Label free process monitoring of 3D bioprinted engineered constructs via dielectric impedance spectroscopy
Label free process monitoring of 3D bioprinted engineered constructs via dielectric impedance spectroscopy
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通过介电阻抗谱对 3D 生物打印工程结构进行无标记过程监测
DOI：
10.1088/1758-5090/aaccbf
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发表时间：
2018
期刊：
影响因子：
9
通讯作者：
Starly, Binil
中科院分区：
文献类型：
--
作者：
Narayanan, Lokesh Karthik;Thompson, Trevor L;Shirwaiker, Rohan A;Starly, Binil
生物制造过程可以影响构建物中被包裹的细胞的生物质量属性。目前，对所制造的构造物的评估是通过对构造物进行破坏性分析来进行的，这些分析需要染色和切片。这一缺陷限制了生物制造工艺向工业实践的转化。在这项工作中，我们研究了包裹在生物打印的3D水凝胶结构中的活细胞对应用交变电场的介电响应，作为一种无标记的非破坏性监测方法。研究了频谱上的β色散参数(介电常数变化Δε、科尔斜率因子α、临界极化频率fc)与关键蜂窝质量属性之间的关系。结果表明，含有更多活细胞(人脂肪干细胞和骨肉瘤细胞系)的藻酸盐载体具有显著更高的Δε和α(P<；0.05)。当扩展到生物打印时，结果显示随着关键生物打印参数(挤压压力、温度、处理时间)的变化，HASC的增殖和活性的变化显著影响∆ε，α和fc。我们还展示了对生物打印后HASC分布的监测以及生物打印的内侧半月板结构的横截面上增殖随时间的变化。∆ε随时间的变化趋势与整个构建体的AlamarBlue检测结果一致，但这种测量方法提供了被包裹细胞状态的局部读数。这项研究的结果支持使用介电阻抗谱作为一种无标记和非破坏性的方法来表征生物打印构建物的关键质量属性。
Biofabrication processes can affect biological quality attributes of encapsulated cells within constructs. Currently, assessment of the fabricated constructs is performed offline by subjecting the constructs to destructive assays that require staining and sectioning. This drawback limits the translation of biofabrication processes to industrial practice. In this work, we investigate the dielectric response of viable cells encapsulated in bioprinted 3D hydrogel constructs to an applied alternating electric field as a label-free non-destructive monitoring approach. The relationship between β-dispersion parameters (permittivity change—Δε, Cole–Cole slope factor—α, critical polarization frequency—f c) over the frequency spectrum and critical cellular quality attributes are investigated. Results show that alginate constructs containing a higher number of viable cells (human adipose derived stem cells—hASC and osteosarcoma cell line—MG63) were characterized by significantly higher Δε and α (both p< 0.05). When extended to bioprinting, results showed that changes in hASC proliferation and viability in response to changes in critical bioprinting parameters (extrusion pressure, temperature, processing time) significantly affected∆ ε, α, and f c. We also demonstrated monitoring of hASC distribution after bioprinting and changes in proliferation over time across the cross-section of a bioprinted medial knee meniscus construct. The trends in∆ ε over time were in agreement with the alamarBlue assay results for the whole construct, but this measurement approach provided a localized readout on the status of encapsulated cells. The findings of this study support the use of dielectric impedance spectroscopy as a label-free and non-destructive method to characterize the critical quality attributes of bioprinted constructs.
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