β-防御素-2多肽在不同类型牙周炎及健康牙龈组织中的表达

目的:检测人β-防御素-2(humanbetadefensin-2,HBD-2)在牙周病病变牙龈和健康牙龈中的表达。方法:应用SP法检测健康牙龈(HC组,11例)、慢性牙周炎(CP组,18例)和侵袭性牙周炎(AgP组,9例)牙龈中HBD-2蛋白的表达水平,所得数据用SPSS11.5统计分析软件进行单因素方差分析。结果:HBD-2蛋白在所有牙龈标本中均有表达;HBD-2蛋白表达可见于牙龈复层鳞状上皮全层,主要位于棘层以上细胞胞质内。3组的表达水平分别为:健康牙龈组(31.55±12.66)%、慢性牙周炎组(17.31±7.64)%、侵袭性牙周炎组(25.06±8.04)%,健康组表达水平显著高于慢性牙周炎组(P<0.05),其他组间差异无统计学意义。结论:HBD-2多肽在健康牙龈和牙周病牙龈上皮中广泛表达,提示其在维系牙周健康及宿主免疫防御反应中可能发挥重要作用。     

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。     

牙周炎患者牙龈组织中IL-21表达的研究

目的：研究不同类型牙周炎患者牙龈组织中IL-21基因的表达,探讨其在牙周炎发病中的作用。方法：选择慢性牙周炎患者12例,侵袭性牙周炎8例,健康对照组8例,采用实时定量PCR方法定量检测IL-21 mRNA在牙龈组织表达情况。结果：慢性牙周炎、侵袭性牙周炎、健康对照组牙龈组织中均有IL-21 mRNA的表达,慢性牙周炎组和侵袭性牙周炎组IL-21 mRNA相对表达量分别为0.000534±0.000504和0.00602±0.000137,显著高于健康对照组0.000161±0.000352,差异有统计学意义(P<0.05)。慢性牙周炎和侵袭性牙周炎牙龈组织IL-21mRNA表达没有显著性差异（P>0.05）。结论：IL-21在慢性牙周炎发病机制中可能发挥重要作用。     

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。     

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     

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     

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。     

慢性牙周炎病变部位白细胞介素-8表达的初步研究

目的探讨牙龈组织中自细胞介素-8(IL-8)与慢性牙周炎的关系。方法采用免疫组化法检测32例牙龈组织样本(16例健康者，16例慢性牙周炎患者)，分析IL-8的检出率及IL-8分泌细胞在牙龈组织中的分布。结果慢性牙周炎患者中的IL-8的检出率均高于健康者，IL-8阳性细胞分布于牙龈的上皮和结缔组织中。结论研究结果提示IL-8参与了牙周炎的病理过程。     

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。     

慢性牙周炎患者牙龈组织内诱导型一氧化氮合酶表达的研究

目的:研究牙周健康者和慢性牙周炎患者牙龈组织中诱导型一氧化氮合酶的表达强度,探讨一氧化氮在牙周病发病过程中的作用.方法:选择牙周健康组、慢性牙周炎活动期组,慢性牙周炎静止期组各20例,采取免疫组织化学的方法染色,光镜下观察牙龈组织内诱导型一氧化氮合酶的表达强度.结果:慢性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和间质组织的细胞胞浆中阳性表达,正常组表达强度弱于慢性牙周炎静止期组和活动期组,慢性牙周炎静止期组表达强度弱于慢性牙周炎活动期组.结论:一氧化氮参与了慢性牙周炎的发生和发展过程,牙龈组织中诱导型一氧化氮合酶的表达强度与慢性牙周炎的炎症程度密切相关.     

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.     

