Label-Free Quantification of Microscopic Alignment in Engineered Tissue Scaffolds by Polarized Raman Spectroscopy
Label-Free Quantification of Microscopic Alignment in Engineered Tissue Scaffolds by Polarized Raman Spectroscopy
复制标题
通过偏振拉曼光谱对工程组织支架中的微观排列进行无标记定量
DOI：
10.1021/acsbiomaterials.3c00242
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发表时间：
2023
影响因子：
5.8
通讯作者：
Subhash, Ghatu
中科院分区：
文献类型：
--
作者：
Zhou, Hui;Llanes, Janny Piñeiro;Lotfi, Maedeh;Sarntinoranont, Malisa;Simmons, Chelsey S.;Subhash, Ghatu
细胞外基质（ECM）微观结构的监测是必不可少的，在研究结构相关的细胞过程，改善细胞功能，并确保足够的机械完整性，在工程组织。本文描述了一种研究工程组织支架（ETS）中基质微观排列的新方法，这些支架通常由细胞衍生的各种生物大分子组成。首先，通过主成分分析（PCA）从高度对准的天然组织的拉曼光谱导出训练的加载函数，其中显著的变化与特定的拉曼带（例如，1444、1465、1605、1627-1660和1665-1689 cm-1）。这些变化主要是由组织内许多化合物相对于激光偏振的排列基质引起的，包括蛋白质、脂质和碳水化合物。因此，该训练函数被应用于量化由细胞衍生的各种基质组分在ETS内的比对。此外，一个简单的度量称为振幅排列度量（AAM）的相关性的偏振拉曼光谱的ETS的矩阵排列的程度的取向依赖。结果发现，AAM是显着较高的各向异性ETS比各向同性的。PRS方法揭示了一个较低的p值区分这两种类型的ETS之间的对齐相比，在类似的显微镜尺度检测荧光标记的蛋白质基质的显微镜方法。这些结果表明，在工程组织中的复杂基质的各向异性可以评估在微观尺度上使用基于PRS的简单的度量，这是上级的传统的显微镜方法。这种基于PRS的方法可以作为一种辅助工具，用于设计和评估模仿天然基质组织微结构的工程组织。
Monitoring of extracellular matrix (ECM) microstructure is essential in studying structure-associated cellular processes, improving cellular function, and for ensuring sufficient mechanical integrity in engineered tissues. This paper describes a novel method to study the microscale alignment of the matrix in engineered tissue scaffolds (ETS) that are usually composed of a variety of biomacromolecules derived by cells. First, a trained loading function was derived from Raman spectra of highly aligned native tissue via principal component analysis (PCA), where prominent changes associated with specific Raman bands (e.g., 1444, 1465, 1605, 1627–1660, and 1665–1689 cm–1) were detected with respect to the polarization angle. These changes were mainly caused by the aligned matrix of many compounds within the tissue relative to the laser polarization, including proteins, lipids, and carbohydrates. Hence this trained function was applied to quantify the alignment within ETS of various matrix components derived by cells. Furthermore, a simple metric called Amplitude Alignment Metric (AAM) was derived to correlate the orientation dependence of polarized Raman spectra of ETS to the degree of matrix alignment. It was found that the AAM was significantly higher in anisotropic ETS than isotropic ones. The PRS method revealed a lowerp-value for distinguishing the alignment between these two types of ETS as compared to the microscopic method for detecting fluorescent-labeled protein matrices at a similar microscopic scale. These results indicate that the anisotropy of a complex matrix in engineered tissue can be assessed at the microscopic scale using a PRS-based simple metric, which is superior to the traditional microscopic method. This PRS-based method can serve as a complementary tool for the design and assessment of engineered tissues that mimic the native matrix organizational microstructures.