慢性牙周炎龈沟液中白细胞介素-8与硫化物的相关关系

目的：探讨慢性牙周炎龈沟液（gingival crevicular fluid，GCF）中白细胞介素-8（interleukin-8，IL-8）与硫化物（suleus sulphide level，SUL）的关系。方法：采用双抗体夹心ELISA法、金刚牙周诊断仪对龈沟液中IL-8和硫化物的含量和临床指标进行测定。23个牙周健康牙作正常对照组（C），12个慢性牙周炎健康牙作实验组1（T1），30个慢性牙周炎患牙作实验组2（T2）。用标准化滤纸条采集观察牙位GCF样本，记录相应位点30’硫化物浓度，同时记录龈沟出血指数（SBI）、牙周袋探诊深度（PPD）、牙周附着丧失水平（CAL）。结果：①受检位点龈沟液中IL-8总量存在显著性差别，其中慢性牙周炎炎症牙位组IL-8总量高于健康牙位组、正常对照组（p〈0.05），但龈沟液中IL-8浓度无显著性差别。龈沟液中IL-8总量与SBI、PPD、CAL之间有明显相关关系（P〈0.05），但IL-8的浓度与临床指标间相关关系不明显。②慢性牙周炎炎症牙位组的硫化物浓度与健康牙位组、正常牙位组之间均有显著性差别（P〈0.05），慢性牙周炎健康牙位组与正常牙位组之间无显著性差别。③慢性牙周炎炎症牙位组硫化物浓度与临床指标间具正相关关系（P〈0.05），而健康牙位组和正常对照组硫化物浓度与临床指标间无相关关系。GCF中慢性牙周炎炎症牙位组的硫化物浓度与IL-8总量具负相关关系（P〈0．05），而慢性牙周炎健康牙位组和正常对照组的硫化物浓度与IL-8总量之间无相关关系。结论：慢性牙周炎患牙龈沟液中IL-8和硫化物的含量与临床指标之间有相关性。龈沟液中细菌代谢产物所产生的硫化物对IL-8的含量存在一定抑制作用。     

侵袭性牙周炎龈沟液中白介素-8 的检测

目的：检测侵袭性牙周炎（AgP）龈沟液中白细胞介素（IL-8）的总量和浓度并探讨其与牙周临床指标的关系。方法：采用常规滤纸条法收集侵袭性牙周炎实验组患牙（T1）和健康牙（T2）各位点及正常对照组（C）各位点的龈沟液（GCF）样本,用ELISA法检测各样本中IL-8的总量和浓度。结果：3组受检牙龈沟液中IL-8的总量和浓度不同。侵袭性牙周炎患牙组GCF中IL-8总量高于健康牙位组（P〈0．05）及正常对照组;而3组中IL-8浓度差异也有显著性,侵袭性牙周炎患牙组GCF中IL-8的浓度高于健康牙位组（P〈0．05）和正常对照组;虽然GCF中IL-8浓度与牙周探诊深度（PD）、牙龈出血指数（BOP）、附着丧失（AL）无相关关系;但IL-8总量与以上牙周临床指标相关。结论：在侵袭性牙周炎患者龈沟液中IL-8是参与牙周炎症反应的重要调节因子。     

盐酸米诺环素缓释剂对慢性牙周炎龈沟液弹性蛋白酶水平的影响

目的：观察慢性牙周炎应用盐酸米诺环素缓释软膏治疗前后龈沟液中细胞外弹性蛋白酶（elastase in su-pernatant,EA-s）和细胞内弹性蛋白酶（elastase in the pellet,EA-p）水平的变化。方法：35例慢性牙周炎患者,每例口腔左右两个象限随机分为实验组和对照组,基础治疗后实验组使用盐酸米诺环素缓释软膏,1周1次,共4周,对照组不使用盐酸米诺环素缓释软膏,测定两组治疗前后龈沟液中EA-s和EA-p水平。结果：实验组、对照组在治疗后龈沟液中EA-s和EA-p的总量和浓度均明显降低（p〈0.01）。治疗前后实验组与对照组龈沟液中EA-s和EA-p的总量和浓度变化有显著性差异,实验组优于对照组（p〈0.01）。结论：基础治疗和盐酸米诺环素缓释软膏联合应用对慢性牙周炎的治疗在降低龈沟液中EA方面优于单纯的基础治疗。     

