基质金属蛋白酶-9及基质金属蛋白酶抑制剂-1在口腔鳞状细胞癌中的表达研究

目的 研究基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)及基质金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)在口腔鳞状细胞癌中的表达及其相关性,探讨其在口腔鳞状细胞癌发生发展中的作用。方法 取20例正常口腔黏膜、40例口腔鳞癌(高、低分化鳞癌各20例),应用免疫组织化学SP法检测上述标本中MMP-9及TIMP-1的表达和分布。结果 口腔高分化鳞癌、低分化鳞癌组织中MMP-9、TIMP-1的表达均高于正常口腔黏膜组织(P<0·05)。口腔高分化鳞癌、低分化鳞癌、正常口腔黏膜组织中,MMP-9表达强度依次递增,TIMP-1表达强度依次递减。结论 MMP-9可能在口腔鳞状细胞癌的发生发展中起重要作用,其活性受TIMP-1的负性调控。     

LPS对体外培养人牙髓成纤维细胞基质金属蛋白酶抑制剂1,2 表达的影响

目的 :观察LPS对人牙髓成纤维细胞 (HDPF)表达基质金属蛋白酶抑制剂 1,2 (TIMP - 1,2 )的影响。方法 :体外培养HDPF ,用 1、10、10 0 μg/mLLPS分别刺激 2d ,采用免疫组化和图像分析法 ,观察HDPF表达TIMP - 1,2的变化。结果 :不同质量浓度的LPS对TIMP - 1,2的影响不同。与空白组相比 ,10 μg/mL的LPS刺激HDPFTIMP - 1的表达明显增加 (P <0 .0 1) ;LPS质量浓度为 10 0 μg/mL时 ,HDPF表达TIMP - 1明显减弱(P <0 .0 1)。LPS对TIMP - 2的影响呈浓度依赖关系 ;LPS质量浓度≥ 10 μg/mL时 ,TIMP - 2表达显著减弱(P <0 .0 1)。结论 :LPS通过改变牙髓成纤维细胞TIMP - 1,2的表达量 ,影响炎症过程     

实验性牙根吸收组织中基质金属蛋白酶-1及其抑制剂-1的定位表达

目的检测小型猪实验性牙根吸收组织中基质金属蛋白酶(MMP)-1及基质金属蛋白酶抑制剂(TIMP)-1的表达,探讨二者在牙根吸收过程中的作用。方法选用6头小型猪的12颗下颌乳侧切牙,随机分为4组,每组3颗下颌乳侧切牙,分别加力0、0.98、1.96、2.94 N,每2周加力1次。第1次加力后45 d切取标本,应用SABC免疫组化方法检测MMP-1及TIMP-1在根吸收区牙周组织中的定位表达。结果加力0.98、1.96、2.94 N时牙根吸收区牙周组织中MMP-1阳性染色均比不加力强,加力0.98、1.96 N时牙根吸收区牙周组织中TIMP-1阳性染色均比不加力强。结论MMP-1与TIMP-1参与细胞外基质的代谢活动;MMP-1与TIMP-1在根吸收活动中起着重要的作用。     

人冠部牙本质中基质金属蛋白酶组织抑制剂1的表达与分布

目的 观察人正常牙冠部牙本质中基质金属蛋白酶组织抑制剂1(TIMP-1)的分布特点,探讨其在牙本质形成与改建过程中的作用.方法 以21颗人正常牙冠部牙本质为研究对象,应用免疫组织化学染色法观察牙本质中TIMP-1的分布特点,采用Image-Pro Plus 6.0软件分析牙本质不同层的平均吸光度(A)值的差异.结果 人冠部牙本质全层均可见TIMP-1的免疫阳性染色.成牙本质细胞层、前期牙本质层及近釉牙本质界层较强,深层牙本质、中层牙本质和浅层牙本质强度逐渐减弱.结论 TIMP-1存在于牙本质全层,其在牙本质中的分布具有不均衡性,提示TIMP-1在调节牙本质基质的形成、改建及矿化过程中发挥重要的作用.     

