Rheol Acta 36:262-268 (1997)

© Steinkopff Verlag 1997

Jagesh V. Shah Strain hardening of fibrin gels and plasma clots

Paul A. Janmey

Abstract Biological macromole- times the moduli at small strains.

Received: 2 January 1997
Accepted: 22 April 1997
Dedicated to Prof. John D. Ferry
on the occasion of his 85th birthday.
J.V. Shah
Harvard-MiT Division of Health Sciences
and Technology
Experimental Medicine Division
Brigham and Women's Hospital
221 Longwood Avenue
Boston, Massachusetts 02115, USA
Dr. R A. Janmey ( ~ )
Experimental Medicine Divison
Brigham and Women's Hospital
221 Longwood Avenue
Boston, Massachusetts 02115, USA
E-mall: j anmey@fas.harvard.edu
cules have unique theological prop-
erties that distinguish them from
common synthetic polymers.
Among these, fibrin has been stud-
ied extensively to understand the ba-
sic mechanisms of viscoelasticity as
well as molecular mechanisms of
coagulation disorders. One aspect of
fibrin gel theology that is not ob-
served in most polymeric systems is
strain hardening: an increase in
shear modulus at strain amplitudes
above 10%. Fibrin clots and plasma
clots devoid of platelets exhibit
shear moduli at strains of approxi-
mately 50% that are as much as 20
The strain hardening of fibrin gels
was eliminated by the addition of
platelets, which caused a large in-
crease in shear storage modulus in
the low strain linear viscoelastic
limit. The reduction in strain hard-
ening may result from fibrin strand
retraction which occurs when plate-
lets become activated. This interpre-
tation is in agreement with recent
theoretical treatments of semi-flex-
ible polymer network viscoelasticity.
Key words Fibrin - viscoelasticity
- strain hardening - clot retraction -
platelets

