Department of Bioengineering, Rice University, MS-142, PO Box 1892, Houston, TX 77251, USA.
Biomaterials. 2010 Dec;31(34):8911-20. doi: 10.1016/j.biomaterials.2010.07.110. Epub 2010 Aug 24.
In this study, electrospun poly(ɛ-caprolactone) (PCL) microfiber scaffolds, coated with cartilaginous extracellular matrix (ECM), were fabricated by first culturing chondrocytes under dynamic conditions in a flow perfusion bioreactor and then decellularizing the cellular constructs. The decellularization procedure yielded acellular PCL/ECM composite scaffolds containing glycosaminoglycan and collagen. PCL/ECM composite scaffolds were evaluated for their ability to support the chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro using serum-free medium with or without the addition of transforming growth factor-β1 (TGF-β1). PCL/ECM composite scaffolds supported chondrogenic differentiation induced by TGF-β1 exposure, as evidenced in the up-regulation of aggrecan (11.6 ± 3.8 fold) and collagen type II (668.4 ± 317.7 fold) gene expression. The presence of cartilaginous matrix alone reduced collagen type I gene expression to levels observed with TGF-β1 treatment. Cartilaginous matrix further enhanced the effects of growth factor treatment on MSC chondrogenesis as evidenced in the higher glycosaminoglycan synthetic activity for cells cultured on PCL/ECM composite scaffolds. Therefore, flow perfusion culture of chondrocytes on electrospun microfiber scaffolds is a promising method to fabricate polymer/extracellular matrix composite scaffolds that incorporate both natural and synthetic components to provide biological signals for cartilage tissue engineering applications.
在这项研究中,通过在流灌注生物反应器中动态培养软骨细胞,然后对细胞构建体进行脱细胞化,制备了包被软骨细胞外基质 (ECM) 的电纺聚己内酯 (PCL) 微纤维支架。脱细胞化过程产生了含有糖胺聚糖和胶原蛋白的无细胞 PCL/ECM 复合材料支架。使用无血清培养基,评估了 PCL/ECM 复合材料支架在添加或不添加转化生长因子-β1 (TGF-β1) 的情况下,体外支持间充质干细胞 (MSCs) 软骨分化的能力。PCL/ECM 复合材料支架支持 TGF-β1 暴露诱导的软骨分化,表现在聚集蛋白聚糖 (11.6 ± 3.8 倍) 和 II 型胶原 (668.4 ± 317.7 倍) 基因表达上调。单独存在软骨基质可将 I 型胶原基因表达降低至 TGF-β1 处理时的水平。软骨基质进一步增强了生长因子处理对 MSC 软骨形成的影响,表现在培养在 PCL/ECM 复合材料支架上的细胞的糖胺聚糖合成活性更高。因此,在静电纺丝微纤维支架上对软骨细胞进行流灌注培养是一种很有前途的方法,可制备包含天然和合成成分的聚合物/细胞外基质复合材料支架,为软骨组织工程应用提供生物信号。