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通过三维培养的牙周膜细胞在细胞水平上研究最佳正畸力。

Investigation of optimal orthodontic force at the cellular level through three-dimensionally cultured periodontal ligament cells.

作者信息

Li Meile, Yi Jianru, Yang Yan, Zheng Wei, Li Yu, Zhao Zhihe

机构信息

Departments of Orthodontics and.

Oral and Maxillofacial Surgery, State Key Laboratory of Oral Diseases, West China School and Hospital of Stomatology, Sichuan University, Chengdu, China.

出版信息

Eur J Orthod. 2016 Aug;38(4):366-72. doi: 10.1093/ejo/cjv052. Epub 2015 Jul 23.

DOI:10.1093/ejo/cjv052
PMID:26209366
Abstract

OBJECTIVES

Optimal orthodontic force (OOF), important as it is, has generally been addressed at the level of tissue response. This study, for the first time, aimed to investigate its underlying mechanisms at the cellular level.

METHODS

Human periodontal ligament tissue cells (PDLCs) were three-dimensionally (3D) cultured in a thin sheet of poly-lactic-co-glycolic acid (PLGA) scaffold. The 3D cultured PDLCs were treated with static compressive force of 0, 5, 15, and 25g/cm(2) for 6, 24, and 72 hours, respectively. After that, methylthiazolyl tetracolium assay was done to evaluate the cell proliferation. The target gene expression in PDLCs was investigated through real-time PCR analysis. The conditioned media was collected for enzyme-linked immunosorbent assay (ELISA) assay, and also used for the coculture of osteoblasts and osteoclast precursors. Tartrate-resistant acid phosphatase (TRAP) staining was employed to examine osteoclasts.

RESULTS

Compressive force inhibited proliferation of PDLCs in a magnitude-dependent manner. Heavier force upregulated expression of the osteoclastogenesis inducers, including RANKL, COX-2, PTHrP, and IL-11, more rapidly; however in the long run, no significant difference was found among different force magnitudes, either in the expression of osteoclastogenesis inducers by PDLCs, or in the osteoclast formation detected by TRAP staining.

LIMITATIONS

The results regarding specific force magnitude as OOF should be confined to the present specific model, but not be extrapolated, without caution, to different in vitro models, nor even to in vivo studies or clinical application.

CONCLUSIONS

Compared with heavier force, lighter force has similar pro-osteoclastogenic whilst less anti-proliferative effects on PDLCs, which provides a novel interpretation for OOF.

摘要

目的

最佳正畸力(OOF)虽很重要,但通常是在组织反应层面进行探讨。本研究首次旨在从细胞水平研究其潜在机制。

方法

将人牙周膜组织细胞(PDLCs)在聚乳酸-乙醇酸共聚物(PLGA)支架薄片中进行三维(3D)培养。对3D培养的PDLCs分别施加0、5、15和25g/cm²的静态压缩力,持续6、24和72小时。之后,进行甲基噻唑基四唑测定以评估细胞增殖。通过实时PCR分析研究PDLCs中的靶基因表达。收集条件培养基用于酶联免疫吸附测定(ELISA),并用于成骨细胞和破骨细胞前体的共培养。采用抗酒石酸酸性磷酸酶(TRAP)染色检测破骨细胞。

结果

压缩力以剂量依赖方式抑制PDLCs增殖。更大的力更快速地上调破骨细胞生成诱导因子的表达,包括核因子κB受体活化因子配体(RANKL)、环氧化酶-2(COX-2)、甲状旁腺激素相关蛋白(PTHrP)和白细胞介素-11(IL-11);然而从长远来看,在不同力的大小之间,无论是PDLCs中破骨细胞生成诱导因子的表达,还是通过TRAP染色检测到的破骨细胞形成,均未发现显著差异。

局限性

关于作为最佳正畸力的特定力大小的结果应局限于当前特定模型,不应在未经谨慎考虑的情况下外推至不同的体外模型,更不能外推至体内研究或临床应用。

结论

与更大的力相比,较小的力对PDLCs具有相似的促进破骨细胞生成作用,但抗增殖作用较小,这为最佳正畸力提供了一种新的解释。

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