口腔粘膜下纤维性变患者角朊细胞分泌细胞因子水平变化

目的：探讨口腔粘膜上皮角朊细胞（ＫＣ）和细胞因子在口腔粘上纤维性变（ＯＳＦ）发生中的作用。方法：分别从正常及ＯＳＦ组织培养出ＫＣ（ＮＭ－ＫＣ）和ＯＳＦ－ＫＣ０,用放射免疫法和酶联免疫吸附法测定培养细胞上清液中内皮素（ＥＴ）和β１转化生长因子（ＴＧＦβ１）的水平。结果：ＯＳＦ－ＫＣ分泌ＥＴ和ＴＧＦ１水平均高于ＮＭ－ＫＣ,且两者呈正相关关系。     

高压氧对体外培养的内皮细胞和口腔粘膜角朊细胞分泌VEGF的影响

目的探讨高压氧治疗OSF的机理。方法通过体外培养内皮细胞和口腔粘膜角朊细胞，经高压氧处理后收集培养上清，用ELISA方法测定培养上清中的VEGF水平。结果经高压氧处理后，口腔粘膜角朊细胞分泌VEGF的量增加，内皮细胞分泌VEGF的水平无明显变化。结论高压氧对OSF起治疗作用的主要因素之一可能是通过促进口腔粘膜角朊细胞分泌VEGF导致病人口腔粘膜下组织血管新生，似乎与内皮细胞分泌的VEGF无关。     

人口腔粘膜上皮角朊细胞的体外培养

人口腔粘膜上皮角朊细胞的原代培养有许多困难 ,其中主要是上皮细胞接种成活率、产量和成纤维细胞污染的问题。我们比较酶消化法和组织块法建立口腔粘膜上皮角朊细胞体外培养模型 ,并用免疫组织化学法进行培养细胞的定性研究。1.材料 :培养物来源于 5个月～ 10岁唇裂患者修复术中的口内粘膜。切取唇内侧粘膜组织修复剩余物若干 ,每块约0 5ｃｍ× 0 5ｃｍ大小。2 .方法 :①酶消化法 :用Ｄ Ｈａｎｋｓ液冲洗粘膜块 ,剪切至0 2ｃｍ× 0 2ｃｍ大小。配制 0 3ｋｇ/ＬⅠ型胶原酶和 0 4%Ｄｉｓｐａｓｅ消化液的混合液 (1∶1) 5ｍｌ,将粘膜块…     

槟榔提取物对人口腔粘膜上皮角朊细胞分泌内皮素的影响

口腔粘膜下纤维化(ｏｒａｌｓｕｂｍｕｃｏｕｓｆｉｂｒｏｓｉｓ,ＯＳＦ)是一种慢性炎性疾病。大量研究结果表明,ＯＳＦ的发生与咀嚼槟榔习惯密切相关,但咀嚼槟榔诱发ＯＳＦ的机理仍不明确。我们采用角朊细胞(ｋｅｒａｔｉｎｏｃｙｔｅｓ,ＫＣ)体外培养技术,比较ＯＳＦＫＣ与健康者ＫＣ分泌内皮素(ｅｎｄｏｔｈｅｌｉｎ,ＥＴ)的差异,同时观察槟榔提取物(ａｒｅｃａｎｕｔｅｘｔｒａｃｔ,ＡＮＥ)对ＫＣ分泌ＥＴ的影响。旨在对ＫＣ及ＥＴ在ＯＳＦ发生中的作用进行初步探讨,并为阐明咀嚼槟榔习惯与ＯＳＦ的因果关系提供一些实验依据。1材料与方…     

