Centre for Biomedical Technologies, School of Mechanical, Medical and Process Engineering, Faculty of Engineering, Queensland University of Technology (QUT), Brisbane, QLD 4000, Australia.
Centre for Biomedical Technologies, School of Mechanical, Medical and Process Engineering, Faculty of Engineering, Queensland University of Technology (QUT), Brisbane, QLD 4000, Australia.
Acta Biomater. 2021 Dec;136:429-440. doi: 10.1016/j.actbio.2021.09.042. Epub 2021 Sep 24.
Tissue engineering involves the seeding of cells into a structural scaffolding to regenerate the architecture of damaged or diseased tissue. To effectively design a scaffold, an understanding of how cells collectively sense and react to the geometry of their local environment is needed. Advances in the development of melt electro-writing have allowed micron and submicron polymeric fibres to be accurately printed into porous, complex and three-dimensional structures. By using melt electrowriting, we created a geometrically relevant in vitro scaffold model to study cellular spatial-temporal kinetics. These scaffolds were paired with custom computer vision algorithms to investigate cell nuclei, cell membrane actin and scaffold fibres over different pore sizes (200-600 µm) and time points (28 days). We find that cells proliferated much faster in the smaller (200 µm) pores which halved the time until confluence versus larger (500 and 600 µm) pores. Our analysis of stained actin fibres revealed that cells were highly aligned to the fibres and the leading edge of the pore filling front, and we found that cells behind the leading edge were not aligned in any particular direction. This study provides a systematic understanding of cellular spatial temporal kinetics within a 3D in vitro model to inform the design of more effective synthetic tissue engineering scaffolds for tissue regeneration. STATEMENT OF SIGNIFICANCE: Advances in the development of melt electro-writing have allowed micron and submicron polymeric fibres to be accurately printed into porous, complex and three-dimensional structures. By using melt electrowriting, we created a geometrically relevant in vitro model to study cellular spatial-temporal kinetics to provide a systematic understanding of cellular spatial temporal kinetics within a 3D in vitro model. The insights presented in this work help to inform the design of more effective synthetic tissue engineering scaffolds by reducing cell culture time; which is valuable information for the implant or lab-grown-meat industries.
组织工程学涉及将细胞接种到结构支架中，以再生受损或患病组织的结构。为了有效地设计支架，需要了解细胞如何集体感知和对其局部环境的几何形状做出反应。熔融电写入技术的进步使得微米和亚微米级聚合物纤维能够精确地打印成多孔、复杂和三维结构。通过使用熔融电写入，我们创建了一个具有几何相关性的体外支架模型，以研究细胞的时空动力学。这些支架与定制的计算机视觉算法相结合，研究了不同孔径（200-600 µm）和时间点（28 天）的细胞核、细胞膜肌动蛋白和支架纤维。我们发现，细胞在较小的（200 µm）孔中增殖得更快，与较大的（500 和 600 µm）孔相比，达到汇合所需的时间缩短了一半。我们对染色肌动蛋白纤维的分析表明，细胞与纤维和孔填充前缘的前沿高度对齐，并且我们发现前沿后面的细胞没有任何特定的对齐方向。这项研究提供了对 3D 体外模型中细胞时空动力学的系统理解，为组织再生的更有效合成组织工程支架的设计提供了信息。意义声明：熔融电写入技术的进步使得微米和亚微米级聚合物纤维能够精确地打印成多孔、复杂和三维结构。通过使用熔融电写入，我们创建了一个具有几何相关性的体外模型，以研究细胞的时空动力学，从而对 3D 体外模型中的细胞时空动力学有系统的了解。本工作中提出的见解有助于通过减少细胞培养时间来设计更有效的合成组织工程支架，这对于植入物或实验室培养肉行业来说是有价值的信息。
