牙龈癌微血管密度及血管内皮生长因子表达的意义

目的 :研究牙龈鳞状细胞癌组织中微血管密度 (MVD)及血管内皮生长因子 (VGEF)的表达和意义。方法 :采用CD3 4单克隆抗体和VEGF多克隆抗体 ,用免疫组化S -P法对 42例牙龈鳞状细胞癌标本进行免疫组化染色。结果 :牙龈癌组织MVD与正常牙龈组织MVD两者差异有显著性 (P <0 .0 0 5)。有淋巴结转移的牙龈癌MVD与无淋巴结转移的牙龈癌MVD ,两者之间差异有显著性 (P <0 .0 5)。牙龈癌组织中VEGF表达阳性率为 71.43 % ,正常牙龈组织中VEGF表达阳性率为 2 5.0 0 % ,两者之间差异有显著性 (P <0 .0 5)。有淋巴结转移组VEGF阳性率为 83 .3 3 % ,高于无淋巴结转移组VEGF阳性率 (66.67% ) ,两者之间差异无显著性 (P >0 .0 5)。VEGF阳性牙龈癌MVD高于VEGF阴性牙龈癌MVD ,两者之间差异有显著性(P <0 .0 2 )。结论 :肿瘤的血管生成在牙龈癌的发生过程中起一定作用。MVD与牙龈癌淋巴结转移及VEGF表达有密切联系     

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目的 探讨碱性成纤维细胞生长因子(bFGF)基因修饰的组织工程化复合物对牙龈组织缺损再生的影响.方法 本研究于2007年7月至2009年1月在福建省高校口腔医学重点实验室进行.利用bFGF基因转染Beagle 犬牙龈成纤维细胞(GFs),并将其接种于脱细胞真皮基质(ADM)形成组织工程化复合物,植入Beagle犬的人工牙周组织缺损区,通过对治疗前后牙周指标测量分析,评价该组织工程化复合物对牙龈组织再生的影响.结果 实验前后附着丧失差值测定显示GFs与ADM复合物组及转染bFGF基因的GFs复合物组之间无显著性差异,与空白对照组相比均有明显增加(P＜ 0.05).角化龈宽度差值,6周时转染基因组有明显增加,12周时两实验组均比空白对照组明显增加(P＜0.05).结论 复合GFs的ADM可能有助于改善附着丧失的程度及角化龈宽度,而bFGF的作用并不肯定.组织工程技术能有效促进牙龈组织再生.     

血管内皮细胞生长因子、碱性成纤维细胞生长因子在口腔鳞状细胞癌中的表达研究

目的　探讨血管内皮细胞生长因子 (VEGF)、碱性成纤维细胞生长因子 (bFGF)在口腔鳞状细胞癌中的表达及意义。方法　应用免疫组织化学方法对 10例正常口腔黏膜、30例口腔鳞状细胞癌分别进行检测 ,所得结果进行图像分析。结果　VEGF、bFGF在口腔鳞状细胞癌组织中均为高表达 ,明显高于正常口腔黏膜 (P <0 .0 0 1)。结论　VEGF、bFGF的过表达在口腔鳞状细胞癌的发生、发展过程中起着十分重要的作用 ,为口腔鳞状细胞癌的抗血管生成治疗提供理论依据。     

碱性成纤维细胞因子在口腔复发性阿弗他溃疡的表达及意义

目的　观察碱性成纤维细胞因子 (bFGF)在口腔复发性阿弗他溃疡组织 (RAU)中表达强度及分布情况。方法　用免疫组化技术检测bFGF在人正常口腔粘膜及RAU患者病变组织中的bFGF表达变化规律。结果　RAU的组织中有大量炎性细胞浸润。bFGF在正常口腔粘膜的阳性表达主要出现在粘膜上皮基底细胞层的细胞基质及粘膜下基质中 ;bFGF在RAU组织中主要由免疫细胞、成纤维细胞、血管内皮细胞表达 ,表达量较正常粘膜有所增加 (P <0 .0 5 )。结论　RAU中的bFGF的表达含量增加 ,可能是其参与调节免疫反应的表现。这可能与RAU的发病机制有着密切的联系。     

黄芩苷对牙龈成纤维细胞和牙周膜细胞上ICAM-1表达调节的研究

目的 :观察黄芩苷对IL - 1β诱导的牙龈成纤维细胞 (humangingivalfibrobl -asts ,HGF)和牙周膜细胞 (peri odontalligamentcells ,PDLcells)上细胞间粘附分子 - 1表达的影响。方法 :应用体外细胞培养和细胞免疫组化及图像分析方法。结果 :正常牙龈成纤维细胞和牙周膜细胞上细胞间粘附分子 - 1表达阴性或弱阳性 ;1μg/mLIL - 1β能诱导细胞间粘附分子 - 1在牙龈成纤维细胞和牙周膜细胞上强阳性表达 ;0 .1μg/mL黄芩苷对IL - 1β诱导的牙龈成纤维细胞和牙周膜细胞上细胞间粘附分子 - 1的表达有显著抑制作用 (P <0 .0 1)。结论 :黄芩苷可抑制IL -1β诱导的细胞间粘附分子 - 1在牙龈成纤维细胞和牙周膜细胞上表达 ,提示黄芩苷具有抗细胞粘附的作用     

口腔鳞癌组织中bFGF的表达与血管生成的关系

目的:观察口腔鳞癌组织中碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)的表达及其与微血管密度(microvesseldensity,MVD)之间的关系,探讨bFGF对口腔鳞癌血管生成的作用及意义。方法:收集口腔鳞癌标本42例(其中有淋巴结转移者14例)和10例正常口腔黏膜组织,用免疫组织化学染色法探测bFGF和CD34的表达情况并计数微血管密度(MVD)。结果:口腔鳞癌中bFGF表达阳性率为69. 05% (29 /42),显著高于正常口腔组织30% (3 /10),口腔鳞癌中MVD显著高于正常组织,且随病理分化不良而增高(P< 0. 05),有淋巴结转移组MVD显著高于无淋巴结转移组(P< 0. 05),bFGF表达阳性组中MVD明显高于bFGF表达阴性组(P< 0. 05 ),bFGF的表达与MVD呈正相关(P< 0. 01 )。结论:bFGF在口腔鳞癌组织中的高度表达在口腔鳞癌发生发展中起重要作用,并与其血管生成和淋巴结转移有关。     

体外培养人牙周牙髓相关细胞中釉原蛋白的表达

目的:观察釉原蛋白(amelogenin,Am)基因在体外培养的人牙龈上皮细胞及口腔外胚间充质来源细胞(人牙龈成纤维细胞、人牙周膜成纤维细胞和人牙髓细胞)中的表达.方法:采用反转录多聚酶链式反应(RT-PCR)技术,检测培养细胞中釉原蛋白mRNA的表达.采用蛋白质免疫印迹技术检测培养细胞中釉原蛋白的表达.结果:培养的人牙周膜成纤维细胞、牙髓细胞、牙龈成纤维细胞和牙龈上皮细胞中均未检测到釉原蛋白及其mRNA的表达.结论:体外培养的人牙周膜成纤维细胞、牙髓细胞、牙龈成纤维细胞和牙龈上皮细胞不表达釉原蛋白基因.     

bFGF、FGFR1在唾液腺肿瘤和瘤旁组织中的表达

目的:探讨碱性成纤维细胞生长因子(bFGF)和成纤维细胞生长因子受体1(FGFR1)在唾液腺良、恶性肿瘤和瘤旁组织中的表达及意义.方法:利用免疫组织化学法检测了43例唾液腺良、恶性肿瘤和30例良性肿瘤瘤旁组织中bFGF和FGFR1的表达.结果:bFGF和FGFR1在唾液腺良、恶性肿瘤组织和瘤旁组织中的表达均具有显著性差异(p<0.05).结论:bFGF和FGFR1在唾液腺肿瘤的发生,发展过程中发挥了一定的作用.     

COX-2、bFGF在口腔鳞癌发生发展过程中的表达及相互关系

目的:研究环氧合酶-2(cyclooxygenase-2,COX-2)、碱性成纤维细胞生长因子(basic fibroblast growth factors,bFGF)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)的发生、发展过程中的表达情况,进而探求COX-2、bFGF在OSCC发生发展过程中的作用及相互关系。方法:应用COX-2,bFGF多克隆抗体对25例口腔白斑(癌前病变)(oral leukoplakia,OLK)、23例口腔鳞状细胞癌进行检测,另取10例口腔正常颊、舌黏膜组织为对照组,做组织切片进行免疫组化研究。结果:COX-2在正常口腔黏膜不表达,bFGF在正常口腔黏膜仅为弱表达或不表达,COX-2、bFGF在口腔白斑中有一定的表达,染色强度与正常口腔黏膜相比具有显著性差异(P<0.01 ),COX-2、bFGF在口腔鳞癌中高表达,其染色强度强于口腔白斑(P<0.01),口腔鳞癌中COX-2的表达阳性率为82.61%,bFGF的表达阳性率为86.96%,二者共同表达阳性率为78.26%,r=0.8,P<0.05,二者表达呈正相关关系。结论:COX-2、bFGF在口腔鳞癌发生、发展中起着重要作用,二者表达显著相关,在口腔鳞癌的发生发展中有协同作用,COX-2可促进bFGF的形成。     

MC、bFGF在皮肤血管瘤中的表达及意义

目的 :研究皮肤血管瘤中肥大细胞 (MC)、碱性成纤维细胞生长因子 (BasicFibroblastGrowthbFGF)之间的关系 ,以及它们在血管瘤形成中的作用。方法 :颌面部血管瘤标本 68例 ,按Mulliken[1] 分类法将血管瘤病程分为三期 :增殖期 3 2例、退化期 2 4例、退化完成期 12例。选取 5例唇裂患者的唇部皮肤做正常对照。将每个标本连续切片 4张 ,分别行HE染色和免疫组织化学染色 ,并计算肥大细胞数和bFGF阳性率。结果 :①肥大细胞的胞浆内抗胰蛋白酶抗体显色为棕褐色 ,胞核为蓝色 ,多位于小血管管壁及其周围。各期血管瘤中肥大细胞计数分别为 :增殖期 2 1.3± 3 .0SD/HPF ,退化期 8.4± 2 .6SD/HPF ,退化完成期 3 .2± 2 .0SD/HPF。正常皮肤组织中肥大细胞计数 3 .1± 1.3SD/HPF。增殖期与退化期、退化完成相比有高度显著性差异 (P <0 .0 1)。②血管瘤中bFGF位于肥大细胞内或胞外基质中 ,增殖期bFGF染色深 ,棕褐色 ,呈阳性连续片状分布。退化期呈不连续片状分布、染色浅。退化完成期染色更浅 ,呈点状分布 ,阳性反应物少。增殖期bFGF阳性表达率 93 .75 % ,退化期 5 4.16% ,退化完成期 3 3 .3 3 %。增殖期bFGF明显高于退化期、退化完成期 ,有显著性差异 (P <0 .0 5 )。正常对照组中bFGF无表达。③bFGF与MC计数表达呈正     

口腔粘膜下纤维化、白斑及鳞癌上皮细胞内皮素1表达的定量分析

目的 :探讨内皮素 (ET 1)在口腔癌变发病机制中的作用和意义。方法 :采用免疫组化染色SABC法和图象分析技术 ,对人口腔粘膜下纤维化 (OSF) 10例、白斑 (OLK) 9例、鳞状细胞癌 (SCC) 14例及正常口腔粘膜(NOR) 10例的上皮细胞ET 1表达进行定量分析。结果 :①ET 1在OSF、OLK、SCC组织中的表达增强 ,阳性颗粒主要位于上皮棘细胞、基底细胞的胞浆胞膜上。ET 1表达阳性率和含量显著高于正常对照 (P <0 .0 1)。②OLK、SCC上皮细胞ET 1含量呈显著增加趋势 (P <0 .0 5 )。③OSF上皮细胞ET 1含量显著高于OLK(P <0 .0 5 ) ,与SCC相比无显著性差异 (P >0 .0 5 )。结论 :ET 1含量在口腔癌前病变至癌变过程中可能存在一种量变关系 ,OSF中ET 1过量表达可能提示其上皮细胞的癌变潜能 ,ET 1与OSF、OLK、SCC的发生发展密切相关     

