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。     

精氨酸酶在大鼠实验性牙周炎牙周组织中的表达变化

目的:观察精氨酸酶(arginase,Arg)在不同时期、不同严重程度大鼠实验性牙周炎牙周组织中表达的变化,探讨Arg与牙周炎的相关性。方法:选用40只Wistar雄性大鼠,每只大鼠均用丝线结扎右下颌第一磨牙(实验组),左下颌第一磨牙不做处理(对照组),喂以高糖饮食。分别于结扎后1、2、4、6周,经临床和组织学(HE染色)观察确认大鼠牙龈炎、不同程度牙周炎模型建立成功后,取大鼠下颌骨连同受试牙和牙周组织,制备切片,免疫组化染色法检测正常对照组、牙龈炎组、不同程度牙周炎组大鼠牙周组织中Arg的表达,并进行图像定量分析。结果:在正常牙周组织中,偶有表达;而各实验组牙周组织中Arg主要在巨噬细胞和血管内皮细胞中表达,其表达阳性细胞数随炎症程度加重而增加,结扎4周(重度牙周炎)时达到高峰,到慢性牙周炎时期(结扎6周),表达较急性期有所降低,各组间比较,差异均有统计学意义(P<0.05)。结论:牙周组织内Arg的表达与牙周炎炎症程度呈明显正相关。     

程序性死亡配体1负性调控慢性牙周炎的炎症反应

目的 探讨程序性死亡配体1（PD-L1）在慢性牙周炎中是否有表达,以及与不同程度慢性牙周炎的相关关系,为阐明慢性牙周炎的免疫调控机制、临床治疗和预后提供依据。方法 收集健康人和慢性牙周炎患者的牙龈和牙周膜组织。根据临床探测,分为正常对照组、轻度牙周炎组、重度牙周炎组。运用荧光定量聚合酶链反应（PCR）测定不同组牙周组织中PD-L1 mRNA的表达;免疫印迹（Western blot）以及免疫组织化学方法测定不同组牙周组织中PD-L1蛋白的表达。结合临床影像资料,分析PD-L1的差异表达量和不同程度慢性牙周炎的关系。结果 轻度牙周炎组牙周组织中PD-L1的相对表达量显著高于重度牙周炎组（P<0.01）;正常对照组牙周组织中PD-L1的相对表达量与重度牙周炎组牙周组织相比无统计学意义（P>0.05）。结论 PD-L1在牙周组织的表达可负性调控炎症牙周组织损伤。     

内质网应激诱导的细胞凋亡在慢性牙周炎中的作用初探

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。     

TNF-α与牙周炎和慢性肺感染的相关性实验研究

目的:观察肿瘤坏死因子-α(TNF-α)在牙周组织及肺组织中的表达及变化,从细胞因子的角度探讨牙周炎和慢性肺感染的相互关系。方法:将30只昆明小鼠随机平均分为A组和B组,随机选取其中一组(A组)建立小鼠慢性肺感染动物模型后,将A组小鼠随机分为3个亚组,在原有肺感染的基础上叠加建立牙周炎动物模型,即A1组(慢性肺感染+牙周正常组)、A2组(慢性肺感染+轻度牙周炎组)、A3组(慢性肺感染+重度牙周炎组)。B组小鼠亦分为3个亚组,建立牙周炎动物模型,即B1组(正常对照组)、B2组(轻度牙周炎组)、B3组(重度牙周炎组),采用免疫组化染色观察TNF-α在各组牙周组织及肺组织中的表达,应用计算机图像分析系统半定量分析TNF-α的阳性表达水平。结果:牙周袋深度、牙槽骨吸收总量、肺灌洗液淋巴细胞百分数及牙周组织和肺组织中TNF-α的平均光密度值(OD)在A组各亚组间及B组各亚组间的差异均有统计学意义(P<0.05),且A3>A2>A1、B3>B2>B1。而在A1与B1、A2与B2、A3与B3的比较中牙周袋深度、牙槽骨吸收总量及牙周组织中TNF-α平均光密度值差异无统计学意义(P>0.05)。结论:TNF-α可能参与了牙周炎和慢性肺感染的发病过程,且在牙周炎与慢性肺感染的关系中发挥作用。     

