Abstract
Collagen is a widely investigated extracellular matrix material with extensive potentials in the field of tissue engineering. This protocol describes a method to prepare reconstituted collagen that can be ready-to-use, storable and suitable for further in vitro and in vivo investigations. Type I collagen was extracted from rat tail tendons and processed in acetic acid solution to obtain sterile soluble collagen. At first, crude collagen was dissolved in acetic acid, then frozen at −20 °C and lyophilized to obtain a sponge, which could be stored at −80 °C. Lyophilized collagen was then dispersed in acetic acid to obtain a sterile solution of collagen at targeted concentrations. The whole low-cost process from the extraction to the final sterile solution takes around 2–3 weeks. The collagen solution, once neutralized, has the potential to be used to produce gels or scaffolds, to deposit thin films on supports and to develop drug delivery systems.
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References
Berglund, J.D., Mohseni, M.M., Nerem, R.M. & Sambanis, A. A biological hybrid model for collagen-based tissue engineered vascular constructs. Biomaterials 24, 1241–1254 (2003).
Purna, S.K. & Babu, M. Collagen based dressings––a review. Burns 26, 54–62 (2000).
Wallace, D.G. & Rosenblatt, J. Collagen gel systems for sustained delivery and tissue engineering. Adv. Drug Deliv. Rev. 55, 1631–1649 (2003).
Sionkowska, A. & Kaminska, A. Thermal helix-coil transition in UV irradiated collagen from rat tail tendon. Int. J. Biol. Macromol. 24, 337–340 (1999).
Steven, F.S. & Tristram, G.R. The presence of non-protein nitrogen in acetic acid-soluble calf-skin collagen. Biochem. J. 83, 240–244 (1962).
Silver, F.H. & Trelstad, R.L. Type I collagen in solution. Structure and properties of fibril fragments. J. Biol. Chem. 255, 9427–9433 (1980).
Wood, G.C. The heterogeneity of collagen solutions and its effect on fibril formation. Biochem. J. 84, 429–435 (1962).
Gross, J., Highberger, J.H. & Schmitt, F.O. Extraction of collagen from connective tissue by neutral salt solutions. Proc. Natl. Acad. Sci. USA 41, 1–7 (1955).
Habermehl, J. et al. Preparation of a ready-to-use, stockable and reconstituted collagen. Macromol. Biosci. 5, 821–828 (2005).
Boccafoschi, F., Habermehl, J., Vesentini, S. & Mantovani, D. Biological performances of collagen-based scaffolds for vascular tissue engineering. Biomaterials 26, 7410–7417 (2005).
Sionkowska, A. Flash photolysis and pulse radiolysis studies on collagen type I in acetic acid solution. J. Photochem. Photobiol. B 84, 38–45 (2006).
Elsdale, T. & Bard, J. Collagen substrata for studies on cell behavior. J. Cell. Biol. 54, 626–637 (1972).
Bell, E., Ivarsson, B. & Merrill, C. Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc. Natl. Acad. Sci. USA 76, 1274–1278 (1979).
Weinberg, C.B. & Bell, E. A blood vessel model constructed from collagen and cultured vascular cells. Science 231, 397–400 (1986).
Deyl, Z., Miksik, I. & Eckhardt, A. Preparative procedures and purity assessment of collagen proteins. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 790, 245–275 (2003).
Lee, C.H., Singla, A. & Lee, Y. Biomedical applications of collagen. Int. J. Pharm. 221, 1–22 (2001).
Acknowledgements
This work was partially supported by the Natural Science and Engineering Research Council of Canada, the Canadian Institute for Health Research and the Quebec City University Hospital Research Center.
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Rajan, N., Habermehl, J., Coté, MF. et al. Preparation of ready-to-use, storable and reconstituted type I collagen from rat tail tendon for tissue engineering applications. Nat Protoc 1, 2753–2758 (2006). https://doi.org/10.1038/nprot.2006.430
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DOI: https://doi.org/10.1038/nprot.2006.430
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