TGFβ和rhBMP2对人牙周膜成纤维细胞的作用

目的 研究转化生长因子β（TGFβ）和重组人骨形成蛋白2（rhBMP2)对人牙周膜成纤维细胞（HPDLFs)增殖和分化的作用。方法 观察TGFβ和rhBMP2单独、同时和相继应用对培养的HPDLFs增殖、碱性磷酸酶（ALP）活性，骨钙素（OC）合成及矿化结节形成的作用。结果 TGFβ刺激HPDLFs增殖，对ALP活性，OC合成和矿化结节形成无明显影响；rhBMP2对HPDLFs增殖无明显作用。却显著提高其ALP活性，刺激OC合成和矿化结节形成；两者同时应用时对细胞增殖和分化的作用居于单独作用之间；先应用TGFβ对随后rhBMP2的促分化作用无明显影响；先应用rhBMP2再应用TGFβ对HPDLFs的分化具有显著的刺激作用。结论 RhBMP2能刺激HPDLFs表达成骨细胞表型，TGFβ对BMP2诱导的细胞分化具有明显的刺激作用。两者可能在促进HPDLFs向成骨细胞表型分化的不同阶段发挥作用。     

蜂胶对人牙周膜成纤维细胞作用的实验研究

目的:探讨蜂胶水提液对人牙周膜成纤维细胞(HPDLFs)生长增殖、骨向分化的影响,寻找蜂胶促增殖及分化作用的最佳浓度。方法:将不同浓度的蜂胶水提液加入体外培养的HPDLFs中,设置阳性对照和空白对照,通过检测HPDLFs的增殖(MTT法)、碱性磷酸酶活性及骨钙素含量,观察蜂胶水提取液对HPDLFs的增殖和分化的影响。结果:10~150μg/ml浓度的蜂胶水提液对体外培养HPDLFs的增殖均有促进作用,100μg/ml浓度时促增殖作用最明显。10~200μg/ml蜂胶水提液均可提高HPDLFs的碱性磷酸酶活性、促进HPDLFs的骨钙素合成(P<0.05),其中以100μg/ml蜂胶水提液作用最显著(P<0.01)。结论:低浓度的蜂胶水提液能促进HPDLFs增殖和向成骨细胞方向分化。     

碱性成纤维细胞生长因子对人牙周膜细胞增殖与ALP活性表达的影响

目的：探讨碱性成纤维细胞生长因子（bFGF）以人牙周膜细胞的增殖与碱性磷酸酶（ALP）活性表达的影响,为bFGF应用于牙周组织的再生与修复提供实验依据。方法：采用MTT和ALP活性检测法,观察不同浓度（0．1－10ng/ml)的bFGF第24、48和72小时对5％和10％胎牛血清（FBS）体外培养的第3代人牙周膜细胞（PDLCs）的作用。结果：与对照组比较,①10％FBS,5ng／ml和10ng／ml的bFGF在第24、48、72小时,可显著促进PDLCs的增殖（P＜0．01－0．05）,5％,10ng／ml的bFGF在24、48小时,5ng／ml的bFGF在24小时对PDLCs有显著促增殖作用（P＜0．01）。②10％FBS,1ng／ml的bFGF在48、72小时,5ng／ml的bFGF在24、48、72小时,10ng／mlbFGF在48小时对PDLCs有显著增强ALP活性的作用P＜0．01－0．05）;5％FBS,10ng／ml的bFGF在24、72小时对PDLCs有显著增强ALP活性的作用（P＜0．05）。结论：以上结果提示：在一定的作用时间内一定条件下,5ng／ml和10ng／ml的bFGF可促进人PDLCs的生长和分化。     

重组人骨形成蛋白2对人牙周膜成纤维细胞的生物学作用

目的:研究重组人骨形成蛋白2 (rhBMP2) 对人牙周膜成纤维细胞(HPDLFs) 的生物学作用。方法:原代培养HPDLFs ,并观察rhBMP2 对HPDLFs 增殖、碱性磷酸酶(ALP) 活性、骨钙素(OC) 合成及矿化结节形成的作用。结果:0125～2 mgPml rhBMP2 对HPDLFs 的增殖无明显作用,015～2 mgPml rhBMP2 显著提高HPDLFs 的ALP 活性,刺激OC合成和矿化结节形成。结论:rhBMP2 对HPDLFs 的作用具有剂量依赖性。rhBMP2 能够刺激HPDLFs 表达成骨细胞表型并促进其表型成熟。     