EGFR在口腔粘膜鳞癌和癌前病变中的表达及意义

应用免疫组化ＳＰ法对２３例口腔鳞癌（ＳＣＣ）,２０例口腔粘膜白斑（ＬＫ）,２１例口腔粘膜扁平苔藓（ＬＰ）及１０例正常口腔粘膜（ＮＭ）蜡块标本行表皮生长因子受体（ＥＧＦＲ）的定性测定．结果发现：ＳＣＣ的阳性率为８２．６１％,ＬＫ４５．０％,ＬＰ３８．１０％,ＮＭ１１．１１％．ＳＣＣ与ＬＫ、ＬＰ、ＮＭ间均有显著差别,ＬＫ、ＬＰ与ＮＭ间的差别无显著性,提示：ＥＧＦＲ高度表达可能是口腔鳞癌的重要指征之一,对癌变细胞的增殖有重要作用。     

口腔粘膜上皮细胞原代培养影响因素分析

目的：分析影响口腔粘膜上皮细胞原代培养影响因素,探讨理想的培养条件。方法：观察不同的标本处理方法,上皮分离方法以及上皮消化酶对原代培养成功率,细胞融合和活细胞率的影响。结果：标本处理方法,上皮分离方法以及上皮消化酶分别对原代培养污染机率,细胞融合时间和活细胞率有影响。结论：口腔粘膜上皮细胞原代培养按优化方法可获得较高的成功率。     

转化生长因子β1在口腔粘膜下纤维性变角朊细胞中的表达

为探讨转化生长因子β１在口腔粘膜下纤维性变发病机理中的作用，作者应用原位杂交方法，对２５例口腔粘膜下纤维性变（ｏｒａｌｓｕｂｍｕｃｏｕｓｆｉｂｒｏｓｉｓ，ＯＳＦ）、５例正常口腔粘膜（ＮＯＲ）及１０例口腔扁平苔藓（ｏｒａｌｌｉｃｈｅｎｐｌａｎｕｓ，ＯＬＰ）组织角朊细胞中转化生长因子β１（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒβ１，ＴＧＦβ１）ｍＲＮＡ进行了检测。结果：１５例（６０％）ＯＳＦ组织角朊细胞中有ＴＧＦβ１ｍＲＮＡ表达，阳性表达主要分布于早、中期ＯＳＦ组织角朊细胞中；５例ＮＯＲ及１０例ＯＬＰ组织角朊细胞表达呈阴性。结果提示：ＯＳＦ组织角朊细胞可合成分泌ＴＧＦβ１，ＴＧＦβ１在ＯＳＦ的发病机理中可能起重要作用，可能作为一中间介质参与了ＯＳＦ的发病过程。     

槟榔提取物对人类口腔黏膜上皮角朊细胞分泌内皮素的影响

目的通过观察槟榔提取物(areca nut extract,ANE)对体外培养的人类口腔黏膜上皮角朊细胞(keratinocytes,KC)分泌内皮素(endothelin,ET)的影响,初步探讨KC和ET在口腔黏膜下纤维性变(oral submucous     

复方藻朊凝胶治疗口腔扁平苔癣临床观察

口腔扁平苔癣 (ＯＬＰ)是口腔粘膜慢性炎性病变。其发病机理不十分明确 ,研究资料认为 :免疫机制及角朊细胞 (ＫＣ)在ＯＬＰ发病中起重要作用。根据这一机理我们应用复方藻朊凝胶治疗ＯＬＰ并对其临床结果进行观察。1　一般资料 :1 1　一般情况 :　收集我院口腔科 1994年～ 2 0 0 0年门诊就诊 60例ＯＬＰ患者 ,根据发病特点进行分别整理。其中普通型 3 4例 ( 5 6 7% ) ,混合型 2 2例 ( 3 6 7% ) ,糜烂型 4例( 6 6% ) ,见表 1。男性 2 4例 ,女性 3 6例 ,男 女　 2 3。1 2　治疗方法 :1 2 1　复方藻朊凝胶配制 :　取海带 15 0ｇ ,水洗尽。…     

角化囊肿癌变临床病理特征

目的 研究和探讨角化囊肿（ＯＫＣ）癌变的临床与组织病学特征。方法 对２２６例ＯＫＣ临床病理资料进行研究分析，采用增殖细胞核抗原ＰＣＮＡ单克隆抗体对单囊和多囊性ＯＫＣ进行免疫组化反应。结果 ２２６你ＯＫＣ中，６例（２．６５％）ＯＫＣ发生癌变。癌变病例临床特征是：①骨腔充满病变组织，类似实体肿瘤。②癌变病例全部为多囊性，单囊ＯＫＣ未见癌变。③囊壁衬里上皮异常增生，侵袭性生长，可见异常核分裂，呈典型鳞癌     

