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磁性纳米纤维支架通过Wnt/MAPK/NF-κB信号通路诱导人牙髓细胞的牙发生和促血管生成。

Magnetic nanofiber scaffold-induced stimulation of odontogenesis and pro-angiogenesis of human dental pulp cells through Wnt/MAPK/NF-κB pathways.

作者信息

Yun Hyung-Mun, Kang Soo-Kyung, Singh Rajendra K, Lee Jung-Hwan, Lee Hae-Hyoung, Park Kyung-Ran, Yi Jin-Kyu, Lee Deok-Won, Kim Hae-Won, Kim Eun-Cheol

机构信息

Department of Oral and Maxillofacial Pathology, School of Dentistry and Research Center for Tooth & Periodontal Regeneration (MRC), Kyung Hee University, Seoul 130-701, Republic of Korea.

Department of Conservative Dentistry, Kyung Hee University, Seoul, Republic of Korea.

出版信息

Dent Mater. 2016 Nov;32(11):1301-1311. doi: 10.1016/j.dental.2016.06.016. Epub 2016 Sep 12.

Abstract

OBJECTIVE

Magnetic biomaterials have recently gained great attention due to their some intriguing cell and tissue responses. However, little attention has been given to the fields of dental tissue regeneration. In this sense, we aim to investigate the effects of magnetic nanofiber scaffolds on the human dental pulp cell (HDPC) behaviors and to elucidate the underlying signaling mechanisms in the events.

METHODS

Magnetic nanofiber scaffolds incorporating magnetic nanoparticles at varying contents were prepared into nanofibrous matrices to cultivate cells. Cell growth by MTS assay, odontoblastic differentiation by alkaline phosphatase (ALP) activity, mineralization, and the mRNA expression of differentiation-related genes of HDPCs, in vitro angiogenesis by migration and capillary tube formation in endothelial cells on the conditioned medium obtained from HDPSCs in the presence or absence of scaffolds. Western blot analysis and confocal immunofluorescene were used to asses signaling pathways.

RESULTS

The growth of HDPCs was significantly enhanced on the magnetic scaffolds with respect to the non-magnetic counterpart. The odontogenic differentiation of cells was significantly up-regulated by the culture with magnetic scaffolds. Furthermore, the magnetic scaffolds promoted the HDPC-induced angiogenesis of endothelial cells. The expression of signaling molecules, Wnt3a, phosphorylated GSK-3β and nuclear β-catenin, was substantially stimulated by the magnetic scaffolds; in parallel, the MAPK and NF-κB were highly activated when cultured on the magnetic nanofiber scaffolds.

SIGNIFICANCE

This study is the first to demonstrate that magnetic nanofiber scaffolds stimulate HDPCs in the events of growth, odontogenic differentiation, and pro-angiogenesis, and the findings imply the novel scaffolds can be potentially useful as dentin-pulp regenerative matrices.

摘要

目的

磁性生物材料因其一些有趣的细胞和组织反应最近受到了极大关注。然而,牙科组织再生领域却很少受到关注。从这个意义上讲,我们旨在研究磁性纳米纤维支架对人牙髓细胞(HDPC)行为的影响,并阐明这些事件背后的信号传导机制。

方法

将含有不同含量磁性纳米颗粒的磁性纳米纤维支架制备成纳米纤维基质来培养细胞。通过MTS法检测细胞生长,通过碱性磷酸酶(ALP)活性检测成牙本质细胞分化、矿化情况以及HDPCs分化相关基因的mRNA表达,通过在有或无支架存在的情况下从HDPSCs获得的条件培养基上内皮细胞的迁移和毛细管形成来检测体外血管生成。采用蛋白质免疫印迹分析和共聚焦免疫荧光法评估信号通路。

结果

与非磁性支架相比,磁性支架上HDPCs的生长显著增强。磁性支架培养显著上调了细胞的成牙本质分化。此外,磁性支架促进了HDPCs诱导的内皮细胞血管生成。磁性支架显著刺激了信号分子Wnt3a、磷酸化GSK - 3β和核β - 连环蛋白的表达;同时,在磁性纳米纤维支架上培养时,MAPK和NF - κB被高度激活。

意义

本研究首次证明磁性纳米纤维支架在HDPCs的生长、成牙本质分化和促血管生成过程中具有刺激作用,研究结果表明这种新型支架有望作为牙本质 - 牙髓再生基质。

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