碱性成纤维细胞生长因子对成骨细胞和牙周膜成纤维细胞迁移、增殖的影响

目的　明确碱性成纤维细胞生长因子(bFGF)对体外成骨细胞和牙周膜成纤维细胞迁移、增殖的影响,以 探讨在牙种植体组织界面局部应用bFGF诱导类牙周膜形成的可行性。方法　同一只SD大鼠来源的成骨细胞和 牙周膜成纤维细胞经传代培养至第4代,建立体外创伤模型,分别在普通培养基和含bFGF的培养基中培养,观察 细胞迁移情况,四唑盐比色实验(MTT)测定细胞的增殖速度。结果　普通培养基中,成骨细胞迁移速度快于成纤维 细胞。加bFGF培养基中牙周膜成纤维细胞迁移速度明显快于其他各组,同时MTT结果显示加入bFGF能明显促进 两种细胞的增殖。结论　bFGF能明显促进牙周膜成纤维细胞的增殖、移行。     

体外培养碱性成纤维细胞因子对人牙周膜成纤维细胞生物学特性的影响

目的：探讨体外培养过程中,碱性成纤维细胞生长因子（bFGF）对人牙周膜成纤维细胞（hPDLFs）生物学特性的影响。方法：体外分离培养人牙周膜成纤维细胞并鉴定,加入不同浓度bFGF（1ng／ml、10ng／ml、50ng／ml、100ng／ml）培养,MTT方法检测细胞增殖情况;并对细胞进行矿化诱导,检测细胞的碱性磷酸酶活性。结果：hPDLFs呈星形或长梭形,免疫组化波丝蛋白阳性,角蛋白阴性,证实该细胞来源可靠。bFGF在一定浓度范围内,与细胞增殖成正比,而在本实验培养条件下（10％FBS）bFGF最大效应浓度为10ng／ml。矿化诱导条件下,碱性磷酸酶活性明显增加,在联合应用bFGF的情况下,100ng／ml组碱性磷酸酶活性明显高于其他组。结论：不同浓度bFGF对人牙周膜成纤维细胞生物学行为的影响不同,在一定浓度条件下,低浓度bFGF促进人牙周膜成纤维细胞的增殖,而高浓度bFGF作用于人牙周膜成纤维细胞可能更易于分化。     

孕酮上调人牙周膜成纤维细胞中ALP的表达

目的:用碱性磷酸酶染色及碱性磷酸酶活性分析的方法,研究孕酮对人牙周膜成纤维细胞(hPDLCs)中碱性磷酸酶(Alkaline phosphatase,ALP)表达的影响。方法:原代培养hPDLCs,取第4～6代细胞用于实验。分为空白对照组,孕酮组,抑制剂组。检测普通培养液和成骨诱导培养液中各组细胞在孕酮作用后ALP染色的变化。用碱性磷酸酶活性分析法检测不同浓度的孕酮对hPDLCs中ALP表达的影响。结果:孕酮能够促进hPDLCs中ALP染色阳性面积的表达,且能够提高hPDLCs中ALP的活性,具有时间及剂量依赖性,抑制剂组各指标减小。结论:孕酮能够上调hPDLCs中碱性磷酸酶的表达。     

bFGF对人牙髓、牙周膜成纤维细胞生物学效应的研究

目的：了解碱性成纤维细胞生长因子（bFGF）对人牙髓、牙周膜成纤维细胞的生物学效应,为bFGF在牙髓、牙周病中的治疗研究提供新的实验依据。方法：利用3H-TdR掺入法观察在bFGF作用下人牙髓、牙周膜成纤维细胞DNA和胶原蛋白合成的情况。结果：20ng/mL～60ng/mLbFGF可明显促进人牙髓、牙周膜成纤维细胞DNA的合成（P〈0.01）,在40ng/mL浓度时牙髓,牙周膜成纤维细胞DNA合成最高,40ng/mLbFGF作用于牙髓,牙周膜成纤维细胞,24～48h可使细胞DAN合成显著增多,牙髓成纤维细胞在36h时DNA合成达最高峰,牙周膜成纤维细胞在24h时DNA合成达最高峰;bFGF对牙髓,牙周膜成纤维细胞胶原蛋白的合成无明显促进作用（P〉0.05）。结论：人牙髓、牙周膜成纤维细胞是bFGF的靶细胞,其胞膜上可能有bFGF特异性受体的存在,也表明bFGF在牙髓、牙周组织的创伤愈合中可能起重要作用。     