槟榔提取物抑制人类口腔黏膜成纤维细胞生长的实验研究

目的通过比较正常口腔黏膜和口腔黏膜下纤维性变(OSF)组织中成纤维细胞(FB)增殖差异、检测槟榔提取物(ANE)对成纤维细胞增殖的影响,来探讨OSF的发病机理.方法对人类口腔黏膜成纤维细胞进行分离培养,然后用四唑盐(MTT)比色试验法检测OSF患者和正常人口腔黏膜FB增殖状况,并且观察ANE对FB增殖的影响.结果表示增殖水平的OD值在OSF-FB为0.254±0.045,高于NM-FB的OD值0.236±0.012(P<0.05),ANE以浓度-效应依赖关系抑制FB增殖.结论 OSF-FB细胞增殖率较NM-FB高;ANE对口腔黏膜FB有细胞毒作用,提示槟榔及其有效成分不完全是通过直接刺激FB增殖而诱发OSF.     

槟榔提取物刺激口腔黏膜角朊细胞培养上清对成纤维细胞增殖活性的影响

目的：探讨槟榔提取物刺激口腔黏膜角朊细胞在黏膜下纤维性变发病中的作用。方法：采用不同浓度槟榔提取液刺激体外培养的角朊细胞,取细胞培养上清,MTT法观察细胞培养上清对成纤维细胞增殖的影响。结果：一定浓度槟榔提取液刺激的角朊细胞培养上清能促进成纤维细胞增殖;细胞培养上清对成纤维细胞增殖的促进作用存在个体差异。结论：槟榔成分可能通过改变口腔黏膜角朊细胞的活性而导致口腔黏膜下纤维性变的发生。     

The fibroblast population in oral submucous fibrosis

The purpose of the investigation was to compare the morphology of fibroblasts cultured from healthy oral mucosa and mucosa of patients with oral submucous fibrosis (OSF) and to collate the occurrence of cell types of similar morphology. Cells cultured from biopsy specimens from the buccal mucosa of six subjects who did not chew the areca nut and six patients with OSF who chewed areca nut were grown according to standard techniques. Ninety cells per cell line were recorded daily for 8 days, classified into types F1, F2 and F3 according to their morphology, and the results statistically analyzed. We found that there was a relative increase of F3 cells in relation to Fl cells in OSF resulting in the ratio of F3 to F1 cells being significantly larger in OSF than the ratio in the controls. As it has been reported that F3 cells m rat connective tissues produce significantly more collagen types I and III than F1 cells, we concluded that a change of fibroblast population has occurred in OSF and that this relative increase of F3 cells in humans, which could be committed to the production of large quantities of collagen, can be an explanation for the excessive collagen formation in OSF.     

Copper stimulates human oral fibroblasts in vitro: a role in the pathogenesis of oral submucous fibrosis

Copper is implicated in the pathogenesis of several fibrotic disorders. Areca nut has been shown to have a high copper content and areca chewing is associated with oral submucous fibrosis (OSF). The effects of copper on human oral fibroblasts were investigated in vitro. Human oral fibroblasts were incubated with copper chloride (CuCl2) at concentrations ranging from 0.01 microM to 500 microM for 24 h, and in vitro cell proliferation was assayed by incorporation of tritiated-thymidine; soluble and non-soluble collagen synthesis was assayed using tritiated-proline. Addition of copper chloride at concentrations ranging from 0.1 microM to 50 microM increased the collagen synthesis by the oral fibroblasts compared with growth without copper (P<0.05). The addition of copper chloride neither increased the synthesis of non-collagenous proteins by the fibroblasts nor influenced their proliferation rate. We conclude that copper upregulates collagen production in oral fibroblasts. This appears to be concentration dependent, with peak collagen synthesis at 50 microM CuCl2. These in vitro results taken together with the recent findings of copper in oral biopsies from OSF subjects support the hypothesis that copper in areca nut acts as a mediator of OSF.     

