自酸蚀牙本质黏结系统对牙髓组织的生物相容性研究

目的:通过动物实验比较两种自酸蚀黏结剂XenoⅢ(XO)、Adper Prompt(AP)和一种全酸蚀黏结剂single bond 2(SB)3者之间的生物相容性.方法:选取3只成年雄性比格犬,在其牙齿的唇颊面颈部制备直径为2 mm的单面洞,实验组分别涂布3种不同的黏结剂XO、AP、SB,F2000复合体充填,阴性对照组用Dycal盖髓后,玻璃离子充填.分别于实验的第7天,30天与90天处死动物取实验牙,固定、脱钙后病理切片检查,评价3种材料对牙髓的影响.结果:3种黏结剂涂布7 d的病理结果显示牙髓反应相似,差异无统计学意义(P>0.05).30 d与90 d大部分标本的牙髓组织结构已恢复正常,但SB组较AP组、XO组出现更多的反应性牙本质,差异有统计学意义(P<0.05),表明SB组具有更强的牙髓刺激性.而AP组、XO组之间无统计学差异(P>0.05).结论:自酸蚀牙本质黏结剂XO、AP的生物相容性好于全酸蚀牙本质黏结剂SB.     

牙本质粘结剂对人牙髓细胞毒性的体外研究

目的：评价全酸蚀牙本质粘结剂和自酸蚀牙本质粘结剂对人牙髓细胞的毒性作用.方法：用人牙髓细胞为实验细胞,采用MTT比色分析法,对4 种牙本质粘结剂（Prime ＆ Bond NT,Single Bond,Xeno Ⅲ,iBond）进行体外细胞毒性研究.结果：不同浓度的粘结剂稀释液均可使人牙髓细胞的形态有所改变.4 种牙本质粘结剂的细胞毒性有显著性差异,且作用时间和浓度的改变对其细胞毒性有影响.全酸蚀粘结剂比自酸蚀粘结剂的细胞毒性强.结论：4 种牙本质粘结剂在体外对人牙髓细胞均有一定程度的细胞毒性,其中Single Bond的毒性较强,临床使用粘结剂时应合理选择粘结剂和掌握固化时间.     

4种牙本质粘结剂对牙髓成纤维细胞毒性作用的比较

目的:对比研究4种第七代牙本质粘结剂对人牙髓成纤维细胞的毒性作用,初步探讨其生物安全性。方法:组织块培养法原代培养人牙髓成纤维细胞,免疫组织化学染色SP法鉴定细胞来源,采用四甲基偶氮唑盐比色法和浸提液法,双盲观察4种新一代牙本质粘结剂(G-Bond、i-Bond、xeno V、Clearfil S3Bond)的不同体积分数浸提液(12.5%、25%、50%、100%)作用不同时间(24、48、72 h)对人牙髓成纤维细胞的毒性作用。结果:4种牙本质粘结剂不同体积分数浸提液的作用下,人牙髓成纤维细胞形态均发生不同程度的变化。24、48 h时,xeno V细胞毒性影响较小,i-Bond细胞毒性影响较大,G-Bond与Clearfil S3 Bond细胞毒性无明显差异;72 h时,4种牙本质粘结剂对细胞毒性影响无明显差异。结论:4种牙本质粘结剂毒性趋于0～2级,随着作用时间延长,细胞毒性无明显差异。     

小型猪牙髓对牙本质自酸蚀和全酸蚀粘接系统的组织病理学反应

目的 观察小型猪牙髓对牙本质自酸蚀和全酸蚀粘接系统的组织病理学反应,研究不同牙本质粘接剂对牙髓组织的刺激.方法 在小型猪磨牙及前磨牙上备洞,分别涂布4种牙本质粘接剂,玻璃离子充填,术后1d、3d、7d取牙髓组织进行组织病理学观察.结果 术后1d各组均表现为成牙本质细胞排列紊乱、毛细血管扩张;术后3d、7d自酸蚀组牙髓逐渐恢复正常,全酸蚀组仍可见炎症性病理变化.结论 自酸蚀粘接系统对牙髓组织刺激轻微,全酸蚀粘接系统对牙髓组织的刺激较重.     