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。     

牙周炎患者龈沟液中IL—8的含量测定

目的：研究IL－8在牙周炎病程中的变化及与临床指标的关系,方法：采用双抗夹心ABC－ELISA法测定慢性牙周炎（CP）患者,健康对照者以及CP治疗前后患者的龈沟液中IL－8含量,IL－8总量,同时检测临床指标并作相关性检验。结果：CP患者龈沟液中IL－8检出率显著高于健康对照者（P＜0．025）,在CP患者和健康者之间以及CP治疗前后患者的龈沟液中IL－8总量,龈沟液（GCF）量均存在统计学差异（分别为P＜0．01,P＜0．05）,而IL－8含量无统计学差异（P＞0．05）,IL－8总量,GCF量与临床指标存在正相关性（P＜0．01）,结论：IL－8总量在牙周炎病程中呈动态性改变,检测GCF中IL－8的水平对评价牙周炎的程度及指导临床治疗有一定价值。     

慢性牙周炎龈沟液中KGF-1含量检测及其意义

目的：检测慢性牙周炎患者牙周基础治疗前及治疗后不同时期龈沟液中角质细胞生长因子（keratinocytegrowth factor,KGF）-1的质量浓度,探讨KGF-1与牙周炎的关系及其在牙周炎发病机理、病情进展方面的作用。方法：用滤纸条浸润法采集牙周健康者和慢性牙周炎患者治疗前后不同时期的龈沟液样本,用酶联免疫吸附测定法检测样本中KGF-1的质量浓度。结果：慢性牙周炎患者龈沟液中KGF-1的质量浓度高于健康对照组（P〈0.05）,牙周基础治疗后第4、6、8周复查,KGF-1的质量浓度低于治疗前,且治疗后不同时期的复查结果与治疗前相比,均有统计学意义（P〈0.05）,KGF-1的质量浓度与牙周探诊深度（PD）、附着丧失（AL）、龈沟出血指数（SBI）存在正相关。结论：龈沟液中KGF-1水平与牙周炎症密切相关,KGF-1可作为反映牙周状态的客观指标。     

快速进展性牙周炎患者龈沟液中的弹性蛋白酶活性

目的探讨中性多形核白细胞与快速进展性牙周炎的关系。方法检测２例快速进展性牙周炎患者共１０２个位点（其中２８个位点进行治疗前后的对比）的龈沟液弹性蛋白酶活性,将人多形核白细胞弹性蛋白酶特异底物———Ｓ２４８４与龈沟液反应,测吸光度值,以反映龈沟液弹性蛋白酶活性。结果快速进展性牙周炎龈沟液中的弹性蛋白酶活性［（０．６３±０．３８）Ａｂｓ／位点］明显高于健康对照组［（０．０７±０．０５）Ａｂｓ／位点］,差异有显著性;弹性蛋白酶活性的高低与龈沟液体积、探诊深度、附着丧失和出血指数呈正相关关系;治疗后龈沟液体积和各临床指数显著降低,弹性蛋白酶活性也从治疗前的（０．７３±０．３６）Ａｂｓ／位点下降为（０．１±０．１７）Ａｂｓ／位点,差异有显著性。结论快速进展性牙周炎患者的中性多形核白细胞并不是趋化反应不足,而是过度浸润与释放溶酶体酶,起协同致炎作用。     

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目的研究妊娠晚期牙周炎孕妇龈沟液、唾液、血清中白介素-2（IL-2）含量变化以及三者间的关联。方法于2011年4—7月在湖北省妇幼保健院接受口腔检查的妊娠33～38周单胎孕妇中,随机选取20例牙周炎、26例牙龈炎、22例牙周健康孕妇（对照）纳入研究。分别采集其唾液、龈沟液和血清,采用双抗体夹心ELISA检测IL-2质量浓度。结果牙周炎组分娩时孕周（36．5周）明显小于牙龈炎组（38．8周）和对照组（39.3周）,牙周炎组分娩时新生儿体重（3079．9g）低于牙龈炎组（3452．5g）和对照组（3462．4g）,差异均有统计学意义（均P〈0．05）。牙周炎组大专及以上学历、孕前半年行口腔保健的比例明显低于牙龈炎组和对照组（均P〈0．05）。牙周炎组龈沟液、唾液中IL-2质量浓度明显高于牙龈炎组和正常对照组,差异有统计学意义（均P〈0．05）;但3组血清中IL-2质量浓度比较,差异无统计学意义（P〉0．05）。各组孕妇龈沟液、唾液、血清中IL-2质量浓度之间均两两呈线性相关,差异有统计学意义（均P〈0．01）。结论妊娠合并牙周炎孕妇口腔中IL-2含量较高,并与血清中IL-2质量浓度呈直线相关,在牙周炎合并不良妊娠结局（如早产、低体重儿等）的发病机制中起-定作用。重视口腔卫生是预防不良妊娠结局的方法之-。     

