Presence and quantification of mast cells in the gingiva of cats with tooth resorption, periodontitis and chronic stomatitis

Objective

Mast cells are tissue-dwelling granule-containing immune cells that play a pivotal role in the pathogenesis of inflammation and other processes. The three most common orodental disorders in cats are periodontitis, feline resorptive lesions (FRL), and chronic gingivostomatitis (FCGS). The presence and density of mast cells in the gingiva has been established in healthy cats but not in cats affected by FRL, FCGS or periodontitis. The aim of the present study was to document and quantify the presence of mast cells in the gingiva adjacent to teeth affected by FRL, FCGS or chronic periodontitis.

Methods

Samples from the gingiva of 32 cats affected by FRL, FCGS or periodontitis were obtained and compared to samples obtained from 7 specific pathogen-free (SPF) cats. Evaluation of mast cells and the inflammatory infiltrate were performed on hematoxylin and eosin, and toluidine blue stained sections.

Results

Mast cells densities were significantly increased in gingival tissues adjacent to teeth affected by FRL, FCGS or periodontitis in comparison to SPF samples. There were no significant differences between gingival tissues of the FRL, FCGS and periodontitis groups. However, the relative inflammatory score in the FRL group was significantly lower as compared to the FCGS or periodontitis groups, yet with similar density of mast cells.

Conclusion

In the gingiva of cats affected with FRL, FCGS or periodontitis, there is an increase in the number of mast cells. The high number of mast cells in the FRL group and concurrent mild inflammatory reaction suggests the notion that mast cells may potentially play role in the pathogenesis of FRL.     

肥大细胞脱颗粒在牙周炎病理中的作用

目的:探讨肥大细胞脱颗粒在牙周炎病理中的意义。方法:将自愿接受研究的受试者分为3组:轻度慢性牙周炎组(轻度组,17例)、重度慢性牙周炎组(重度组,18例)、正常对照组(15例)。取牙龈活检组织制作连续切片,HE染色在光镜下观察各组牙周组织的组织学改变;甲苯胺蓝染色法观察各组肥大细胞的数量及肥大细胞脱颗粒状况;天狼猩红染色法观察各组牙周组织纤维化的程度。结果:与正常对照组相比,各慢性牙周炎组的牙周组织中肥大细胞显著增多(P<0.01);重度组与轻度组相比较肥大细胞数量和脱颗粒率均明显升高(P<0.01),牙周组织纤维化程度更加严重(P<0.05)。结论:肥大细胞的募集、脱颗粒和纤维化程度与牙周炎的严重程度相关,提示肥大细胞可能在牙周炎的发病和疾病进程中起着重要的作用。     

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。     

Immunohistochemical expression of mast cell tryptase in giant cell fibroma and inflammatory fibrous hyperplasia of the oral mucosa

This study analysed the immunohistochemical expression of mast cell tryptase in giant cell fibromas (GCFs). In addition, the possible interaction of mast cells with stellate giant cells, as well as their role in fibrosis and tumour progression, was investigated. For this purpose, the results were compared with cases of inflammatory fibrous hyperplasia (IFH) and normal oral mucosa. Thirty cases of GCF, 30 cases of IFH and 10 normal mucosa specimens used as control were selected. Immunoreactivity of mast cells to the anti-tryptase antibody was analysed quantitatively in the lining epithelium and in connective tissue. In the epithelial component (p = 0.250) and connective tissue (p = 0.001), the largest mean number of mast cells was observed in IFHs and the smallest mean number in GCFs. In connective tissue, the mean percentage of degranulated mast cells was higher in GCFs than in IFHs and normal mucosa specimens (p < 0.001). Analysis of the percentage of degranulated mast cells in areas of fibrosis and at the periphery of blood vessels also showed a larger mean number in GCFs compared to IFHs and normal mucosa specimens (p < 0.001). The percent interaction between mast cells and stellate giant cells in GCFs was 59.62%. In conclusion, although mast cells were less numerous in GCFs, the cells exhibited a significant interaction with stellate giant cells present in these tumours. In addition, the results suggest the involvement of mast cells in the induction of fibrosis and modulation of endothelial cell function in GCFs.     

