Ultrathin self-assembled fibrin sheets.
Fibrin polymerizes into the fibrous network that is the major structural component of blood clots and thrombi. We demonstrate that fibrin from three different species can also spontaneously polymerize into extensive, molecularly thin, 2D sheets. Sheet assembly occurs in physiologic buffers on both hydrophobic and hydrophilic surfaces, but is routinely observed only when polymerized using very low concentrations of fibrinogen and thrombin. Sheets may have been missed in previous studies because they may be very short-lived at higher concentrations of fibrinogen and thrombin, and their thinness makes them very difficult to detect. We were able to distinguish fluorescently labeled fibrin sheets by polymerizing fibrin onto micro-patterned structured surfaces that suspended polymers 10 microm above and parallel to the cover-glass surface. We used a combined fluorescence/atomic force microscope system to determine that sheets were approximately 5 nm thick, flat, elastic and mechanically continuous. Video microscopy of assembling sheets showed that they could polymerize across 25-microm channels at hundreds of microm(2)/sec (approximately 10(13) subunits/s x M), an apparent rate constant many times greater than those of other protein polymers. Structural transitions from sheets to fibers were observed by fluorescence, transmission, and scanning electron microscopy. Sheets appeared to fold and roll up into larger fibers, and also to develop oval holes to form fiber networks that were "pre-attached" to the substrate and other fibers. We propose a model of fiber formation from sheets and compare it with current models of end-wise polymerization from protofibrils. Sheets could be an unanticipated factor in clot formation and adhesion in vivo, and are a unique material in their own right.
O'Brien
E Tim
ET
Department of Physics and Astronomy, University of North Carolina, Chapel Hill, NC 27599-3255, USA. etobrien@email.unc.edu
Falvo
Michael R
MR
Millard
Daniel
D
Eastwood
Brian
B
Taylor
Russell M
RM
Superfine
Richard
R
D000818:Animals; D001777:Blood Coagulation; D002645:Chickens; D005337:Fibrin / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D005340:Fibrinogen / Q000494:pharmacology; D005898:Glass; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology
D011108:Polymers; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin