Online quantitative monitoring of live cell engineered cartilage growth using diffuse fiber-optic Raman spectroscopy.
Online quantitative monitoring of live cell engineered cartilage growth using diffuse fiber-optic Raman spectroscopy.
复制标题
DOI：
10.1016/j.biomaterials.2017.06.015
复制
发表时间：
2017-09
期刊：
影响因子：
14
通讯作者：
Stevens MM
中科院分区：
文献类型：
--
作者：
Bergholt MS;Albro MB;Stevens MM
组织工程（TE）有可能改善骨关节炎（OA）患者的结局。该技术作为治疗的一部分的成功临床转化需要体外测量工程化组织的细胞外基质（ECM）产生的能力，以确保质量控制并提高植入后组织存活的可能性。用于评估工程化软骨的ECM含量的常规技术，例如生化测定和组织学染色固有地具有破坏性。另一方面，拉曼光谱代表了用于原位生物化学表征的非侵入性技术。在这里，我们概述了当前TE中平移拉曼光谱的障碍，并介绍了一个全面的工作流程，旨在非破坏性地监测和量化大（>3 mm）活细胞TE构建体中的ECM生物分子。在56天的培养期内，从活细胞软骨TE构建体测量漫射近红外光纤拉曼光谱。我们开发了一个多变量曲线分辨率模型，使定量生化分析的TE结构。拉曼光谱能够非侵入性地定量ECM组分，并且显示出与用于测量胶原蛋白（R2 = 0.84）和糖胺聚糖（GAG）（R2 = 0.86）的生化测定的极好的相关性。我们进一步证明了该技术用于活细胞TE构建体的在线前瞻性分析的稳健性。在这项工作中开发的光纤拉曼光谱策略提供了在线非破坏性监测构建生长的能力，并且可以适应再生医学中广泛的TE应用，以控制临床转化。
Tissue engineering (TE) has the potential to improve the outcome for patients with osteoarthritis (OA). The successful clinical translation of this technique as part of a therapy requires the ability to measure extracellular matrix (ECM) production of engineered tissues in vitro, in order to ensure quality control and improve the likelihood of tissue survival upon implantation. Conventional techniques for assessing the ECM content of engineered cartilage, such as biochemical assays and histological staining are inherently destructive. Raman spectroscopy, on the other hand, represents a non-invasive technique for in situ biochemical characterization. Here, we outline current roadblocks in translational Raman spectroscopy in TE and introduce a comprehensive workflow designed to non-destructively monitor and quantify ECM biomolecules in large (>3 mm), live cell TE constructs online. Diffuse near-infrared fiber-optic Raman spectra were measured from live cell cartilaginous TE constructs over a 56-day culturing period. We developed a multivariate curve resolution model that enabled quantitative biochemical analysis of the TE constructs. Raman spectroscopy was able to non-invasively quantify the ECM components and showed an excellent correlation with biochemical assays for measurement of collagen (R2 = 0.84) and glycosaminoglycans (GAGs) (R2 = 0.86). We further demonstrated the robustness of this technique for online prospective analysis of live cell TE constructs. The fiber-optic Raman spectroscopy strategy developed in this work offers the ability to non-destructively monitor construct growth online and can be adapted to a broad range of TE applications in regenerative medicine toward controlled clinical translation.
登录
查看更多内容
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
影响因子：
3.5
作者：
Ebert, Daniel W.;Roberts, Cynthia;Bertone, Alicia L.
通讯作者：
Bertone, Alicia L.
影响因子：
6.2
作者：
Palukuru, Uday P.;Hanifi, Arash;Pleshko, Nancy
通讯作者：
Pleshko, Nancy
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：
2.5
作者：
Gamsjaeger, Sonja;Klaushofer, Klaus;Paschalis, Eleftherios P.
通讯作者：
Paschalis, Eleftherios P.