成人牙周炎龈组织内浆细胞的超微结构观察

为了探讨牙周炎龈组织的免疫反应机理，对２０例牙周炎龈组织进行了透射电镜观察。结果表明，在炎性浸润细胞中以浆细胞（ｐｌａｓｍａｃｅｌ，ＰＣ）数目最多且有变性，根据浆细胞胞核、质膜及粗面内质网腔的变性程度，将浆细胞分为４期。浆细胞核中染色质从正常形态逐渐趋于浓聚、边集、固缩和破碎，质膜从完整清晰、部分缺失到全部丧失其完整性；粗面内质网腔从狭窄、扩张、断裂至囊泡化，其腔内容物的电子密度由低至高再降低；卢氏体（Ｒｕｓｓｅｌ′ｓｂｏｄｉｅｓ，Ｒｂ）从偶见到数目增多再减少；变性浆细胞最终发生凋亡。我们的观察结果提示，成人牙周炎龈组织反应有体液免疫参与。     

Release of cytokines by stimulated peripheral blood mononuclear cells in chronic periodontitis

Objective

The emergence of periodontal medicine increased interest in defining the behaviour of peripheral blood cells in periodontitis subjects in comparison with healthy group. The aim of this study was to evaluate the levels of interleukin (IL)-8, tumour necrosis factor-α (TNF-α), IL-6 and IL-10 released by Escherichia coli lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMC) obtained from the peripheral blood of chronic periodontitis subjects.

Design

PBMC samples were isolated from 19 systemically healthy donors, divided into generalized chronic periodontitis (n = 10) and healthy (n = 9) subjects. Cells were incubated for 24–48 h in 500 μL wells containing RPMI 1640 and stimulated with 1.0 ng/mL of E. coli LPS. Supernatants were used to quantify the amounts of IL-8, TNF-α, IL-6 and IL-10 released using enzyme-linked immunosorbent assay (ELISA).

Results

PBMC cells from periodontitis subjects released higher levels of TNF-α and IL-6 than those from healthy subjects (P < 0.05). Conversely, the supernatants of the stimulated PBMC cells obtained from healthy subjects presented higher amounts of IL-8 than those from periodontitis (P < 0.05). No differences were observed in the levels of IL-10 (P > 0.05) between groups.

Conclusion

In conclusion, the results of the present study showed that E. coli LPS-stimulated PBMC from subjects with periodontitis present a different pattern of cytokine release when compared to PBMC from healthy subjects. This phenomenon could have implications locally, in periodontitis, as well as in systemic diseases.     

The presence of IL-17+/FOXP3+ double-positive cells in periodontitis

Increasing evidence suggests that distinct inflammatory cytokines convert forkhead box protein P3 (FOXP3(+)) regulatory T-cells (Tregs) into IL-17-producing cells (Th17 cells) in vitro. However, this functional plasticity has not been examined in the pathogenesis of periodontal disease. In this study, we analyzed the IL-17A(+)FOXP3(+) cells present in periodontitis lesions to determine the association between Treg conversion and the pathogenesis of periodontitis. The immunohistochemical analysis of gingival tissues demonstrated that the numbers of Th17 cells (IL-17A(+)FOXP3(-)) and Tregs (IL-17A(-)FOXP3(+)) were greater in periodontitis lesions than in gingivitis lesions. We further identified a small number of IL-17A(+)FOXP3(+) cells in periodontitis lesions but not in gingivitis lesions. The flow cytometry analysis of CD4(+) T-cell lines established from gingival tissues and the peripheral blood of periodontitis patients showed that the proportion of Tregs was reduced and the proportion of IL-17A(+)FOXP3(+) cells among all FOXP3(+) cells was elevated in gingival tissue T-cell lines relative to the proportions in peripheral blood T-cell lines. Our findings indicate that Treg-Th17 conversion may occur in periodontitis lesions. Further studies addressing the role of Treg conversion during inflammatory responses against periodontopathic bacteria are needed.     