Introduction constituent of mammalian blood clots. The rheology of

Many types of biological macromolecules form gels
that exhibit viscoelastic properties differing in several
respects from those of synthetic polymer networks and
gels. The unique viscoelasticity of biopolymers is fun-
damental to their biological function and the mainte-
nance of normal physiology. Defects that alter the vis-
coelasticity of biopolymers such as those comprising
the cytoskeleton or the extracellular matrix are impor-
tant in a number of disease states. In addition to their
biological function, biopolymers have uniquely macro-
scopic size, length and time scales making them useful
for probing fundamental theories of viscoelasticity and
polymer dynamics (e.g., Wang and Bloomfield, 1991;
Perkins et al., 1994; Yanagida et al., 1984; Gittes et al.,
1993; K~s et al., 1994; Kishino and Yanagida, 1988).
Perhaps the best characterized and most informative of
the gel-forming protein polymers is fibrin, the major
fibrin clots has been studied for over 50 years (Wag-
reich and Tarlov, 1945; Ferry and Morrison, 1947), in
work pioneered by Ferry and his coworkers (Ferry,
1983).
Initial work established that the network strands in a
fibrin gel form by linear polymerization of bi-functional
monomers activated by the enzyme thrombin. Mono-
mers combine by non-covalent interactions to form pro-
tofibrils of 10 nm diameter by a staggered overlapping
arrangement of symmetric, elongated subunits. Such
protofibrils can associate laterally with other protofibrils
to form wider, branching fibers (see (Mosesson, 1990)
and (Hermans and McDonagh, 1982) for review). The
width of these fibers depends on pH and ionic strength,
suggesting a dominant role for electrostatic interactions.
By modulating fiber width, gels with elastic elements
ranging from single protofibrils (fine clots) to fibers of
diameter > 300 nm (coarse clots) can be prepared in vi-
tro. During and after gelation (Bale et al., 1984) cova-
 J.V. Shah and E A. Janmey
Strain hardening of clots
lent bonds between subunits either within or between
protofibrils can be introduced by the enzymatic activity
of Factor XIlIa in a reaction that depends on both
thrombin and mM Ca 2÷ concentrations (Shen and Lor-
and, 1983). The ability to vary the mass to length ratio
of the network strands and to introduce covalent bonds
between strands has enabled a thorough study of the
importance of these factors on the rheology of fibrin
clots. Such studies have been relevant both to elucidat-
ing the physical origin of fibrin viscoelasticity, and to
providing insight into the clinical complications arising
when some of these reactions are compromised in dis-
ease (Schreiber and Schmer, 1984).
Among the unusual features of fibrin rheology is a
very large degree of strain hardening: an increase in the
elastic modulus measured under shear as the magnitude
of the deformation is increased. Such strain hardening
was apparent in early studies showing plots of stress vs.
shear strain with a distinct upward curvature for strains
above 10% (Roberts et al., 1973). The dependence of
dynamic shear moduli measured during oscillatory de-
formation at moderate to large strain amplitudes was
later studied systematically using fine fibrin clots either
with or without covalent ligation (Janmey et al., 1983;
Bale and Ferry, 1988). These studies report an increase
by a factor of more than 20 in the differential shear
storage modulus (dG'/dy) when the shear strain, y, in-
creases from 1% to 50%. In the present report, rheolo-
gic studies of coarse fibrin clots and clots of human
plasma both with and without platelets are compared.
The addition of platelets to human plasma and fibrin in-
creases gel stiffness and eliminates strain hardening.
Experimental considerations
Human plasma
Human blood, obtained from volunteers upon informed
consent, was drawn into a 0.1 volume of citrate-based
anticoagulant (8.5 mM sodium citrate, 69 mM citric
acid, 20 mg/ml glucose, pH 4.6). The samples were
centrifuged at ll0xg for 15 rain. The platelet-rich plas-
ma (PRP) was further centrifuged at 1000×g for 15 rain
to yield platelet-poor plasma (PPP). Gel-filtered plate-
lets were prepared from platelet-rich plasma as de-
scribed in (Hartwig et al., 1995), stored at 37°C in
Platelet Buffer (145raM NaC1, 5raM KC1, 2raM
MgC12, 0.5 mM Na2HPO4, 10 mM HEPES, 10 mM glu-
cose, 0.3% albumin, pH 7.4) and used within 2 h. Lyo-
philized bovine thrombin was obtained from Sigma (St.
Louis, MO), diluted in deionized water to 100 NIH U/
ml and frozen. Thrombin aliquots were thawed and
used immediately.
263
Human fibrinogen
Human fibrinogen was partially purified from platelet-
poor plasma (Mosher and Blout, 1973). One volume sa-
turated ammonium sulfate was added to three volumes
platelet-poor plasma and incubated at 4°C for 1 h with
occasional mixing. The precipitate was collected by
centrifugation at 1000xg for 30 rain, and the pellet re-
suspended in Tris-buffered saline (150raM NaC1,
10 mM Tris, pH 7.4). Fibrinogen content was assessed
by optical density (E= 1.52 at 280 nm) and SDS-PAGE.
Fibrinogen yield varied from 2 to 2.5 mg protein per
one ml of plasma.
Rheologic analysis
Shear deformation of clot samples was carded out on a
Rheometrics RFS II Fluids Spectrometer (Piscataway,
NJ) equipped with a cone and plate geometry. The prin-
ciple of operation of this instrument is described by
Janmey (1988). All samples were placed within the in-
strument immediately after the clotting agent was added
to allow the clot to form between the instrument plates.
Clotting of plasma was initiated by addition of 20 mM
CaC12 and 1 NIH U/ml thrombin. Fibrin clots were in-
itiated by addition of 3 mM CaC12 and 1 NIH U/ml
thrombin. The clots were monitored over time until the
storage modulus reached a steady value. The plateau in
storage modulus took no longer than 1 h for all condi-
tions considered. The clot was then subjected sequen-
tially to frequency dependence and strain dependence
measurements. Since the storage (G') and loss (G")
moduli are calculated from the in-phase and out-of-
phase components, respectively, of an oscillating stress
measured after imposition of a calibrated oscillatory
strain, these values are strictly accurate only in the limit
of linear viscoelasticity. As the moduli become strain-
dependent, the stress response is no longer perfectly fit
by a sinusoidal function, and therefore a quantitative in-
accuracy will occur. Most previous oscillatory measure-
ments (Bale and Ferry, 1988; Janmey et al., 1983) were
made with a torsion pendulum in which small oscilla-
tions (1%) were applied to a sample held at a steady,
larger strain to obtain a differential shear modulus
(dG'/dT). Control experiments measuring the same
samples by both methods, however, confirm that the
differences between measurements are minor, although
the magnitude of strain hardening may be slightly un-
derestimated by the types of forced oscillatory measure-
ments done in this study. All rheologic studies were
performed at 23°C.
Light microscopy
Clots and platelets were imaged by phase contrast mi-
croscopy on a Nikon Diaphot 300 microscope with a
264 Rheologica Acta, Vol. 36, No. 3 (1997)
© Steinkopff Verlag 1997

