碱性成纤维细胞生长因子对牙周细胞生物学活性的影响

目的：观察基因重组入碱性成纤维细胞生长因子（rh-bFGF)牙龈成纤维细胞（GF）、人牙周韧带成纤维细胞（PDLF）及人牙槽骨细胞（ABC）的增殖、碱性磷酸酶活性、总蛋白含量及对3种细胞矿化结节形成能力的影响。方法：采用细胞培养、MTT比色测定、碱性磷酸酶测定法、考马宙蓝法及茜素红染色法。结果bFGF能促进3种细胞的增殖,但对PDLF和ABC的ALP活性,蛋白含量及矿化结节的形成有抑制作用。结论bFGF可促进细胞的增殖,抑制细胞的分化成熟,从而促进牙周再生。     

牙骨质基质提取物促进牙周结缔组织细胞在牙根表面上增殖的实验研究

目的：证实牙骨质基质提取物可促进牙周结缔组织细胞在牙根表面上增殖。方法：用细胞培养法和增殖细胞记数法。结果：加入牙骨质基质提取物的实验组无论是牙龈成纤维细胞还是牙周膜细胞的增殖能力均有显著的提高。结论：牙骨质基质提取物可促进牙龈成纤维细胞和牙周膜成纤维细胞在牙根表面上的增殖，牙骨质基质提取物对牙龈成纤维细胞的促有丝分裂作用强于对牙周膜成纤维细胞的作用     

牙骨质基质提取物促进牙周结缔组织细胞在牙根表面上增？…

目的：证实牙骨质基质提取物可是牙周结缔组织细胞在牙根表面上增殖。方法：用细胞培养法和增殖细胞记数法。结果：加入牙骨质基质提取物的实验组无认是牙龈成纤维细胞还是牙周膜细胞的增殖能力均有显著的提高。结论：牙骨质基质提取物可促进牙龈成纤维细胞和牙周膜成纤维细胞在牙根表面上的增殖，牙骨质基质提取物对牙龈成纤维细胞的促有丝分裂作用强于对牙周膜成纤维细胞的作用。     

bFGF对成骨细胞和成纤维细胞生物学特性的影响

目的：明确bFGF埘体外成骨细胞和牙周膜成纤维细胞迁移、矿化相关蛋白合成的影响，以探讨bFGF促进牙周组织再生的内任机制以及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大鼠米源的两种细胞经传代培养至第四代，建立细胞迁移模型，分别在普通培养基和含bFGF的培养基中培养，观察测量细胞迁移速度，并分别测试两种细胞在普通培养基和含bFGF的培养基中的碱性磷酸酶（ALP）活性歧Ⅰ型胶原的含量。结果：普通培养基培养时，成骨细胞迁移速度快于成纤维细胞；但牙周膜成纤维细胞加bFGF培养辽移速度明显高于其他各组；bFGF抑制成骨细胞的Ⅰ型胶原的合成和ALP的活性。结论：bFGF能日月硅促进牙周膜成纤维细胞的移行；bFGF能抑制成骨细胞的碱性磷酸酶活性。     

Nd：YAG激光照射对提高人牙周膜成维细胞增殖力的作用

目的观察Nd：YAG激光对人牙周膜成纤维细胞增殖和碱性磷酸酶（ALP）活性的影响。方法采用酶消化组织块法原代培养人牙周膜成纤维细胞，并进行波形蛋白及角蛋白的免疫细胞化学染色，对所培养的细胞进行鉴定。将牙周膜成纤维细胞分组，接受不同能量密度的Nd：YAG激光照射。测定牙周膜成纤维细胞碱性磷酸酶活性并观察其生长情况。结果5个实验组的细胞增殖速度及碱性磷酸酶含量均高于对照组，其中能量密度为60j／cm^2组、80j／cm^2组、100i／cm^2组与对照组相比有统计学的差异。结论低能量密度，短时间的Nd：YAG激光照射能促进牙周膜成纤维细胞的增殖，提高碱性磷酸酶活性。     