转化生长因子-β1和碱性成纤维细胞生长因子对人牙周膜成纤维细胞增殖的影响

目的观察转化生长因子．晶和碱性成纤维细胞生长因子单独或联合应用对人牙周膜成纤维细胞增殖的影响。方法采用组织块培养技术进行人牙周膜成纤维细胞的体外培养，对照组采用含小牛血清的DMEM培养液，实验组分别在培养液中加入不同浓度的转化生长因子-β1或碱性成纤维细胞生长因子或两者联合加入，通过四唑盐比色法观察细胞增殖情况。结果转化生长因子-β1或碱性成纤维细胞生长因子单独作用后，人牙周膜成纤维细胞较对照组有显著的增殖，而两者联合作用后的增殖较各自单独作用时明显（P〈0．05）。结论转化生长因子-β1和碱性成纤维细胞生长因子两者联合具有促进人牙周膜成纤维细胞增殖的作用。     

碱性成纤维细胞生长因子对牙周膜细胞作用的研究进展

碱性成纤维细胞生长因子(bFGF)是具有多种功能的多肽生长因子,在正常生理和病理过程中参与生长发育和组织损伤的修复过程.bFGF能促进创伤愈合和组织修复,促进组织再生,对神经元也有促进分化和营养作用.牙周膜细胞在牙周组织重建过程中起着重要作用.目前关于bFGF对牙周膜细胞作用的研究已取得了一定的进展,本文就此作一综述.     

成纤维细胞生长因子对牙周膜细胞群及克隆钙化的作用

目的观察碱基成纤维细胞生长因子（ｂａｓｉｃｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ，ｂＦＧＦ）对牙周膜（ｐｅｒｉｏｄｏｎｔａｌｌｉｇａｍｅｎｔ，ＰＤＬ）细胞克隆体外钙化的作用。方法对３颗前磨牙的牙周膜细胞进行细胞克隆化，观察ｂＦＧＦ对ＰＤＬ细胞克隆钙化能力的影响，测定细胞牙骨质附着蛋白（ｃｅｍｅｎｔｕｍｄｅｒｉｖｅｄａｔａｃｈｍｅｎｔｐｒｏｔｅｉｎ，ＣＡＰ）的结合力及碱性磷酸酶（ａｌｋａｌｉｎｅｐｈｏｓｐｈａｔａｓｅ，ＡＬＰ）的表达。结果ｂＦＧＦ使ＰＤＬ细胞克隆钙化率增加了１１％。ｂＦＧＦ依赖性钙化克隆与非ｂＦＧＦ依赖性钙化克隆相比具有较低的ＡＬＰ表达率和较高的ＣＡＰ结合力。结论ｂＦＧＦ可在体外促进ＰＤＬ细胞克隆钙化的形成，ｂＦＧＦ依赖与非依赖性钙化克隆是不同的细胞种群。     

胰岛素样生长因子-I对培养人牙周膜成纤维细胞生物学活性的影响

胰岛素样生长因子(IGF)可通过直接作用于成骨细胞或通过甲状旁腺素间接影响骨代谢过程而参与骨的改建。本文利用细胞培养技术观察了IGF-I对培养人牙周膜成纤维细胞(PDLFs)增殖、碱性磷酸酶(ALP)活性变化以及蛋白质含量改变的影响。结果显示,IGP-I对PDLFs有明显的促增殖作用,对PDLFs的ALP活性、蛋白质合成也具有较强的促进作用,表明IGF-I可能通过PDLFs发挥其调节牙槽骨改建的作用,从而在牙齿移动过程中起重要作用。     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10-¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodiflerentiation of PDL cells into mineralized tissue forming cells.     

