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通过细胞分离仪评估基底形貌对 hFOB 细胞行为和初始细胞黏附的影响。

The effect of substrate topography on hFOB cell behavior and initial cell adhesion evaluated by a cytodetacher.

机构信息

Institute of Oral Medicine, National Cheng Kung University, Tainan, 701 Taiwan, ROC.

出版信息

J Mater Sci Mater Med. 2011 Apr;22(4):1027-36. doi: 10.1007/s10856-011-4255-1. Epub 2011 Feb 18.

Abstract

This study examined human fetal osteoblast (hFOB) cell morphology, adhesion force, and proliferation on a titanium-coated grooved surface. V-shaped grooves with a depth of 2.4 μm (T1) or 4.8 μm (T2) were produced in silicon wafers using photolithography and wet etching techniques. The grooved substrates were coated with a 200-nm-thick layer of titanium using a sputtering system. Smooth Ti-coated Si wafers were used as control surfaces. Analysis of the scanning electron microscopy observations shows that the cells responded to the micropattern by spreading out and becoming elongated. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay indicated that the grooved specimens had a significantly larger number of cells than did the control group after 5- and 15-day cultures. The cytocompatibility of specimens was quantitatively evaluated by a cytodetacher, which directly measures the detachment shear force of an individual cell to the substrate. After 30-min culture, the cell adhesion forces were 48.4, 136.6, and 103.3 nN for the smooth specimen, the T1 specimen, and the T2 specimen, respectively. The cell adhesion strengths were 294, 501, and 590 Pa for the smooth specimen, the T1 specimen, and the T2 specimen, respectively. The cell adhesion force and cell adhesion strength indicate the quality of cell adhesion, explaining the largest number of cells on grooved specimens. The experimental results suggest that the grooved patterns affect the cell shape and cytoskeletal structure, and thus influence the cell proliferation and cell adhesion force. The cytodetachment test with nanonewton resolution is a sensitive method for studying cell-biomaterial interaction.

摘要

本研究考察了人胎儿成骨细胞(hFOB)在钛涂层的沟槽表面的细胞形态、黏附力和增殖情况。使用光刻和湿法刻蚀技术在硅片上制作了深度为 2.4μm(T1)或 4.8μm(T2)的 V 形沟槽。使用溅射系统在沟槽基底上涂覆了 200nm 厚的钛层。光滑的 Ti 涂层 Si 晶片用作对照表面。扫描电子显微镜观察分析表明,细胞对微图案的反应是展开并变得细长。MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)测定表明,在 5 天和 15 天培养后,沟槽标本的细胞数量明显多于对照组。通过细胞分离仪对标本的细胞相容性进行了定量评估,细胞分离仪直接测量单个细胞对基底的分离剪切力。在 30 分钟的培养后,光滑标本、T1 标本和 T2 标本的细胞黏附力分别为 48.4、136.6 和 103.3nN。光滑标本、T1 标本和 T2 标本的细胞黏附强度分别为 294、501 和 590Pa。细胞黏附力和细胞黏附强度表明细胞黏附的质量,解释了在沟槽标本上有最多的细胞。实验结果表明,沟槽图案影响细胞形状和细胞骨架结构,从而影响细胞增殖和细胞黏附力。具有纳牛顿分辨率的细胞分离测试是研究细胞-生物材料相互作用的敏感方法。

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