骨形成蛋白和（或）甲状旁腺激素对牙周膜细胞碱性磷酸酶活性…

利用对硝基苯酚法，在体外ＢＭＰ作用的牙周膜细胞和牙龈成纤维细胞进行碱性磷酸酶活性的测定。结果表明，牙周膜细胞的碱性磷酸酶活性明显高于牙龈成纤维细胞。经骨形成蛋白作用后，牙周膜细胞的碱性磷酸酶活性有所升高，在甲状腺素的作用下，经ＢＭＰ作用过的牙周膜细胞的碱性磷酸酶活性有较大幅度地下降，而牙龈成纤维细胞无明显变化，提示牙周膜细胞中可能存在有骨形成蛋白的靶。     

雌激素对牙周膜细胞生物学性质的影响

目的：明确雌激素对体外牙周膜成纤维细胞迁移、增殖和碱性磷酸酶的影响，以探讨雌激素对牙周组织改建的影响。方法：SD大鼠来源的牙周膜成纤维细胞经传代培养至第四代，建立体外创伤模型，分别在普通培养基和含雌激素的培养基中培养，观察测量细胞迁移情况，运用MTT、酶联检测仪测定细胞的增殖速度，对细胞的ALP进行提取和测定。结果：MTT结果硅示加入雌激素对该细胞的增殖无明显影响；在含雌激素的培养基中成纤维细胞的迁移速度加快；同时牙周膜成纤维细胞的ALP表达娃著提高。结论：雌激素能明显促进牙周膜成纤维细胞的迁移，促进其分化。     

人牙周膜成纤维细胞在纯钛表面附着的电镜观察

目的：研究人牙周膜成纤维细胞(PDLF)在纯钛金属表面的结合形式和超微结构。方法：将纯钛试件放在12孔培养板内，取生长良好的第五代人牙周膜成纤维细胞接种在试件表面，培养72h后取出，原位包埋法制作透射电镜标本，透射电镜观察。结果：人牙周膜成纤维细胞在纯钛表面附着形式不同于上皮细胞在金属表面的附着形式，细胞与金属结合界面中未观察到典型半桥粒结构。人牙周膜成纤维细胞胞浆中，细胞器发达，富含与蛋白质合成、代谢及增殖等生物学功能有关的细胞超微结构，如粗面内质网、线粒体、核糖体、细胞核等。结论：人牙周膜成纤维细胞在钛金属表面的附着形式，不同于上皮细胞的附着形式，表现为蛋白合成旺盛，可能为细胞直接附着，其具体附着形式还待进一步研究。     

骨形成蛋白和（或）甲状旁腺激素对牙周膜细胞碱性磷酸酶活性的影响

利用对硝基苯酚法，在体外ＢＭＰ作用的牙周膜细胞和牙龈成纤维细胞进行碱性磷酸酶活性的测定。结果表明，牙周膜细胞的碱性磷酸酶活性明显高于牙龈成纤维细胞。经骨形成蛋白作用后，牙周膜细胞的碱性磷酸酶活性有所升高；在甲状旁腺素的作用下，经ＢＭＰ作用过的牙周膜细胞的碱性磷酸酶活性有较大幅度地下降，而牙龈成纤维细胞无明显变化，提示牙周膜细胞中可能存在有骨形成蛋白的靶细胞。     

