Mahsa Khatami Seyedeh, Parivar Kazem, Naderi Sohi Alireza, Soleimani Masoud, Hanaee-Ahvaz Hana
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Stem Cell Technology Research Center, Tehran, Iran.
Tissue Cell. 2020 Aug;65:101363. doi: 10.1016/j.tice.2020.101363. Epub 2020 Apr 11.
Construction of scaffolds which are similar to natural niches regarding both biochemical composition and mechanical characteristics has gained great attention in the field of tissue engineering. However, application of natural polymers, such as hyaluronic acid, is challenging in construction of scaffolds due to physicochemical properties, difficult to use in electrospinning and low cell adhesion rate. In this study, HA was acetylated to make it soluble in high polarity solvent and blended with PCL for construction of nanofibrous composite (ac-HA/PCL) scaffolds. Chondroinductivity of the constructed scaffolds was investigated using human mesenchymal stem cells (hADSCs). The presence of acetyl groups, as well as morphology and biocompatibility of the composite scaffolds were characterized by HNMR, FTIR, SEM and MTT assay respectively. Expression of cartilage-specific genes (SOX9, Col II and Aggrecan) was monitored by Real-time PCR. Significant increase in expression of Sox9 and Col II as the markers of chondrogenic differentiation as well as the results of Alcian blue staining, indicated the chondro-inductive potential of HA/PCL nanofibrous scaffolds. Acetylated HA was biocompatible with chondroinductivity features, therefore it not only had the positive characteristics of natural HA, but also enhanced the cellular attachment and application potential.
在组织工程领域，构建在生化组成和力学特性方面都与天然微环境相似的支架受到了极大关注。然而，由于物理化学性质，天然聚合物（如透明质酸）在支架构建中应用具有挑战性，难以用于静电纺丝且细胞粘附率低。在本研究中，对透明质酸进行乙酰化处理使其可溶于高极性溶剂，并与聚己内酯共混以构建纳米纤维复合（ac-HA/PCL）支架。使用人间充质干细胞（hADSCs）研究构建支架的软骨诱导性。分别通过核磁共振氢谱（HNMR）、傅里叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）和MTT法对复合支架中乙酰基的存在以及形态和生物相容性进行表征。通过实时聚合酶链反应（Real-time PCR）监测软骨特异性基因（SOX9、II型胶原和聚集蛋白聚糖）的表达。作为软骨分化标志物的Sox9和II型胶原表达显著增加以及阿尔新蓝染色结果，表明HA/PCL纳米纤维支架具有软骨诱导潜力。乙酰化透明质酸具有生物相容性和软骨诱导特性，因此它不仅具有天然透明质酸的积极特性，还增强了细胞附着和应用潜力。
