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生物反应器培养条件可调节组织工程软骨的组成和力学性能。

Bioreactor cultivation conditions modulate the composition and mechanical properties of tissue-engineered cartilage.

作者信息

Vunjak-Novakovic G, Martin I, Obradovic B, Treppo S, Grodzinsky A J, Langer R, Freed L E

机构信息

Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge 02139, USA.

出版信息

J Orthop Res. 1999 Jan;17(1):130-8. doi: 10.1002/jor.1100170119.

Abstract

Cartilaginous constructs have been grown in vitro with use of isolated cells, biodegradable polymer scaffolds, and bioreactors. In the present work, the relationships between the composition and mechanical properties of engineered cartilage constructs were studied by culturing bovine calf articular chondrocytes on fibrous polyglycolic acid scaffolds (5 mm in diameter, 2-mm thick, and 97% porous) in three different environments: static flasks, mixed flasks, and rotating vessels. After 6 weeks of cultivation, the composition, morphology, and mechanical function of the constructs in radially confined static and dynamic compression all depended on the conditions of in vitro cultivation. Static culture yielded small and fragile constructs, while turbulent flow in mixed flasks yielded constructs with fibrous outer capsules; both environments resulted in constructs with poor mechanical properties. The constructs that were cultured freely suspended in a dynamic laminar flow field in rotating vessels were the largest, contained continuous cartilage-like extracellular matrices with the highest fractions of glycosaminoglycan and collagen, and had the best mechanical properties. The equilibrium modulus, hydraulic permeability, dynamic stiffness, and streaming potential correlated with the wet-weight fractions of glycosaminoglycan, collagen, and water. These findings suggest that the hydrodynamic conditions in tissue-culture bioreactors can modulate the composition, morphology, mechanical properties, and electromechanical function of engineered cartilage.

摘要

软骨构建物已通过使用分离的细胞、可生物降解的聚合物支架和生物反应器在体外培养而成。在本研究中,通过在三种不同环境中,将牛犊关节软骨细胞培养在纤维状聚乙醇酸支架(直径5毫米、厚2毫米、孔隙率97%)上,研究了工程化软骨构建物的组成与力学性能之间的关系。这三种环境分别是:静态培养瓶、混合培养瓶和旋转容器。培养6周后,在径向受限的静态和动态压缩条件下,构建物的组成、形态和力学功能均取决于体外培养条件。静态培养产生的构建物小且易碎,而混合培养瓶中的湍流产生的构建物带有纤维状外膜;这两种环境产生的构建物力学性能都很差。在旋转容器中动态层流场中自由悬浮培养的构建物最大,含有连续的软骨样细胞外基质,糖胺聚糖和胶原蛋白的比例最高,并且具有最佳的力学性能。平衡模量、水力渗透率、动态刚度和流动电位与糖胺聚糖、胶原蛋白和水的湿重分数相关。这些发现表明,组织培养生物反应器中的流体动力学条件可以调节工程化软骨的组成、形态、力学性能和机电功能。

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