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Isolation of intact elastin fibers devoid of microfibrils. 2005

Willeke F Daamen, and Theo Hafmans, and Jacques H Veerkamp, and Toin H van Kuppevelt
Department of Biochemistry, NCMLS, Radboud University Medical Centre, Nijmegen, The Netherlands.

Purification protocols for elastin generally result in greatly damaged elastin fibers and this likely influences the biological response. We here describe a novel protocol for the isolation of elastin whereby the fibers stay intact, and introduce the term "elastin fiber" for intact elastic fibers with elastin as their sole component. As opposed to elastic fibers, elastin fibers do not contain any microfibrils or associated molecules. Elastin fibers were isolated from equine elastic ligaments according to various protocols and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, amino acid quantification, immunofluorescence assay, transmission/scanning electron microscopy, and cellular reactivity in vivo. The optimal protocol comprised several extraction steps and trypsin digestion. Elastin fibers were free of contaminants and had a smooth, regular appearance. The cellular response to purified, intact elastin fibers was different in comparison with purified, but affected, fibers and to contaminated fibers. Intact fibers consisting only of elastin may be important for both fundamental and applied research, for example, tissue engineering, which need well-defined preparations to study the cellular biological effect of individual components.

UI MeSH Term Description Entries
D008022 Ligaments Shiny, flexible bands of fibrous tissue connecting together articular extremities of bones. They are pliant, tough, and inextensile. Interosseal Ligament,Interosseous Ligament,Interosseal Ligaments,Interosseous Ligaments,Ligament,Ligament, Interosseal,Ligament, Interosseous
D004549 Elastin alpha-Elastin,kappa-Elastin,alpha Elastin,kappa Elastin
D006736 Horses Large, hoofed mammals of the family EQUIDAE. Horses are active day and night with most of the day spent seeking and consuming food. Feeding peaks occur in the early morning and late afternoon, and there are several daily periods of rest. Equus caballus,Equus przewalskii,Horse, Domestic,Domestic Horse,Domestic Horses,Horse,Horses, Domestic
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001672 Biocompatible Materials Synthetic or natural materials, other than DRUGS, that are used to replace or repair any body TISSUES or bodily function. Biomaterials,Bioartificial Materials,Hemocompatible Materials,Bioartificial Material,Biocompatible Material,Biomaterial,Hemocompatible Material,Material, Bioartificial,Material, Biocompatible,Material, Hemocompatible
D020894 Microfibrils Components of the extracellular matrix consisting primarily of fibrillin. They are essential for the integrity of elastic fibers. Microfibril
D066298 In Vitro Techniques Methods to study reactions or processes taking place in an artificial environment outside the living organism. In Vitro Test,In Vitro Testing,In Vitro Tests,In Vitro as Topic,In Vitro,In Vitro Technique,In Vitro Testings,Technique, In Vitro,Techniques, In Vitro,Test, In Vitro,Testing, In Vitro,Testings, In Vitro,Tests, In Vitro,Vitro Testing, In

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