不同糖蛋白B基因型人巨细胞病毒感染与慢性牙周炎的关系

目的探讨不同糖蛋白B（gB）基因型人巨细胞病毒（HCMV）感染与慢性牙周炎（CP）的关系。方法采用套式聚合酶链反应（nPCR）检测65例CP患者龈下菌斑标本和24名牙周健康者龈沟液标本中HCMV的gB基因，并进行限制性片段长度多态性分析（RFLP）对gB基因进一步分型。分析HCMV及其不同gB基因型感染与牙周炎严重程度的关系。结果以位点计，CP的HCMV感染率59．23％（154／260），明显高于牙周健康者的感染率32．29％（31／96）（P〈0．01）。cP的HCMV感染者中gBⅠ型、gBⅡ型、gBⅠ和Ⅱ混合型分别占11．7％（18／154）、80．5％（124／154）和7．8％（12／154）；牙周健康的HCMV感染者中gBⅠ型占45．2％（14／31），gBⅡ型占38．7％（12／31），gBⅠ和Ⅱ型混合感染占16．1％（5／31）。与牙周健康者相比，CP患者感染HCMV以gBⅡ型明显占多数（P〈0．01）。不同gB基因型HCMV感染在不同临床附着丧失、探诊深度和牙龈指数的位点中的分布差异均无统计学意义（P〉0．05）。结论HCMV感染与cP密切相关，但与牙周炎严重程度无直接关系。gBⅡ型可能是与CP相关的HCMV优势基因型。     

双黄补缓释药条治疗慢性牙周炎的临床研究

目的 评价局部应用双黄补缓释药条对控制牙周菌斑微生物及牙同炎症的作用。方法选择中、重度慢性牙周炎患者31例，采用同一口腔内病情相同牙齿的自身对照设计，将口腔内左右两侧共120个受试牙位点配对分组。基线前须完成全口龈上洁治及口腔卫生宣教。基线时受试牙住点刮取龈下菌斑涂片，记录螺旋体比例，并记录探诊深度、附着丧失和探诊后出血等临床指数，然后行龈下刮治术，用药组每周放两次双黄补缓释药条，共放4次，对照组不放药。第14天和第41天检查各项指数(同基线)。结果 用药组和对照组的各项临床指标、龈下菌斑中螺旋体比例均较基线时下降，且用药组下降程度更明显。结论 双黄补缓释药条可有效改善牙周菌斑微生物的组成，控制牙周炎症。     

实验性牙周炎中调节性T细胞的检测及免疫状态

目的：通过口腔涂抹牙周致病菌加腹腔注射抗原的方法建立一种模拟人类牙周病发病的动物模型,分析调节性T细胞在牙周炎小鼠中的表达情况。方法：采用实体显微镜评价牙周骨吸收的程度,HE染色法观察牙周组织病变的炎细胞浸润情况;FACS检测调节性T细胞在牙周炎慢性炎症期间的表达;实时定量PCR检测牙龈组织中TGF-β1、IL-10mRNA的表达。采用SPSS 17.0软件包对数据进行统计学分析。结果：第4周后,牙周炎组与正常对照组相比,釉牙骨质界至牙槽嵴顶的距离显著增加(P<0.05);牙周组织中大量的炎性细胞浸润,牙周袋加深;TGF-β1、IL-10mRNA的表达水平显著降低(P<0.01);脾脏中调节性T细胞的比率及数量显著少于正常对照组(P<0.05)。结论：牙周炎小鼠中调节性T细胞的数量及功能存在异常,可能与其分泌的免疫抑制性细胞因子表达降低有关。     

