Migration of Periodontal Ligament Fibroblasts on Nanometric Topographical Patterns: Influence of Filopodia and Focal Adhesions on Contact Guidance
Migration of Periodontal Ligament Fibroblasts on Nanometric Topographical Patterns: Influence of Filopodia and Focal Adhesions on Contact Guidance
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DOI：
10.1371/journal.pone.0015129
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发表时间：
2010-12-01
期刊：
影响因子：
3.7
通讯作者：
Mittler, Silvia
中科院分区：
文献类型：
--
作者：
Hamilton, Douglas W.;Oates, Christine J.;Mittler, Silvia
人类牙周膜的再生被认为是牙科的“圣杯”，但它仍然是一个主要的临床问题。通常使用 EDTA 凝胶或激光来去除细菌生物膜。这些治疗方案的一个副作用是牙齿表面纳米形貌的蚀刻。然而，牙周膜成纤维细胞对这些特征的反应却很少受到关注。 通过激光干涉光刻，我们制造了具有连续或不连续纳米凹槽的精确定义的形貌，以评估 PDL 成纤维细胞的粘附、扩散和迁移。 PDL 成纤维细胞粘附并在所有测试表面上扩散，在不连续的纳米凹槽上初始扩散和粘着斑形成较慢。细胞的两个平面面积明显较小 与非图案对照上的细胞相比，连续和不连续的纳米凹槽。接种后 24 小时，两种类型纳米凹槽上的细胞均平行于凹槽长轴高度伸长。延时视频显微镜显示，PDL 成纤维细胞运动在两种类型的凹槽上受到引导，但迁移速度与在非图案对照上培养的细胞没有显着差异。分析 使用延时视频显微镜和纽蛋白和肌动蛋白标记的丝状伪足形成显示，在纳米凹槽上，丝状伪足在细胞两端高度对齐，但随着时间的增加，丝状伪足和膜突起在垂直于细胞长轴的细胞一侧形成。我们得出结论，牙周膜成纤维细胞对纳米拓扑结构敏感 深度85-100μm，可用于牙周膜再生。
Considered to be the "holy grail'' of dentistry, regeneration of the periodontal ligament in humans remains a major clinical problem. Removal of bacterial biofilms is commonly achieved using EDTA gels or lasers. One side effect of these treatment regimens is the etching of nanotopographies on the surface of the tooth. However, the response of periodontal ligament fibroblasts to such features has received very little attention. Using laser interference lithography, we fabricated precisely defined topographies with continuous or discontinuous nanogrooves to assess the adhesion, spreading and migration of PDL fibroblasts. PDL fibroblasts adhered to and spread on all tested surfaces, with initial spreading and focal adhesion formation slower on discontinuous nanogrooves. Cells had a significantly smaller planar area on both continuous and discontinuous nanogrooves in comparison with cells on non-patterned controls. At 24 h post seeding, cells on both types of nanogrooves were highly elongated parallel to the groove long axis. Time-lapse video microscopy revealed that PDL fibroblast movement was guided on both types of grooves, but migration velocity was not significantly different from cells cultured on non-patterned controls. Analysis of filopodia formation using time-lapse video microscopy and labeling of vinculin and F-actin revealed that on nanogrooves, filopodia were highly aligned at both ends of the cell, but with increasing time filopodia and membrane protrusions developed at the side of the cell perpendicular to the cell long axis. We conclude that periodontal ligament fibroblasts are sensitive to nanotopographical depths of 85-100 mu m, which could be utilized in regeneration of the periodontal ligament.