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The Kinetics of Cell Adhesion to Solid Scaffolds: An Experimental and Theoretical Approach
Journal of Biological Physics ( IF 1.8 ) Pub Date : 2008-09-02 , DOI: 10.1007/s10867-008-9108-x
I O Doagă 1 , T Savopol , M Neagu , A Neagu , E Kovács
Affiliation  

The process of cell seeding on biocompatible scaffolds has a major impact on the morphological evolution of an engineered tissue because it involves all the key factors of tissue formation: cells, matrix, and their mutual interactions. In order to characterize the efficiency of cell seeding techniques, mainly static parameters are used such as cell density, cell distribution, and cell viability. Here, we present an experimental model that incorporates an optical density meter providing real-time information on the cell seeding velocity, a relevant dynamic parameter of cell–matrix interaction. Our setup may be adapted to fit various cell seeding protocols. A modified fluorimetric cuvette is used as bioreactor culture flask. The optical density of the magnetically stirred cell suspension is recorded by a digital optoelectronic device. We performed calibration experiments in order to prove that, in our experimental conditions, optical density depends linearly on the number of cells in the unit volume of suspension. Control studies showed that, during the time course of a typical experiment (up to 10 h), the cells (murine 3T3 fibroblasts) neither aggregated nor adhered significantly to the walls of the cuvette. Hence, our setup yields the number of cells attached to the scaffold as a function of time. In order to analyze the experimental seeding curves, we built a kinetic model based on Langmuir’s adsorption theory, which was extended to include a preliminary step of integrin function recovery. We illustrate the proposed approach by two sets of experiments that involved trypsin–EDTA or only EDTA treatment (no trypsin) used to detach the cells from the culture flasks. The data indicate that in both cases cell–matrix adhesion has a sequential, two-step dynamics, but kinetic parameters and attachment site availability depend on the experimental protocol.

中文翻译:

细胞粘附在固体支架上的动力学:一种实验和理论方法

在生物相容性支架上接种细胞的过程对工程组织的形态演变有重大影响,因为它涉及组织形成的所有关键因素:细胞、基质及其相互作用。为了表征细胞接种技术的效率,主要使用静态参数,例如细胞密度、细胞分布和细胞活力。在这里,我们提出了一个实验模型,该模型结合了光密度计,可提供有关细胞接种速度的实时信息,这是细胞 - 基质相互作用的相关动态参数。我们的设置可以适应各种细胞接种协议。改进的荧光比色皿用作生物反应器培养瓶。磁力搅拌的细胞悬浮液的光密度由数字光电设备记录。我们进行了校准实验,以证明在我们的实验条件下,光密度与单位体积悬浮液中的细胞数量呈线性关系。对照研究表明,在典型实验的时间过程中(长达 10 小时),细胞(鼠 3T3 成纤维细胞)既没有聚集也没有明显粘附到比色皿的壁上。因此,我们的设置产生附着在支架上的细胞数量作为时间的函数。为了分析实验接种曲线,我们建立了基于朗缪尔吸附理论的动力学模型,并将其扩展为包括整合素功能恢复的初步步骤。我们通过两组实验来说明所提出的方法,这些实验涉及用于将细胞从培养瓶中分离的胰蛋白酶-EDTA 或仅 EDTA 处理(无胰蛋白酶)。
更新日期:2008-09-02
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