釉基质蛋白对牙周细胞覆盖体外创面的影响

目的：评价体外创面模型环境中,釉基质蛋白对牙周膜细胞、牙龈成纤维细胞覆盖创面能力的影响。方法：利用在盖玻片上形成的人牙周膜和牙龈成纤维细胞的融合培养物,以机械方法去除7mm宽的细胞层条带,形成体外创面模型。受损培养物在含有10％胎牛血清的培养液中继续培养2、6、9d,同时以100μg／ml的釉基质蛋白分别刺激牙周膜、牙龈成纤维细胞,并与不加釉基质蛋白者作对照。玻片经同定后作甲紫染色。以计算机辅助图像分析系统对细胞覆盖体外创面的面积进行百分比定量,采用SAS6、12软件包进行样本均数的t检验。结果：对照组组内牙周膜、牙龈成纤维细胞覆盖创面的面积存在差异,牙龈成纤维细胞覆盖创面面积在创面形成后第9天显著高于牙周膜细胞,差异有显著性（P〈0.05）。在加用100μg／ml釉基质蛋白的条件下,牙周膜细胞覆盖体外创面的面积较对照组明显增加．创面形成后第6、9天,牙周膜、牙龈成纤维细胞覆盖体外创面面积均无显著差异（P〉0．05）。结论：牙龈成纤维细胞覆盖创面的能力高于牙周膜细胞。釉基质蛋白有增进创面愈合,特别是牙周膜细胞创面覆盖的作用。     

胰岛素样生长因子-Ⅱ对培养人牙周膜成纤维细胞生物学活性的影响

目的：研究ＩＧＦ－ＩＩ对培养人牙周膜成纤维细胞（ＰＤＬＦ）生物学活性的影响,方法：接种一定量培养至第５代的人ＰＤＬＦ,用ＭＴＴ法和碱性磷酸酶比色法观察ＩＧＦ－ＩＩ作用下,ＰＤＬＦ的增殖和碱性磷酸酶活性的变化。结果：ＩＧＦ－ＩＩ在实验浓度范围内,对ＰＤＬＦ有较明显的促增殖作用,对ＰＤＬＦ的ＡＬＰ活性具有较强的促进作用。结论：ＩＧＦ－ＩＩ对ＰＤＬＦ的生物学活性有一定的影响,揭示其可能通过ＰＤＬＦ发挥其     

胶原基纳米骨的遗传毒性及对体外培养细胞影响的研究

目的：体外研究胶原基纳米骨的遗传毒性,及对原代培养的人牙周膜成纤维样细胞、兔成骨细胞的影响。方法：采用Ames致突变试验、MTT法及碱性磷酸酶（ALP）检测法,测定不同浓度胶原基纳米骨（nHAC）的浸提液对鼠伤寒沙门菌的致突变比值的影响和对人牙周膜成纤维样细胞及兔成骨细胞的影响。结果：nHAC的浸提液各剂量组对鼠伤寒沙门菌的致突变比值均小于2,nHAC不会引起鼠伤寒沙门菌的回复突变数增加。不同时间点用不同浓度浸提液培养的人牙周膜成纤维样细胞正常增殖,浸提液不影响兔成骨细胞的功能表达。结论：胶原基纳米骨无遗传毒性,不影响人牙周膜成纤维样细胞的增殖活性和兔成骨细胞的成骨活性,是一种组织工程骨支架的良好材料。     

牛血浆FN对人牙髓成纤维细胞骨形成蛋白和碱性磷酸酶分泌和表达的影响

目的：观察纤维粘连蛋白（ＦＮ）与牙髓细胞分化的关系．方法：将传代的人牙髓细胞接种于９６孔板,分别用不同浓度牛血浆ＦＮ（１０,２０,４０,８０μｇ／ｍｌ）作用３天,进行骨形成蛋白（ＢＭＰ）和碱性磷酸酶（ＡＬＰ）的免疫组化染色,图象分析仪半定量测定,并测定培养上清液中ＡＬＰ活性．结果表明：四种浓度的ＦＮ组的牙髓细胞ＢＭＰ和ＡＬＰ表达无明显差异,ＦＮ（２０,４０μｇ／ｍｌ）组培养上清液中ＡＬＰ活性较强．结论：ＦＮ在牙髓细胞分化过程中的作用是有限的．     

