Research Center for Experimental Orthopaedics, Orthopaedic University Hospital Heidelberg, Heidelberg, Germany.
Tissue Eng Part A. 2010 Dec;16(12):3697-707. doi: 10.1089/ten.TEA.2009.0817. Epub 2010 Sep 1.
Natural cartilage shows column orientation of cells and anisotropic direction of collagen fibers. However, matrices presently used in matrix-assisted autologous chondrocyte implantation do not show any fiber orientation. Our aim was to develop anisotropic scaffolds with parallel fiber orientation that were capable to support a cellular cartilaginous phenotype in vitro. Scaffolds were created by flock technology and consisted of a membrane of mineralized collagen type I as substrate, gelatine as adhesive, and parallel-oriented polyamide flock fibers vertically to the substrate. Confocal laser scan microscopy demonstrated that mesenchymal stem cells (MSCs) adhered and proliferated well in the scaffolds and cell vitality remained high over time. Articular chondrocytes seeded in a collagen type I gel into flock scaffolds deposited increasing amounts of proteoglycans and collagen type II over time. MSC-seeded flock scaffold constructs under chondrogenic conditions deposited significantly more proteoglycans and collagen type II than MSC collagen type I gel constructs only. Biomechanical testing revealed higher initial hardness of flock scaffolds than that of a clinically applied collagen type I/III scaffold combined with superior relaxation and an increasing hardness in MSC-loaded flock biocomposites during chondrogenesis. In conclusion, flock technology allows fabrication of scaffolds with anisotropic fiber orientation that mediates superior biomechanical and biochemical composition of tissue engineering constructs for cartilage repair.
天然软骨表现出细胞的柱状排列和胶原纤维的各向异性方向。然而,目前用于基质辅助自体软骨细胞移植的基质并不显示任何纤维取向。我们的目的是开发具有平行纤维取向的各向异性支架,能够在体外支持细胞软骨样表型。支架通过 flock 技术制备,由矿化 I 型胶原作为基底的膜、明胶作为粘合剂以及垂直于基底的平行取向的聚酰胺 flock 纤维组成。共聚焦激光扫描显微镜显示,间充质干细胞(MSCs)在支架中很好地黏附和增殖,并且细胞活力随时间保持较高水平。关节软骨细胞在 flock 支架中的 I 型胶原凝胶中接种后,随着时间的推移,不断地分泌更多的蛋白聚糖和 II 型胶原。在软骨生成条件下,与仅含有 MSC 的 I 型胶原凝胶支架相比,MSC flock 支架构建体显著增加了蛋白聚糖和 II 型胶原的沉积。生物力学测试表明,flock 支架的初始硬度高于临床应用的 I/III 型胶原支架,并且在软骨生成过程中,MSC 负载的 flock 生物复合材料的松弛度更高,硬度逐渐增加。总之,flock 技术允许制备具有各向异性纤维取向的支架,这种支架可以调节组织工程软骨修复构建体的优越生物力学和生化组成。