High-throughput investigation of osteoblast response to polymer crystallinity: influence of nanometer-scale roughness on proliferation
High-throughput investigation of osteoblast response to polymer crystallinity: influence of nanometer-scale roughness on proliferation
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DOI：
10.1016/j.biomaterials.2003.08.043
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发表时间：
2004-03-01
期刊：
影响因子：
14
通讯作者：
Amis, EJ
中科院分区：
文献类型：
--
作者：
Washburn, NR;Yamada, KM;Amis, EJ
提出了一种用于分析聚合物材料中细胞对结晶度的响应的高通量方法。结晶度的变化会导致纳米长度尺度上的表面粗糙度变化，并且研究表明细胞对这些变化非常敏感。采用热处理温度梯度法在聚L-乳酸薄膜上制备了高分子结晶度梯度材料。使用原子力显微镜对所得的形貌进行表征。在单个样品上创建0.5至13 nm范围内的均方根（rms）粗糙度值。MCM-E1成骨细胞培养1、3和5 d后，用自动荧光显微镜检测细胞数量。它示出的膜的光滑区域上的增殖率是远远大于粗糙区域上的，并且观察到作为粗糙度的函数的速率的单调变化。临界均方根粗糙度约为1.1 nm，高于此值时，增殖率会出现统计学显著降低。免疫染色细胞上的荧光显微镜测量表明，细胞面积、形成的粘连的数量或类型或肌动蛋白聚合的程度没有显著变化。从酶联免疫荧光试验的结果表明，没有检测到的变化，粘附蛋白的可及性，表明细胞直接响应基板地形。使用梯度库的方法产生的功能依赖的细胞增殖纳米级的粗糙度，并给出了一个敏感的估计的临界粗糙度，减少增殖观察。(C)2003爱思唯尔有限公司。保留所有权利。
A high-throughput method for analyzing cellular response to crystallinity in a polymer material is presented. Variations in crystallinity lead to changes in surface roughness on nanometer length scales, and it is shown that cells are exquisitely sensitive to these changes. Gradients of polymer crystallinity were fabricated on films Of poly(L-lactic acid) using a gradient in annealing temperature. The resultant morphologies were characterized using an atomic force microscope. Root-mean-square (rms) roughness values ranging from 0.5 to 13 nm were created on a single sample. MCM-E1 osteoblastic cells were cultured for 1, 3 and 5 d, and the number of cells was measured using automated fluorescence microscopy. It is shown that the rate of proliferation on the smooth regions of the films is much greater than that on the rough regions, and a monotonic variation in rate is observed as a function of roughness. The critical rms roughness, above which a statistically significant reduction in rate of proliferation occurs, was approximately 1.1 nm. Fluorescence microscopy measurements on immunostained cells indicate there is no significant change in cell area, the number or type of adhesions formed, or the degree of actin polymerization. Results from enzyme-linked immunofluorescence assays indicated that there was no detectable change in adhesion protein accessibility, suggesting the cells directly respond to substrate topography. The use of the gradient library approach yielded the functional dependence of cell proliferation on nanometer-scale roughness and gave a sensitive estimate of the critical roughness for which a decrease in proliferation is observed. (C) 2003 Elsevier Ltd. All rights reserved.