硬化牙本质粘接界面的激光扫描共聚焦显微镜观察

目的使用激光扫描共聚焦显微镜（CLSM）观察牙颈部硬化牙本质在全酸蚀和自酸蚀粘接系统形成粘接界面的超微结构。方法选用12颗因牙周病拔除的具有典型楔状缺损的上颌前磨牙为实验组;12颗新鲜拔除的无龋人上颌前磨牙为对照组,制备人造楔状缺损。使用Single Bond （SB,全酸蚀单瓶系统）、Clearfil SE Bond（CB,自酸蚀底胶系统）、Xeno Ⅲ（XB,自酸蚀一步粘接系统）粘接系统处理牙面,以罗丹明B异硫氰酸盐为荧光素,使用CLSM观察粘接界面的混合层与树脂突的微观结构。结果双因素方差分析表明粘接剂种类、牙本质类型对粘接界面树脂突长度、混合层厚度有显著影响（P<0.05）。无论是正常牙本质还是硬化牙本质,全酸蚀粘接剂（SB）产生的树脂突长度、混合层厚度均大于自酸蚀粘接剂（CB、XB）,并且其差异具有统计学意义（P<0.05）;而CB和XB间树脂突长度、混合层厚度相差不多,二者间无统计学意义。结论硬化牙本质相对于正常牙本质形成的混合层较薄或者没有,树脂突短、少。全酸蚀粘接系统与自酸蚀粘接系统作用同类型的牙本质上,自酸蚀比全酸蚀形成的混合层薄,树脂突短。     

五种牙本质粘接剂体外细胞毒性研究

目的 对5种牙本质粘接剂的体外细胞毒性进行对比研究,为临床应用提供参考依据.方法 采用体外细胞培养技术和四甲基偶氮唑盐(MTT)比色法,观察第7代牙本质粘接剂Clearfil tri-S Bond和4种第6代牙本质粘接剂Clearfil SE Bond、Adper Prompt、ONE UP BOND F、XenoⅢ粘接剂浸提液对L929小鼠成纤维细胞生长的影响,测定各组吸光度值(OD值),计算L929小鼠成纤维细胞的相对增殖率,用5级毒性分类法评级,并进行统计分析.结果 各组细胞生长均较好,Clearfil tri-S Bond组和Clearfil SE Bond组的OD值显著高于其它各组的OD值.结论 5种牙本质粘接剂的体外细胞毒性均较弱,其中第7代牙本质粘接剂Clearfil tri-S Bond和第6代牙本质粘接剂Clearfil SE Bond的细胞毒性最低.     

5种牙本质粘接剂的细胞毒性与单体双键转化率的关系

目的 比较5种牙本质粘接剂的细胞毒性,探讨影响其差异的相关因素及其与单体双键转化率的关系。方法 选择5种临床常用的商品牙本质粘接剂OptiBond FL（FL）、Single Bond 2（SB）、Prime&Bond NT（PB）、Clearfil SE Bond（SE）和Adper Easy One（EO）,采用MTT法检测材料的细胞毒性,运用荧光染色法观察细胞形态。采用傅里叶红外光谱测量5种牙本质粘接剂的单体双键转化率。结果 牙本质粘接剂浸提1 d时,MTT检测得到的OD值从大到小依次为SE > C ≈ FL ≈ EO ≈ PB > SB,其中SB组的吸光度值明显低于对照组,且差异具有统计学意义（P<0.05）;牙本质粘接剂浸提3 d和5 d时,SB组和EO组的吸光值远低于对照组,呈现明显的细胞毒性。牙本质粘接剂单体双键转化率值从大到小依次为PB > EO > FL > SB > SE,相邻两组间差异有统计学意义（P<0.05）。但各组分本质粘接剂的细胞毒性和单体双键转化率的相关性无统计学意义（P>0.05）。结论 不同牙本质粘接剂的细胞毒性不同,与其单体双键转化率不存在线性关系,两者均受到多种因素的影响。     

新型牙本质黏结剂体外细胞毒性研究

目的：评价一种新型牙本质黏结剂的细胞毒性，并与商品化的两种黏结剂细胞毒性进行比较。方法：采用琼脂覆盖法和四甲基偶氮唑盐(MTT)比色法，评价新型牙本质黏结剂优邦(UB)和商品化黏结剂Adper Prompt自酸蚀黏结剂(AP)与Prime＆Bond NT(PB)在人牙本质片上使用后，对L929小鼠成纤维细胞的毒性作用。结果：MTT实验中三种牙本质黏结剂细胞毒性均为1级；琼脂覆盖实验中，三种材料两种厚度牙本质片均出现脱色区和部分细胞溶解，细胞毒性评价均为1级。结论：新型牙本质黏结剂具有轻做的细胞毒性，UB对牙髓组织的影响还有待进一步研究。     

