表皮生长因子对牙囊细胞增殖活性的影响

目的：研究表皮生长因子（EGF）对体外培养的大鼠牙囊细胞增殖活性的影响。方法：原代培养Wistar大鼠牙囊细胞,选择生长良好的第3代细胞,分别加入不同浓度的EGF（0.5、1、5、10、50 ng/mL）与牙囊细胞共孵育5 d,MTT法对比观察不同浓度的EGF对牙囊细胞增殖活性的影响。数据采用SPSS 13.0软件包进行方差分析。结果：在EGF浓度为5～10 ng/mL时,牙囊细胞的增殖活性显著增高（P〈0.05）,10 ng/mL时达到最高（P〈0.01）。结论：EGF可作用于牙囊细胞,合适浓度的EGF能显著增加牙囊细胞的增殖活性。     

表皮生长因子在牙齿萌出通道形成中的作用研究

目的:研究表皮生长因子(EGF)在牙齿萌出过程中,是否参与软、硬组织通道的形成。方法:①免疫组化法检测表皮生长因子及其受体在出生后13、15 d以及成年小鼠下颌第一磨牙萌出部位口腔黏膜的表达变化;②原代培养Wistar大鼠牙囊细胞,选择生长良好的第3代细胞,MTT法筛选EGF作用于牙囊细胞的较佳效应浓度。将第3代牙囊细胞以1×105/孔接种到培养皿中,加入最佳浓度的EGF孵育0.5、1、3、6 h后,Trizol一步法分别提取总RNA,采用反转录聚合酶链反应(RT-PCR)检测同一浓度的EGF作用不同时间后,牙囊细胞MCP-1 mRNA的表达变化。结果:①牙萌出时,EGF在萌出牙齿冠方黏膜的上皮层呈弱阳性表达,表皮生长因子受体(EGFR)在口腔上皮全层呈强阳性表达。而牙萌出后,EGF的表达集中于口腔黏膜的固有层,EGFR的表达集中于上皮基底层;②在EGF浓度为5～10 ng/mL时,对牙囊细胞的增殖具有明显促进作用(P<0.05),其中10 ng/mL促进作用最强。牙囊细胞与10 ng/mL的EGF共同孵育0.5、1、3、6 h均能明显促进牙囊细胞MCP-1 mRNA的表达(P<0.05),其中3 h时促进作用最强,以后逐渐恢复,但仍比对照组高(P<0.05)。结论:EGF及其受体可能促进萌出牙齿冠方实性上皮团的形成;适当浓度的EGF能显著增加牙囊细胞的增殖活性,并上调牙囊细胞中MCP-1 mRNA的表达。     

单核细胞趋化蛋白-1与口腔疾病关系的研究进展

单核细胞趋化蛋白-1是趋化因子CC亚族成员之一,能趋化和激活单核细胞、巨噬细胞等,参与调节多种生理、病理和免疫应答过程。下面就单核细胞趋化蛋白-1的结构及其与口腔常见疾病,如牙髓炎、根尖周炎、牙周炎和口腔鳞癌等的关系作一综述。     

单核细胞趋化蛋白-1和细胞间黏附分子-1与动脉粥样硬化和牙周病的关系

以动脉粥样硬化为病理基础的冠心病是目前主要的致死性疾病之一,而牙周病与动脉粥样硬化之间存在着一定的相关性。单核细胞趋化蛋白(MCP)-1和细胞间黏附分子(ICAM)-1在动脉粥样硬化和牙周病的发生发展中起着重要的作用。本文就MCP-1和ICAM-1以及二者与动脉粥样硬化、牙周病间的关系作一综述。     

单核细胞趋化蛋白-1和细胞间黏附分子-1与动脉粥样硬化和牙周病的关系

以动脉粥样硬化为病理基础的冠心病是目前主要的致死性疾病之一，而牙周病与动脉粥样硬化之间存在着一定的相关性。单核细胞趋化蛋白（MCP）-1和细胞间黏附分子（ICAM）-1在动脉粥样硬化和牙周病的发生发展中起着重要的作用。本文就MCP-1和ICAM-1以及二者与动脉粥样硬化、牙周病间的关系作一综述。     

炎症组织中CD40/CD40L与白细胞介素-8和单核细胞趋化蛋白-1的相关性

CD40/CD40L相互作用可促进多种细胞前炎症细胞因子和趋化因子的产生,如白细胞介素(IL)-8和单核细胞趋化蛋白(MCP)-1,而IL-8和MCP-1可趋化炎症细胞向炎症部位聚集,从而调节炎症的发生和发展.下面就CD40/CD40L与IL-8和MCP-1在炎症组织中的相关性以及CD40/CD40L诱导IL-8和MCP-1的调节因素等作一.     