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。     

骨质疏松症与重度慢性牙周炎之间的相关因素探讨

目的:探讨骨质疏松症与慢性牙周炎之间的相关性。方法:对126例骨质疏松症病人和143例骨密度正常者进行牙周组织检查,根据诊断标准,将诊断为牙周健康者以及轻、中、重度慢性牙周炎病人进行统计分析。结果:慢性牙周炎在骨质疏松症病人与非骨质疏松症者中的患病率无显著性差异(P>0.05),重度慢性牙周炎在骨质疏松症病人与非骨质疏松症者中的患病率有显著性差异(P<0.05)。结论:骨质疏松症可能是重度慢性牙周炎的危险因素。     

龈沟液中神经肽P物质与慢性牙周炎关系临床研究

目的:探讨龈沟液中神经肽P物质(SP)与慢性牙周炎发生发展的关系.方法:研究组48 例,对照组45 例,研究组口内各选1 个牙周炎牙位、对照组各选1 个健康牙位收集龈沟液样本,采用放射免疫分析技术分别测定2 组龈沟液SP含量;记录牙龈指数(GI)、牙周袋探诊深度(PD)、附着丧失(AL)、牙槽骨吸收(ABL)情况,并进行与SP的相关性分析.结果:研究组SP含量显著高于对照组(t＝9.92,P<0.01);轻度牙周炎SP含量及牙周临床指标与对照组相比,均有显著差异(P<0.01);轻度牙周炎龈沟液SP含量与GI无显著相关性,但与PD、AL、ABL之间,以及中、重度牙周炎龈沟液SP含量与GI之间,均呈显著性相关(P＜0.05);中、重度牙周炎SP含量与PD、AL、ABL之间均呈非常显著性相关(P<0.01).结论:龈沟液中SP含量与慢性牙周炎的发生发展密切相关,可以反映牙周组织的破坏程度;测定患者龈沟液中SP含量,对于判断病情、指导治疗具有重要意义.     

自体富血小板纤维蛋白治疗重度慢性牙周炎临床效果观察

目的观察自体富血小板纤维蛋白(PRF)用于重度慢性牙周炎手术治疗中的临床效果。方法选择2009年1月至2011年8月需要进行翻瓣术的重度慢性牙周炎患者18例(18颗患牙),分为试验组和对照组各9例。行常规翻瓣术,将PRF植入牙周组织缺损处(试验组)或仅行翻瓣术(对照组),术前及术后4个月测量探诊深度(PD)与临床附着水平(CAL)。结果两组各自术前与术后4个月的PD和CAL值比较,差异均有统计学意义(P<0.05)。试验组术前、术后的PD和CAL差值分别与对照组比较,差异均有统计学意义(P<0.05)。结论将PRF用于重度慢性牙周炎手术治疗中,可促进患牙牙周组织修复、改善牙周状况。     

Mast Cell Degranulation in Human Periodontitis

Background: Mast cells are tissue‐resident immune cells that participate in a variety of allergic and inflammatory conditions. Limited attention has been given to the role of mast cells in periodontal diseases, and the effects of mast cell degranulation on the chronic stages of non‐allergic inflammation, particularly in periodontitis, are not known. The present study analyzes the relationship between the mast cell degranulation and human periodontal disease progression. Methods: A total of 50 clinical specimens including moderate periodontitis (n = 17), advanced periodontitis (n = 18), and healthy control tissues (n = 15) were used in this study. All specimens were fixed in 10% buffered formalin and stained with hematoxylin and eosin for histopathology, with toluidine blue for identifying mast cells, and by immunohistochemistry for the expressions of mast cell tryptase in periodontal tissues. The total and degranulated mast cell densities (per high‐power field) were quantified in the specimens. Results: Compared with healthy controls, there were significantly increased both total and degranulated mast cell densities in human moderate (P <0.01) and advanced (P <0.01) periodontitis groups by toluidine blue staining, and there were significantly higher densities of both total and degranulated tryptase‐positive mast cell subpopulation in the moderate periodontitis group (P <0.01) and even significantly higher subpopulation densities in the advanced periodontitis group by immunohistochemical staining, in which both total and degranulated mast cell densities were significantly higher in the advanced periodontitis group than those in the moderate periodontitis group (P <0.01) by both toluidine blue staining and immunohistochemical staining. There was significantly more severe periodontal inflammatory pathology in the advanced periodontitis group than in the moderate periodontitis group (P <0.01). Conclusion: These findings indicate a significant correlation among tryptase‐positive mast cell density, the degree of their degranulation, and the human periodontitis severity, and the results of this study further indicate that mast cell degranulation appears to be associated with human periodontal disease.     