Correlation of basic fibroblast growth factor expression with the invasion and the prognosis of oral squamous cell carcinoma

BACKGROUND: The aim of this study was to evaluate the relationship between the expression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in cancer cells and fibroblasts at the invasive front of oral squamous cell carcinoma (OSCC), and the pathologic and clinical characteristics. METHODS: Sections of 61 biopsy specimens of primary OSCC were immunostained to assess the expression of bFGF and FGFR-1 in cancer cells and fibroblasts at the invasive front. RESULTS: The bFGF and FGFR-1 expressions in the cancer cells were evident in all specimens, whilst, in fibroblasts, they were detected in 41 (67%) of 61 specimens. These expressions in the fibroblasts occurred notably more often in high-invasive OSCC specimens than low-invasive OSCC specimens. The prevalence of bFGF and FGFR-1 expressions in cases with lymph node metastasis was significantly higher (P < 0.05) than in cases without metastasis. Moreover, these expressions were well correlated with patient prognosis. CONCLUSION: This study concludes that bFGF and FGFR-1 expressions in fibroblasts at the invasive front are linked to the mode of invasion and the prognosis in OSCC.     

Ultrastructure of the gingiva in cardiac patients treated with or without calcium channel blockers

OBJECTIVES: In the last few years, several studies have suggested that periodontal diseases are related to the development of atherosclerosis and its complications. Our objective was to study the ultrastructural morphology of the gingiva from cardiac patients, some of whom were treated and some not with calcium channel blockers compared to a control group. MATERIAL AND METHODS: Fifty-five patients were studied and grouped in the following way: (a) healthy group (HG) (n=12) healthy patients with at least two pockets between 3 and 5 mm; (b) cardiac group (CG) (n=12) patients with cardiac disease untreated with calcium channel blockers; (c) diltiazem group (DG) (n=13) cardiac patients treated with diltiazem; (d) nifedipine group (NG) (n=18) cardiac patients treated with nifedipine. RESULTS: Ultrastructural studies in the CG showed inflammatory cells, collagen fibers disruption and a more extended morphologically compromised fibroblast mitochondria. Morphometric studies in CG showed mitochondria that were impaired in number but increased in volume, suggesting metabolic cell suffering. In DG and NG, morphometric data were similar to HG. The presence of myofibroblasts and collagen neosynthesis was detected in DG and NG. CONCLUSIONS: Our data showed differences in the ultrastructure of the gingival fibroblasts between the studied groups; the DG and NG showed features that could be interpreted as an attempt to restore the cellular metabolic function.     

The role of basic fibroblast growth factor in oral submucous fibrosis pathogenesis

Background:  Oral submucous fibrosis (OSF) is a chronic fibrotic disease of oral mucosa and oropharynx, induced by betel quid chewing often resulting in restricted mouth opening. The principal cells implicated as a source of extracellular matrix in areas of fibrosis are fibroblasts. Accumulation of connective tissue matrix is secondary to factors such as cytokines and growth factors. The contribution of basic fibroblast growth factor (bFGF) in disease progression and the consequent stromal changes with increase in the severity of OSF was studied.
Methods:  A case series analysis of 30 cases of OSF was carried out for bFGF expression using immunohistochemistry. Connective tissue changes in these cases were corroborated using aldehyde fuchsin and Verhoeff's hematoxylin special stains.
Results:  bFGF immunoreactivity was found to be increased in fibroblasts and in endothelial cells in early OSF cases, while the expression of bFGF in stroma increased notably in advanced fibrosis.
Conclusion:  Increased bFGF expression in early stages of the disease was explainable to an initial injury phase because of areca consumption, followed by cellular activation by chemotactic cytokines and other growth factors with eventual fibrosis occurring as a result of molecular alteration at the cellular level.     