人牙囊细胞中表皮生长因子的表达以及流体静压力下的改变

目的：研究体外培养的人牙囊细胞EGF的表达以及在不同流体静压力作用下对细胞分泌．EGF的影响,探讨EGF在牙齿萌出过程中的作用。方法：取年龄为12岁男孩埋藏阻生牙齿的牙囊进行人牙囊细胞培养,将传代的培养细胞应用原位杂交的方法表达EGF,RT-PCR定量分析,分别在50、100kPa流体静压力作用1h后RT-PCR方法观察细胞EGF的浓度变化。结果：培养的人牙囊细胞呈梭形,主要为成纤维样细胞,原位杂交染色显示人牙囊细胞中EGF的表达呈阳性,定位于细胞的胞质中,50kPa流体静压力表达略有增加,100kPa压力表达明显降低,说明流体静压力在一定力值范围内对人牙囊细胞中有EGF的表达具有空间性。结论：培养的人牙囊细胞具有合成与分泌EGF的能力．流体静压力可以影响体外培养的人牙囊细胞EGF的分泌。     

IGF-1对小鼠牙囊细胞生物学特性的影响

目的:探讨胰岛素样生长因子-1(IGF-1)对小鼠牙囊细胞增殖、蛋白含量及分化能力影响。方法:将第3代小鼠牙囊细胞与不同浓度IGF-1(0.005、0.01、0.05、0.1、0.5 mg/L)共孵育后,测定IGF-1对细胞增殖、碱性磷酸酶活性、总蛋白含量的影响;并用Von Kossa法检测0.05和0.1 mg/L的IGF-1对牙囊细胞矿化结节形成能力的影响。结果:在0.05~0.1 mg/L的浓度范围内,IGF-1能显著促进牙囊细胞的增殖,使ALP活性及总蛋白含量增加(P<0.01),但三者的最佳效应浓度有所差异,当浓度升高至0.5 mg/L时,促进效应下降,与对照组无显著性差异(P>0.05);0.05和0.1 mg/L的IGF-1均能显著促进牙囊细胞矿化结节的形成(P<0.05,与对照组相比)。结论:合适浓度的IGF-1能促进牙囊细胞的增殖,增加蛋白含量及向成骨细胞(成牙骨质细胞)方向分化的能力。     

单核细胞趋化蛋白-1在口腔黏膜下纤维性变中的表达及意义

目的：探讨单核细胞趋化蛋白-1（MCP-1）在口腔黏膜下纤维性变（OSF）组织中的表达及其意义。方法：应用免疫组织化学SP法检测45例OSF和15例正常口腔黏膜组织中MCP-1的表达及分布情况。结果：OSF组织中MCP-1的表达较正常组织增加,差异有统计学意义（P〈0.01）,MCP-1在OSF早期组织与中、晚期组织中的表达有显著性差异（P〈0.05）。结论：MCP-1可能通过趋化、激活免疫和炎症细胞,在OSF发生发展中发挥重要作用。     

