Raman spectroscopic imaging for quantification of depth-dependent and local heterogeneities in native and engineered cartilage.
Raman spectroscopic imaging for quantification of depth-dependent and local heterogeneities in native and engineered cartilage.
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DOI：
10.1038/s41536-018-0042-7
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发表时间：
2018
影响因子：
7.2
通讯作者：
Stevens MM
中科院分区：
文献类型：
--
作者：
Albro MB;Bergholt MS;St-Pierre JP;Vinals Guitart A;Zlotnick HM;Evita EG;Stevens MM
关节软骨具有显著的、机械稳健的细胞外基质（ECM），其被组织并分布在整个组织中以抵抗生理应变并在关节运动期间提供低摩擦。表征软骨ECM的组成和分布的能力对于理解关节软骨经历疾病相关变性的过程和开发新的组织修复策略以恢复组织功能至关重要。然而，定量测量软骨ECM成分在整个组织中的空间分布的能力仍然是一个主要的挑战。在这项实验研究中，我们评估了拉曼显微光谱成像的分析能力，半定量测量软骨组织中主要ECM成分的分布。获得两种不同软骨组织类型的拉曼光谱图像，所述软骨组织类型在其整个深度上具有大的空间ECM梯度：天然关节软骨外植体和大的工程化软骨组织构建体。通过多变量曲线分辨率处理光谱采集，将“指纹”范围光谱（800-1800 cm-1）分解为GAG、胶原和水的组分光谱，得到整个组织中每种组分的深度依赖性浓度曲线。这些拉曼光谱获得的配置文件表现出强烈的协议，通过空间组织切片的直接生化测定独立获得的配置文件。此外，我们利用这种光谱技术来评估通过软骨的深度局部异质性。这项工作代表了一个强大的分析验证拉曼光谱成像测量的生物组织中的生化成分的空间分布的准确性，并表明它可以被用作一个有价值的工具，用于定量测量的分布和组织的ECM成分在本地和工程软骨组织标本。结合成像和统计方法可以更好地了解骨关节炎如何发展，并改善软骨工程策略。伦敦帝国理工学院的Molly Stevens及其同事使用拉曼光谱成像技术定量测量了天然和工程软骨中细胞外基质（ECM）成分的空间分布，这些成分在组织的不同部分之间存在差异。在拉曼光谱学中，来自照射在组织样本上的激光的光子被与它们相互作用的分子以不同程度反射。该团队测量了这些反射，并使用称为多元曲线分辨率的统计技术将它们彼此区分开来。该方法成功地测量了两种类型软骨的ECM中糖胺聚糖链、胶原和水的分布。该方法可以帮助研究人员更多地了解组织退行性疾病，并开发更好的修复和替换组织工程的策略。
Articular cartilage possesses a remarkable, mechanically-robust extracellular matrix (ECM) that is organized and distributed throughout the tissue to resist physiologic strains and provide low friction during articulation. The ability to characterize the make-up and distribution of the cartilage ECM is critical to both understand the process by which articular cartilage undergoes disease-related degeneration and to develop novel tissue repair strategies to restore tissue functionality. However, the ability to quantitatively measure the spatial distribution of cartilage ECM constituents throughout the tissue has remained a major challenge. In this experimental investigation, we assessed the analytical ability of Raman micro-spectroscopic imaging to semi-quantitatively measure the distribution of the major ECM constituents in cartilage tissues. Raman spectroscopic images were acquired of two distinct cartilage tissue types that possess large spatial ECM gradients throughout their depth: native articular cartilage explants and large engineered cartilage tissue constructs. Spectral acquisitions were processed via multivariate curve resolution to decompose the “fingerprint” range spectra (800–1800 cm−1) to the component spectra of GAG, collagen, and water, giving rise to the depth dependent concentration profile of each constituent throughout the tissues. These Raman spectroscopic acquired-profiles exhibited strong agreement with profiles independently acquired via direct biochemical assaying of spatial tissue sections. Further, we harness this spectroscopic technique to evaluate local heterogeneities through the depth of cartilage. This work represents a powerful analytical validation of the accuracy of Raman spectroscopic imaging measurements of the spatial distribution of biochemical components in a biological tissue and shows that it can be used as a valuable tool for quantitatively measuring the distribution and organization of ECM constituents in native and engineered cartilage tissue specimens. A combined imaging and statistical approach could lead to a better understanding of how osteoarthritis develops and to improved cartilage engineering strategies. Molly Stevens and colleagues at Imperial College London used Raman spectroscopic imaging to quantitatively measure the spatial distribution of extracellular matrix (ECM) components in natural and engineered cartilage, which varies from one part of the tissue to another. In Raman spectroscopy, photons from a laser shined on a tissue sample are reflected in varying degrees by the molecules they interact with. The team measured these reflections and distinguished them from one another using a statistical technique called multivariate curve resolution. The method successfully measured the distribution of glycosaminoglycan chains, collagen and water in the ECM of both types of cartilage. The method could help researchers learn more about tissue degenerative diseases and develop better strategies for engineering tissues for repair and replacement.
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影响因子：
18.2
作者：
Bergholt, Mads S.;St-Pierre, Jean-Philippe;Offeddu, Giovanni S.;Parmar, Paresh A.;Albro, Michael B.;Puetzer, Jennifer L.;Oyen, Michelle L.;Stevens, Molly M.
通讯作者：
Stevens, Molly M.
影响因子：
7
作者：
Bian L;Angione SL;Ng KW;Lima EG;Williams DY;Mao DQ;Ateshian GA;Hung CT
通讯作者：
Hung CT
短暂暴露于 TGF-β3 可改善装载 MSC 的透明质酸水凝胶的功能性软骨形成。
DOI：
10.1016/j.jmbbm.2012.03.006
发表时间：
2012-07
影响因子：
3.9
作者：
Kim M;Erickson IE;Choudhury M;Pleshko N;Mauck RL
通讯作者：
Mauck RL
DOI：
10.1016/0304-4165(86)90306-5
发表时间：
1986-09-04
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
FARNDALE, RW;BUTTLE, DJ;BARRETT, AJ
通讯作者：
BARRETT, AJ
影响因子：
3.5
作者：
Kim, M;Bi, XH;Camacho, NP
通讯作者：
Camacho, NP