60X oil immersion objective and a Sanyo CCD camera 1000 A i i i

with a 2X image coupler. Images were captured to a
Macintosh computer equipped with a Scion video cap-
ture card and NIH Image software. Plasma clots for mi-
croscopy were initiated by recalcification alone, fibrin lOO
clots were initiated by addition of 3 mM CaC12 and m-
0.1 NIH U/ml thrombin. Slides were prepared by addi-
tion of the clotting agent directly to a small volume of
sample on a glass slide. Samples were sealed with nail & lO
polish and then allowed to sit for at least 1 h before
being visualized. All microscopic visualizations were
performed at 23°C.
1
0 1000 2000 3000 4000
Time (s)
Results

When thrombin is added to either purified fibrinogen 1000

under coarse clotting conditions or whole blood plasma, B
• . ." ~_--
the shear storage modulus (GI), measured at constant
[I_
strain amplitude (2%) and frequency (1 rad/s), increases
as the gel forms and is strengthened by increasing fi- E~
brin polymerization, as shown in Fig. 1 A. The magni- -I

tude of the shear modulus for platelet poor plasma is lOO 0

0
within a factor of 3 of that of an equivalent concentra-
tion of polymerized fibrinogen purified from that plas- Q

ma, but the modulus of platelet-rich plasma is approxi- o

mately 5-10 times greater. This large difference in (,9
shear moduli is most apparent in the limit of small
strain amplitudes (<10%) and disappears when the 10 I I 1 I I I r I I I I r T i i i i i i r

strain amplitude of oscillatory deformation exceeds 1 10 100

40%. Figure 1 B shows a large increase in G / at strains
above 15% for both fibrin and PPP gels, but a nearly
strain-independent modulus for PRP clots. Moreover,
the shear moduli of both fibrin and PPP gels at 40%
maximal strain are very near those of PRP clots at that
deformation.
The frequency dependence of both the shear storage
(G ~) and shear loss modulus (G r~) for the samples in
Fig. 1 are shown in Fig. 2. Over a broad range of fre-
quencies the storage modulus (A) is nearly independent
of deformation frequency for all samples. However, the
loss modulus (B) is independent of frequency only for
the platelet-rich plasma clot. Both the platelet-poor plas-
ma and fibrin gels display a decline in G ~ with decreas-
ing frequency, with fibrin exhibiting a stronger depen-
dence. The similarity in the frequency dependence of
the storage moduli suggests that all samples are exten-
sively interconnected either by direct fibrin-fibrin con-
tacts or through fibrin-platelet bonds, and that the pres-
ence of platelets does not qualitatively alter the broad
plateau modulus at small strain characteristic of fibrin
gels. The decreased frequency dependence of G ~t in
PRP clots suggests that some modes of relaxation, per-
haps by internal motions within fibrin strands, are di-
minished by the action of the platelets.
Strain (%)
Fig. 1 Time-course and strain dependence of plasma and fibrinogen
clots. The time-courses of clot formation (A) for platelet-rich plasma
(O), platelet-poor plasma (O) and purified fibrinogen (El). The in-
creased storage modulus of platelet-rich plasma, over that of platelet-
poor plasma and purified polymerized fibrinogen is apparent. Panel B
shows the dependence of G' on the magnitude of deformation. An
increased degree of strain hardening for platelet-poor plasma and fi-
brin compared to platelet-rich plasma is observed. Time-course mea-
surements (A) were made at a strain amplitude of 2% and frequency
of 1 rad/s. Strain sweep measurements (B) were made at 1 rad/s
Figure 3 shows structural differences of PPP (A) and
PRP (B) clots observed by phase contrast microscopy.
While the general appearance of individual fibers is
similar in the two samples, the network of strands in
the PRP clot is notably more heterogeneous, with clus-
ters of aligned fibrils bridging foci where activated
platelets have aggregated and contracted. A large differ-
ence in crosslink density is not observed.
The large effect of platelets on the rheology of plasma
clots is further evident in Fig. 4, where the strain-depen-
dent moduli of three homologous plasma samples differ-
ing only in the density of platelets are compared. Similar
to the results in Fig. 1, as increasing numbers of platelets
 J.V. Shah and R A. Janmey 265
Strain hardening of clots

