表达明显增强,两组间有显著性差异(P<0.05)。结论:慢性牙周炎病人牙龈组织细胞发生凋亡,且通过激活细胞凋亡信号传导途径中的Caspase-3而导致慢性牙周炎发生。     

牙周炎患者血清、龈沟中的IL-8检测及其局部病变组织的免疫组织化学研究

目的 :探讨龈沟液中白细胞介素 8(IL - 8)的来源及IL - 8与成人牙周炎的关系。方法 :用ELISA法检测了30份血清样本 (13例牙周健康者 ,17例成人牙周炎患者血清 )和 2 7份龈沟液样本 (9例牙周健康者 ,18例成人牙周炎患者 )中IL - 8的水平。用免疫组织化学的方法检查了IL - 8分泌细胞在牙龈组织中的分布 (7例健康牙龈组织 ,18例成人牙周炎牙龈组织 )。结果 :成人牙周炎患者中血清IL - 8的检出率 (4 7% ,8 18)显著高于健康人血清中IL- 8的检出率 (7.7% ,1 13) ,P <0 .0 5 ;患者血清与患者龈沟液 ,健康者血清与健康者龈沟液之间 ,血清中IL - 8的含量均显著低于龈沟液 (P <0 .0 1)。IL - 8阳性细胞分布于牙龈的口腔龈上皮、结缔组织及沟内的上皮中 ;且结缔组织及沟内上皮中的IL - 8阳性细胞数量 ,成人牙周炎组显著高于健康对照组。结论 :龈沟液中的IL - 8主要来源于局部牙龈组织。IL - 8参与了牙周炎的病理过程。     

糖尿病型牙周炎牙龈组织中炎症因子表达及与细胞凋亡关系

目的:探讨糖尿病型牙周炎(diabetes associated periodontitis,DAP)牙龈组织中凋亡细胞发生及与IL-1β和TNFα表达的关系.方法:纳入DAP患者和健康龈(H)受试者各20例,用HE染色和Tunnel法观察牙龈细胞凋亡,透射电镜观察凋亡细胞超微结构;用免疫组化(IHG)检测牙龈组织炎症因子IL-1β和TNFα表达.结果:DAP组牙龈上皮棘细胞层和基底细胞层见明显细胞凋亡,固有层凋亡细胞较少;DAP组牙龈上皮棘细胞和基底细胞凋亡百分率高于H组(P＜0.01);IHC染色发现DAP组IL-1β和TNFα表达明显高于H组,主要阳性表达细胞为巨噬细胞、浆细胞和淋巴细胞.结论:DAP患者牙龈组织中IL-1β和TNFα在细胞凋亡中起重要作用.     

Ⅱ型糖尿病伴牙周炎病人牙龈组织中VEGF表达及MVD计数的研究

目的:探讨Ⅱ型糖尿病伴牙周炎病人牙龈组织中血管内皮生长因子(VEGF)的表达水平及其作用。方法:选取Ⅱ型糖尿病伴重度牙周炎(DP)病人、单纯重度慢性牙周炎(CP)病人、健康对照者(N)各15例,分别切取牙龈组织,用免疫组化染色方法检测牙龈组织中VEGF表达和MVD计数。结果:DP组牙龈组织中VEGF表达和MVD计数均显著高于CP组和N组(P<0.05);CP组高于N组(P<0.05)。结论:Ⅱ型糖尿病伴重度慢性牙周炎病人牙龈组织中VEGF表达水平和MVD计数明显升高。     

基质金属蛋白酶-2在人慢性牙周炎肥大细胞上表达的研究

目的观察基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)在慢性牙周炎牙龈组织中肥大细胞上的表达,探讨MMP-2-tryptase双阳性肥大细胞(mast cells,MCs)在慢性牙周炎发病机制中的作用。方法将45例参试者依据慢性牙周炎的病变程度分成3组:健康对照组15例;轻度牙周炎组15例;重度牙周炎组15例。牙龈标本经10%福尔马林液固定48 h;制作牙龈组织连续切片,HE染色,光学显微镜下观察组织学改变;采用免疫荧光双染色法,荧光显微镜下观察牙龈组织中MMP-2-tryptase双阳性MCs的表达情况。结果与健康对照组相比,轻度和重度慢性牙周炎组牙龈组织MMP-2-tryptase双阳性MCs密度均显著升高(P<0.01);重度慢性牙周炎组牙龈组织MMP-2-tryptase双阳性MCs密度显著高于轻度牙周炎组(P<0.05)。结论慢性牙周炎牙龈组织MMP-2-tryptase双阳性MCs密度与牙周炎病变程度的趋势相一致,MMP-2-tryptase双阳性MCs在牙周炎的进展中可能有促进作用。     