Role of Smoking and Type 2 Diabetes in the Immunobalance of Advanced Chronic Periodontitis

Background : This study evaluates the tissue levels of interleukin (IL)‐17⁺, IL‐15⁺, Foxp3⁺ cells, fibrosis, and plasma B‐cell infiltration in sites with chronic periodontitis in smokers and subjects with type 2 diabetes. Methods: Gingival biopsies were harvested from the following groups: systemically and periodontally healthy subjects (healthy group, n = 10); non‐smokers and subjects with advanced periodontitis and without diabetes (non‐risk factor/periodontitis group, n = 10); heavy smokers with advanced periodontitis and without diabetes (smoking/periodontitis group, ≥20 cigarettes per day for at least the past 5 years, n = 10); and non‐smoking poorly controlled subjects with diabetes (glycated hemoglobin levels ≥9%) with advanced periodontitis (diabetes mellitus/periodontitis group [DMP], n = 10). The number of IL‐17⁺, IL‐15⁺, and Foxp3⁺ cells was analyzed by immunohistochemistry, whereas the amount of fibrosis and plasma B‐cell infiltration in gingival tissue was analyzed by histomorphometry. Results: The number of Foxp3⁺ cells was significantly higher in the periodontitis groups compared to the healthy group (P <0.05). The DMP group presented higher levels of Foxp3⁺ cells than other periodontitis groups (P <0.05). The levels of IL‐15⁺ and IL‐17⁺ cells and the amount of fibrosis were higher in the DMP group than in the other groups (P <0.05). There was a trend for a decreased B‐cell infiltration in the DMP group (P >0.05). There was a slightly significant negative correlation between B‐cell infiltration and the amount of fibrosis (P <0.05). Conclusion: Upregulation of IL‐17⁺, IL‐15⁺, and Foxp3⁺ cells and increased amounts of fibrosis were observed in chronic periodontitis sites in subjects with type 2 diabetes, suggesting that periodontitis development in these subjects may be influenced by the T helper 17/T regulatory axis.     

Role of receptors of advanced glycation end‐products (RAGE) in type 2 diabetic and non‐diabetic individuals with chronic periodontal disease: an immunohistochemical study

Aim: The relationship between diabetes and periodontal disease is well established. It has been shown that advanced glycation end‐products might exert noxious effects on several tissues of the body through its receptor. Evidence for the role of receptors of advanced glycation end‐products in periodontal disease for diabetes is limited, and their presence in human gingival tissues has been demonstrated in few studies. In this study, we demonstrate the presence of receptors of advanced glycation end‐products in patients with chronic periodontitis, with and without type 2 diabetes. Methods: Gingival biopsies from 19 patients with both type 2 diabetes and chronic periodontitis, and 18 healthy controls with chronic periodontitis, were immunohistochemically stained for receptors of advanced glycation end‐products. Results: On immunohistochemical analysis, positive staining for receptors of advanced glycation end‐products was seen in the endothelium and the basal and spinous layers of the inflamed gingival epithelium in both type 2 diabetes and non‐diabetes tissue, with a statistically‐significant difference between both groups (P < 0.05). Conclusions: There was a significant difference in receptors of advanced glycation end‐product immune reactivity between both groups. Receptors of advanced glycation end‐product increase in type 2 diabetes gingival tissue might indicate possible involvement of this receptor in periodontal destruction in individuals with type 2 diabetes.     

