Enhanced depth-independent chondrocyte proliferation and phenotype maintenance in an ultrasound bioreactor and an assessment of ultrasound dampening in the scaffold.
Enhanced depth-independent chondrocyte proliferation and phenotype maintenance in an ultrasound bioreactor and an assessment of ultrasound dampening in the scaffold.
复制标题
超声生物反应器中增强的与深度无关的软骨细胞增殖和表型维持以及支架中超声阻尼的评估。
DOI：
10.1016/j.actbio.2014.07.013
复制
发表时间：
2014
影响因子：
9.7
通讯作者：
Subramanian,Anuradha
中科院分区：
文献类型：
--
作者：
GuhaThakurta,Sanjukta;Kraft,Mikail;Viljoen,HendrikJ;Subramanian,Anuradha
在超声(US)辅助的生物反应器中培养软骨细胞种植的支架，该反应器为细胞提供共振频率(∼5.0兆赫兹)附近的声能。选择不同孔隙率的聚碳酸酯(BM)、壳聚糖(CS)和壳聚糖正丁醇(CSB)为支架材料，采用以下US方案：15kPa和60kpa，每次5min，每天6次，共21天。非刺激性支架作为对照。对于BM支架，与对照组相比，US刺激显著影响细胞增殖和与深度无关的细胞密度。与对照组相比，经美国处理的BM支架上的COL2A1/COL1A1比率和Acan mRNA水平最高。在美国处理的细胞接种CS和CSB支架上也注意到了类似的趋势，尽管COL2A1/COL1A1比率明显低于BM支架。在超声作用下，SOX-9的表达也升高，且与COL2A1/COL1A1的比值平行。作为原创性贡献，基于Biot理论建立了一个简化的数学模型来理解入射超声波在支架中的传播，并将模型分析与细胞响应联系起来。支架结构影响了US场的分布，其中US场是BM支架中衰减最小的，从而将更多的机械能耦合到细胞内，导致细胞活性增加。
Chondrocyte-seeded scaffolds were cultured in an ultrasound (US)-assisted bioreactor, which supplied the cells with acoustic energy around resonance frequencies (∼5.0 MHz). Polyurethane-polycarbonate (BM), chitosan (CS) and chitosan–n-butanol (CSB) based scaffolds with varying porosities were chosen and the following US regimen was employed: 15 kPa and 60 kPa, 5 min per application and 6 applications per day for 21 days. Non-stimulated scaffolds served as control. For BM scaffolds, US stimulation significantly impacted cell proliferation and depth-independent cell population density compared to controls. The highest COL2A1/COL1A1 ratios and ACAN mRNA were noted on US-treated BM scaffolds compared to controls. A similar trend was noted on US-treated cell-seeded CS and CSB scaffolds, though COL2A1/COL1A1 ratios were significantly lower compared to BM scaffolds. Expression of Sox-9 was also elevated under US and paralleled the COL2A1/COL1A1 ratio. As an original contribution, a simplified mathematical model based on Biot theory was developed to understand the propagation of the incident US wave through the scaffolds and the model analysis was connected to cellular responses. Scaffold architecture influenced the distribution of US field, with the US field being the least attenuated in BM scaffolds, thus coupling more mechanical energy into cells, and leading to increased cellular activity.