Development of an in vivo mouse model to study oral submucous fibrosis

BACKGROUND: Epidemiological data have shown an association of areca nut chewing with oral submucous fibrosis (OSF). Experimental evidence to confirm this has been limited. Fibrosis-promoting activity of areca nut was tested in an animal model. METHOD: Buccal mucosa of a group of 20 female BALB/c strain mice, 10-12 weeks of age, was treated twice daily 6 days per week with topical application of aqueous areca nut extracts for 300-600 days. A control group (n = 20) was treated with 50 mM NaCl. The influence of areca nut on the oral epithelium and connective tissue was recorded semiquantitatively by light microscopy. RESULTS: The areca nut-treated oral epithelium showed progressive changes in epithelial thickness leading to atrophy, increased cellularity of fibroblasts, fibrosis of connective tissue, focal infiltration of inflammatory cells and muscle atrophy. On killing after 600 days of treatment, the scores on cellularity, inflammation and muscle atrophy were significantly different to the control group (P = 0.03). CONCLUSION: The study provides further evidence that areca nut contributes to the development of OSF in treated animals. The model has the potential to test synergism of areca nut with other carcinogens and any therapeutic interventions.     

槟榔致癌物质与口腔癌

槟榔为一级致癌物,咀嚼槟榔引起口腔癌缘于槟榔中的槟榔碱（ARC）、槟榔鞣质、槟榔特异性亚硝胺（ASNA）和活性氧（ROS）等具有细胞毒性、遗传毒性、致突变性和致癌性。ARC可诱导口腔成纤维细胞、角质形成细胞和人脐静脉内皮细胞程序性死亡。槟榔鞣质有否遗传毒性和致突变性至今仍有争议,不同类型的短期筛选试验结果差异很大,但含鞣质的槟榔多酚是槟榔的主要致癌成分。3-甲基亚硝氨基内醛可诱发人颊黏膜角质形成细胞的DNA链断裂和DNA蛋白交联。3-甲基亚硝氨基丙腈为强致癌剂,可诱发试验动物肿瘤,靶器官包括鼻腔、食管、舌等。槟榔咀嚼过程中可产生大量的ROS,造成DNA氧化性损伤和激活癌基因的方式促使癌症的发生。相对分子质量为3．0×10^4-10．0×10^4的槟榔提取物组分中一种新发现的蛋白聚糖通过增加胞内ROS水平及一系列信号级联放大,上调口腔癌细胞低氧诱导因子-1d的表达,最终诱导细胞自噬。细胞自噬有利于保护癌细胞免遭ARC诱导的程序性细胞死亡,促进口腔癌的发展。槟榔提取物还可能通过ROS增强舌鳞状上皮细胞癌细胞株刺激血小板聚集的效应,从而促进舌癌转移。     

Experimental study on aqueous areca nut extracts inducing oral submucous fibrosis in rats. II. Effection of mast cells on collagen metabolism]

The effection of mast cells (MC) on collagen metabolism of oral submucous fibrosis (OSF) induced by aqueous areca nut extracts (AANE) in rats was studied by transmission electron microscope, histochemical and biochemical analyses. The results revealed that there was a close relationship between MC and fibroblats (FB) in the pathogensis of OSF; MC which appeared in the buccal mucosa in the model groups increased significantly and became more active in the function. A great deal of collagen compacted in the buccal mucosa in the model groups. The contents of tissue hydroxyproline (Hyp) in the model groups were obviously higher than those in the control groups. Moreover, the longer the time of irritation continued, the higher the contents were. The number of MC had a higher positive correlation with the contents of tissue Hyp at the same time. Total serum Hyp had no significant difference among all the groups and any stages. The results indicated that some aqueous components of areca nuts might disturb collagen metabolism by accumulation and activation of MC.