Tumor necrosis factor-alpha expression and detection of apoptosis at the site of chronic periodontitis in AIDS patients

BACKGROUND: Apoptosis, or programmed cell death, is associated with the regulation of the life cell cycle of leukocytes in healthy and diseased states. OBJECTIVES: In the present study, we investigated the presence of apoptosis of mononuclear inflammatory cells in the periodontal lesion from adult periodontitis in healthy control patients and AIDS patients. MATERIALS AND METHODS: Tissue samples adjacent to a 5-6 mm gingival sulcus, measured with a periodontal probe, were obtained during routine periodontal surgical procedures. The direct immuno-peroxidase of digoxigenin-labeled genomic DNA method was used for in situ detection of apoptosis in gingival tissues. RESULTS: Many tumor necrosis factor-alpha (TNFalpha)-positive cells, detected by immunohistochemistry method, were observed in gingival samples of both groups of patients. In addition, a significant lower number ( p < 0.05) of mononuclear apoptotic cells were observed in AIDS patients when compared with healthy control patients. CONCLUSION: These data suggested an important role of the apoptosis of mononuclear cells in the pathogenesis of chronic adult periodontitis in AIDS patients.     

内质网应激诱导的细胞凋亡在慢性牙周炎中的作用初探

目的:初步探讨内质网应激诱导的细胞凋亡在慢性牙周炎中的作用。方法:利用光学显微镜和透射电镜观察牙周组织中的凋亡细胞,免疫组化染色法检测慢性牙周炎患者及牙周健康者牙周组织中GRP78和CHOP的分布及含量变化。结果:慢性牙周炎组内浆细胞呈现凋亡状态,且GRP78和CHOP在慢性牙周炎组中的阳性表达明显高于正常组(P<0.05)。结论:内质网应激参与了慢性牙周炎的病理生理过程。     

Preparation and characterization of human gingival cells

A procedure was developed for the preparation of single cell suspensions from gingival and periodontal tissues using collagenase digestion followed by gentle mechanical disruption. The procedure was evaluated on rat gingival tissues. One-hour incubation was found to produce the greatest number of cells with the highest viability, and the largest recovery of mononuclear cells. Collagenase did not appear to effect recovery, viability or B cell surface markers on human peripheral blood cells. Human gingival and periodontal tissues were obtained from 35 patients: 20 with adult periodontitis; 8 with juvenile periodontitis; 7 with normal gingivae or chronic gingivitis. Scaling, followed by probing measurements and then surgery of the affected sites was routinely performed. In single cell suspensions from tissue obtained from surgery, significantly fewer mononuclear cells were recovered from the tissues of normal/chronic gingivitis patients than cells/mg of tissue recovered from adult periodontitis or juvenile periodontitis groups. The maximum contribution of blood mononuclear cells to the gingival cells averaged 0.5+0.2%. This method is useful for recovering gingival (periodontal) cells for phenotypic and functional studies.     

Tissue localization of Actinobacillus actinomycetemcomitans in human periodontitis. I. Light, immunofluorescence and electron microscopic studies

Invasion of periodontal tissues by different bacterial morphotypes has been reported in human periodontitis; however, limited information is available as to prevalence, localization and the bacterial species involved. The present study determined prevalence and gingival localization of Actinobacillus (Haemophilus) actinomycetemcomitans in periodontal lesions of juvenile periodontitis patients. Thirty-five gingival biopsies were obtained from 12 juvenile periodontitis patients at the time of periodontal therapy. One additional control biopsy was obtained from each of two adult periodontally healthy subjects, one adult periodontitis patient and one periodontally healthy monkey (Macaca fosibolius). The biopsies were carefully processed to avoid mechanical introduction of bacteria into the tissues and were examined using light and electron microscopy. Rabbit antisera specific for the three A. actinomycetemcomitans serotypes were used for immunofluorescence microscopic localization of A. actinomycetemcomitans antigens in the gingival sections. Immunofluorescence microscopy showed A. actinomycetemcomitans specific antigens in the gingival tissues of 11 of the 12 juvenile patients examined. None of the control specimens showed evidence of A. actinomycetemcomitans antigens in the gingival connective tissue. One specimen from a periodontally healthy subject and the monkey biopsy, however, showed A. actinomycetemcomitans antigens in bacterial plaque on the surface of the crevicular epithelium. Transmission electron microscopic examination showed microcolonies of small gram-negative rods in the connective tissue, as well as single bacterial cells between collagen fibers and in areas of cell debris. In addition to these extracellular bacterial cells, evidence of bacterial cells was also found within gingival connective tissue phagocytic cells. The data from the present study suggest that the gingival tissue in juvenile periodontitis lesions harbors A. actinomycetemcomitans.     

