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采用三维细胞培养模型评价 Biodentine 的细胞相容性。

Cytocompatibility of Biodentine using a three-dimensional cell culture model.

机构信息

Department of Endodontics, School of Dentistry, Grande Rio University (UNIGRANRIO), Rio de Janeiro, Brazil.

Department of Endodontics, Piracicaba School of Dentistry, Campinas State University (UNICAMP), Piracicaba, Brazil.

出版信息

Int Endod J. 2016 Jun;49(6):574-80. doi: 10.1111/iej.12485. Epub 2015 Jul 8.

Abstract

AIM

To evaluate the cytotoxic effects of Biodentine, using a three-dimensional (3D) cell culture associated with an in situ root-end filling experimental model. White mineral trioxide aggregate (MTA) and zinc oxide cement were used as reference for comparison. IL-1α and TNF-α cytokine production were also evaluated.

METHODOLOGY

The root canals of 24 human maxillary incisor teeth were prepared using a single-file reciprocating technique. After root filling, a 3-mm root-end resection was performed and 3 mm of gutta-percha was removed from the canal. The teeth were randomly distributed to receive one of the following root-end filling materials: Biodentine, white MTA or zinc oxide cement (positive control group). In the negative control group, the root canal was not retro-filled. The cytocompatibility of the materials was evaluated using the methyl-thiazol-diphenyl-tetrazolium (MTT) assay in an in situ root-end filling experimental model. Balb/c 3T3 fibroblasts, cultured in rat tail collagen type I 3D scaffold, were exposed to the root apex for 24 h, and cell viability was measured by means of reduction MTT salt. IL-1α and TNF-α production were analysed using enzyme-linked immunosorbent assay. One-way analysis of variance was performed and, when the F-ratios were significant, data were compared by Duncan's multiple-range test. The alpha-type error was set at 0.05.

RESULTS

Biodentine and MTA groups had similar cell activity to the negative control group (P > 0.05), indicating low cytotoxicity for both materials. The stronger cytotoxicity effect was identified on the zinc oxide cement (P < 0.05). Zinc oxide cement caused a significant up-regulation in IL-1α and TNF-α (P < 0.05). No significant differences amongst MTA, Biodentine and the negative control group were observed for TNF-α (P > 0.05); however, both MTA and Biodentine were associated with overproduction of IL-1α when compared to the control group (P < 0.05).

CONCLUSIONS

Biodentine and MTA had similar cytocompatibility in a 3D cell culture model associated with an in situ root-end filling model. The methodology could be used as an alternative to assess the cytocompatibility of endodontic cements because it is more closely related to the in vivo situation.

摘要

目的

使用与原位根管填充实验模型相关的三维(3D)细胞培养来评估 Biodentine 的细胞毒性作用。将白色矿物三氧化物聚合体(MTA)和氧化锌水泥用作比较的参考。还评估了白细胞介素-1α(IL-1α)和肿瘤坏死因子-α(TNF-α)细胞因子的产生。

方法

使用单文件往复技术对 24 个人上颌切牙的根管进行预备。根管填充后,进行 3mm 的根尖切除,并从根管中去除 3mm 的牙胶。牙齿随机分配接受以下根管填充材料之一:Biodentine、白色 MTA 或氧化锌水泥(阳性对照组)。在阴性对照组中,根管未进行逆行填充。在原位根管填充实验模型中,使用噻唑蓝(MTT)比色法评估材料的细胞相容性。在大鼠尾胶原 I 型 3D 支架中培养的 Balb/c 3T3 成纤维细胞,暴露于根尖 24 小时,通过还原 MTT 盐测量细胞活力。使用酶联免疫吸附试验分析白细胞介素-1α(IL-1α)和肿瘤坏死因子-α(TNF-α)的产生。进行单因素方差分析,当 F 比值显著时,通过 Duncan 多重范围检验比较数据。α 型错误设置为 0.05。

结果

Biodentine 和 MTA 组的细胞活性与阴性对照组相似(P>0.05),表明两种材料的细胞毒性较低。氧化锌水泥组的细胞毒性作用更强(P<0.05)。氧化锌水泥导致白细胞介素-1α和肿瘤坏死因子-α(TNF-α)显著上调(P<0.05)。MTA、Biodentine 与阴性对照组之间 TNF-α 无显著差异(P>0.05);然而,与对照组相比,MTA 和 Biodentine 均与白细胞介素-1α(IL-1α)的过度产生相关(P<0.05)。

结论

在与原位根管填充模型相关的 3D 细胞培养模型中,Biodentine 和 MTA 具有相似的细胞相容性。该方法可作为评估根管水泥细胞相容性的替代方法,因为它更接近体内情况。

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