自酸蚀粘接剂后牙复合树脂充填术后敏感性的临床研究

目的　评价自酸蚀粘接剂后牙复合树脂充填术后的敏感性。方法　 70例中深龋磨牙分为两组 ,实验组 36例采用可乐丽菲露自酸蚀粘接剂 ,对照组 34例使用登士伯磷酸全酸蚀牙本质粘接剂 ,充填材料为各自厂商配套的复合树脂。临床观察时间为术后 1周、6个月及 1年。结果　实验组在术后 1周复查时均无术后敏感 ,A级率 10 0 %。对照组在术后 1周复查时有 1例一过性敏感 (洞深 6mm) ,并且持续了 5个月后消失 ,A级率 97% ,两组之间无显著性差异 (P >0 .0 5 )。实验组和对照组在 6个月及 1年的复查时均无术后敏感 ,A级率 10 0 % ,牙髓电活力测试正常。结论　自酸蚀粘接剂和磷酸全酸蚀牙本质粘接剂复合树脂充填术后的敏感性无显著差异 ,但使用自酸蚀粘接剂的效果更好     

Adper Prompt自酸蚀粘结剂对人牙龈成纤维细胞的毒性分析

目的 分析Adper Prompt(AP)自酸蚀粘结剂对人牙龈成纤维细胞的毒性作用。方法 用MTT法测定人牙龈成纤维细胞在Adper Prompt自酸蚀粘结剂不同浓度稀释液中培养4 d、7 d的吸光度(A)值,计算细胞相对增殖率(RGR),并结合流式细胞仪测定不同浓度稀释液中培养4 d后的人牙龈成纤维细胞不同增殖周期时相的DNA百分含量,计算细胞增殖指数百分比,来评估AP的细胞毒性。结果 随作用时间和浓度的改变,Adper Prompt对细胞的毒性发生变化。时间越长,浓度越大,细胞相对增殖率(RGR)越小,细胞增殖指数百分比越小。结论 Adper Prompt自酸蚀粘结剂在体外对人牙龈成纤维细胞有一定程度的细胞毒性,在操作过程中应采取措施保护牙龈组织。     

The induction of oxidative stress, cytotoxicity, and genotoxicity by dental adhesives.

OBJECTIVES: Polymerized dental resin materials release residual monomers that may interact with pulp tissues. We hypothesized that dental adhesives might cause cytotoxicity in pulp cells via the generation of reactive oxygen species (ROS), which may also contribute to genotoxic effects in vitro. METHODS: For cytotoxicity testing, transformed human pulp-derived cells were exposed to extracts of primers and bonding agents of Clearfil SE bond, Clearfil Protect bond, AdheSE, Prompt L-Pop, and Excite for 24h. The cytotoxicity of the same materials was also analyzed in a dentin barrier test device using three-dimensional pulp cell cultures. The generation of ROS in monolayer cultures was measured after a 1h exposure period by flow cytometry (FACS), and genotoxicity as indicated by the formation of micronuclei was determined in V79 cells after a 24h exposure period. RESULTS: The dentin primers and bonding agents decrease cell survival in a dose-related manner. Cytotoxicity of bonding agents based on concentrations which caused 50% cell death (EC50) were ranked as follows: Excite (0.16 mg/ml)>AdheSE bond (0.30 mg/ml)>Clearfil Protect bond (0.35 mg/ml)>Clearfil SE bond (0.37 mg/ml), and Prompt L-Pop bond (0.68 mg/ml). Dentin primers were about 10-fold less effective. In contrast, no cytotoxic effects of the dental adhesives were observed in a dentin barrier test device. Yet, all dental adhesives increased the amounts of ROS about fivefold in pulp cells in a dose-related manner, and, again, the bonding agents were more efficient than the dentin primers. Finally, the number of micronuclei was increased about sixfold by extracts of the AdheSE primer. SIGNIFICANCE: Our results suggest that the cytotoxic potencies demonstrated by these materials might be of clinical relevance, since all dental adhesives disturbed the cellular redox state of pulp cells in monolayer cultures. As a result, the concentrations of biologically active ingredients of some of the agents may be high enough to modify pulp cell metabolism when the materials are used in deep cavities or directly contact pulp tissue.     

Polymerized bonding agents and the differentiation in vitro of human pulp cells into odontoblast-like cells.