BMP-2对体外培养人牙髓细胞TIMP-1、TIMP-2表达的影响

目的：观察BMP－2对HDPF表达TIMP－1、TIMP－2的影响。方法：用4ng/ml、40ng/ml、400ng/ml BMP-2分别作用于HDPF 2d，利用免疫组化和图像分析法法半定量观察BMP－2对HDPF表达TIMP－1、TIMP－1、TIMP－2表达显著增强，结论：BMP－2通过改变HDPF表达TIMP－1、－2的量来影响胶原的代谢。     

Syndecan-1、基质金属蛋白酶-9在口腔黏膜鳞状细胞癌组织中表达的相关性及其临床意义

目的研究口腔黏膜鳞状细胞癌组织中Syndecan-1和基质金属蛋白酶(MMP)-9的表达,并探讨其与口腔黏膜癌变侵袭转移的关系。方法采用免疫组织化学SP法检测Syndecan-1和MMP-9在50例口腔黏膜鳞癌、8例不典型增生组织、8例正常口腔黏膜组织中的表达。结果口腔黏膜鳞状细胞癌组织中Syndecan-1表达的阳性率为54%(27/50),显著低于不典型增生组和正常黏膜组织组;MMP-9表达的阳性率为92%(46/50),显著高于不典型增生组和正常黏膜组织组;Syndecan-1和MMP-9的表达与口腔黏膜鳞状细胞癌患者的性别和年龄无明显关系,与组织分化程度、临床分期和淋巴结转移有关。Syndecan-1与MMP-9表达在口腔癌呈负相关(r=-0.492,P<0.01)。结论Syndecan-1与MMP-9的表达与口腔黏膜鳞状细胞癌的增殖、浸润和转移有关,检测口腔黏膜鳞状细胞癌中Syndecan-1与MMP-9的表达对判断肿瘤进展及恶性程度具有重要意义。     

基质金属蛋白酶2及其抑制剂在口腔鳞癌中的表达

我们应用逆转录聚合酶链反应(RT PCR)及免疫组织化学法对口腔鳞癌及癌旁组织中的基质金属蛋白酶2 (matrixmatalloproteinases 2 ,MMP 2 )及其抑制剂(tissueinhibitorofmatalloproteinase 2 ,TIMP 2 )的mRNA及蛋白表达进行半定量、定性及定位研究,探讨其与口腔鳞癌临床特征的关系。1.材料与方法:(1)组织标本:取自4 0例原发性口腔鳞癌患者的癌组织、癌旁2cm及癌旁≥5cm组织各2份。病理证实均为鳞状细胞癌。临床分期按国际抗癌协会分期标准。(2 )研究方法:①RT PCR :取用异硫氰酸胍 酚 氯仿一步法提取的组织总RNA合成cDNA。应用96 0…     

基质金属蛋白酶抑制剂-1在口腔疣状癌中的表达研究

目的观察基质金属蛋白酶抑制剂(tissue inhibitor of metalloproteinase,TIMP)-1在口腔疣状癌(OVC)中的表达。方法选择15例口腔疣状癌、10例正常口腔黏膜、20例口腔鳞癌标本(高、低分化鳞癌各10例),应用免疫组化S-P法检测TIMP-1的表达和分布。结果口腔疣状癌组织中TIMP-1阳性表达率为80%(12/15),高于高分化鳞癌(60%,6/10)和低分化鳞癌(40%,4/10)(P<0.05);平均染色强度高于高、低分化鳞癌(P<0.05);口腔疣状癌、口腔高分化鳞癌、口腔低分化鳞癌组织中TIMP-1的表达均高于正常口腔黏膜组织(P<0.05)。结论OVC是一种不同于口腔鳞状细胞癌的独立类型的恶性肿瘤,TIMP-1可以抑制肿瘤的浸润与转移。     

凝血酶受体在人牙髓组织和细胞中表达的免疫组化研究

目的：探讨凝血酶受体在人正常、炎症牙髓组织及体外培养的牙髓成纤维细胞中的表达及其意义。方法：体外培养人牙贿成纤维细胞,收集正常和炎症牙髓组织,采用免疫组织化学方法观察凝血酶受体在牙髓组织和细胞中的表达和分布。结果：体外培养的牙髓成纤维细胞可见受体强阳性表达,正常及炎症牙髓组织中成牙本质细胞层、牙髓细胞、血管内皮细胞均呈广泛的阳性表达,炎症区淋巴细胞、单核细胞、中性粒细胞染色为强阳性。结论：人牙髓组     

