Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis
Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis
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用于成骨的新型纳米振动生物反应器和培养器皿的设计、构建和表征
DOI：
10.1101/543660
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Campsie P
中科院分区：
文献类型：
--
作者：
Campsie P
在再生医学中，控制干细胞谱系承诺的技术是一个迅速发展的领域。最近，对间充质干细胞（MSCs）的纳米级机械刺激已被证明可以激活刺激2D和3D培养成骨的机械转导通路。这有可能通过在不使用化学诱导的情况下从自体或异体间充质干细胞来源制造细胞移植物材料来彻底改变骨移植手术。随着对细胞机械刺激的兴趣的增加和临床应用的巨大潜力，很明显，研究人员和临床医生需要一种可扩展的生物反应器系统，该系统可以通过简单的统包方法提供一致的可重复性结果。提出了一种新型生物反应器系统，该系统包括：一个生物反应器振动板，针对1 kHz的纳米振动进行校准和优化，一个电源单元，提供1 kHz正弦波信号，以产生大约30 nm的振动幅值，以及定制的6孔培养器，每个孔的底部都装有环形磁铁，用于与生物反应器的磁性振动板共形附着。采用有限元分析方法设计了培养器和振动板，确定了模态响应和谐波响应，并通过干涉测量进行了验证。这有助于确保振动板和培养皿，以及胶原蛋白和间充质干细胞都作为一个刚体运动，避免振动板共振频率附近的大变形和共振以外的振动阻尼。在系统进行初始生物学实验后，进行成骨蛋白表达评估以确认MSCs的分化，并在振动刺激时对3D凝胶构建进行原子力显微镜检查以验证凝胶没有发生应变硬化。这表明细胞分化是由生物反应器单独提供的纳米振动刺激的结果，而其他细胞分化因素，如胶原凝胶的硬化，没有贡献。
In regenerative medicine, techniques which control stem cell lineage commitment are a rapidly expanding field of interest. Recently, nanoscale mechanical stimulation of mesenchymal stem cells (MSCs) has been shown to activate mechanotransduction pathways stimulating osteogenesis in 2D and 3D culture. This has the potential to revolutionise bone graft procedures by creating cellular graft material from autologous or allogeneic sources of MSCs without using chemical induction. With the increased interest in mechanical stimulation of cells and huge potential for clinical use, it is apparent that researchers and clinicians require a scalable bioreactor system that provides consistently reproducible results with a simple turnkey approach. A novel bioreactor system is presented that consists of: a bioreactor vibration plate, calibrated and optimised for nanometre vibrations at 1 kHz, a power supply unit, which supplies a 1 kHz sine wave signal necessary to generate approximately 30 nm of vibration amplitude, and custom 6-well cultureware with toroidal shaped magnets incorporated in the base of each well for conformal attachment to the bioreactor’s magnetic vibration plate. The cultureware and vibration plate were designed using finite element analysis to determine the modal and harmonic responses, and validated by interferometric measurement. This helps ensure that the vibration plate and cultureware, and thus collagen and MSCs, all move as a rigid body, avoiding large deformations close to the resonant frequency of the vibration plate and vibration damping beyond the resonance. Assessment of osteogenic protein expression was performed to confirm differentiation of MSCs after initial biological experiments with the system, as well as atomic force microscopy of the 3D gel constructs during vibrational stimulation to verify that strain hardening of the gel did not occur. This shows that cell differentiation was the result of the nanovibrational stimulation provided by the bioreactor alone, and that other cell differentiating factors, such as stiffening of the collagen gel, did not contribute.
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影响因子：
5.5
作者：
Gabriel D. Pemberton;Peter G. Childs;S. Reid;H. Nikukar;P. Tsimbouri;N. Gadegaard;A. Curtis;M. Dalby
通讯作者：
Gabriel D. Pemberton;Peter G. Childs;S. Reid;H. Nikukar;P. Tsimbouri;N. Gadegaard;A. Curtis;M. Dalby
影响因子：
17.1
作者：
Nikukar, Habib;Reid, Stuart;Dalby, Matthew J.
通讯作者：
Dalby, Matthew J.
影响因子：
--
作者：
Wehland, Markus;Ma, Xiao;Grimm, Daniela
通讯作者：
Grimm, Daniela
用于刺激人类间充质干细胞的纳米级振动的产生。
影响因子：
2.9
作者：
Nikukar H
通讯作者：
Nikukar H
DOI：
10.3390/ma10101115
发表时间：
2017-09-21
期刊：
Materials (Basel, Switzerland)
影响因子：
--
作者：
Neděla O;Slepička P;Švorčík V
通讯作者：
Švorčík V