OBJECTIVES: Odontoblasts are highly differentiated post-mitotic cells, which under pathological conditions such as carious lesions and dental injuries may degenerate and be replaced by other pulp cells. We have recently shown that this physiological event can be reproduced in an in vitro assay system, but is highly modified by the presence of unpolymerized resinous monomers. Our hypothesis was that the photopolymerization of the monomers in the bonding agents might abolish these negative effects. The purpose of this study was to evaluate the effects of polymerized dentin bonding agents, through dentin slices, on odontoblast differentiation in vitro. METHODS: Pulp cells were obtained from human third molars. They were used to study the effects of four dentin bonding agents through 0.7 mm dentin slices which served as a barrier between the bonding agents and the culture medium. The media containing the bonding agents' extracts were added at non-toxic concentrations onto the cultured cells. Immunohistochemistry was performed to study the differentiation of pulp fibroblasts into odontoblasts under these conditions by evaluating the expression of several odontoblast specific genes. RESULTS: Pulp fibroblasts cultivated under these conditions synthesized type I collagen, osteonectin, dentin sialoprotein and nestin at the same level as in control cultures. Moreover, pulp cells synthesized a mineralized nodular extracellular matrix. Expression of these proteins was higher in the cells contributing to the nodule formation. In addition, except nestin, all these proteins were expressed in the mineral nodules. SIGNIFICANCE: This work shows the lack of effects of photopolymerized bonding agents, through dentin slices, on cytodifferentiation of secondary odontoblasts.     

应用牙器官培养模型评价粘结剂的生物相容性

目的:应用大鼠全牙器官培养模型对两种牙科粘结剂进行生物相容性评价。方法:在新鲜拔除的4周龄SD大鼠切牙唇面制洞,随机分成3组:对照组(不涂粘结剂);AP组(涂Adper Prompt粘结剂)和PB组(涂Prime&Bond NT粘结剂),3组洞壁做相应处理后光照10 s后填入树脂,分别于术后即刻和体外全牙培养1周两个时间点,40 g/L多聚甲醛(PBS)固定,100 g/L甲酸脱矿,常规切片,HE染色进行组织学观察,并采集图像进行细胞计数及统计学分析。结果:术后即刻与对照组相比,PB组制洞部位下方的牙髓细胞发生聚集,AP组则表现为成牙本质细胞的细胞数减少,两种细胞均发生形态改变。术后培养1周时与对照组相比,AP组制洞部位下方的牙髓内牙髓细胞坏死,而PB组则表现为成牙本质细胞的细胞数目减少,两种细胞均发生形态改变。结论:两种牙科粘结剂均对牙髓组织有毒性作用,早期AP的毒性大于PB。     

人前磨牙牙髓牙本质复合体层粘连蛋白受体的免疫组织化学研究

目的研究层粘连蛋白受体在牙髓牙本质复合体内的表达,探讨其分布与功能的关系.方法收集新鲜健康人前磨牙60个,固定,脱钙,石蜡包埋,层粘连蛋白受体SABC免疫组织化学染色.结果层粘连蛋白受体免疫反应物(LNR-IR)普遍存在于牙髓牙本质复合体的神经纤维膜、血管内皮细胞、成牙本质细胞及成纤维细胞上,以牙髓的神经纤维膜、血管内皮细胞最为丰富.结论层粘连蛋白受体一般见于牙髓牙本质复合体的各种细胞膜上,它与层粘连蛋白一起在支持、固定牙髓牙本质复合体各结构的正常位置、形态和功能的发挥上可能起了重要作用.     

Cytotoxicity of three dentin bonding agents on human dental pulp cells

OBJECTIVES: Dentin bonding agents (DBA) have been widely used in operative restoration to prevent leakage and promote bonding strength in the resin-dentin interface. However, DBA may exert potentially harmful effects to the dental pulp. In the present study, differential cytotoxicity of three DBA (Syntac Sprint, SP; Prime and Bond 2.1, PB; and Single Bond, SB) on the pulp cells was tested. METHODS: Three DBA were diluted with the culture medium by a ratio of 1:1000, 1:2000 and 1:4000 (v/v). Pulp cells (5 x 10(4) cells/well) were then exposed to culture medium containing different diluents of three DBA for 12, 24h and 3 days. Cytotoxicity was measured with a modified 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. RESULTS: A 12h experiment revealed that SP was the strongest cytotoxic agent, followed sequentially by SB and PB. Exposure of pulp cells to 1:4000 (v/v) dilution of SP, PB and SB for 24h reduced the cell number by 23, 6 and 45%, respectively. A 1:2000 (v/v) of DBA diluents reduced the cell number for 32, 13 and 65%, respectively, by SP, PB and SB. Dilution of DBA by 1000-fold of culture medium further enhanced the cytotoxic response. Cell number decreased by 89, 65 and 72%, respectively, by SP, PB and SB. Similar to the 12h-cytotoxicity data, SB is more toxic at high dilution condition, whereas, at low dilution condition, SP is the most toxic agent to pulp cells. Similar cytotoxicity was noted when pulp cells were exposed to DBA for 3 days. Toxicity of DBA was concomitant with marked retraction and rounding of dental pulp cells. SIGNIFICANCE: These results indicate that DBA exerts potential harmful effects to the pulp. Differential toxic effects of DBA on the pulp cells should be considered during selection of a suitable DBA for operative restoration.     

