Russlies M, Behrens P, Wünsch L, Gille J, Ehlers E M
Department of Orthopaedic Surgery, University of Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
Ann Anat. 2002 Jul;184(4):317-23. doi: 10.1016/S0940-9602(02)80045-0.
Chondrocytes in monolayer cultures lose their phenotype and capability to express type-II collagen, they dedifferentiate into a fibroblastic cell type. Using three-dimensional culture systems a redifferentiation of these cells may occur. In the present study we investigated the morphology and biosynthetic activity of human articular chondrocytes seeded on porous matrices of type I/III collagen (Chondrogide, Geistlich Biomaterials, Wolhusen, Switzerland). Microscopical examinations showed that chondrocytes adhere firmly to a collagen-I/III-membrane exhibiting their characteristic spherical cell shape. Cell numbers after enzymatic digestion of the membrane showed a 93% recovery of seeded cells. Immunohistological examination revealed positive staining for type-II collagen in some areas. The generated biocomposite withstands mechanical stress, keeps its size and design and does not shrink in culture. It is therefore easy to handle, can be sutured, glued or fixed with pins. This study shows, that in vitro production of autologous cartilage-like tissue could be established using a bilayer collagen type I/III fleece. This biocomposite carries active chondrocytes and is currently being evaluated in vivo in a sheep model as well as in a clinical trial for the repair of localized cartilage defects in the knee.
单层培养的软骨细胞会丧失其表型以及表达II型胶原蛋白的能力,它们会去分化为成纤维细胞类型。使用三维培养系统,这些细胞可能会发生再分化。在本研究中,我们调查了接种于I/III型胶原蛋白多孔基质(Chondrogide,Geistlich Biomaterials,瑞士沃尔胡森)上的人关节软骨细胞的形态和生物合成活性。显微镜检查显示,软骨细胞牢固地附着在I/III型胶原膜上,呈现出其特有的球形细胞形态。对膜进行酶消化后的细胞数量显示,接种细胞的回收率为93%。免疫组织学检查显示某些区域有II型胶原蛋白的阳性染色。生成的生物复合材料能够承受机械应力,保持其大小和设计,在培养中不会收缩。因此,它易于操作,可以缝合、粘贴或用针固定。本研究表明,使用双层I/III型胶原绒可以在体外建立自体软骨样组织的生产。这种生物复合材料携带活性软骨细胞,目前正在绵羊模型中进行体内评估,以及在一项修复膝关节局部软骨缺损的临床试验中进行评估。