慢性牙周炎牙周基础治疗前后血清超敏C反应蛋白与单核细胞趋化蛋白-1变化研究

目的    研究慢性牙周炎对血清超敏C反应蛋白（hs-CRP）和单核细胞趋化蛋白-1（MCP-1）浓度的影响。方法    选取2015—2016年于中国人民解放军陆军总医院口腔科门诊就诊的80例牙周炎患者（牙周病组）和40名无牙周炎症状的志愿者（对照组）。初诊记录探诊出血（BOP）、牙周探诊深度（PD）和临床附着丧失（CAL）情况,收集空腹静脉血,检测血清超敏C反应蛋白（hs-CRP）和单核细胞趋化蛋白-1（MCP-1）。对牙周病组进行为期3个月的牙周基础治疗,治疗结束3个月后,再次收集空腹静脉血,检测血清hs-CRP和MCP-1。结果    牙周病组与对照组基线条件无明显差异。对照组hs-CRP和MCP-1浓度为（1.48 ± 1.1）mg/L和（42.3 ± 4.9）mg/L,牙周病组分别为（3.89 ± 2.8）mg/L和（88.7 ± 5.8）mg/L,两组差异有统计学意义（P＜0.05）。牙周治疗结束3个月后,牙周病组hs-CRP和MCP-1浓度为（2.04 ± 1.5）mg/L和（64.2 ± 5.1）mg/L,与治疗前比较差异有统计学意义（P＜0.05）。结论    　慢性牙周炎可引起低滴度菌血症,造成血清炎症标志物hs-CRP和MCP-1浓度增高,有效的牙周基础治疗可降低血清hs-CRP和MCP-1。     

bFGF对体外培养人牙囊细胞VEGF表达的影响

目的：研究碱性成纤维细胞生长因子（bFGF）对体外培养人牙囊细胞血管内皮生长因子（VEGF）表达的影响。方法：选取生长状态良好的人牙囊细胞,与10ng／mlbFGF孵育不同时间后,采用反转录聚合酶链反应（RT—PCR）及酶联免疫吸附试验（ELISA）分别检测人牙囊细胞VEGFmRNA表达及上清液中VEGF蛋白分泌变化。结果：bFGF作用1、3、6、12h后人牙囊细胞VEGFmRNA表达均较0h组显著增强（p〈0．01）,其中bFGF作用6h后VEGFmRNA表达最强;细胞培养上清液中VEGF蛋白分泌量随时间延长逐渐增加,但bFGF处理组VEGF蛋白分泌量高于对照组（p〈0．01）。结论：bFGF能够促进体外培养人牙囊细胞VEGF的表达。     

大鼠牙囊细胞集落刺激因子1受体的研究

目的 研究不同浓度集落刺激因子1(colony—stimulating factor-1，CSF-1)、白细胞介素1α(interleukin—1α，IL—1α)对大鼠牙囊细胞CSF—1受体基因表达的影响；明确CSF—1自分泌作用的抑制机制，为研究细胞因子在牙齿萌出和牙槽骨吸收中的作用奠定基础。方法 间接免疫酶标法检测体外培养牙囊细胞，以及出生后不同时期牙囊组织中CSF—1受体蛋白表达。逆转录多聚酶链反应法(RT—PCR)检测在细胞培养液中分别加入不同浓度CSF—1、IL—1α对牙囊细胞CSF—1受体基因表达的影响。结果 间接免疫酶标法染色显示CSF—1受体蛋白在生后早期牙囊中染色明显，10d后染色不明显或者完全无染色，高浓度CSF—1作用下，牙囊细胞CSF—1受体mRNA表达明显消失。不同浓度IL—1α作用后，CSF—1受体mRNA水平保持不变。结论 CSF—1受体蛋白在出生后早期牙囊中表达明显，随后逐渐消失。高浓度CSF—1抑制CSF—1受体基因的表达，不同浓度IL—1α对牙囊细胞CSF—1受体mRNA表达无影响。     

Effects of epidermal growth factor and transforming growth factor-beta on insulin-induced differentiation in rat dental pulp cells.

An established pulp cell line (RPC-C2A) was used to study the regulatory effect of insulin on dentinogenesis. Insulin increased alkaline phosphatase activity and the incorporation of [2,3-3H]-proline into collagenase-digestible protein, whereas [3H]-thymidine incorporation by the cells was inhibited by insulin. The enhancing effect of insulin on alkaline phosphatase activity was inhibited by epidermal growth factor (EGF) or transforming growth factor-beta (TGF-beta). The stimulatory effect of insulin on collagen synthesis was also inhibited when insulin was combined with EGF, but was accelerated by the addition of TGF-beta. Inhibitory effects of insulin on the [3H]-thymidine incorporation were potentiated by EGF, though EGF alone strongly increased the effect; whereas the addition of TGF-beta had no significant effect on the insulin action. These findings suggest that insulin may be concerned with the differentiation of pulp cells in dentinogenesis and that EGF or TGF-beta regulate the insulin effects.     

表皮生长因子与碱性成纤维细胞生长因子对人牙髓干细胞分化的影响

目的探讨表皮生长因子(epidermal growth factor,EGF)和碱性成纤维细胞生长因子(basic fibroblastgrowth factor,bFGF)对人牙髓干细胞(human dental pulp stem cells,hDPSCs)碱性磷酸酶(alkaline phosphatase,ALP)活性的影响。方法确定EGF与bFGF最大效应浓度后,对细胞分组,根据EGF和bFGF单独或联合应用,分为4组:EGF组、bFGF组、EGF联合bFGF组、不加任何生长因子的对照组。作用1、3、5、7、14 d后,采用ALP活性检测法(酶动力法)检测hDPSCs细胞ALP活性。结果 1～14 d bFGF组ALP活性与对照组相比差异无统计学意义(P>0.05);在5、7、14 d,EGF组和EGF联合bFGF组ALP活性显著高于对照组(P<0.05);EGF联合bFGF组的ALP活性明显高于bFGF组(P<0.05),但EGF联合bFGF组ALP活性与EGF组相比差异无统计学意义(P>0.05)。结论 bFGF单独应用不能诱导hDPSCs分化,EGF在hDPSCs的分化中发挥作用,EGF和bFGF无明显协同作用。     

Effects of transforming growth factor-beta 1 on cultured dental follicle cells from rat mandibular molars.

Analysis of the total proteins secreted by cultured dental follicle cells revealed that transforming growth factor-beta 1 (TGF-beta 1) stimulated them to secrete more extracellular matrix proteins into a serum-free medium than did follicle cells not exposed to the growth factor. Electrophoresis and scanning densitometry showed that secretion of all the major proteins was increased by exposure to the growth factor but the amounts ranged from a 66% increase for one of the procollagen chains to a 7% increase for fibronectin. Immunofluorescence using anti-type I collagen and anti-fibronectin showed that the intracellular concentration and intracellular localization of the antibodies was not changed by incubating the cells with the growth factor. The growth factor did not cause an increase in cell number but did modify the association of the cells in the culture, causing them to aggregate into clusters whereas the control cells formed a confluent monolayer. These results suggest that TGF-beta 1 may signal the fibroblasts of the dental follicle to secrete the extracellular matrix needed for its development into a periodontal ligament.     

肿瘤坏死因子α对体外培养人牙囊细胞血管内皮生长因子表达的影响

目的研究肿瘤坏死因子α(TNF-α)对体外培养人牙囊细胞血管内皮生长因子(VEGF)表达的影响。方法原代培养人牙囊细胞,传代至第4代,采用反转录聚合酶链反应(RT-PCR)及酶联免疫吸附试验(ELISA)分别检测TNF-α对体外培养人牙囊细胞VEGF mRNA表达及上清液中VEGF含量改变的浓度和时间效应。结果①不同浓度TNF-α作用1h后,TNF-α处理组人牙囊细胞VEGF mRNA表达均较对照组增强(P<0.05),最佳效应浓度为25ng/ml;不同浓度TNF-α作用12h后,除1ng/ml组外,其余各组上清液中VEGF蛋白含量明显高于对照组(P<0.05)。②在时间效应上,25ng/mlTNF-α作用1h后人牙囊细胞VEGF mRNA表达最强,然后随时间延长其表达逐渐下降,但仍强于对照组(P<0.05);细胞培养上清液中VEGF蛋白含量随时间延长逐渐增加,但TNF-α处理组VEGF蛋白含量高于对照组(P<0.05)。结论TNF-α能够促进体外培养人牙囊细胞合成并分泌VEGF。     

Effects of transforming growth factor-beta and epidermal growth factor on clonal rat pulp cells

