Effects of a biophysically defined environment on cells of different developmental origin: a contribution for the development of versatile biomaterials

与生物化学因素类似，生物物理线索控制多种细胞功能。在这方面，在医学和牙科中，仍然没有通过其材料固有的生物物理特性控制治疗相关细胞功能的携带细胞的生物材料。在这种情况下，根本的问题还没有得到回答，是否一个定义的生物物理线索有类似的影响细胞形态和表型，即使在不同的发育起源的细胞，如果是真的，如果这种影响可能是由相同的信号分子处理。在这种情况下，可以使用相同的生物物理学线索来控制不同发育起源的细胞。在该项目中，中胚层和外胚层上皮牙龈角质形成细胞的间充质基质（MSC）将在支架上培养。这些市售支架将由申请人关于固有支架（i）纤维厚度和（ii）纤维密度和所得基底刚度来定义和表征。将在两种细胞实体中分析这些生物病理学定义的细胞外信号对细胞形态、增殖和分化以及与此相关的信号分子激活的影响，也包括机制方法。我们将集中在信号分子，出现从局灶性粘连（FA）和细胞间接触介导的钙粘蛋白系统，这两个基本上是参与细胞内处理的生物物理线索。分析将基于既定的标准化方法，以量化细胞形态、生物功能标志物的基因和蛋白质表达、信号分子的活化/磷酸化以及统计学比较。将通过使用特异性抑制剂和RNAi进行机制分析。材料特性的表征包括扫描电子显微镜和纳米压痕。总之，所得到的数据将有助于回答该提案的中心问题，如果相同的生物物理线索激活不同发育起源的细胞中的相同信号分子，并在此介导可比的效果。所获得的知识可以在生物制药定制的生物材料的开发中实施，以控制治疗用细胞的治疗相关细胞功能。

Similar to biochemical factors, biophysical cues govern multiple cell functions. In this regard, in medicine and dentistry, cell-carrying biomaterials, which control therapy-relevant cell functions through their material-inherent biophysical properties, are still not available. In this context, the fundamental question has not yet been answered, whether a defined biophysical cue has similar effects on cell morphology and phenotype, even in cells of different developmental origin, and, if true, if this effect is possibly processed by the same signaling molecules. In this case, the same defined biophysical cue could be used to control cells of different developmental origin. Within the project, mesenchymal stromal (MSCs) of the mesoderm and ectodermal epithelial gingival keratinocytes will be cultivated on nanofiber scaffolds. These commercially available scaffolds will be defined and characterized by the applicants in regard to the innate scaffold (i) fiber thickness and (ii) fiber densities and the resulting substrate stiffness. The effects of these biophysically defined extracellular cues on cell morphology, proliferation and differentiation as well as the herewith connected activation of signaling molecules will be analyzed in both cell entities, also including mechanistic approaches. We will focus on signaling molecules, emerging from focal adhesions (FAs) and the cell-to-cell contact-mediating cadherin system, both of which are essentially involved in the intracellular processing of biophysical cues. The analyses will be based on established, standardized methods to quantify cell morphology, gene and protein expression of biofunctional markers, activation/phosphorylation of signaling molecules, and statistical comparisons. Mechanistic analyses will be conducted by using specific inhibitors and RNAi. Characterization of material properties includes scanning electron microscopy and nanoindentation. Taken together, the resulting data will facilitate answering the central question of the proposal, if the same biophysical cues activate the same signaling molecules in cells of different developmental origin, and herein mediate comparable effects. The knowledge gained can be implemented in the development of biophysically-tailored biomaterials to control therapy-relevant cell functions for therapeutically utilized cells.