吸烟与非吸烟者慢性牙周炎龈下福赛类杆菌的检测

目的：利用聚合酶链反应（PCR）检测吸烟与非吸烟者龈下福赛类杆菌（Bf）水平,通过Bf的检出率进一步探讨吸烟与牙周炎的关系。方法：健康者与慢性牙周炎伴吸烟与非吸烟患者共75例,用纸捻法提取每位观察者龈沟液,PCR检测。结果：慢性牙周炎组Bf检出率（59．5％）高于健康组（24．2％）,具有显著性差异（P〈0．01）;吸烟者慢性牙周炎组Bf检出率（72．0％）高于非吸烟者健康组、吸烟者健康组、非吸烟者慢性牙周炎组（15．8％、35．7％、41．2％）,P〈0．05。结论;Bf与吸烟及慢性牙周炎有密切关系。     

慢性牙周炎患者唾液中miR-146a的表达及其与龈沟炎症、MMP-8/TIMP-1水平的关系

目的： 探讨慢性牙周炎患者唾液中miR-146a的表达及其与龈沟炎症、基质金属蛋白酶8（MMP-8）、基质金属蛋白酶抑制剂1（TIMP-1）水平的关系。方法： 选择2015年3月—2017年1月间收治的慢性牙周炎患者68例作为慢性牙周炎组,同期在本院进行体检的健康志愿者50例作为正常对照组。检测2组研究对象唾液中miR-146a的表达量,龈沟液中炎症因子、MMP-8/TIMP-1的水平及牙周临床症状指标。采用SPSS24.软件中的Pearson检验评估慢性牙周炎患者唾液中miR-146a的表达量与病情严重程度的相关关系。结果： 慢性牙周炎患者唾液中miR-146a的表达量显著高于正常对照组（P<0.05）,龈沟液中炎症因子（IL-1β、IL-6、IL-35、TNF-α）水平、牙周临床症状指标（PD、AL、PLI、BI）以及龈沟液中MMP-8、TIMP-1的水平显著高于正常对照组（P<0.05）。Pearson检验发现,慢性牙周炎患者唾液中miR-146a表达量与龈沟炎症程度、牙周临床症状严重程度及MMP-8/TIMP-1水平呈正相关。结论： 慢性牙周炎患者唾液中miR-146a表达量异常增高,且与龈沟炎症程度、牙周损伤程度一致。     

Expression of intracellular elastase activity in peripheral neutrophils from patients with adult periodontitis

This study aimed to investigate whether circulating neutrophils from patients with periodontitis contain more catalytically active elastase than neutrophils from healthy controls. The amount of IL-1beta in these cells was also analyzed and the correlation with elastase activity was tested. The periodontitis group consisted of 15 subjects with marked periodontal destruction. The healthy control group consisted of 15 subjects with no clinical signs of periodontal destruction. The elastase activity and the IL-1beta content in the cells were measured with flow cytometry using a specific substrate and antibodies, respectively. The plasma concentration of IL-1beta and the total content of antigenic elastase in the crushed cells were measured with an ELISA. The elastase activity per neutrophil was significantly higher in the patients than in the controls, while the total antigenic elastase content did not differ. The % of cells positively stained for IL- 1beta was somewhat higher in the patient group. Significantly higher amounts of IL-1beta per sample, estimated by multiplying the % of stained cell with the amount of staining per cell, were found in the patient group. A significant correlation between IL-Ibeta and elastase activity was noted in the patient group (R=0.6, p=0.001), but not in the control group. In conclusion, peripheral neutrophils from patients with adult periodontitis express more active elastase and total amounts of IL-1. The similar amounts of antigenic elastase suggests that this higher activity is possibly due to some kind of priming/activation already in the circulation.     

