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     

不同牙周状态正畸力诱导破骨细胞分化因子RANK在牙周组织的表达

目的 研究不同牙周状态正畸力作用下破骨细胞分化因子RANK在牙周组织的表达变化,为牙周病正畸治疗提供参考.方法 建立大鼠牙周炎静止期、牙周炎活动期模型,以50克力移动牙周正常组、牙周炎静止期组、牙周炎活动期组的上颌磨牙.分别于牙齿移动的第3和第7天处死大鼠.采用免疫组化和实时荧光定量PCR定性定量分析各组RANK在牙周组织的表达.结果 正常牙周移动组RANKmRNA的表达高于正常对照组(P＜0.05).静止期牙齿移动组RANKmRNA在牙周组织的表达高于正常牙周移动组(P＜0.01).较活动期移动组显著减小(P＜0.01).牙周炎活动期牙齿移动组RANKmRNA的表达均高于其他各组(P＜0.01).结论 RANK参与调节正常牙周及牙周炎正畸牙齿移动.牙周炎静止期正畸力可诱导RANK的表达增加,但显著小于炎症活动期.此时要密切控制牙周易感因素.     

不同牙周状态正畸力诱导IL-23在牙周组织表达的实验研究

目的 研究不同牙周状态正畸力作用下IL-23在牙周组织的表达变化,为牙周病正畸治疗提供参考.方法 建立大鼠牙周炎静止期、牙周炎活动期模型,以50克力移动牙周正常组、牙周炎静止期组、牙周炎活动期组的上颌磨牙.分别于牙齿移动的第3和7天处死大鼠.采用免疫组化和实时荧光定量PCR定性定量分析各组IL-23在牙周组织的表达.结果 正常牙周移动组IL-23mRNA的表达较正常对照组差异无显著性（P＞0.05）.静止期牙齿移动组IL-23mRNA在牙周组织的表达较正常牙周移动组增高,差异有显著性（P＜0.05）;较活动期移动组显著减小（P＜0.01）.牙周炎活动期牙齿移动组IL-23mRNA的表达较其他各组均高,且差异有高度显著性（P＜0.01）.结论 IL-23参与调节牙周炎正畸牙齿移动.牙周炎静止期正畸治疗不会导致IL-23的过度高表达,但要密切控制牙周易感因素.     

慢性牙周炎与健康牙龈组织中MMP-1的表达和意义

目的:通过检测基质金属蛋白酶-1(MMP-1)在慢性牙周炎牙龈组织中的表达和分布特征,探讨MMP-1在慢性牙周炎发病中的作用和临床意义。方法:收集8例因非牙周疾病而需拔牙患者健康牙龈及24例慢性牙周炎患者牙龈组织,按健康对照、牙周袋深度≤4mm、4~6mm、>6mm分A、B、C、D四组,利用免疫组化方法检测其中MMP-1的表达。结果:正常牙龈组织上皮及固有层MMP-1弱表达,慢性牙周炎患者牙龈组织MMP-1表达明显增高,两者间有显著性差异(P<0.05)。并且MMP-1表达与牙周袋深度间显著正相关(r=0.623,P<0.01)。结论:MMP-1参与慢性牙周炎病变牙周组织的破坏过程,并且其表达随牙周袋深度加深有增加趋势。     

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。     

巨噬细胞移动抑制因子在慢性牙周炎患者血清及牙龈组织中的表达分析

目的    观察并分析慢性牙周炎患者血清及牙龈组织中巨噬细胞移动抑制因子（macrophage migration inhibitory factor,MIF）表达及其临床意义。方法    选择2016年6月至2017年6月就诊于中国医科大学附属口腔医院牙周科及口腔外科的患者,收集重度慢性牙周炎患者（牙周炎组,18例）及牙周健康者（健康对照组,18例）血清,酶联免疫吸附试验检测MIF及单核细胞趋化蛋白-1（monocyte chemotactic protein-1,MCP-1）水平。收集未经治疗（CP组,8例）及牙周基础治疗后1个月（SRP组,11例）的重度慢性牙周炎患者、牙周健康者（H组,18例）的牙龈组织,采用免疫组化法检测MIF及MCP-1在牙龈中的表达。采用SPSS 24.0软件t检验比较血清中MIF及MCP-1表达的差异,单因素方差分析比较CP组、SRP组及H组牙龈组织中MIF及MCP-1表达的差异。结果    慢性牙周炎患者血清中MIF及MCP-1浓度较牙周健康者明显升高（P < 0.05）。MIF在牙龈上皮呈阳性表达,且在未经治疗的CP组表达水平最高,经牙周基础治疗后的SRP组MIF表达水平显著下降,但仍高于H组（P < 0.05）;MCP-1在牙龈组织的上皮层和结缔组织中均有弱表达,表达水平在3组间差异无统计学意义（P > 0.05）。结论    MIF作为一种炎症因子参与慢性牙周炎的发展进程,其表达水平可作为炎症指标辅助慢性牙周炎活动性及炎症程度的判断。     

糖尿病对大鼠牙周组织及诱导型一氧化氮合酶分布的影响

目的：观察糖尿病对大鼠牙周炎牙周组织形态结构的影响及诱导型一氧化氮合酶（iNOS）在牙龈组织中的分布。方法：建立大鼠糖尿病牙周炎模型,制作组织切片,进行HE及免疫组织化学染色,观察牙周组织形态结构的破坏情况及iNOS在牙龈组织中的分布。结果：糖尿病牙周炎组的结缔组织附着丧失量及牙槽骨高度丧失量均明显高于牙周炎组、糖尿病组及正常组。差异有显著性（P〈0．05）。糖尿病牙周炎组和糖尿病组的免疫组化染色均为阳性或强阳性。结论：糖尿病加重了牙周炎牙周组织的破坏程度。     

氨基胍干预对创伤性面瘫大鼠外周面神经及周围组织一氧化氮合酶表达的影响

目的　研究诱导型一氧化氮合酶抑制剂氨基胍对创伤性面瘫大鼠外周面神经及周围组织一氧化氮合酶表达的影响。方法　通过大鼠面瘫前后腹膜内小剂量给予氨基胍,在伤后各个时间点切取损伤面神经和软组织, 采用兔抗大鼠一氧化氮合酶(NOS)抗血清免疫组织化学ABC法对面神经和肌肉软组织内NOS表达的变化进行研究。结果　氨基胍组面神经和肌肉软组织伤后诱导型一氧化氮合酶免疫反应性明显降低。结论　氨基胍慢性干预明显抑制面神经和肌肉软组织内诱导型一氧化氮合酶的表达,为面神经再生与组织创伤修复创造了有利条件。     

唾液中一氧化氮含量与牙周炎的相关性研究

目的:研究牙周炎患者唾液中一氧化氮(NO)含量与牙周炎各种临床指标的相关性,探讨一氧化氮在牙周炎发展过程中可能发挥的作用.方法:选择牙周健康者28人(对照组),慢性牙周炎32人(实验组),用Griess反应测定牙周基础治疗前后唾液中亚硝酸盐含量,间接反映NO水平,分析牙周临床指标与NO含量的关系.结果:实验组唾液中NO含量较对照组显著增加,牙周炎患者唾液NO含量与牙周探诊深度,附着水平之间有显著正相关.结论:NO的产生与牙周炎症过程有关.牙周基础治疗后,随着局部炎症逐渐减轻,唾液NO含量呈下降趋势.     

β-防御素基因在牙龈组织中的表达

目的:检测人β-防御素(HBD-1,-2,-3)基因在牙周炎病变和健康牙龈组织中的表达。方法:应用反转录多聚合酶链反应(RT-PCR)技术检测健康牙龈(HC组,11例)、慢性牙周炎(CP组,12例)和侵袭性牙周炎(AgP组,9例)牙龈组织中HBD-1、HBD-2和HBD-3mRNA表达水平。结果:HBD-1,-2,-3在所有牙龈组织样本中均有mRNA表达;HBD-3mRNA在HC组、CP组、AgP组的表达水平分别为0.53±0.12,0.30±0.17和0.40±0.17,3组间差异有统计学意义(P<0.01),健康牙龈中HBD-3mRNA表达相对强度明显高于慢性牙周炎组;HBD-2和HBD-3基因的mRNA表达水平呈正相关(P<0.01,r=0.48)。结论:牙龈上皮表达的β-防御素(HBD-1,-2,-3),尤其是HBD-3在健康牙龈组织较高水平的mRNA表达,提示其在牙周宿主免疫防御反应中可能发挥重要作用。     