1000 , i i i i,iJ I , i i i I,L, I , i i LILll I

(3-

"5 100
0 0 0 0 '
0

o
c0

10 ~I i i i liliil t l i lillll r I I lllf~l

0.1 I I 0 I O0
Frequency (rad/s)

100 :'t g ' ' ' '''"1 ' ' ' ' .... I ' ' ' ' ' ' " -

-ff
(3_

10

13
0

~9
O0
0
--.I

0.1 ~1 i ~ r ~ 1 ~ t r r~t~l ~ ~ ~ r~l~[

0.1 1 10 1 O0
Frequency (rad/s) Fig. 3 Plasma clots visualized by phase contrast microscopy. Plate-
let-poor plasma clots (A) and platelet-rich plasma clots (B) are visua-
Fig. 2 Frequency dependence of plasma and fibrinogen clots. Fre- lized in phase contrast microscopy. Note the same general fiber width
quency dependence of both the shear storage modulus (A) and shear but increased heterogeneity of the network near the retracted platelets
loss modulus (B) for platelet-rich plasma (O), platelet-poor plasma
(C)) and purified fibrinogen ([]). The storage modulus is independent
of frequency for all samples. The loss moduli of platelet-poor plasma moduli are compared for fibrinogen polymerized in the
and fibrinogen have a moderate dependence on frequency, whereas presence or absence of gel-filtered platelets. The plate-
the platelet-rich plasma has frequency-independent loss modulus. All
measurements where made at a strain amplitude of 2% let-fibrinogen mixture exhibits a dramatic increase in G /
compared to fibrinogen alone (A). This increase is simi-
lar to that seen in the PRP clot shown in Fig. 1 A. The
are left in the plasma after differential centrifugation, the platelet-fibrin mixture also exhibits a very small degree
modulus at small strain increases strongly (A), but the of strain hardening when compared to the fibrin sample
maximal modulus at moderate strains is only slightly al- (C). Again, this is similar to the finding for PRP clots
tered (B). The shear modulus of the PPP gel increases at (Fig. 1B), where the PRP clots had virtually no strain
strains above 15% and reaches a maximum at 100% that hardening when compared to PPP or fibrin. The strain
is near the level of the PRP clot at small strains, which hardening seen in the platelet-fibrin mixture is likely
exhibits no strain hardening (B). Moreover, the strain at due to dilution of the platelet concentration from physio-
which rupture of the clots occurs, as evidenced by a de- logic PRP values as a result of gel filtration and mixing
crease in modulus, decreases with increasing platelet with fibrinogen. Fig. 5 B shows that G / for the platelet-fi-
count. brinogen mixture is independent of frequency, again par-
The importance of activated platelets in determining alleling the PRP clot frequency dependence seen in
the shear modulus of purified fibrin clots is shown in Fig. 2. The finding that the relatively low modulus of
Fig. 5, where the time-course (A), frequency dependence the fibrin gel at low strain is increased to the levels of
(B) and the strain dependence (C) of the shear storage PRP clots simply by the addition of platelets confirms
266 Rheologica Acta, Vol. 36, No. 3 (1997)
© Steinkopff Verlag 1997

1000 1000 L-- , I I I

A A
13-
v
cE 100
L~ lOO m

Z3
0

"0
0 10

10 o
(D
i

0 1000 2000 3000 4000

Time (s)
~ , I i i , .X_ , , F I , f ~
1 --'
1000 '1 ' * ' ' .... t . . . . . . . . ~ ' ' ~ ..... I
0 1000 2000 3000 4000
n
B
T i m e (s) v

1000 ~i ........ ~ ~" ....... I

B -o 100
0

03
o o
0

~ 100 10 L I F itTlll I I I lITIlI I 1 I plllrt

O. 1 10 100
Frequency (rad/s)
1000 .... i .......................
0

C
10 . 100
1 10 100
Strain (%) Q

Fig. 4 Time-course and strain dependence for a variety of platelet C~

10
concentrations. Low speed ceutrifugation, 100×g, yielded platelet-rich
plasma (O), centrifugation at 300×g resulted in a lower platelet content
co
plasma (qualitatively assessed by turbidity) (©), and centrifugation at
1000xg resulted in clear platelet-poor plasma (r~). The time-course of 1 i t i1[ i t i i illll i i ~ Iiiitl i i i ItlIL