Interleukin-8 and granulocyte elastase in gingival crevicular fluid in relation to periodontopathogens in untreated adult periodontitis

BACKGROUND: This study aimed to determine the relationships among interleukin (IL)-8 and granulocyte elastase levels in gingival crevicular fluid (GCF) and the concomitant presence of periodontopathogens in untreated adult periodontitis. METHODS: GCF and subgingival plaque samples were collected from 16 patients with untreated adult periodontitis and 10 healthy control subjects. IL-8 levels were determined by enzyme-linked immunosorbent assay (ELISA). Granulocyte elastase was analyzed with a neutrophilic granulocyte-specific, low molecular weight and chromogenic substrate, L-pyroglutamyl-L-prolyl-L-valine-p-nitroanilide, and the maximal rate of elastase activity (MR-EA) was calculated. Five DNA probes were used to detect the presence of A. actinomycetemcomitans (A.a.), B. forsythus (B.f.), P. gingivalis (P.g.), P. intermedia (P.i.), and T. denticola (T.d.). RESULTS: Lower IL-8 concentrations and higher granulocyte elastase activities were found in patients than in healthy controls as well as in diseased conditions co-infected with B.f., P.g., P.i., and T.d. as compared to healthy conditions without the target species (P <0.05). IL-8 concentrations were positively correlated with MR-EA levels in the periodontitis conditions co-infected with B.f., P.g., P.i., and T.d. (P <0.05). A wide range of IL-8 concentrations was found among 15 patients when the periodontitis condition was characterized by co-infection with B.f., P.g., P.i., and T.d. MR-EA levels in the high IL-8 group of subjects were significantly higher than those in the low IL-8 group of subjects (P <0.01). CONCLUSIONS: The present study shows that the local host-bacteria interactions in untreated periodontitis are diverse in terms of the intensity of inflammatory responses measured by IL-8-related granulocyte elastase activity in GCF. This might reflect different phases of the inflammatory response due to shifts in host-bacteria interactions and therefore be indicative of a range of periodontal disease activity levels.     

牙周治疗对血清IFN-γ及IL-4水平的影响

目的:探讨牙周基础治疗对牙周炎患者血清干扰素-γ(IFN-γ)及白细胞介素-4(IL-4)水平的影响。方法:检测慢性牙周炎患者牙周基础治疗前和治疗后4～6周内,牙龈探诊深度、出血指数;采用双抗体夹心ELISA法,通过绘制标准曲线求出标本中IFN-γ和IL-4的浓度,对检测结果进行t检验。结果:慢性牙周炎患者血清IFN-γ、IL-4显著高于对照组(P<0.01);慢性牙周炎患者经牙周基础治疗后,其探诊深度、出血指数、血清IFN-γ浓度均显著降低(P<0.01),血清IL-4浓度无显著变化(P>0.05)。结论:IFN-γ、IL-4与牙周组织炎症的发病有关,控制牙龈炎症可降低血清IFN-γ水平,但IL-4浓度不因牙龈炎症的控制而下降。     

Shh蛋白与慢性牙周炎炎症程度的相关性研究

目的:探讨Sonic Hedgehog(Shh)信号通路中Shh蛋白与慢性牙周炎炎症程度的相关性。方法:选择健康对照20例(健康对照组),轻度牙周炎患者20例(轻度牙周炎组),中重度牙周炎患者20例(中重度牙周炎组),收集其龈沟液样本。采用ELISA法检测龈沟液中Shh蛋白、白细胞介素-6(IL-6)及白细胞介素-10(IL-10)的水平。结果:Shh蛋白、IL-6在慢性牙周炎组的水平均高于健康对照组(P<0.05),且中重度慢性牙周炎组高于轻度慢性牙周炎组(P<0.05);IL-10在轻度慢性牙周炎组的水平高于健康对照组及中重度慢性牙周炎组(P<0.05),且健康对照组高于中重度慢性牙周炎组(P<0.05);Shh蛋白、IL-6水平与出血指数(BI)、探诊深度(PD)均呈正相关(P<0.05)。结论:Shh信号通路可能参与了慢性牙周炎的炎症反应过程。     

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。     

Levels of interleukin-17 in gingival crevicular fluid and in supernatants of cellular cultures of gingival tissue from patients with chronic periodontitis

