Preparation and evaluation of molecularly-defined collagen-elastin-glycosaminoglycan scaffolds for tissue engineering.

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.

[1]  D. Courtman,et al.  The role of crosslinking in modification of the immune response elicited against xenogenic vascular acellular matrices. , 2001, Journal of biomedical materials research.

[2]  I. Yannas,et al.  Design of an artificial skin. II. Control of chemical composition. , 1980, Journal of biomedical materials research.

[3]  Antonios G. Mikos,et al.  Growth Factor Delivery for Tissue Engineering , 2000, Pharmaceutical Research.

[4]  A Oosterhof,et al.  Preparation and characterization of porous crosslinked collagenous matrices containing bioavailable chondroitin sulphate. , 1999, Biomaterials.

[5]  D. Ducassou,et al.  In vitro association of type III collagen with elastin and with its solubilized peptides. , 1991, Biomaterials.

[6]  J. Feijen,et al.  Cross-linking of dermal sheep collagen using a water-soluble carbodiimide. , 1996, Biomaterials.

[7]  Anuj Singla,et al.  Effect of elastin on the calcification rate of collagen-elastin matrix systems. , 2002, Journal of biomedical materials research.

[8]  J. Veerkamp,et al.  Immunoquantification of type I, III, IV and V collagen in small samples of human lung parenchyma. , 1995, The international journal of biochemistry & cell biology.

[9]  W. Nichols McDonald's Blood Flow in Arteries , 1996 .

[10]  J. Bancroft,et al.  Theory and Practice of Histological Techniques , 1990 .

[11]  J. Veerkamp,et al.  Comparison of five procedures for the purification of insoluble elastin. , 2001, Biomaterials.

[12]  L. Debelle,et al.  Elastin: molecular description and function. , 1999, The international journal of biochemistry & cell biology.

[13]  F J Schoen,et al.  Founder's Award, 25th Annual Meeting of the Society for Biomaterials, perspectives. Providence, RI, April 28-May 2, 1999. Tissue heart valves: current challenges and future research perspectives. , 1999, Journal of biomedical materials research.

[14]  A A Poot,et al.  Improved endothelialization of vascular grafts by local release of growth factor from heparinized collagen matrices. , 1998, Journal of controlled release : official journal of the Controlled Release Society.

[15]  G. Goissis,et al.  Preparation and characterization of collagen-elastin matrices from blood vessels intended as small diameter vascular grafts. , 2000, Artificial organs.

[16]  Yugyung Lee,et al.  Biomedical applications of collagen. , 2001, International journal of pharmaceutics.

[17]  M. Aprahamian,et al.  A new reconstituted connective tissue matrix: preparation, biochemical, structural and mechanical studies. , 1987, Journal of biomedical materials research.

[18]  A A Poot,et al.  Binding and release of basic fibroblast growth factor from heparinized collagen matrices. , 2001, Biomaterials.

[19]  Beat Steinmann,et al.  Connective tissue and its heritable disorders —Molecular, genetic and medical aspects , 1993 .

[20]  J. Veerkamp,et al.  The biochemical and structural maturation of human skeletal muscle cells in culture: the effect of the serum substitute Ultroser G. , 1991, Experimental cell research.

[21]  H. Watanabe,et al.  Roles of aggrecan, a large chondroitin sulfate proteoglycan, in cartilage structure and function. , 1998, Journal of biochemistry.

[22]  R. Iozzo Matrix proteoglycans: from molecular design to cellular function. , 1998, Annual review of biochemistry.

[23]  S. Selleck,et al.  Order out of chaos: assembly of ligand binding sites in heparan sulfate. , 2002, Annual review of biochemistry.

[24]  J. Veerkamp,et al.  Development of tailor-made collagen-glycosaminoglycan matrices: EDC/NHS crosslinking, and ultrastructural aspects. , 2000, Biomaterials.

[25]  J. Veerkamp,et al.  Loading of collagen-heparan sulfate matrices with bFGF promotes angiogenesis and tissue generation in rats. , 2002, Journal of biomedical materials research.