慢性牙周炎维护治疗期病程纵向观察

目的:了解慢性牙周炎维护治疗期临床指标变化规律。方法:对牙周非手术基础治疗后进入维护治疗期的22名慢性牙周炎患者进行9个月的纵向观察。每3个月给予口腔卫生宣教,龈上下洁刮治和根面平整,并在基线和每次复查时记录临床检查指标,包括:探诊牙周袋深度(PPD)、探诊附着水平(PAL)和探诊出血(BOP)。结果:患者平均PPD减少0.44 mm,PAL增加0.38 mm,病情的好转主要集中在观察期的前3个月。4mm以上的牙周袋较浅袋更易随时间延长发生牙周袋深度增加,后牙较前牙有更多位点出现牙周袋加深。结论:非手术基础治疗配合定期维护,在较短观察期内可使大多数位点临床指标保持稳定甚至好转。     

Middle zone of the root surface integument of human teeth affected by chronic periodontitis

This study has confirmed the presence on the root surface of teeth affected by chronic periodontitis of a zoned organic integument analogous to that on enamel. Measurements showed a highly significant correlation between the levels of the bounding structures of this zone, that is, the apical plaque border and the coronal border of residual periodontal ligament, for both shallow and deep pockets. The middle zone between these two was on average 1 mm wide.     

Elevation of white blood cells and platelet counts in patients having chronic periodontitis

Background and aimMany risk factors that might contribute to the pathogenesis of atherosclerosis have been proposed, including chronic inflammation and infection. Furthermore, systemic inflammatory responses to periodontal bacteria have been suggested as a pathogenetic link between periodontal disease and atherosclerosis. The purpose of this study was to estimate the white blood cell (WBC) and platelet counts in chronic periodontitis patients.Materials and methodsFifty patients with chronic periodontitis and 50 patients with healthy periodontium were included in this study. Oral hygiene status, pocket probing depth (PPD) and clinical attachment level (CAL) were measured. During clinical evaluation, venous blood samples were taken to analyze the WBC and platelet counts. Statistical analysis was utilized to compare differences across groups.ResultsPeriodontitis patients demonstrated a significantly higher WBC count (7.22 ± 1.42 × 10⁹ cells/L) than that of control patients (5.64 ± 1.56 × 10⁹ cells/L; P < 0.001). The platelet count of patients with chronic periodontitis (290.73 ± 56.56 × 10⁹ cells/L) was also significantly higher compared to the healthy group (223.37 ± 50.27 × 10⁹ cells/L; P < 0.001).ConclusionLevels of WBCs and platelets are elevated in periodontitis patients compared to healthy controls.     

Quantification of mast cells in different stages of human periodontal disease

OBJECTIVE AND METHODS: Among the cells involved in immune and inflammatory responses in periodontal disease, mast cells have been shown to be capable of generating a large number of biologically active substances. The present study was undertaken to identify and quantify the presence of mast cells in different stages of human periodontal disease using histochemical (toluidine blue) and immunohistochemical (tryptase-positive mast cells) techniques. RESULTS: Mast cell densities (cells per mm(2)) were significantly increased in chronic periodontitis/gingivitis lesions compared with clinically healthy gingival tissues (Health) uniquely by immunohistochemical technique. Interestingly, mast cells were distributed specially in close apposition to mononuclear cells. CONCLUSIONS: In human periodontal disease there is an increase in the number of mast cells that may be participating either in the destructive events or in the defense mechanism of periodontal disease via secretion of cytokines, including perpetuation of the Th2 response, and cellular migration and healing processes.     