BACKGROUND AND AIMS: Interleukin-17 (IL-17) is a T-cell-derived cytokine that may play an important role in the initiation or maintenance of the pro-inflammatory response and has recently been found to stimulate osteoclastic resorption. The purpose of the present study was to determine the presence of IL-17 in gingival crevicular fluid (GCF) samples and in the culture supernatants of gingival cells from patients with chronic periodontitis. METHOD: GCF samples were collected during 30 s from two sites in 16 patients from periodontally affected sites (probing depth > or =5 mm, attachment loss > or =3 mm). The comparison with healthy controls was carried out by collecting GCF samples from eight healthy volunteers. GCF was collected using a paper strip and ELISA was performed to determine the total amount of IL-17. Supernatant cellular cultures of gingival cells were obtained from periodontal biopsies taken from 12 periodontitis patients and from eight healthy control subjects during the surgical removal of wisdom teeth. Spontaneous and phytohaemagglutinin (PHA)-stimulated levels of IL-17 were determined by ELISA. RESULTS: The total amount of cytokine IL-17 was significantly higher in the periodontitis group than the control group (45.9 versus 35.6 pg, p=0.005). Significantly higher GCF volume and amount of total proteins were obtained from periodontitis patients as compared with control subjects (0.98 versus 0.36 microl, p=0.0005; 0.12 versus 0.05 microg, p=0.0005, respectively). A higher concentration of IL-17 was detected in culture supernatants from periodontitis patients compared with healthy subjects, either without stimulation (36.28+/-8.39 versus 28.81+/-1.50 microg/ml, p=0.011) or with PHA stimulation (52.12+/-14.56 versus 39.00+/-4.90 microg/ml, p=0.012). Treatment with PHA induced a significant increase in the production of IL-17 in healthy subjects and periodontitis patients (p=0.001 and 0.003). CONCLUSIONS: The total amount of cytokine IL-17 in GCF samples and in the culture supernatants of gingival cells are significantly increased in periodontal disease.     

Increased amounts of laminin in GCF from untreated patients with periodontitis

BACKGROUND/AIMS: Our aim was to compare the levels of laminin and interleukin-8 (IL-8) in GCF from inflamed shallow (GP) and deep pockets (PP) in patients with periodontitis to these levels in GCF from inflamed shallow pockets (GG) in subjects with gingivitis alone. METHOD: Lactoferrin was used as a marker for the number of neutrophils in the sites sampled. The periodontitis group consisted of 13 subjects, having at least 6 sites with a pocket depth > or = 5 mm. The healthy control group consisted of 12 subjects with no clinical signs of periodontal destruction. GCF was collected with paper strips and the volume was measured immediately after sampling. Laminin, lactoferrin and IL-8 were measured using specific antibodies with an ELISA. RESULTS: The total amount of laminin was significantly higher in PP than in GG, but no significant difference was seen between GP and PP. The concentration and the total amounts of lactoferrin were similar in the three groups. The ratio between laminin and lactoferrin was higher in the samples from the patient, suggesting that neutrophils in patients are more activated and degrades more of the membrane per cell. The total amounts of IL-8 were very similar in the 3 groups, while the concentration also tended to be higher in the GP. CONCLUSIONS: Our study showed higher amounts of laminin in GCF from patients with periodontitis suggesting the presence of hyperactive neutrophils during the transmigration process through the endothelium/epithelium.     

Correlation between salivary IL-1beta levels and periodontal clinical status

OBJECTIVE: To assess the concentration of the proinflammatory cytokine IL-1beta in saliva of periodontally diseased and healthy patients and their relationship with the periodontal status. DESIGN: Unstimulated whole saliva samples from patients with chronic periodontitis (n=30), aggressive periodontitis (n=18) and healthy controls (n=18) were obtained for the study. The periodontal status of each subject was assessed by criteria based on probing depth, clinical attachment loss and the extent/severity of periodontal breakdown. The levels of IL-1beta were measured in saliva samples with a high sensitivity enzyme-linked immunosorbent assay (ELISA). RESULTS: Although no significant difference (P=0.624) was found for salivary IL-1beta levels between periodontitis groups, they were significantly greater (P<0.01) than those detected for healthy controls. Furthermore, Spearman correlation analysis showed statistically significant correlations (P<0.01) between data from salivary IL-1beta levels and clinical measurements. CONCLUSION: The findings of the present study reemphasize the importance of whole saliva as sampling method in terms of immunological purposes in periodontal disease and suggest that the elevated IL-1beta concentration may be one of the host-response components associated to the clinical manifestations of periodontal disease.