牙本质支架体外诱导兔骨髓干细胞分化

目的：探讨将人牙本质作为组织工程支架的可行性,采用不同的方式处理牙本质片,观察牙本质片处理后诱导兔骨髓基质干细胞贴壁附着生长的规律及成骨分化潜能。方法：将兔的骨髓基质干细胞分别接种于经机械去除前期牙本质加EDTA-柠檬酸脱矿处理（A组）,标准根管预备-EDTA处理（B组）的牙本质片上,体外培养1周,扫描电镜观察细胞与牙本质片的附着情况,碱性磷酸酶、茜素红及Vonkossa染色观察兔骨髓基质干细胞成骨分化。结果：兔骨髓基质细胞附着在2种方式处理的牙本质片上生长良好,贴附紧密。B组牙本质片上细胞碱性磷酸酶表达阳性率高达80％,茜素红、Vonkossa染色观察到钙结节数量多于A组。结论：经标准根管预备一EDTA处理的人牙本质能有效促进骨髓基质干细胞增殖及成骨分化,可能成为一种理想的成骨生物支架,并为牙体牙髓疾病的治疗提供了新的思路。广西科学研究与技术开发计划项目（桂科攻1012400lA一42）     

Cytotoxicity of a dentine bonding agent on four different cell-lines.

The aim of this study was to compare the cytotoxicity of a recently available dentine bonding agent on four different cell-lines (three human dental pulp fibroblast cell-lines and one mouse 3T3 fibroblast cell-line). METHODOLOGY: Three human dental pulp cell-lines from 3 different donors and one established 3T3 mouse cell-line were grown and sub-cultured. Cell viability following exposure to Scothbond was then compared to a similar number of controls using the MTT assay. RESULTS: Scotchbond 1 was cytotoxic to all four cell-lines. 3T3 cells showed a survival rate of about 60% as compared to two of the human dental pulp cells which showed a significantly lower survival rate (p<0.05, Kruskal-Wallis Multiple-Comparison Test). CONCLUSION: These findings indicated that is cytotoxic to both human pulp and 3T3 cell-lines. In general, the human pulp cell-lines showed higher sensitivity than the 3T3 cell-lines. CLINICAL SIGNIFICANCE: Scotchbond 1 cannot be recommended for direct pulp capping techniques and care should be taken when using this dentine bonding agent in cavities where the remaining dentine layer is minimal.     

Cytotoxicity of dentine-bonding agents on human pulp cells in vitro

AIM: To investigate the cytotoxicity of five different dentine-bonding agents on human pulp cells in vitro. METHODOLOGY: Set specimens from Clearfil SE Bond (CB), Heliobond (HB), Prime & Bond NT (PB), Single Bond (SB), and Syntac Single Component (SC) were eluted with culture medium for 2 and 5 days. Cytotoxicity was judged using tetrazolium bromide reduction assay on human primary pulp cells. RESULTS: Elutes from five dentine-bonding agents were cytotoxic to primary human pulp cells (P < 0.05). CB was the least toxic sealer amongst the chemicals tested. The cytotoxic response decreased in an order of SB > PB > SC > HB > CB. CONCLUSIONS: The influence of the cytotoxicity depended on the materials tested. Dentine-bonding agents have significant potential for pulpal toxicity.     

牙本质黏结剂对牙髓直接影响的动物实验

目的：观察两种牙本质黏结剂对活体牙髓组织的影响，并与氢氧化钙直接盖髓结果进行对比。方法：选用54个犬牙，机械穿髓后，无菌蒸馏水冲洗，控制穿髓孔处的出血和渗出。使用优邦(UB)和Adper Prompt Self-etching Adhesive(AP)两种牙本质黏结剂盖髓，Dycal作对照。于7、30、90d拔除实验牙，脱钙切片，HE染色、Mallory三色法染色和Gram快速细菌染色检查?结果：所有处理组在短期都出现了轻到中度的炎症反应，在长期的观察中，UB和AP两组出现持续的慢性炎症反应，均未发现牙本质桥肜成。Dycal组出现更多的修复性牙本质，并有牙本质桥形成(P＜0．05)结论：这两种牙本质黏结剂会引起牙髓的慢性炎症反应，在穿髓孔不能形成完整的牙本质桥，临床上这两种黏结剂不能直接与牙髓接触。