Apoptotic genes are differentially expressed in aged gingival tissue

Cellular and molecular changes of the periodontium associated with a higher prevalence of oral diseases (e.g., chronic periodontitis) in aged populations have received little attention. Since impaired apoptosis during aging appears to be related to chronic inflammatory disorders, we hypothesized that the expression of genes associated with apoptotic processes are altered in aged healthy and periodontitis-affected gingival tissue. Ontology analysis of 88 genes related to apoptotic pathways was performed in gingival biopsies of healthy and periodontitis sites from young, adult, and aged non-human primates (Macaca mulatta), using the GeneChip? Rhesus Macaque Genome Array. Lower expression of anti-apoptotic and higher expression of pro-apoptotic genes were associated with healthy gingival tissue from young compared with aged animals. Few differences in gene expression were observed in healthy gingival tissue between adult and aged animals. Comparison between healthy and periodontitis gingival tissues showed that the up- or down-regulated apoptotic genes in diseased gingival tissue are different in adults compared with aged animals. These results suggest that apoptotic events normally occurring in gingival tissues could be reduced in aging,and unique aspects of apoptotic pathways are potentially involved in the pathophysiology of periodontal disease in adult vs. aged gingival tissues.     

IL-1βmRNA、TNF-αmRNA在成人牙周炎患者牙龈组织中表达的研究

目的:检测白细胞介素1B(IL-1B)mRNA、肿瘤坏死因子A( TNF-A)mRNA在成人牙周炎患者牙龈组织中表达, 探讨IL-1B和TNF-A与牙周炎致病机理的关系。方法:采用RT- PCR方法检测IL-1BmRNA、TNF-AmRNA在19例成人牙周炎患者炎症牙龈和9例牙周健康者牙龈组织中表达。结果:IL-1BmRNA表达强度在成人牙周炎病变牙龈 (0.819?01045)显著高于正常牙龈(0.306?0.087)(t=3.71,P<0.01)。TNF-AmRNA表达强度在成人牙周炎病变牙龈(0.696?0.098)显著高于正常牙龈(0)(t=7.09,P<0.01)。结论:IL-1B和TNF-A作为炎症和骨吸收介质可能在牙周炎的发生发展中起一定的作用。     

Immunohistochemical evaluation of Ki-67 expression and apoptosis in cyclosporin A-induced gingival overgrowth

BACKGROUND: This study was planned to evaluate cell division rate and apoptosis by immunohistochemical and in situ hybridization techniques in cyclosporin A (CsA)-induced gingival overgrowth tissue samples to determine whether these processes played a role in the pathogenesis of this condition. METHODS: Fourteen CsA-induced overgrowth tissues from renal transplant recipients, 10 control tissues from patients with plaque-induced gingivitis, and 14 control tissues from systemically and periodontally healthy subjects were evaluated. In patient groups, clinical periodontal recordings and tissue sampling were performed before initiation of any periodontal intervention. Numbers of Ki-67-positive cells/field and apoptotic cells/field in formalin-fixed/paraffin-embedded tissue sections were determined. Data were evaluated by one-way analysis of variance, post hoc Sidak test with modified Bonferroni correction, and Pearson correlation analysis. Three phenytoin- and five nifedipine-induced overgrowth tissues were also processed in the same way, and findings in these tissue specimens were evaluated as case series. RESULTS: The number of keratinocytes was significantly greater in the CsA-induced gingival overgrowth group than in the healthy control group (P <0.05). Cells labeled by in situ end labeling, namely the apoptotic cells, were significantly fewer in the CsA group than in the gingivitis and healthy control groups (P <0.01). Overall, statistically significant positive correlations were found between the numbers of Ki-67-positive cells and probing depth and hyperplastic, bleeding, and plaque indices (P <0.01). Phenytoin and nifedipine samples exhibited obviously higher expression of Ki-67-positive cells than the CsA, gingivitis, and healthy control groups. CONCLUSION: Our findings suggest that decreased apoptosis may have a more prominent role than increased cell division in the pathogenesis of CsA-induced gingival overgrowth.