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Bioorthogonal Modification of Cell Derived Matrices by Metabolic Glycoengineering
ACS Biomaterials Science & Engineering ( IF 5.8 ) Pub Date : 2018-03-13 00:00:00 , DOI: 10.1021/acsbiomaterials.8b00264
Marcus Gutmann 1 , Alexandra Braun 1 , Jürgen Seibel 2 , Tessa Lühmann 1
Affiliation  

Cell-derived matrices (CDMs) emerged as an attractive biomaterial in regenerative medicine. Here we present a strategy for site-specific decoration of CDMs with bioactive molecules deploying metabolic glycoengineering. NIH 3T3 fibroblasts were cultured in the presence of a tetra acetylated azide bearing monosaccharide to metabolically incorporate the synthetic sugar into the glycan structure of extracellular matrix proteins. Glycoengineered CDMs were isolated and analyzed for fibronectin as one of the most abundant ECM species by western blotting and fluorescence labeling of the azide-monosaccharides deploying bioorthogonal chemistries. Glycoengineered CDMs were biocompatible for incorporated and reseeded NIH 3T3 fibroblasts, respectively. Successful modification of glycoengineered CDMs was demonstrated by a therapeutic peptide inhibitor against myostatin. This study details a site-specific, effective, and biocompatible strategy for the decoration of glycan structures within CDMs for future tissue engineering application.

中文翻译:

代谢糖工程技术对细胞衍生基质的生物正交修饰

细胞衍生基质(CDM)在再生医学中成为一种有吸引力的生物材料。在这里,我们提出了利用生物活性分子进行代谢糖工程技术对CDM进行位点修饰的策略。在带有单糖的四乙酰化叠氮化物的存在下培养NIH 3T3成纤维细胞,以将合成糖代谢整合到细胞外基质蛋白的聚糖结构中。通过蛋白质印迹和部署生物正交化学的叠氮化物-单糖的荧光标记,分离了糖工程化的CDM,并分析了纤连蛋白作为最丰富的ECM物种之一。糖工程化的CDM对掺入和再播种的NIH 3T3成纤维细胞具有生物相容性。抗肌生长抑制素的治疗性肽抑制剂证明了糖基改造的CDM的成功修饰。这项研究详细介绍了针对CDM内聚糖结构装饰的针对特定地点,有效且具有生物相容性的策略,以供将来的组织工程应用。
更新日期:2018-03-13
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