Department of Phoniatrics, ENT University Hospital Graz, Medical University of Graz, Graz, Austria.
PLoS One. 2019 Mar 14;14(3):e0213788. doi: 10.1371/journal.pone.0213788. eCollection 2019.
Vocal fold fibroblasts (VFF) constitute the main cell type of the vocal fold's lamina propria, produce the extracellular matrix and thereby determine the tissue characteristics. To study VFF behavior under in vitro conditions it is important to mimic the dynamic environment of the in vivo state. The aim of our study was to develop and validate a novel phonomimetic bioreactor system mainly based on commercially available components. The use of cell culture dishes with flexible silicone bottoms in combination with a suitable loudspeaker made it possible to expose the cells to various kinds of phonatory stimuli. The fundamental vibration characteristics of silicone membranes were investigated with and without cell culture medium by laser Doppler vibrometry. Human VFF were seeded in flexible-bottomed plates and placed in a custom-made housing containing a loudspeaker. After the cells were exposed to a predefined audio stimulation protocol, cell viability was assessed and gene as well as protein expression levels were compared to static controls. Laser Doppler vibrometry revealed that addition of cell culture medium changed the resonance frequencies of vibrating membranes. Gene expression of hyaluronan synthase 2, collagen III, fibronectin and TGFβ-1 was significantly upregulated in VFF exposed to vibration, compared to static control. Vibration also significantly upregulated collagen I gene and protein expression. We present a new type of phonomimetic bioreactor. Compared to previous models, our device is easy to assemble and cost-effective, yet can provide a wide spectrum of phonatory stimuli based on the entire dynamic range of the human voice. Gene expression data of VFF cultured in our phonomimetic bioreactor show a significant effect of vibration on ECM metabolism, which illustrates the efficacy of our device.
声带成纤维细胞（VFF）构成声带固有层的主要细胞类型，产生细胞外基质，从而决定组织特征。为了在体外条件下研究 VFF 的行为，模拟体内状态的动态环境非常重要。我们的研究旨在开发和验证一种主要基于市售组件的新型声学仿生生物反应器系统。使用具有柔性硅树脂底部的细胞培养皿与合适的扬声器相结合，使细胞能够暴露于各种发声刺激下。通过激光多普勒测振仪研究了有和没有细胞培养基的硅树脂膜的基本振动特性。将人 VFF 接种在柔性底部培养皿中，并将其置于包含扬声器的定制外壳中。在细胞暴露于预定的音频刺激方案后，评估细胞活力，并将基因和蛋白质表达水平与静态对照进行比较。激光多普勒测振仪显示，添加细胞培养基会改变振动膜的共振频率。与静态对照相比，暴露于振动下的 VFF 中透明质酸合酶 2、胶原 III、纤连蛋白和 TGFβ-1 的基因表达明显上调。振动还显著上调了胶原 I 基因和蛋白质的表达。我们提出了一种新型声学仿生生物反应器。与以前的模型相比，我们的设备易于组装且具有成本效益，但可以根据人类声音的整个动态范围提供广泛的发声刺激。在我们的声学仿生生物反应器中培养的 VFF 的基因表达数据显示，振动对 ECM 代谢有显著影响，这说明了我们设备的功效。
