应用于三维细胞培养的亚大气压生物反应器。

Robert P Wilkes, Amy K McNulty, Teri D Feeley, Marisa A Schmidt, Kris Kieswetter
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引用次数: 20

摘要

真空辅助封闭(VAC)负压伤口治疗(NPWT)是一种非常成功和广泛应用的伤口愈合治疗方式,尽管目前还没有仪器来监测亚大气压在体外应用的效果。这样的装置是需要的,以便更好地了解该疗法的生物学效应并可能在其基础上进行改进。本文描述了一种允许此类研究的新型生物反应器的开发和验证。由含有纤维蛋白凝块的三维成纤维细胞组成的组织类似物在现成的一次性细胞培养插入物和集成到生物反应器模块中的多孔板中培养。用于伤口治疗的商业化负压敷料置于培养物的顶部,并对敷料施加亚大气压。以控制生理性伤口渗出液流速的培养基灌注培养物。该生物反应器的设计允许在明场和荧光模式下使用倒置显微镜观察培养,并在5%二氧化碳的气氛中对系统进行持续培养。该封闭系统模拟了真空NPWT下的伤口微环境。当亚大气压力将修整材料向下拉时,就会发生基质压缩。在接触区,含有成纤维细胞的组织类似物在24 h时表面波动明显,似乎与敷料微观结构相对应。生物反应器的设计,由可消毒的机械塑料和一次性实验室用具组成,可以很容易地扩展到多个单元。验证实验表明,该系统中的细胞存活率与静态组织培养中的细胞存活率相当。应用VAC NPWT后,细胞形态发生变化,细胞变厚,有组织的肌动蛋白细胞骨架。这种新的培养系统的开发和验证为体外研究亚大气压对组织的应用建立了一个稳定的平台。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Bioreactor for application of subatmospheric pressure to three-dimensional cell culture.

Vacuum-assisted closure (VAC) negative pressure wound therapy (NPWT) is a highly successful and widely used treatment modality for wound healing, although no apparatus exists to monitor the effects of subatmospheric pressure application in vitro. Such an apparatus is desirable to better understand the biological effects of this therapy and potentially improve upon them. This article describes the development and validation of a novel bioreactor that permits such study. Tissue analogues consisting of 3-dimensional fibroblast-containing fibrin clots were cultured in off-the-shelf disposable cell culture inserts and multi-well plates that were integrated into the bioreactor module. Negative pressure dressings, commercialized for wound therapy, were placed on top of the culture, and subatmospheric pressure was applied to the dressing. Cultures were perfused with media at controlled physiologic wound exudate flow rates. The design of this bioreactor permits observation of the culture using an inverted microscope in brightfield and fluorescence modes and sustained incubation of the system in a 5% carbon dioxide atmosphere. This closed-system mimics the wound micro-environment under VAC NPWT. Matrix compression occurs as the subatmospheric pressure draws the dressing material down. At the contact zone, surface undulations were clearly evident on the fibroblast-containing tissue analogues at 24 h and appeared to correspond to the dressing microstructure. The bioreactor design, consisting of sterilizable machined plastics and disposable labware, can be easily scaled to multiple units. Validation experiments show that cell survival in this system is comparable with that seen in cells grown in static tissue culture. After application of VAC NPWT, cell morphology changed, with cells appearing thicker and with an organized actin cytoskeleton. The development and validation of this new culture system establishes a stable platform for in vitro investigations of subatmospheric pressure application to tissues.

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来源期刊
Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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