采用磁拉力加载的工程三维牙周韧带模型

Journal of dental research Pub Date : 2024-09-01 Epub Date: 2024-08-26 DOI:10.1177/00220345241264792
P Mulimani, N A Mazzawi, A J Goldstein, A M Obenaus, S M Baggett, D Truong, T E Popowics, N J Sniadecki
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引用次数: 0

摘要

体外模型是解构牙周韧带(PDL)生物复杂性的宝贵工具。模型系统可近似再现原生组织中细胞-细胞和细胞-基质相互作用的三维(3D)结构,从而提供与生理相关的见解。然而,目前还缺乏包含机械负荷的 PDL 三维模型。因此,我们开发了一种模型,通过在一对细长的硅胶柱之间悬浮的胶原凝胶中铸造 PDL 细胞来制作牙周组织构建体 (PTC),从而实现磁性拉伸加载。具体来说,其中一个硅胶柱是刚性的,另一个硅胶柱是柔性的,其顶端嵌入了磁铁,这样 PTC 就能承受外部磁铁的拉伸负荷。此外,柔性支柱的偏转可用于测量 PTC 中 PDL 细胞的收缩力。在拉伸加载之前,对 PTC 中的胶原纤维进行二次谐波生成分析,结果显示 PDL 细胞的加入导致了胶原重塑。通过拉伸加载对 PTC 进行的生物力学测试显示,4 小时后出现弹性反应,1 天后出现永久变形,1 周后出现蠕变伸长。随后,PDL 细胞在拉伸负荷下的收缩力大大低于 PTC。免疫荧光分析表明,拉伸负荷导致 PDL 细胞数量增加,表达更高水平的 F-肌动蛋白和 α 平滑肌肌动蛋白,并与拉伸轴对齐。二次谐波生成分析表明,随着时间的推移,PTCs 中的胶原纤维会随着拉伸负荷而逐渐重塑。基因表达分析也证实了张力介导的 F-actin/Rho 通路和成骨基因的上调。我们的模型在三维环境中展示了导致细胞介导的 PDL 组织重塑的机械生物学行为,具有新颖性。因此,它可以成为开发牙周炎、牙周再生和正畸疗法的重要工具。
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Engineered 3D Periodontal Ligament Model with Magnetic Tensile Loading.

In vitro models are invaluable tools for deconstructing the biological complexity of the periodontal ligament (PDL). Model systems that closely reproduce the 3-dimensional (3D) configuration of cell-cell and cell-matrix interactions in native tissue can deliver physiologically relevant insights. However, 3D models of the PDL that incorporate mechanical loading are currently lacking. Hence, we developed a model where periodontal tissue constructs (PTCs) are made by casting PDL cells in a collagen gel suspended between a pair of slender, silicone posts for magnetic tensile loading. Specifically, one of the posts was rigid and the other was flexible with a magnet embedded in its tip so that PTCs could be subjected to tensile loading with an external magnet. Additionally, the deflection of the flexible post could be used to measure the contractile force of PDL cells in the PTCs. Prior to tensile loading, second harmonics generation analysis of collagen fibers in PTCs revealed that incorporation of PDL cells resulted in collagen remodeling. Biomechanical testing of PTCs by tensile loading revealed an elastic response at 4 h, permanent deformation by 1 d, and creep elongation by 1 wk. Subsequently, contractile forces of PDL cells were substantially lower for PTCs under tensile loading. Immunofluorescence analysis revealed that tensile loading caused PDL cells to increase in number, express higher levels of F-actin and α-smooth muscle actin, and become aligned to the tensile axis. Second harmonics generation analysis indicated that collagen fibers in PTCs progressively remodeled over time with tensile loading. Gene expression analysis also confirmed tension-mediated upregulation of the F-actin/Rho pathway and osteogenic genes. Our model is novel in demonstrating the mechanobiological behavior that results in cell-mediated remodeling of the PDL tissue in a 3D context. Hence, it can be a valuable tool to develop therapeutics for periodontitis, periodontal regeneration, and orthodontics.

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