These factors influence proliferation and differentiation in various cell types. Their effects on a clonal cell line (RPC-C2A) having high ALPase activity were examined by assay of [3H]-thymidine incorporation and ALPase activity. Neither factor (at a dose of 0.5 ng/ml) altered the shape of the pulp cells. DNA synthesis was not affected by transforming growth factor-beta either in growing cells or in those nearly confluent, but epidermal growth factor, in doses ranging from 0.5 to 10 ng/ml, stimulated the incorporation of [3H]-thymidine in nearly confluent cells. Both factors inhibited ALPase activity in a dose-dependent manner. Indomethacin did not affect this inhibition, suggesting that this effect of growth factors may not be mediated by prostaglandin synthesis. Inhibitory effects of ALPase antagonists (L-phenylalanine, L-homoarginine, levamisole) were not affected by transforming growth factor-beta. Thus epidermal growth factor stimulates DNA synthesis and both transforming growth factor-beta and epidermal growth factor inhibit ALPase activity of clonal rat pulp cells, suggesting that both factors may act as regulators of biological function, including cell differentiation, in pulp cells.     

Synthesis and secretion of MCP-1 by dental follicle cells--implications for tooth eruption

The monocyte chemotactic protein-1 (MCP-1) gene is expressed in the dental follicle, a loose connective tissue sac that must be present for eruption to occur. The role of MCP-1 may be to recruit mononuclear cells (monocytes) to the dental follicle, where these cells, in turn, fuse to form osteoclasts to resorb alveolar bone for the formation of an eruption pathway. Thus, it was the aim of this study to determine if MCP-1 is secreted by dental follicle cells in culture and if its secretion is enhanced by potential tooth eruption molecules. Western blotting and a two-site capture enzyme-linked immunoabsorbent assay demonstrated that MCP-1 was synthesized and secreted into the medium by the follicle cells. Incubation of the cells with either transforming growth factor-beta one (TGF-beta 1) or interleukin-one alpha (IL-1 alpha) enhanced the secretion of MCP-1 by the cells. Measurement of the chemotactic ability of the conditioned medium to attract mouse monocytes demonstrated that the chemotaxis of the medium was increased if the cells had previously been incubated in IL-1 alpha, although there appears to be a threshold concentration of MCP-1 above which chemotaxis is not enhanced. These combined results suggest that the critical initial cellular event of tooth eruption, an influx of mononuclear cells into the dental follicle at an early post-natal age, may be initiated by the secretion of MCP-1 by the dental follicle cells.     

白细胞介素-1α和集落刺激因子-1对大鼠牙囊细胞骨保护素的影响

目的:研究白细胞介素(IL)-1α和集落刺激因子(CSF)-1对体外培养大鼠牙囊细胞骨保护素(OPG)表达的影响。方法:组织块结合酶消化法体外培养原代牙囊细胞并传代。取50 ng/ml IL-1α或CSF-1刺激第4代牙囊细胞,western blot检测第0、1、3、6、12 h OPG表达。采用不同浓度(0、4、10、50、250 ng/ml)的IL-1α或CSF-1孵育第4代牙囊细胞12 h,western blot检测OPG表达。结果:western blot结果显示随着IL-1α刺激时间和浓度的增加,OPG条带逐渐加深。随着CSF-1刺激时间的增加,OPG条带变浅但高于对照组;各CSF-1浓度组OPG条带无明显变化趋势,但高于对照组。结论:IL-1α、CSF-1均可上调大鼠牙囊细胞内OPG蛋白水平。     

PDGF BB and bFGF stimulate DNA synthesis and upregulate CSF-1 and MCP-1 gene expression in dental follicle cells

CSF-1 and MCP-1, released by dental follicle cells, stimulate the influx of monocytes into the follicle sac and enhance the formation of osteoclasts that, in turn, resorb alveolar bone for the eruption pathway. PDGF and bFGF, released by cells adjacent to the follicle or by activated monocytes, are prime candidates that may regulate CSF-1 and MCP-1 gene expression. The present study demonstrates that PDGF and bFGF are mitogens for dental follicle cells and stimulate CSF-1 and MCP-1 mRNA, but with different time course kinetics. Peak induction of CSF-1 mRNA was observed at 6-8h, while maximal MCP-1 induction was observed at 2h. These findings suggest that MCP-1 is an early chemotactic signal for monocytes and that subsequent release of CSF-1 may act synergistically with MCP-1 to enhance monocyte influx. Further understanding of the molecular mechanisms by which cytokines regulate CSF-1 and MCP-1 may lead to more effective treatment regimens for disorders associated with abnormal tooth eruption.