Daamen W F, van Moerkerk H Th B, Hafmans T, Buttafoco L, Poot A A, Veerkamp J H, van Kuppevelt T H
Department of Biochemistry 194, NCMLS, University Medical Centre Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
Biomaterials. 2003 Oct;24(22):4001-9. doi: 10.1016/s0142-9612(03)00273-4.
Extracellular matrix components are valuable building blocks for the preparation of biomaterials involved in tissue engineering, especially if their biological, chemical and physical characteristics can be controlled. In this study, isolated type I collagen fibrils, elastin fibres and chondroitin sulphate (CS) were used for the preparation of molecularly-defined collagen-elastin-glycosaminoglycan scaffolds. A total of 12 different scaffolds were prepared with four different ratios of collagen and elastin (1:9, 1:1, 9:1 and 1:0), with and without chemical crosslinking, and with and without CS. Collagen was essential to fabricate coherent, porous scaffolds. Electron microscopy showed that collagen and elastin physically interacted with each other and that elastin fibres were enveloped by collagen. By carbodiimide-crosslinking, amine groups were coupled to carboxylic groups and CS could be incorporated. More CS could be bound to collagen scaffolds (10%) than to collagen-elastin scaffolds (2.4-8.5% depending on the ratio). The attachment of CS increased the water-binding capacity to up to 65%. Scaffolds with a higher collagen content had a higher tensile strength whereas addition of elastin increased elasticity. Scaffolds were cytocompatible as was established using human myoblast and fibroblast culture systems. It is concluded that molecularly-defined composite scaffolds can be composed from individual, purified, extracellular matrix components. Data are important in the design and application of tailor-made biomaterials for tissue engineering.
细胞外基质成分是制备组织工程生物材料的宝贵构建单元，尤其是当它们的生物学、化学和物理特性能够得到控制时。在本研究中，分离出的I型胶原纤维、弹性纤维和硫酸软骨素（CS）被用于制备分子定义的胶原-弹性蛋白-糖胺聚糖支架。总共制备了12种不同的支架，胶原和弹性蛋白的比例有四种（1:9、1:1、9:1和1:0），有化学交联和无化学交联，有CS和无CS。胶原蛋白对于制造连贯的多孔支架至关重要。电子显微镜显示，胶原蛋白和弹性蛋白相互物理作用，弹性纤维被胶原蛋白包裹。通过碳二亚胺交联，胺基与羧基偶联，CS可以被掺入。与胶原-弹性蛋白支架（根据比例为2.4-8.5%）相比，更多的CS可以与胶原支架结合（10%）。CS的附着使水结合能力提高到65%。胶原蛋白含量较高的支架具有较高的拉伸强度，而添加弹性蛋白则增加了弹性。使用人类成肌细胞和成纤维细胞培养系统证实，支架具有细胞相容性。结论是，分子定义的复合支架可以由单个、纯化的细胞外基质成分组成。这些数据对于组织工程定制生物材料的设计和应用具有重要意义。
