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Heat denaturation of fibrinogen to develop a biomedical matrix
Author(s) -
Marx Gerard,
Mou Xiaode,
HotovelySalomon Anna,
Levdansky Lilia,
Gaberman Elena,
Belenky Dimitry,
Gorodetsky Raphael
Publication year - 2008
Publication title -
journal of biomedical materials research part b: applied biomaterials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.665
H-Index - 108
eISSN - 1552-4981
pISSN - 1552-4973
DOI - 10.1002/jbm.b.30842
Subject(s) - fibrinogen , denaturation (fissile materials) , fibrin , chemistry , epitope , polyclonal antibodies , biophysics , biochemistry , microbiology and biotechnology , antibody , biology , immunology , nuclear chemistry
Abstract Native and heat denatured fibrinogen are the basis for various matrices used to establish hemostasis as well as for constructing biomedical devices. For example, fibrin microbeads (FMB) prepared by a heated (∼70°C) oil emulsion process were reported to be attractive to mesenchymal‐type cells, such as fibroblasts, endothelial and smooth muscle cells, and useful for isolating mesenchymal stem cells from bone marrow. Here, we examined the solution properties of fibrinogen subjected to heat (47–60°C). Fibrinogen exhibited maximal stability of pH max stab = 6.8. At physiologically relevant concentrations, Ca(II) stabilized and Zn(II) destabilized fibrinogen against heat denaturation. Scanning electron micrographs (SEM) of precipitated, heat denatured, fibrinogen showed globular structures (∼400 nm diameter), composed of aggregates of >3000 fibrinogen monomers. Monoclonal antibodies (MAb) to various regions of fibrinogen, as well as two polyclonal antibody (Ab) to haptotactic peptides (Haptides) equivalent to or near the C‐termini of β and γ‐chains (β 463–483 and γ 372–391/411 ), were used to monitor epitopic changes of fibrinogen bound to and heated on plastic ELISA plates. The pattern of altered Ab binding indicated that fibrinogen heat denaturation on plastic exposed the C‐terminal epitope γ 397‐411 as well as Haptide epitopes (β 463–483 and γ 372–391 ). Immuno‐staining of FMB prepared by a heated (below 75°C) oil emulsion process, also presented many exposed Haptide epitopes, which probably helped to attract cells. Our results indicated that moderately heat‐denatured fibrinogen, in the form of FMB, could be used for cell culturing and biomedical applications. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008