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不同三维培养模型对受压缩应变的人牙周膜成纤维细胞表达谱的评估

An Evaluation of Different 3D Cultivation Models on Expression Profiles of Human Periodontal Ligament Fibroblasts with Compressive Strain.

作者信息

Schröder Agnes, Schöniger Ricarda, Oeldemann Juliane, Spanier Gerrit, Proff Peter, Jantsch Jonathan, Kirschneck Christian, Ullrich Niklas

机构信息

Department of Orthodontics, University Medical Centre of Regensburg, 93053 Regensburg, Germany.

Department of Oral and Maxillofacial Surgery, University Medical Centre of Regensburg, 93053 Regensburg, Germany.

出版信息

Int J Mol Sci. 2022 Feb 12;23(4):2029. doi: 10.3390/ijms23042029.

Abstract

The effects of compressive strain during orthodontic treatment on gene expression profiles of periodontal ligament fibroblasts (PDLFs) have mostly been studied in 2D cell culture. However, cells behave differently in many aspects in 3D culture. Therefore, the effect of pressure application on PDLFs in different 3D structures was investigated. PDLFs were either conventionally seeded or embedded into different 3D structures (spheroids, Mebiol gel, 3D scaffolds) and exposed to compressive force or incubated without pressure. For one 3D scaffold (POR), we also tested the effect of different compressive forces and application times. Expression of an angiogenic gene (), a gene involved in extracellular matrix synthesis (), inflammatory genes (, ), and genes involved in bone remodelling (, ) were investigated by RT-qPCR. Depending on the used 3D cell culture model, we detected different effects of compressive strain on expression profiles of PDLFs. was downregulated in all investigated 3D culture models. Angiogenetic and proinflammatory genes were regulated differentially between models. In 3D scaffolds, regulation of bone-remodelling genes upon compressive force was contrary to that observed in 3D gels. 3D cell culture models provide better approximations to in vivo physiology, compared with conventional 2D models. However, it is crucial which 3D structures are used, as these showed diverse effects on the expression profiles of PDLFs during mechanical strain.

摘要

正畸治疗期间的压缩应变对牙周膜成纤维细胞(PDLFs)基因表达谱的影响大多是在二维细胞培养中进行研究的。然而,细胞在三维培养中的许多方面表现不同。因此,研究了对不同三维结构中的PDLFs施加压力的影响。将PDLFs常规接种或嵌入不同的三维结构(球体、Mebiol凝胶、三维支架)中,使其受到压缩力作用或在无压力条件下培养。对于一种三维支架(POR),我们还测试了不同压缩力和施加时间的影响。通过逆转录定量聚合酶链反应(RT-qPCR)研究血管生成基因()、参与细胞外基质合成的基因()、炎症基因(,)以及参与骨重塑的基因(,)的表达。根据所使用的三维细胞培养模型,我们检测到压缩应变对PDLFs表达谱有不同影响。在所有研究的三维培养模型中均下调。血管生成基因和促炎基因在不同模型之间的调控存在差异。在三维支架中,压缩力对骨重塑基因的调控与在三维凝胶中观察到的情况相反。与传统的二维模型相比,三维细胞培养模型能更好地模拟体内生理情况。然而,使用哪种三维结构至关重要,因为这些结构在机械应变期间对PDLFs的表达谱显示出不同的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e077/8876762/e08d9358f5b6/ijms-23-02029-g001.jpg

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