牙龈组织炎症程度与COX-2蛋白表达的关系

目的:运用免疫组化技术观察牙龈组织中COX-2的表达,探讨COX-2与牙周病的关系。方法:首先对选用组织采用普通苏木素-伊红(HE)制片,观察炎症细胞浸润程度,根据炎症水平高低,选择两组进一步进行免疫组化检测,结果与正常组织相比较。结果:实验组COX-2蛋白表达要明显高于对照组(P<0.01)。结论:COX-2表达与炎症程度有关,炎症程度越高,COX-2表达也越高,两者呈正相关。     

Vascular endothelial growth factor in gingival tissues and crevicular fluids of diabetic and healthy periodontal patients

BACKGROUND: Periodontal disease is one of the major oral problems encountered in patients with diabetes mellitus (DM). Vascular changes, neutrophil dysfunction, altered collagen synthesis, and genetic predisposition observed in DM may contribute to periodontitis; and the vascular alterations observed in such patients may depend on vascular endothelial growth factor (VEGF) actions. Few reports are available about the mechanism of neovascularization and the angiogenic factors that contribute to the periodontal pathology and the role of VEGF in periodontal diseases. The aim of this study is to compare VEGF expression in healthy and periodontally diseased tissues with gingival crevice fluid (GCF) of healthy persons and diabetic patients. METHODS: Gingival tissue and GCF samples were collected from sites of periodontitis in 10 healthy subjects and in 10 type 2 diabetic patients, and from the sites of healthy gingiva within the same groups. Therefore, each patient became his/her own control. Additionally, 10 people without any systemic or periodontal diseases were enrolled, forming a negative control group. Thus, a total of 50 tissue and 50 GCF samples were provided. RESULTS: No VEGF staining was observed in the negative control group or in the systemically healthy people's healthy tissue samples, whereas four samples of diabetic patients showed positive staining (P < 0.05). However, VEGF was revealed in two tissue samples of periodontal sites of systemically healthy people and in six samples of the diabetic patients (P > 0.05). In all test groups, GCF VEGF levels were higher in periodontal sites (P < 0.05) than in healthy sites. CONCLUSION: The results of this study showed that VEGF is increased in all periodontal tissues of both groups and in the healthy sites of diabetic patients. Additionally, GCF VEGF values increased in periodontal sites of all test groups.     

Expression of toll-like receptors 2, 4 and 9 is increased in gingival tissue from patients with type 2 diabetes and chronic periodontitis

Rojo‐Botello NR, García‐Hernández AL, Moreno‐Fierros L. Expression of toll‐like receptors 2, 4 and 9 is increased in gingival tissue from patients with type 2 diabetes and chronic periodontitis. J Periodont Res 2012; 47: 62–73. © 2011 John Wiley & Sons A/S Background and Objective: Broad evidence indicates that diabetes both increases the risk and hastens the progression of periodontal disease. Likewise, chronic inflammation or infections seem to provoke insulin resistance and thereby contribute to the development of diabetes and its complications. Innate immune responses, which appear to be altered in individuals with diabetes, are usually mediated by the recognition of pathogens through toll‐like receptors (TLRs). The constitutive expression of some TLRs has been reported in healthy human gingival tissue. Interestingly, the expression of TLRs 2 and 4 is increased with the severity of periodontal disease. Considering that the inflammatory reaction is exacerbated in individuals with diabetes and periodontitis, we suspected that the expression of some TLRs might be increased in gingival tissue in these patients. Material and Methods: In this study, we analyzed, by immunofluorescence, the expression of TLRs 2, 3, 4 and 9 in gingival tissues from healthy individuals and from periodontal patients with or without type 2 diabetes. Results: We found that the expression levels of TLRs 2, 3, 4 and 9 were higher in all periodontal patients than in healthy individuals. The expression of some TLRs was increased in subjects with periodontitis and diabetes relative to subjects with periodontitis but without diabetes; this increase in expression was found particularly in TLR2 and TLR9 in the connective tissue and in TLR4 at the epithelial region. Conclusion: These data suggest that the expression of these TLRs 2, 3, 4 and 9 in gingival tissue is higher in individuals with diabetes because its inflammatory reaction is exacerbated. Additionally, the expression of these TLRS is positively regulated with the severity of periodontal disease.     

