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首页> 外文期刊>Acta biomaterialia >Enzymatically cross-linked gelatin-phenol hydrogels with a broader stiffness range for osteogenic differentiation of human mesenchymal stem cells
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Enzymatically cross-linked gelatin-phenol hydrogels with a broader stiffness range for osteogenic differentiation of human mesenchymal stem cells

机译：具有更大硬度范围的酶交联明胶-酚水凝胶，用于人间充质干细胞的成骨分化

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An injectable hydrogel system, composed of gelatin-hydroxyphenylpropionic acid (Gtn-HPA) conjugates chemically cross-linked by an enzyme-mediated oxidation reaction, has been designed as a biodegradable scaffold for tissue engineering. In light of the role of substrate stiffness on cell differentiation, we herein report a newly improved Gtn hydrogel system with a broader range of stiffness control that uses Gtn-HPA-tyramine (Gtn-HPA-Tyr) conjugates to stimulate the osteogenic differentiation of human mesenchymal stem cells (hMSCs). The Gtn-HPA-Tyr conjugate was successfully synthesized through a further conjugation of Tyr to Gtn-HPA conjugate by means of a general carbodiimide/active ester-mediated coupling reaction. Proton nuclear magnetic resonance and UV-visible measurements showed a higher total phenol content in the Gtn-HPA-Tyr conjugate than that content in the Gtn-HPA conjugate. The Gtn-HPA-Tyr hydrogels were formed by the oxidative coupling of phenol moieties catalyzed by hydrogen peroxide (H 2O 2) and horseradish peroxidase (HRP). Rheological studies revealed that a broader range of storage modulus (G′) of Gtn-HPA-Tyr hydrogel (600-26,800 Pa) was achieved using different concentrations of H 2O 2, while the G′ of the predecessor Gtn-HPA hydrogels was limited to the range of 1000 to 13,500 Pa. The hMSCs on Gtn-HPA-Tyr hydrogel with G′ greater than 20,000 showed significantly up-regulated expressions of osteocalcin and runt-related transcription factor 2 (RUNX2) on both the gene and protein level, with the presence of alkaline phosphatase, and the evidence of calcium accumulation. These studies with the Gtn-HPA-Tyr hydrogel with G′ greater than 20,000 collectively suggest the stimulation of the hMSCs into osteogenic differentiation, while these same observations were not found with the Gtn-HPA hydrogel with a G′ of 13,500.
机译：由可通过酶介导的氧化反应化学交联的明胶-羟苯基丙酸（Gtn-HPA）共轭物组成的可注射水凝胶系统已被设计为可生物降解的组织工程支架。考虑到底物刚度在细胞分化中的作用，我们在此报告了一种新的改良的Gtn水凝胶系统，该系统具有更大范围的刚度控制，该系统使用Gtn-HPA-酪胺（Gtn-HPA-Tyr）共轭物刺激人的成骨分化间充质干细胞（hMSCs）。通过一般的碳二亚胺/活性酯介导的偶联反应将Tyr进一步偶联至Gtn-HPA偶联物，成功地合成了Gtn-HPA-Tyr偶联物。质子核磁共振和紫外可见测量显示，Gtn-HPA-Tyr共轭物中的总酚含量高于Gtn-HPA共轭物中的总酚含量。 Gtn-HPA-Tyr水凝胶是由过氧化氢（H 2O 2）和辣根过氧化物酶（HRP）催化的酚部分氧化偶联形成的。流变学研究表明，使用不同浓度的H 2O 2可获得更大范围的Gtn-HPA-Tyr水凝胶（600-26,800 Pa）储能模量（G'），而以前的Gtn-HPA水凝胶的G'受限制GMSC大于20,000的Gtn-HPA-Tyr水凝胶上的hMSC在基因和蛋白质水平上均显着上调了骨钙素和与矮子相关的转录因子2（RUNX2）的表达。碱性磷酸酶的存在，以及钙积累的证据。这些对G'大于20,000的Gtn-HPA-Tyr水凝胶的研究共同表明，刺激hMSCs成骨分化，而对G'为13,500的Gtn-HPA水凝胶没有发现相同的观察结果。

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