碱性成纤维细胞生长因子对人牙髓细胞增殖和分化的作用

目的 探讨碱笥成纤维细胞生长因子（ｂａｓｉｃ ｆｉｂｒｏｂｌａｓｔ ｇｒｏｗｔｈ ｆａｃｔｏｒ,ｂＦＧＦ）对培养人牙髓细胞增殖和分化的效应。方法 采用四唑盐法、^3Ｈ－ＴｄＲ掺入法、图象分析,检测重组人ｂＦＧＦ（ｈｂＦＧＦ）对细胞ＤＮＡ、胶原蛋白、纤维为连蛋白、碱笥磷酸酶、骨形成蛋白合成和凝集素表达的影响。结果 ｈｂＦＧＦ浓度在１～１０μｇ／Ｌ时显著促进细胞增殖,浓度为１～１００μｇ／Ｌ）,Ⅰ型胶原     

Adverse effects of arecoline and nicotine on human periodontal ligament fibroblasts in vitro

BACKGROUND, aims: The habit of betel nut chewing impinges on the daily lives of approximately 200 million people. Betel quid chewers have a higher prevalence of periodontal diseases than non-chewers. This study examined the pathobiological effects of arecoline, a major component of the betel nut alkaloids, on human periodontal ligament fibroblasts (PDLF) in vitro. METHOD: Cell viability, proliferation, protein synthesis, and cellular thiol levels were used to investigate the effects of human PDLF exposed to arecoline levels of 0 to 200 microg/ml. In addition, nicotine was added to test how it modulated the effects of arecoline. RESULTS: Arecoline significantly inhibited cell proliferation in a dose-dependent manner. At concentrations of 10 and 30 microg/ml, arecoline suppressed the growth of PDLF by 20% and 50% (p < 0.05), respectively. Arecoline also decreased protein synthesis in a dose-dependent manner during a 24-h culture period. A 100 microg/ ml concentration level of arecoline significantly inhibited protein synthesis to only 50% of that in the untreated control (p < 0.05). Moreover, arecoline significantly depleted intracellular thiols in a dose-dependent manner. At concentrations of 25 microg/ml and 100 microg/ml, arecoline depleted about 18% and 56% of thiols (p < 0.05), respectively. This suggests that arecoline itself might augment the destruction of periodontium associated with betel nut use. Furthermore, the addition of nicotine acted with a synergistic effect on the arecoline-induced cytotoxicity. At a concentration of 60 microg/ml, arecoline suppressed the growth of PDLF by about 33% and 5 mM nicotine enhanced the arecoline-induced cytotoxic response to cause about 66% cell death. CONCLUSION: During thiol depletion, arecoline may render human PDLF more vulnerable to reactive agents within cigarettes. Taken together, people who combine habits of betel nut chewing with cigarette smoking could be more susceptible to periodontium damage than betel nut chewing alone.     

白细胞介素-10对体外培养人牙囊细胞增殖和碱性磷酸酶活性的影响

目的:研究白细胞介素-10(Interleukin-10,IL-10)对体外培养人牙囊细胞增殖和牙囊细胞碱性磷酸酶活性的影响。方法:体外培养人牙囊细胞,取生长状态良好的第5代细胞,用四甲基偶氮唑蓝法(MTT法)检测IL-10对细胞增殖的作用;用碱性磷酸酶试剂盒检测IL-10及其抑制剂对细胞碱性磷酸酶活性的作用。结果:0、1、10、25、50、100ng/mLIL-10对人牙囊细胞的增殖影响无显著性差异(P>0.05);10ng/mLIL-10作用0、1、3、5、7d对人牙囊细胞的增殖影响无显著性差异(P>0.05)。10ng/mLIL-10作用3～7d降低人牙囊细胞的碱性磷酸酶活性(P<0.05),10ng/mLIL-10抑制剂作用5～7d增加人牙囊细胞的碱性磷酸酶活性(P<0.05)。结论:IL-10对人牙囊细胞的增殖无影响。IL-10降低人牙囊细胞的碱性磷酸酶活性,说明IL-10抑制人牙囊细胞向成骨方向分化。     