IL-1β对体外培养人牙髓成纤维细胞中基质金属蛋白酶-2的影响

目的:探讨白细胞介素-1β(interleukin-β,IL-1β)对体外培养的人牙髓成纤维细胞中基质金属蛋白酶-2(matrix metallproteinase-I,MMP-2)的影响。方法:利用免疫组化和明胶酶谱法,检测正常和经IL-1β刺激后的牙髓成纤维细胞中MMP-2的表达、分泌和活性。结果:MMP-2在正常和经IL-1β刺激后的牙髓成纤维细胞中均有表达,后者表达明显强于前者。明胶酶谱分析显示,与对照组相比,经IL-1β刺激的牙髓成纤维细胞中MMP-2的水平在48 h后持续显著升高。结论:IL-1β可能参与调节牙髓成纤维细胞合成和分泌MMP-2,从而促进牙髓炎症的发生。     

Immunohistochemical detection of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in ameloblastomas

BACKGROUND: To evaluate the roles of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in tumor progression, expression of MMP-1, -2 and -9 and TIMP-1 and -2 was analyzed in ameloblastomas as well as tooth germs. METHODS: Frozen tissue sections of seven tooth germs and 22 ameloblastomas were immunohistochemically examined using anti-MMP-1, -2 and -9 and anti-TIMP-1 and -2 antibodies. RESULTS: MMP-1, -2 and -9 and TIMP-1 and -2 were expressed strongly in mesenchymal components of tooth germs, and stromal cells of ameloblastomas. Immunoreactivity for MMP-9 in stromal cells of ameloblastomas was significantly stronger than in mesenchymal cells of dental follicles and dental papillae. Dental laminae showed weak MMP-2 expression in six tooth germs, MMP-9 expression in two tooth germs and TIMP-1 expression in six tooth germs. Some tumor cells showed weak MMP-2 expression in 19 ameloblastomas, MMP-9 expression in four ameloblastomas and TIMP-1 expression in all cases. TIMP-2 reactivity was prominently found in basement membrane zones of dental laminae in tooth germs, and tumor cell islands or nests in ameloblastomas. CONCLUSION: Expression of MMPs and TIMPs was considered to be associated with interactions between epithelial cells and mesenchymal components in normal and neoplastic odontogenic tissues; these molecules might play a role in regulation of tumor progression in ameloblastomas as well as regulation of developmental processes in tooth germs.     

S—100蛋白和胶质酸性蛋白在人牙髓组织中表达的研究

目的：观察Ｓ－１００蛋白和胶质酸性蛋白（ＧＡＦＰ）的阳染神经纤维在人第三磨牙的分布。方法：免疫组化染色。结果：Ｓ－１００蛋白的阳性神经纤维广泛分布于牙髓组织，在牙髓中央区其表现为神经束，或环绕于血管壁，在牙髓的周边形成神经网，少许神经末稍进入成牙本质细胞层或前期牙本质。在龋坏牙本质下的成牙本质细胞下层有较多的神经分布。ＧＡＦＰ的阳性纤维分布类似于Ｓ－１００蛋白阳性纤维。结论：Ｓ－１００蛋白和ＧＡＦＰ阳染可显示人牙髓神经纤维支配。     

Langerhans cells express matrix metalloproteinases 9 and 2 and tissue inhibitors of metalloproteinases 1 and 2 in healthy human gingival tissue and in periodontitis

BACKGROUND: As antigen-presenting cells, Langerhans cells may play an important role in the initiation and maintenance of periodontal disease. This study is the first report that extends our knowledge of the expression of matrix metalloproteinases and their endogenous tissue inhibitors by Langerhans cells in healthy and diseased gingival tissues. METHODS: Single and double immunolabeling procedures were carried out using monoclonal antibodies against CD1a, matrix metalloproteinases 2 and 9, and tissue inhibitors of matrix metalloproteinases 1 and 2, and analyzed by conventional and confocal microscopes. RESULTS: Langerhans cells expressed matrix metalloproteinases 2 and 9, and tissue inhibitors of matrix metalloproteinases 1 and 2 in healthy and diseased gingival tissues. The tissue inhibitors of matrix metalloproteinase-positive Langerhans cells were mainly observed in the upper epithelial layers. Matrix metalloproteinase 9-positive Langerhans cells were observed especially during periodontitis and in the basal epithelial layer or crossing the basement membrane. CONCLUSION: During periodontal disease, changes in the expression of matrix metalloproteinases and their tissue inhibitors by gingival Langerhans cells could be implicated in the migration of the cells towards the connective tissue.     