Evaluation of gingival and periodontal conditions following causal periodontal treatment in patients treated with nifedipine and diltiazem*

Abstract It is established that phenytoin, cyclosporin and some calcium antagonists produce gingival overgrowth, but it is not known how this condition may respond to causal periodontal treatment. In order to find out, a longitudinal study was carried out, over a year, comparing a group of patients who were given nifedipine (NG, n= 18) and another group who were given diltiazem (DG. n= 13) with 2 others: one comprised cardiopathic patients who took no calcium antagonists (CG, n= 12) and the other contained patients who were medically healthy. with moderate periodontitis (HG, n= 12). On their basal visit, they were examined and instructed in oral hygiene, and then given causal periodontal treatment, being seen again at 4 and 8 months, when hygiene instructions were reinforced. They were seen for the last time at 12 months, when they were again examined. Groups NG and DG, on their basal visit, showed larger gum size than groups HG and CG. which was statistically significant; on their final visit, these differences remained only at the interproximal level. The number of patients with gingival overgrowth-taking the average of group HG as a minimal value-was much higher in groups CG (92%). DG (100%) and NG (89%) on the basal visit; on the final visit, the differences remained only in groups DG (85%) and NG (83%). The probing pocket depth reduction was much greater in groups HG and CG than in DG and NG. basically due to a greater gaining on clinical attachment level. The % of sites in which the pocket depth improved by more than 2 mm was 39.8% in HG, 54.5% in CG, 23.7% in DG and 28.7% in NG. The % of sites where the attachment gain by more than 2 mm was 46.2% in HG, 55.5% in CG, 22.8% in DG and 21.4% in NG. The amount of plaque and bleeding on probing, which was similar in all groups on the basal visit, decreased throughout the study, especially between the basal and 2nd visit in groups HG and CG. We have demonstrated that patients that take nifedipine and diltiazem show a larger gum size and their response to causal periodontal treatment is poorer than in the healthy and the cardiac groups.     

Cytokinetic studies of oral cancer cells using bromodeoxyuridine labelling in relation to factors influencing prognosis

Cytokinetics of 27 untreated oral squamous cell carcinomas were evaluated by in vitro bromodeox- yuridine (BrdU) labelling. A mononuclear cell suspension was prepared for the labelling using collagenase and DNase treatment. The labelled cells were visualised by immunofluorescence staining with an anti-BrdU monoclonal antibody. The labelling index (LI%) was calculated by determining the percentage of BrdU labelled cells. The LIs of the carcinomas ranged from 3.8 to 19.2%. The LIs of poorly-differentiated (HG3) tumours were 2–3-fold higher compared with those of well-differentiated (HG1) and moderately-differentiated (HG2) carcinomas. Results also showed statistically significant (P < 0.0005) increases in LIs from nuclear grade 1 (NG1) to nuclear grade 3 (NG3) carcinomas. Higher LIs were observed in stage III/IV (10.82 ± 4.62; 10.36 ± 4.90) than those in stage I (6.87 ± 2.09) and II (7.14 ± 1.87) carcinomas. A significant (P < 0.0005) difference in LI values was found between the patients with positive and negative lymph nodes. Good correlation (r = 0.77) was exhibited between the LI values and mitotic counts (MC) of the specimens. These results on oral cancer cell proliferation seem to have prognostic implications.     

口腔黏膜癌前病变及口腔鳞癌中bFGF的表达及意义

目的　探讨口腔黏膜癌前病变及口腔鳞癌的发生、发展过程中bFGF的表达及意义。方法　应用免疫组织化学方法对 10例正常口腔黏膜、2 7例口腔扁平苔藓、2 4例口腔白斑及 3 0例口腔鳞癌分别进行检测。结果　口腔鳞癌组织中bFGF高表达 ,明显高于正常口腔黏膜、口腔扁平苔藓和口腔白斑组织 (P <0 .0 5 ) ;口腔扁平苔藓和口腔白斑组织中bFGF表达高于正常口腔黏膜 (P <0 .0 5 )。结论　bFGF的过表达在口腔鳞癌的发生、发展过程中起着十分重要的作用 ,可以将其作为预测口腔黏膜恶变潜能的重要标志物     

Exendin-4 relieves the inhibitory effects of high glucose on the proliferation and osteoblastic differentiation of periodontal ligament stem cells