牙周炎龈组织中IL-1α基因的表达

白细胞介素１是由多种细胞分泌的一种免疫调节因子，在炎症过程中可介导多种病理损害。本实验通过反转录－聚合酶链反应技术检测龈组织ＩＬ－１α基因的表达，初步探讨ＩＬ－１α与牙周炎发产现机理的关系。     

硝酸盐对牙龈局部炎症程度影响的实验研究

目的检测分析牙周组织局部滴注硝酸盐后,一氧化氮（NO）含量变化与局部炎症发生和发展的关系,初步探讨硝酸盐在牙周炎症预防与治疗中的作用。方法建立大鼠实验性牙周炎动物模型,将70只健康SD大鼠随机分为正常对照组（N组）,牙龈炎组（P组）和药物对照组（PP组）,其中牙龈炎组又分为1000uM／L硝酸钾（KNO3）滴注组和非KNO3滴注组,所有动物分别于术后1、4周处死,采用酶法检测实验动物牙龈组织中NO2^-和NO3^-的含量,并用肉眼和组织切片观察牙龈组织中炎症的变化情况。结果正常对照组与牙龈炎组中未滴注1000uM／LKNO3溶液的牙龈组织中NO含量有高度显著性差异（P〈0．01）,在牙龈炎组中,牙龈局部给予KNO3溶液滴注与未给予KNO3溶液滴注的牙龈组织中NO含量无显著性差异（P〉0．01）,药物对照组与正常对照组牙龈组织中NO含量有高度显著性差异（P〈0．01）,药物对照组（PP组）与P组未滴注KNO3溶液的牙龈组织中NO含量有显著性差异（P〈0．01）。结论牙周局部给予硝酸盐滴注后能够增加NO的产生,提高局部NO的浓度,从而改善微循环,增强牙周局部抵抗能力,减轻和预防牙周炎症的发生。     

Herpesviruses in periodontal pocket and gingival tissue specimens

Human cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1) are frequently detected in crevicular fluid of deep periodontal pockets, but little or no information is available on occurrence of herpesviruses in gingival tissue. This investigation studied the presence of herpesviruses in periodontal pockets and the corresponding gingival tissues from 11 periodontally healthy and 14 periodontitis sites. A nested-polymerase chain reaction was employed to identify the presence of HCMV, EBV-1, EBV-2, herpes simplex virus, human herpesvirus (HHV)-6, HHV-7 and HHV-8 in each test sample. In healthy periodontal sites, HCMV was detected in 1 (9%) and EBV-1 in 2 (18%) pocket samples, and HCMV was detected in 2 (18%) and EBV-1 in 3 (27%) gingival tissue samples. In periodontitis lesions, HCMV was detected in 9 (64%) pocket samples and in 12 (86%) gingival tissue samples, and EBV-1 was detected in 6 (43%) pocket samples and in 11 (79%) gingival tissue samples. HHV-6 and HHV-8 were detected exclusively in gingival tissue samples. The present findings confirm the frequent presence of HCMV and EBV-1 in periodontitis lesions and suggest using gingival tissue specimens for detecting periodontal HHV-6, HHV-7 and HHV-8.     

Immunohistochemical expression of RANKL, RANK and OPG in gingival tissue of patients with periodontitis

Abstract Objectives. To evaluate the expression of the receptor activator of NF-κB (RANK), the receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG), in the gingival tissue of patients with periodontitis. Materials and methods. Gingival tissue was obtained from 14 systemically healthy subjects with chronic periodontitis during conventional periodontal surgery. Immunohistochemistry was used to detect the expression of RANK, RANKL and OPG in the oral and periodontal pocket epithelium as well as in the connective tissue cells. Results. RANKL was negatively expressed in both oral and periodontal pocket epithelium. OPG was also negative or weakly positive in the whole epithelium. RANK showed moderate/strong positive staining mainly in the basal and suprabasal layer of oral and periodontal pocket epithelium. In most of the cases, more than 60% of the inflammatory cell infiltrate stained for RANK and RANKL. In these cases the intensity of the stained cells ranged from moderate-to-strong. In less than half of the cases, OPG was positive in more than 60% of the stained cells of the inflammatory cell infiltrate. Conclusion. The RANK, RANKL and OPG proteins are differentially expressed in periodontal tissues and may play a major role in the bone loss occurring in 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.