慢性牙周炎龈下菌斑中福赛斯坦纳菌的检测

目的检测慢性牙周炎患者和牙周健康者龈下菌斑中福赛斯坦纳菌的数量和所占比例,探讨福赛斯坦纳菌与牙周炎发生发展的关系。方法采集经常规聚合酶链反应（PCR）法检测福赛斯坦纳菌为阳性的61例慢性牙周炎患者和12例牙周健康者的龈下菌斑,应用TaqMan实时荧光定量PCR法对菌斑的细菌总量和福赛斯坦纳菌的数量进行定量检测,构建含有福赛斯坦纳菌和真细菌靶基因的重组质粒,建立定量标准。结果本研究设计的引物和探针具有良好的特异性和敏感性;慢性牙周炎患者病变位点的福赛斯坦纳菌数量和细菌总量均高于牙周健康者的健康位点,且福赛斯坦纳菌在龈下菌斑中的比例也比健康位点高（P<0.05）;龈下菌斑的细菌数量与探诊深度呈正相关（P<0.001）;龈下菌斑中福赛斯坦纳菌所占比例在不同的探诊深度间无统计学差异（P>0.05）。结论龈下菌斑中福赛斯坦纳菌的数量与牙周状况有密切关系,实时荧光定量PCR法是研究牙周病病因及治疗方法的有效手段。     

牙周炎对慢性阻塞性肺疾病大鼠肺损伤的作用研究

目的:应用SD大鼠建立慢性牙周炎(chronic periodontitis,CP)和慢性阻塞性肺疾病(chronic obstructive pulmonarydiseases,COPD)动物模型,探讨CP在COPD发生发展过程中对肺损伤的作用。方法:将40只SD大鼠雌雄各半随机分成4组:正常对照组,CP组,COPD组,COPD伴CP组,每组10只。各组按照实验设计施加相应建模因素,实验10周末分别检查大鼠的龈沟出血指数(SBI)、肺功能状况,ELISA法检测血清中TNF-α和IL-1含量,然后处死大鼠,取材观察牙槽骨吸收程度(ABL)、牙周组织和肺部组织病理学改变,对数据进行统计分析。结果:COPD伴CP组大鼠的肺功能明显降低,龈沟出血指数和牙周组织改变与CP组相似,实验组血清IL-1和TNF-α水平明显高于对照组(P<0.05)。结论:CP在COPD大鼠肺损伤的发生发展过程中具有促进作用。     

Immunohistochemical analysis of lymphatic vessel density and mast cells in oral tongue squamous cell carcinoma

The aim of this study was to analyze lymphangiogenesis and the presence of mast cells in oral tongue squamous cell carcinoma (OTSCC), correlating the findings with clinicopathological parameters (clinical stage, tumor size, nodal metastasis, histological grade of malignancy, local recurrence, and clinical outcome). Fifty-six cases of primary OTSCC were selected. Lymphatic vessels and mast cells were identified by immunostaining with anti-podoplanin (D2-40) and anti-tryptase antibody, respectively. Lymphatic vessel density (LVD) and mast cell density (MCD) were determined in the intratumoral and peritumoral areas. Intratumoral LVD was higher in advanced clinical stages (III/IV) when compared to early-stage (p = 0.017) and in metastatic cases compared to non-metastatic tumors (p = 0.013). Peritumoral LVD and intratumoral or peritumoral MCD did not differ significantly according to the clinicopathological parameters of OTSCCs (p > 0.05). No significant correlations between LVD and MCD were observed at the intratumoral (r = ?0.014; p = 0.918) or peritumoral level (r = 0.156; p = 0.251). Our findings suggest that intratumoral lymphatic vessels, compared to peritumoral lymphatic vessels, appear to be more related to the progression of OTSCC. MCD alone does not seem to be determinant for lymphangiogenesis or for the biological behavior of OTSCC, indicating multiple pro- and antitumor effects of these inflammatory cells.     

2型糖尿病患者慢性牙周炎细菌学研究

目的 :研究 2型糖尿病患者慢性牙周炎的龈下菌群以及菌群变化与血糖、糖化血红蛋白的关系。方法 :细菌学厌氧培养、PCR检测技术。结果 :2型糖尿病患者慢性牙周炎的龈下菌群以厌氧菌为主。产黑菌、二氧化碳噬纤维菌数量与糖尿病患者空腹血糖、糖化血红蛋白之间存在正相关关系。产黑菌、二氧化碳噬纤维菌数量随患者空腹血糖、糖化血红蛋白的升高而增加。结论 :2型糖尿病慢性牙周炎患者龈下菌斑相关菌与血糖、糖化血红蛋白变化密切相关     

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。