淫羊藿素对 SD 大鼠骨髓基质细胞增殖与成骨分化的影响

目的：研究淫羊藿素（ICT）对体外培养 SD 大鼠骨髓基质细胞（rBMSCs）增殖与成骨分化的影响。方法：体外分离培养 rBMSCs,传代至第4代作多向分化鉴定。分别以10－9、10－8、10－7、10－6、10－5 mol／L ICT 刺激 rBMSCs 后3、6、9 d,分别用 cck-8及碱性磷酸酶 ALP 试剂盒检测 rBMSCs 的增殖及 ALP 活性;10－9 mol／L ICT 处理 rBMSCs 后21 d 作茜素红（AR）染色以判断钙结节的形成。结果：原代培养的 rBMSCs 贴壁生长、呈梭形,能多向分化;ICT 明显抑制了 rBMSCs 的增殖;但增高其 ALP 活性、钙结节形成。结论：ICT 以剂量依赖方式抑制 rBMSCs 的增殖但促进其分化和矿化。     

Effects of enamel matrix derivative on proliferation and differentiation of primary osteoblasts

OBJECTIVE: The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on proliferation, protein synthesis, and mineralization in primary mouse osteoblasts. STUDY DESIGN: Osteoblasts were obtained from mouse calvaria by enzymatic digestion and grown in monolayer together with EMD (2-100 microg/ml). Metabolic activity and cell proliferation were determined by tetrazolium salt assay (MTT) and by 5-bromo-2'-deoxyuridine (BrdU) incorporation. For differentiation studies, a 3-dimensional organoid culture system was used. Osteoblastic differentiation was estimated by alkaline phosphatase (ALP) activity and calcium content. Collagen synthesis was assessed by [(3)H]-proline incorporation. Morphologic observations were made by electron microscopy. RESULTS: EMD treatments increased metabolic cell activity and BrdU incorporation. In the organoid cultures, ALP activity and calcium accumulation were enhanced by EMD treatment, but [(3)H]-proline incorporation was not. Morphologically, an increased deposition of mineralized nodules was found. CONCLUSIONS: EMD treatment enhanced cellular activities of primary osteoblasts and might support the regeneration of periodontal bony defects.     

Characterization of fibroblasts derived from human periodontal ligament and gingiva

Growth characteristics and macromolecular synthesis of fibroblasts derived from human periodontal ligament (PDLF) and gingiva (GF) have been compared in cell culture. Cells were isolated from explants and plated at 500,000 cells/100 mm culture dish (day 0) with daily changes of culture medium. DNA histograms were obtained by flow microfluorimetric analysis to confirm the growth state of the cell cultures. Human PDLF cultures became confluent at day 6 as determined by cell number and cell cycle analysis while GF were confluent by day 4. Initially, DNA content of logarithmically growing cells was significantly greater in GF cultures; however, when confluent, DNA content and cell number was greater in PDLF cultures. Total protein content in GF was slightly greater than PDLF until day 7 but this difference was not significant. Analysis of collagen and noncollagen protein synthesis revealed a greater trend in noncollagen protein synthesis in the GF cultures compared to PDLF cultures. Analysis of glycosaminoglycans in the culture medium of GF and PDLF revealed similar distributions of components. In the cellular fraction, GF had greater amounts of hyaluronic acid and heparin and lesser amounts of chondroitin sulfates A and C than PDLF cultures. The results indicate that the growth characteristics of PDLF and GF, although similar in many respects, do exhibit specific differences in proliferative rates and macromolecular synthesis. The differences observed in these parameters may be important during in vivo events, such as guided tissue regeneration, where significant functional differences are observed between gingival connective tissue and periodontal ligament connective tissue.