Matrix metalloproteinases and their inhibitors in gingival mast cells in persons with and without human immunodeficiency virus infection

BACKGROUND: Mast cells are a prominent cell type in the gingival infiltrate in periodontitis. In this study we examined the expression by gingival mast cells of matrix metalloproteinases, MMP-1, MMP-2, MMP-8 and the tissue inhibitors of metalloproteinases, TIMP-1 and TIMP-2. METHODS: Gingival specimens from 12 human immunodeficiency virus-negative (HIV-) and 15 HIV-positive (HIV+) patients with chronic marginal periodontitis (CMP), and from 10 HIV- and four HIV+ controls with clinically healthy gingiva (HG) were examined after double immunofluorescence staining for mast cell tryptase, combined with antibodies for MMP-1, MMP-2, MMP-8 or their inhibitors TIMP-1 and TIMP-2. RESULTS: In the HIV+CMP, HIV+HG and HIV-CMP groups, all mast cells expressed MMP-1 and MMP-8, whereas a smaller proportion (40-60%) in the HIV-HG controls displayed such staining. The former groups also displayed a significantly higher proportion (39-64%) of mast cells expressing MMP-2 as compared with the HIV-HG group (21-31%). All groups displayed similar proportions of TIMP-1 expressing mast cells (86-100%), whereas significantly increased proportions of TIMP-2+ mast cells were seen in the HIV+CMP, HIV+HG and HIV-CMP groups (18-25%) as compared with the HIV-HG group (8-13%). Mast cells were the cell type that most prominently expressed MMP-1 and MMP-8. MMP-2 expression was also strong in mast cells, but was also similarly expressed in other cell types. CONCLUSION: The chronically inflamed periodontal lesions in the present study appeared with little evidence of mast cell degranulation. The results show, however, that mast cells in inflamed gingiva have the potential to degrade extracellular matrix if appropriately triggered.     

不同程度慢性牙周炎患者病情的二年自然进展

目的　观察不同程度慢性牙周炎的自然进展规律。方法　纵向观察 16 9例轻、中、重度慢性牙周炎患者在 2年中的疾病自然进展 ,检查除第三磨牙外的全口牙 ,每颗牙 6个位点 ,以探诊深度、附着丧失为指标 ,两次检查之间附着丧失加重≥ 3mm的牙位定为活动性进展。结果　活动性发生率依基线时疾病的轻、中、重程度而逐渐增高 ,按位点活动性的发生率分别为 0 14 %、0 39%及0 73% ,按个体活动性的发生率分别为 15 5 6 %、2 9 89%及 4 3 2 4 % ,差异有统计学意义。结论　重度牙周炎患者是牙周破坏活动性进展的高危人群。     

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.     

可溶性髓样细胞触发性受体-1在不同牙周状态的龈沟液中表达研究

目的　分析比较可溶性髓样细胞触发性受体-1（soluble triggering receptor expressed on myeloid cells,sTREM-1）在不同牙周状态的龈沟液（gingival crevicular fluid,GCF）中表达情况,探讨sTREM-1在慢性牙周炎发生发展过程中的作用。方法　选取2017年1月至2018年6月于福建医科大学附属口腔医院牙周科就诊的18 ~ 60岁患者85例。根据牙周临床检查,将所有患者分成5组：健康对照组（牙周组织健康者18例）、慢性牙龈炎组（18例）、轻度慢性牙周炎组（14例）、中度慢性牙周炎组（15例）和重度慢性牙周炎组（20例）。采用酶联免疫吸附法（ELISA）检测比较不同牙周状态的GCF中sTREM-1的表达,并与患者的牙龈指数（GI）、牙周袋探诊深度（PD）及附着丧失（CAL）情况进行相关性分析。结果　慢性牙龈炎组患者及轻度、中度和重度慢性牙周炎组患者GCF中sTREM-1的表达水平均高于健康对照组,差异均有统计学意义（均P < 0.05）。与慢性牙龈炎组相比,中度和重度慢性牙周炎组患者GCF中sTREM-1的表达水平均显著增高,差异均有统计学意义（均P < 0.05）;重度慢性牙周炎组患者GCF中sTREM1的表达水平显著高于轻度慢性牙周炎组,差异有统计学意义（P < 0.05）;其余组间两两比较,差异均无统计学意义（均P > 0.05）。此外,sTREM-1的表达水平与牙周临床指标GI、PD及CAL均呈正相关。结论　GCF中sTREM-1的表达水平与牙周组织的炎症程度密切相关。检测GCF中sTREM-1的含量对判断牙周组织的炎症状态具有潜在应用价值。