BackgroundWith the impaired regenerative potential in patients with diabetes mellitus (DM), Periodontal ligament stem cells (PDLSCs) are regarded as an attractive source of stem cells for periodontal cytotherapy. Recent studies have shown that Exendin-4 (Ex-4) exerts cell-protective effects and bone remodeling ability on many types of cells. The aim of this study was to investigate whether Ex-4 alleviates the inhibition of high glucose on the proliferation and osteogenic differentiation of PDLSCs.MethodsPDLSCs were incubated in medium supplemented with 5.5 mM d-glucose (NG), 30 mM d-glucose (HG), NG plus Ex-4, and HG plus different concentration (1, 10, 20, 100 nM) of Ex-4 respectively. Cell proliferation was detected by CCK-8 assay and cell cycle analysis. Osteogenesis was assessed by Alizarin Red S staining and evaluation of the mRNA expression of Runx2, ALP and Osx at day 7, 14 and 21. Intracellular level of reactive oxygen species (ROS) was detected using 5-(and-6)-chloromethyl-2′,7′-dichlorodihydro-fluorescein diacetate (CMH2DCF-DA).ResultsThe proliferation ability, mineralized nodules forming capacity and the mRNA expression of Runx2, ALP and Osx of PDLSCs in HG group were decreased, the ROS level was increased compared to NG group. With the treatment of Ex-4, the HG-inhibited proliferation ability and osteogenic differentiation ability of PDLSCs were significantly reversed, the HG-increased ROS level could be down-regulated. Moreover, Ex-4 enhanced the osteogenic differentiation of normal PDLSCs.ConclusionsEx-4 alleviates the inhibitory effect of HG on the proliferation and osteoblastic differentiation of PDLSCs, and has a significant enhance in the osteoblastic differentiation of normal PDLSCs, giving new insights into the possible therapeutic method of diabetic periodontitis.     

PTTG和bFGF在口腔鳞癌中的表达及意义

目的：探讨垂体肿瘤转化基因（pituitary tumor transforming gene，PTFG）和碱性成纤维细胞生长因子（basic fibroblast growth factor，bFGF）在口腔鳞癌中的表达及相互关系，研究它们的表达与肿瘤临床病理指标的联系。方法：应用SP染色法检测PTTG蛋白和bFGF在55例口腔鳞癌组织、10例正常口腔黏膜组织中的阳性率。结果：在口腔鳞癌中PTTG和bFGF的阳性表达率分别为78．2％和67．3％，其阳性率及表达等级均显著高于正常对照组（P〈0．05）。PTTG在中一低分化组和有淋巴结转移组中的表达显著高于高分化组和无淋巴结转移组（P〈0．05）。PTTG表达与bFGF表达成等级正相关（r=0．382，P〈0．05）。结论：PTFG或bFGF与口腔鳞癌生物学行为及预后有密切关系，二者的联合检测，有助于口腔鳞癌恶性程度和预后的判断。     

Aggressive and Chronic Periodontitis Correlate With Distinct Cellular Sources of Key Immunoregulatory Cytokines

Background: Chronic periodontitis (CP) and aggressive periodontitis (AP) are inflammatory diseases and the main cause of dental loss in adults. We aimed to investigate the expression of adhesion molecules and the source of proinflammatory and anti‐inflammatory cytokines in circulating mononuclear cells from patients with CP and AP. Methods: Peripheral blood mononuclear cells from healthy controls and CP or AP patients were collected. The expression of the cell adhesion molecules CD11a and CD11b, and the cellular sources of interleukin (IL)‐4, IL‐10, IL‐12, interferon‐γ, and tumor necrosis factor‐α by distinct subpopulations of circulating leukocytes were determined using flow cytometry. Results: The expression of CD11a, but not CD11b, was significantly higher within the CD4⁺ and CD8⁺ T cells in CP and AP than in healthy controls. The frequencies of tumor necrosis factor‐α–expressing CD4⁺ T cells and CD14⁺ cells were higher in AP and CP, compared to healthy controls, respectively. Moreover, the frequency of IL‐10 expressing CD14⁺ cells was higher in CP, but not AP, compared to healthy controls CD4⁺ T cells committed to IL‐4 production was higher in CP than in healthy controls. Conclusion: These results suggest the participation of CD11a in the pathogenesis of periodontal lesions and show distinct cellular sources of immunoregulatory cytokines in AP versus CP.