Cell-scaffold interactions in artificial bone: regulation of collagen-I networks and focal adhesion complexes by biochemical and environmental cues

人造骨中细胞支架的相互作用：生化和环境因素对 I 型胶原蛋白网络和粘着斑复合物的调节

• 批准号: 410498501
• 负责人: Professor Dr. Oliver Friedrich
• 金额: --
• 依托单位: Lehrstuhl für Medizinische Biotechnologie (MBT)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/410498501?language=en
• 关键词: Cell; scaffold; interactions; artificial; bone

Collagen-I is the major fibrous extracellular component of bone responsible for its ultimate tensile strength. In tissue engineering (TE) one of the most important issues is to get cells inter-connected via a strong and functional extracellular matrix (ECM) mimicking as closely as possible naturally occurring ECM geometry. What is still missing is (i) a versatile, high-resolution and non-invasive online method to evaluate and quantify different aspects of engineered ECM and (ii) deeper insights into the mechanism as to why cellular ECM production is enhanced in 3D cell-scaffold composites, putatively via enhanced focal adhesion (FA) linkages, unlike in the 2D setting. The goal within this project is to develop such methods with collagen-I producing and bone forming cells based on multiphoton imaging (MPI) technologies, to use those techniques to compare collagen-I production and organization capabilities in 2D- and 3D-seeded constructs, to link them to the number, extension and distribution of FA complexes and to monitor changes due to biomechanical treatment of the constructs. Our major work hypothesis is that biomechanical stability of engineered bone tissue constructs is directly encoded by the angular distribution and fibrillary geometry of ECM collagen fibers, and high stability occurs within more isotropic interwoven fibrils as a result of activation of increased FA complex formation with ascorbic acid playing a critical role in the regeneration of collagen-I with optimum fiber stability.

胶原-I是骨的主要纤维细胞外组分，负责其极限拉伸强度。在组织工程（TE）中，最重要的问题之一是通过尽可能接近天然存在的ECM几何形状的强大且功能性的细胞外基质（ECM）使细胞相互连接。仍然缺少的是（i）一种多功能，高分辨率和非侵入性的在线方法来评估和量化工程ECM的不同方面，以及（ii）更深入地了解为什么细胞ECM的产生在3D细胞-支架复合物中增强，通过增强的粘着斑（FA）连接来增强，而不是在2D环境中。本项目的目标是开发基于多光子成像（MPI）技术的I型胶原生产和骨形成细胞的方法，使用这些技术比较2D和3D接种结构中I型胶原的生产和组织能力，将其与FA复合物的数量、延伸和分布联系起来，并监测结构生物力学处理引起的变化。我们的主要工作假设是，工程化骨组织结构的生物力学稳定性是直接编码的角分布和几何形状的ECM胶原纤维，和高稳定性发生在更各向同性交织的原纤维作为一个结果，激活增加FA复合物的形成与抗坏血酸发挥关键作用，在再生胶原-I与最佳的纤维稳定性。