Pernagallo Salvatore, Unciti-Broceta Asier, Díaz-Mochón Juan José, Bradley Mark
School of Chemistry, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, UK.
Biomed Mater. 2008 Sep;3(3):034112. doi: 10.1088/1748-6041/3/3/034112. Epub 2008 Aug 15.
A quantitative and qualitative analysis of cellular adhesion, morphology and viability is essential in understanding and designing biomaterials such as those involved in implant surfaces or as tissue-engineering scaffolds. As a means to simultaneously perform these studies in a high-throughput (HT) manner, we report a normalized protocol which allows the rapid analysis of a large number of potential cell binding substrates using polymer microarrays and high-content fluorescence microscopy. The method was successfully applied to the discovery of optimal polymer substrates from a 214-member polyurethane library with mouse fibroblast cells (L929), as well as simultaneous evaluation of cell viability and cellular morphology. Analysis demonstrated high biocompatibility of the binding polymers and permitted the identification of several different cellular morphologies, showing that specific polymer interactions may provoke changes in cell shape. In addition, SAR studies showed a clear correspondence between cellular adhesion and polymer structure. The approach can be utilized to perform multiple experiments (up to 1024 single experiments per slide) in a highly reproducible manner, leading to the generation of vast amounts of data in a short time period (48-72 h) while reducing dramatically the quantities of polymers, reagents and cells used.
对细胞黏附、形态和活力进行定量和定性分析，对于理解和设计生物材料（如植入物表面材料或组织工程支架材料）至关重要。作为一种以高通量（HT）方式同时进行这些研究的手段，我们报告了一种标准化方案，该方案允许使用聚合物微阵列和高内涵荧光显微镜对大量潜在的细胞结合底物进行快速分析。该方法已成功应用于从小鼠成纤维细胞（L929）的214种聚氨酯文库中发现最佳聚合物底物，以及同时评估细胞活力和细胞形态。分析表明结合聚合物具有高生物相容性，并能识别几种不同的细胞形态，表明特定的聚合物相互作用可能引发细胞形状的变化。此外，构效关系研究表明细胞黏附与聚合物结构之间存在明显的对应关系。该方法可用于以高度可重复的方式进行多个实验（每张载玻片最多可进行1024个单实验），从而在短时间内（48 - 72小时）生成大量数据，同时大幅减少聚合物、试剂和细胞的用量。