正常人、牙龈炎和牙周炎患者龈沟液内一氧化氮含量的检测

目的：检测正常人和牙周病患者龈沟液中NO含量,探讨NO在牙周病发病过程中的作用。方法：选择牙周健康组20例,牙龈炎组22例,慢性牙周炎组32例,分别采集龈沟液标本,免疫荧光法检测龈沟液内NO的含量。结果：慢性牙周炎患者和牙龈炎患者龈沟液内NO含量与牙周健康组相比均有高度显著性差异（P〈0．01）,慢性牙周炎患者龈沟液内NO含量与牙龈炎组相比有高度显著性差异（P〈0．01）。结论：牙周健康者、牙龈炎患者、慢性牙周炎患者龈沟液中能检测出NO的存在,NO参与了慢性牙周炎的发展过程,龈沟液内NO含量与慢性牙周炎炎症程度密切相关。     

慢性牙周炎龈组织中Smac和Bax的表达及其对细胞凋亡的作用

目的:通过检测Smac和Bax蛋白在慢性牙周炎龈组织中的表达,旨在探讨二者的相关性及其在细胞凋亡中与牙周炎发生、发展的关系。方法:慢性牙周炎测试组27人,健康对照组27人,利用光镜和透射电镜观察凋亡细胞形态结构,流式细胞仪测定线粒体膜电位。免疫组化检测Smac、Bax在龈组织不同区域的表达。结果:光镜观察慢性牙周炎龈组织中存在有凋亡细胞;电镜观察凋亡细胞中线粒体形态结构改变;慢性牙周炎龈组织线粒体膜电位降低,与健康组比较,两组间有显著性差异(P<0.05);慢性牙周炎组中Smac和Bax在上皮组织、结缔组织的表达增加,与健康组相比具有显著性差异(P<0.05);Smac、Bax二者的表达在慢性牙周炎龈组织具有明显正相关(P<0.05)。结论:慢性牙周炎龈组织中有凋亡细胞存在;Smac、Bax的表达在慢性牙周炎组中呈明显正相关,表明在细胞凋亡过程中起协同作用,这与牙周组织破坏有关。     

Inducible nitric oxide synthase expression in periodontitis

Recently, nitric oxide (NO) has been shown to be vital in inflammatory processes. Nitric oxide synthase (NOS) exists in three different isoforms, two constitutively produced with physiological roles, and an inducible form, iNOS, which is involved in inflammation. This study examined the localisation of iNOS in biopsies from patients with periodontitis using immunohistochemistry, and compared these with healthy tissue biopsies. Biopsies were obtained from 16 periodontitis patients undergoing periodontal surgery and from clinically healthy tissues of 5 patients having crown lengthening procedures. The periodontitis diseased tissue demonstrated a greater level of iNOS expression than the healthy tissue. The source of iNOS in the periodontal tissues was determined by our monoclonal antibody to be the macrophage, with the endothelial cells also contributing. A role for NO in the inflammatory response of periodontal tissues is suggested, but the precise role requires further elucidation.     

The in vivo expression of membrane-bound CD14 in periodontal health and disease

BACKGROUND: Membrane-bound CD14 (mCD14) is a myeloid differentiation antigen expressed on monocytes/macrophages and neutrophils. It is a key molecule responsible for the innate recognition of bacteria by host cells and functions as an important receptor for bacterial lipopolysaccharide. This study investigated the in vivo expression profile and levels of mCD14 in healthy and diseased gingival tissues. METHODS: Gingival biopsies were obtained from 24 patients with chronic periodontitis, including 22 periodontal pocket tissues, 13 clinically healthy tissues, and 18 inflamed connective tissues (i.e., granulation tissues). Gingival biopsies from seven periodontally healthy subjects were used as controls. mCD14 was detected by immunohistochemistry. RESULTS: mCD14 was detected in 21 of 22 periodontal pocket tissues and all other categories of tissues. The mCD14-positive cells were mainly confined to the gingival epithelium-connective tissue interface. The expression levels in periodontally healthy subjects were significantly higher than in the patients. Within the patients, clinically healthy tissues showed greater levels of mCD14 than periodontal pocket tissues and granulation tissues. CONCLUSIONS: mCD14 was commonly expressed in both healthy and diseased gingival tissues and was predominantly confined to the epithelium-connective tissue interface. The positive relationship observed between mCD14 expression levels and periodontal health may imply that mCD14 is associated with favorable host responses to bacterial challenge and contributes to maintaining periodontal homeostasis.     

