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用于填充关节软骨损伤的生物复合材料的研发。在I/III型胶原海绵上培养的绵羊软骨细胞的光学、扫描和透射电子显微镜观察。

Development of a biocomposite to fill out articular cartilage lesions. Light, scanning and transmission electron microscopy of sheep chondrocytes cultured on a collagen I/III sponge.

作者信息

Ehlers E M, Fuss M, Rohwedel J, Russlies M, Kühnel W, Behrens P

机构信息

Institut fur Anatomie, Medizinischen Universität zu Lübeck, Germany.

出版信息

Ann Anat. 1999 Dec;181(6):513-8. doi: 10.1016/S0940-9602(99)80055-7.

Abstract

The regenerative capacity of hyaline articular cartilage is limited. Thus, lesions of this tissue are a proarthrotic factor, and up to now the conservative treatment of cartilage lesions and arthrosis does not yield satisfying results. Therefore, autologous transplantation of articular chondrocytes is being investigated in a variety of different assays. The aim of our study was to create a mechanically stable cell-matrix implant with viable and active chondrocytes which could serve to fill out articular lesions created in the knees of sheep. For this purpose, articular cartilage was collected from knee lesions, chondrocytes were liberated enzymatically and seeded in culture flasks and cultured till confluency. Cells were then trypsinized and grown on a type I/III collagen matrix (Chondro-Gide, Geistlich Biomaterials, Wolhusen, Switzerland) for 3, 6 and 10 days before being fixed and embedded for electron microscopy by routine methods. Scanning electron microscopy was performed after dehydration in acetone, critical point drying and sputter-coating with gold-paladium. Light microscopically, clusters of chondrocytes can be seen on the surface of the matrix with a few cells growing into the matrix. Transmission electron microscopic photographs yield a rather differentiated chondrocyte-like appearance, which is evidence of a matrix-induced redifferentiation after dedifferentiation during the growth period in the culture flasks. Scanning electron microscopic results show large, flattened chondrocytes without signs of differentiation on plastic, whereas chondrocytes grown on the Chondro-Gide sponge show a more roundish aspect wrapping firmly around the collagen fibrils, exhibiting numerous contacts with the matrix. This cell-matrix biocomposite can now serve to fill out articular cartilage lesions created in the knees of sheep.

摘要

透明关节软骨的再生能力有限。因此,这种组织的损伤是一个促关节病变因素,到目前为止,软骨损伤和关节炎的保守治疗效果并不理想。因此,正在通过各种不同的试验研究自体关节软骨细胞移植。我们研究的目的是创建一种机械稳定的细胞 - 基质植入物,其中含有存活且活跃的软骨细胞,可用于填充绵羊膝关节中形成的关节损伤。为此,从膝关节损伤处采集关节软骨,酶解分离出软骨细胞,接种到培养瓶中培养至汇合。然后将细胞用胰蛋白酶处理,并在I/III型胶原基质(Chondro - Gide,Geistlich Biomaterials,瑞士沃尔胡森)上培养3天、6天和10天,之后通过常规方法固定并包埋用于电子显微镜检查。在丙酮中脱水、临界点干燥并用金 - 钯溅射镀膜后进行扫描电子显微镜检查。在光学显微镜下,可以看到基质表面有软骨细胞簇,有一些细胞生长到基质中。透射电子显微镜照片显示出相当分化的软骨细胞样外观,这证明在培养瓶中生长期间去分化后,基质诱导了再分化。扫描电子显微镜结果显示,在塑料上的大而扁平的软骨细胞没有分化迹象,而在Chondro - Gide海绵上生长的软骨细胞呈现出更圆的形态,紧紧包裹在胶原纤维周围,与基质有大量接触。这种细胞 - 基质生物复合材料现在可用于填充绵羊膝关节中形成的关节软骨损伤。

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