胰岛素样生长因子和碱性成纤维细胞生长因子对人牙周膜细胞增殖的影响

目的：探讨重组人胰岛素样生长因子(rhIGF）和碱性成纤维细胞生长因子（bFGF)单独及联合应用对人牙周膜细胞（PDLC）增殖的影响。方法：采用细胞培养技术,噻唑盐比色测定（MTT）法。结果：rhIGF-1和bFGF对PDLC的促增殖效应较对照组有明显提高,rhIGF-1在浓度为3.125μg/L时对PDLC作用非常显著。bFGF浓度在1－10μg/L之间时对PDLC有较强的促进增殖作用,其起效浓度是1μg/L。rhIGF和bFGF联合作用时其促增殖作用较两种因子在相同浓度时单独应用相差显著。结论：rhIGF和bFGF单独或联合应用时有促进PDLC的增殖的作用。     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10⁻¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodifferentiation of PDL cells into mineralized tissue forming cells.     

Effects of connective tissue growth factor on human periodontal ligament fibroblasts

ObjectiveThe aim of this study was to evaluate the effects of different concentrations of connective tissue growth factor (CTGF) on human periodontal ligament fibroblasts(HPLFs).DesignHPLFs were cultured and identified. Then, different concentrations of CTGF (1, 5, 10, 50, 100 ng/ml) were added to the HPLF culture. Next, CCK-8 assays, alkaline phosphatase (ALP) assays, hydroxyproline determination, alizarin red staining methods, Transwell chambers and real-time PCR methods were applied to observe the effects of CTGF on the proliferation, ALP activity, synthesis of collagen, formation of mineralized nodules and migration. We also studied expression of ALP, fiber link protein (FN), integrin-binding sialoprotein (IBSP), osteocalcin (OC), and integrin beta 1 (ITGB1) mRNA by HPLFs. Statistical significance was assumed if P < 0.05 or P < 0.01.ResultsThe addition of CTGF (1, 5, 10 ng/ml) remarkably promoted the proliferation and collagen synthesis of HPLFs compared with controls. CTGF (1, 5, 10, 50 ng/ml) improved ALP activity of HPLFs, and at all concentrations, CTGF (1, 5, 10, 50, 100 ng/ml) improved the expression of ALP, FN, IBSP and ITGB1 mRNA. In addition, CTGF (1, 5, 10, 50, 100 ng/ml) promoted the migration of HPLFs, which was dose-dependent, with maximal promotion in the 10 ng/ml group (P < 0.05 or P < 0.01).ConclusionsThus, in a certain range of concentrations, CTGF can promote the biological effects, including proliferation, migration and collagen synthesis of HPLFs, to promote the differentiation of HPLFs in the process of osteogenesis.     

碱性成纤维细胞生长因子对人牙周膜成纤维细胞内核心蛋白多糖基因表达的影响

目的研究碱性成纤维细胞生长因子（bFGF）对人牙周膜成纤维细胞内核心蛋白多糖（decorin）的影响,探讨bFGF在牙周再生中的意义。方法体外原代培养人牙周膜成纤维细胞,用外源性bFGF刺激细胞,半定量RTPCR法检测牙周膜成纤维细胞内decorin基因表达的变化。结果电泳结果表明,bFGF抑制人牙周膜成纤维细胞内decorin的mRNA合成,并且随着质量浓度的增加抑制作用减弱。结论decorin具有很多生物学功能,bFGF对decorin的抑制作用很可能是牙周炎损伤修复过程中一个重要的调节因素,为bFGF在牙周组织再生中的作用提供理论基础。     

人牙周膜成纤维细胞增殖与压力关系的初步观察

目的：在体外培养环境下观察压力对人牙周膜成纤维细胞（HPLF）增殖的影响。方法：取第4－6代培养的HPLF,应用可控压力细胞加载装置分别间断性施加30、60、90kPa的压力,分别在培养3、5、7d后用MTT法测定各组的A值。结果：HPLF在30、60、90kPa的压力培养环境下MTT反应的A值显著小于对照组,压力值越大,A值越小。结论：30、60、90kPa间断性压力显著抑制PLF细胞增殖,该抑制作用随压力值增大而增强。