Biosystems & Biomaterials Division, National Institute of Standards & Technology, 100 Bureau Drive, Gaithersburg, MD 20899, USA.
Biosystems & Biomaterials Division, National Institute of Standards & Technology, 100 Bureau Drive, Gaithersburg, MD 20899, USA.
Biomaterials. 2014 Mar;35(9):2558-67. doi: 10.1016/j.biomaterials.2013.12.092. Epub 2014 Jan 15.
Many scaffold systems have evolved for tissue engineering and in vitro tissue models to provide a 3D (three-dimensional) microenvironment that enables cells to behave more physiologically. We hypothesized that cells would adopt morphologies with more 3D character during culture in scaffolds as compared to planar substrates. Cell shape and function are tightly linked and effects of scaffold niche properties on cell shape and dimensionality are important for directing cell function. Herein, primary human bone marrow stromal cells (hBMSCs) were cultured in 6 different scaffolds and on a planar control substrate. hBMSCs were imaged using 3D confocal microscopy, and 3D image analyses were used to assess hBMSC shape and dimensionality. A characteristic gyration tensor ellipsoid was calculated for hBMSCs in the different scaffolds which enabled hBMSC dimensionality to be classified based on shape. A "Dimensionality Matrix" was developed that showed that hBMSC shape and dimensionality were influenced by scaffold properties, and that scaffolds could drive hBMSCs into 1D, 2D or 3D shapes. In addition, the hBMSC Z-Depth was measured to determine if hBMSCs became less flat during culture in scaffolds. Z-Depth results showed that all 6 scaffolds caused an increase in cell Z-Depth compared to the 2D planar substrate. These results demonstrate that hBMSCs take on morphologies with greater 3D character in scaffolds than on a planar substrate and that scaffold properties can be adjusted to modify cell dimensionality. In addition, biomaterialists can use this measurement approach to assess and compare scaffold design modifications as they strive to create optimal cell niches that provide a 3D microenvironment.
许多支架系统已经发展起来，用于组织工程和体外组织模型，以提供一个 3D（三维）微环境，使细胞能够更具生理活性。我们假设，与平面基底相比，细胞在支架中的培养过程中会采用更具 3D 特征的形态。细胞的形状和功能密切相关，支架小生境特性对细胞形状和维度的影响对于指导细胞功能非常重要。在此，原代人骨髓基质细胞（hBMSCs）在 6 种不同的支架和平面对照基质上培养。使用 3D 共聚焦显微镜对 hBMSCs 进行成像，并使用 3D 图像分析评估 hBMSCs 的形状和维度。计算了 hBMSCs 在不同支架中的特征回旋张量椭球，使 hBMSCs 的维度能够基于形状进行分类。开发了一个“维度矩阵”，表明支架特性会影响 hBMSC 的形状和维度，并且支架可以将 hBMSCs 驱动成 1D、2D 或 3D 形状。此外，还测量了 hBMSC 的 Z 深度，以确定 hBMSCs 在支架中培养时是否变得更平坦。Z 深度结果表明，与 2D 平面基底相比，所有 6 种支架都导致细胞 Z 深度增加。这些结果表明，与平面基底相比，hBMSCs 在支架中呈现出更具 3D 特征的形态，并且可以调整支架特性来改变细胞的维度。此外，生物材料学家可以使用这种测量方法来评估和比较支架设计修改，因为他们努力创造提供 3D 微环境的最佳细胞小生境。
