Delakowski Axel J, Posselt Jared D, Wagner Christopher T
Department of Biomedical Engineering, The College of New Jersey, Ewing, NJ 08628, USA.
Bioengineering (Basel). 2022 Mar 21;9(3):127. doi: 10.3390/bioengineering9030127.
Functional tissue-engineered tendons and ligaments remain to be prepared in a reproducible and scalable manner. This study evaluates an acellular 3D extracellular matrix (ECM) scaffold for tendon/ligament tissue engineering and their ability to support strain-induced gene regulation associated with the tenogenesis of cultured mesenchymal stromal cells. Preliminary data demonstrate unique gene regulation patterns compared to other scaffold forms, in particular in Wnt signaling. However, the need for a robust bioreactor system that minimizes process variation was also evident. A design control process was used to design and verify the functionality of a novel bioreactor. The system accommodates 3D scaffolds with clinically-relevant sizes, is capable of long-term culture with customizable mechanical strain regimens, incorporates in-line load measurement for continuous monitoring and feedback control, and allows a variety of scaffold configurations through a unique modular grip system. All critical functional specifications were met, including verification of physiological strain levels from 1-10%, frequency levels from 0.2-0.5 Hz, and accurate load measurement up to 50 N, which can be expanded on the basis of load cell capability. The design process serves as a model for establishing statistical functionality and reliability of investigative systems. This work sets the stage for detailed analyses of ECM scaffolds to identify critical differentiation signaling responses and essential matrix composition and cell-matrix interactions.
功能性组织工程化肌腱和韧带仍有待以可重复和可扩展的方式制备。本研究评估了一种用于肌腱/韧带组织工程的脱细胞三维细胞外基质（ECM）支架及其支持与培养的间充质基质细胞肌腱生成相关的应变诱导基因调控的能力。初步数据显示，与其他支架形式相比，其具有独特的基因调控模式，尤其是在Wnt信号通路方面。然而，显然需要一个强大的生物反应器系统来最小化工艺变化。采用设计控制流程来设计和验证一种新型生物反应器的功能。该系统可容纳具有临床相关尺寸的三维支架，能够通过可定制的机械应变方案进行长期培养，集成在线负载测量以进行连续监测和反馈控制，并通过独特的模块化夹具系统允许各种支架配置。所有关键功能规格均得到满足，包括验证1%至10%的生理应变水平、0.2至0.5Hz的频率水平以及高达50N的精确负载测量，可根据称重传感器的能力进行扩展。该设计过程为建立研究系统的统计功能和可靠性提供了一个模型。这项工作为详细分析ECM支架以确定关键的分化信号反应、基本的基质组成和细胞-基质相互作用奠定了基础。