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。     

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??Objective    To investigate the expression of macrophage migration inhibitory factor??MIF??in the serum and gingival tissues of patients with chronic periodontitis??and to explore its role in clinical prognosis. Methods    Serum samples were collected from chronic periodontitis patients??18 cases??and healthy controls??18 cases????MIF and monocyte chemotactic protein-1??MCP-1??levels were assayed by enzyme-linked immunosorbent assay. Gingival tissues samples were collected from chronic periodontitis patients??before and after periodontal initial therapy??and periodontally healthy individuals. Then the MIF and MCP-1 expression were detected with immunohistochemical method. The data was analyzed with SPSS 24.0 software package. Results     Serum MIF and MCP-1 were significantly higher in periodontitis patients compared with healthy controls??P < 0.05??. MIF protein was expressed in gingival epithelia??and the MIF level was highest in periodontitis gingiva??its level was decreased by periodontal initial therapy??but still higher than the expression of MIF in healthy gingiva ??P < 0.05????MCP-1 protein was also expressed in both gingival epithelia and connective tissues??but there was no significant difference between the periodontitis patients and healthy controls??P > 0.05??. Conclusion          MIF is involved in the progression of chronic periodontitis as an inflammatory factor??and its level may be used as auxiliary indicators for the evaluation of the activity and degree of periodontitis.     

Expression of vascular endothelial growth factor in human periodontal tissues: comparison of healthy and diabetic patients

BACKGROUND: Vascular endothelial growth factor (VEGF) induces proliferation of endothelial cells, stimulates angiogenesis, and increases vascular permeability, but information about its role in periodontal lesions is limited. The aim of this study is to determine the association between VEGF expression in healthy and periodontally diseased tissues of healthy and diabetic patients. METHODS: Ten systemically healthy and 10 Type 2 diabetic patients (DM) all diagnosed with periodontitis were enrolled into the study. Gingival samples were collected from both periodontal and healthy sites in all patients. Each patient served as his/her own control. Additionally, 10 people without any systemic or periodontal diseases were enrolled as a negative control group. RESULTS: In the negative control group tissue samples, no VEGF expression was observed. Among the 10 systemically healthy people, no evidence of VEGF was observed in healthy gingival samples, but was found in diseased tissues in 2 cases. In the diabetic patients, VEGF was observed in 4 healthy gingival tissues and in 6 periodontal sites. VEGF was intensely present in monocytes and macrophages. CONCLUSION: The results of this study show that VEGF is increased in gingival tissues of diabetic patients, especially those with periodontal disease.     

Arginine–Nitric Oxide–Polyamine Metabolism in Periodontal Disease

Background: Arginine is converted to nitric oxide (NO) via NO synthase and to ornithine via arginase. Ornithine decarboxylase (ODC) catalyzes the conversion of ornithine to polyamines. Arginase can inhibit NO production, and NO can inhibit ODC activity as part of an early inflammatory response. This study examines the arginine‐NO‐polyamine pathway alteration in saliva and gingival biopsy samples of patients with gingivitis or periodontitis and healthy controls and evaluates the response to periodontal treatment. Methods: This study includes nine gingivitis patients, 15 chronic periodontitis patients, and 11 healthy age‐matched controls. Periodontal clinical measurements, gingival biopsies, and saliva samples were obtained before treatment (BT) and 1 month after periodontal treatment (AT). Arginase and ODC activities and NO levels were determined spectrophotometrically. Results: The BT salivary and gingival NO levels were found to be highest in the gingivitis group, followed by the healthy and the periodontitis groups, respectively. Salivary NO levels significantly increased in the periodontitis group and decreased in the gingivitis group AT (P <0.05). Gingival NO levels decreased significantly in the periodontitis and the gingivitis groups AT (P <0.05). Arginase levels were detected highest in the gingivitis group and lowest in the periodontitis group, both in saliva and gingiva. Only gingival arginase levels significantly increased AT (P <0.05). ODC activity was highest in saliva, and lowest in the gingiva of the periodontitis patients BT. It was found to be significantly higher in the periodontitis group AT (P <0.05). Conclusions: In this study, regarding arginine‐NO‐polyamine metabolism, gingival tissue seems to be more informative about periodontal pathogenesis than saliva. At early phase of periodontal inflammation, NO arginase and ODC levels were measured as higher than at an established lesion of periodontitis.