In situ hybridization study on tissue inhibitors of metalloproteinases (TIMPs) mRNA-expressing cells in human inflamed gingival tissue

This study presents the exact cell types and localization of tissue inhibitors of metalloproteinases (TIMPs) production sites in periodontal diseased gingiva by means of in situ hybridization. Gingival tissue specimens were fixed, embedded and hybridized in situ with specific digoxigenin-labeled cRNA probes (386 and 496 bp). TIMP-1 and -2 mRNAs were expressed on macrophages, mononuclear cells, capillary endothelial cells and some fibroblasts throughout the gingival tissue. In periodontitis, TIMP-1 and -2 mRNA-expressing cells showed significantly different localization. TIMP-1 mRNA was broadly observed in the gingival connective tissue while TIMP-2 mRNA was predominantly expressed in the connective tissue adjacent to the pocket epithelium (p < 0.01). Fewer TIMPs mRNA were observed in minimal gingivitis than in periodontitis, especially in the middle zone of gingival tissue. Thus, TIMP-1 and TIMP-2 mRNA was detected differentially and site-specifically in periodontal diseased gingival tissue.     

Cell proliferation in developing human dental pulp A combined flow cytometric and immunohistochemical study

Information concerning cell proliferation and differentiation in dental pulp may be important to understand tooth response to exogenous stimuli. Since few data concerning human dental pulp are available, we have investigated the growth fraction and the localization of proliferating cells in pulp tissue of third molars of young adult human males and females, using flow cytometry and immunohistochemistry. Flow cytometric analysis demonstrates a low proliferative activity of pulp tissue that appears to be confined to radicular pulp, as revealed by immunohistochemical detection of proliferating cells. No polyploid or aneuploid cell populations could be identified, and G2-blocked cells, if any. represented a negligible cell population. Odontoblasts, cells of the sub-odontoblastic layer, and cells of coronal pulp were found to be not proliferating under normal conditions. These data provide the basis for future investigations on proliferative activity and regenerative potentiality of human pulp cells in experimental and clinical situations.     

人牙髓细胞诱导牙本质再生的实验观察

目的:观察人成体牙髓细胞体内诱导牙髓组织修复反应的能力.方法:在矿化诱导液作用下,将一定数量级的人牙髓细胞与β-TCP生物陶瓷颗粒进行复合,植入免疫缺陷鼠磨牙穿髓孔处,7 d、14 d后分别取材进行组织学观察.对照组采用氢氧化钙(Oycal)和空白对照组.结果:组织学观察表明,盖髓术后7 d,各组均出现了牙髓细胞向穿髓孔处迁移、聚集.术后14 d,牙髓细胞组炎症反应仅局限于穿髓孔处,有明显的修复性牙本质桥形成;Dycal组炎症反应涉及到少量冠髓,有部分矿化的纤维性屏障形成;空白对照组炎症反应涉及了大部分冠髓,仅有弥散的骨样牙本质形成.结论:在矿化诱导液作用下,牙髓细胞具有向成牙本质细胞样细胞定向分化的能力,将牙髓细胞植入鼠磨牙的穿髓孔处,显示其具有良好的维持牙髓活力和诱导修复性牙本质形成的能力.     

人牙髓细胞在离体牙髓腔内向成牙本质细胞样细胞分化的研究

目的观察人牙髓细胞在离体牙髓腔内向成牙本质细胞样细胞分化的潜能。方法将原代培养的人牙髓细胞接种至处理过的离体牙髓腔内,2周后固定、脱钙、包埋、切片、染色,显微镜下观察其生长及分化情况。结果牙髓细胞在牙本质表面生长良好,部分细胞伸出胞质突伸入牙本质小管中,表现出成牙本质细胞样细胞的形态。结论牙本质可以诱导牙髓细胞向成牙本质细胞样细胞分化,可为组织工程化牙髓的研制提供实验依据。