polymerization demonstrates that increasing number of platelets in- 1 10 1 O0 1000

creases the value of the storage modulus (A) at low (2%) strain. Plate-
let-poor plasma was measured for only 20 rain, but allowed to sit for 1
before strain sweep measurements were conducted. However, strain
dependence measurements (B) show that increasing platelet concentra-
tions decreases the degree of strain hardening seen. Time-course mea-
surements were performed at a strain amplitude of 2% and a frequency
of 1 rad/s. Strain dependence was also measured at 1 rad/s
that the large difference between plasma clots and pure
fibrin results mainly from the presence of platelets and
not from other proteins such as fibronectin (Kamykows-
ki et al., 1981; Hansen, 1984) or albumin (Galanakis et
al., 1987; Torbet, 1986) that may add to or alter clot struc-
ture.
Strain (%)
Fig. 5 Time-course, frequency and strain dependence of fibrin and
platelet-fibrin mixtures. Time-course of polymerization (A) is shown
for a gel-filtered platelet-fibrinogen mixture (O) and an equivalent
fibrinogen concentration alone (O). Time-course indicates the in-
crease in storage modulus created by addition of platelets to the fibri-
nogen. Both samples have storage moduli which are independent of
frequency (B). The addition of platelets decreases the degree of strain
hardening seen at moderate strains (C). The small strain hardening
seen in the platelet-fibrin mixture is likely to be due to dilution of the
platelets both during gel filtration and preparation of the mixture.
Time-course measurements were made at a strain amplitude of 2%
and a frequency of 1 rad/s. Frequency dependence was measured at
2% strain. The strain dependence was performed at a frequency of
1 rad/s
 J.V. Shah and E A. Janmey
Strain hardening of clots
Discussion
The large increase in the shear modulus of fibrin gels
that occurs at strains between 15% and 100% is one of
the most remarkable rheologic properties of these poly-
meric systems (Bale and Ferry, 1988; Janmey et at.,
1983). In contrast, the rheologic properties of most
common gels composed of flexible polymers are inde-
pendent of strain or show a decrease in shear modulus
over the range where fibrin exhibits strain hardening.
Even aqueous gels composed of relatively stiff poly-
mers such as polysaccharides exhibit either a very
broad linear viscoelastic range, or become weaker at
strains between 10% and 100% (Ross-Murphy and
Shatwell, 1993). Some protein polymer gels, however,
such as those formed by cytoskeletal actin filaments
(Janmey et al., 1988) and intermediate filaments (Le-
terrier et al., 1996; Janmey et al., 1991) show similar
degrees of strain hardening.
The molecular origin of the strain hardening seen in
fibrin is unknown, but thought to be related to the rod-
like or semi-flexible nature of the fibrin strands. Initial
studies of strain hardening (Bale and Ferry, 1988; Jan-
mey et at., 1983) were interpreted in terms of a theory
that considered the increase in steric contacts within
isotropic systems of long rods as these systems were
deformed in shear (Doi and Kuzuu, 1980). In this mod-
el the increase in shear modulus is due to an increased
number of elastically active network junctions. This in-
terpretation accounted qualitatively for the increase in
G r at moderate strain. Collapse at larger strains could
be due to either collapse of filaments into separated
bundled domains or breakage of the strands between
junction points. The finding that strain hardening did
not depend on the presence of covalent ligation (Bale
and Ferry, 1988) supported the concept that steric inter-
actions play an important role.
Previous theoretical treatments, however, have not
been able to explain the quantitative aspects of strain
hardening in fibrin. More recently a model has been
proposed to explain strain hardening in networks com-
prised of the cytoskeletal polymer actin that could ap-
ply equally to fibrin. In this model, the elasticity of
semi-flexible polymer networks derives from the reduc-
tion of entropy that results from the stretching of semi-
flexible chains with a thermal distribution of transverse
undulations (MacKintosh et al., 1995). As a network of
such filaments is sheared, some elastically active junc-
tions, either steric or covalent, become separated, and
the strands between them are stretched. The resistance
increases as the filaments lose transverse undulations,
and a limit, corresponding to the yield point, is reached
when the distance between junctions equals the contour
length of the polymer.
267
The large effect of fibrin gel retraction by platelets to
increase the modulus at low strain and to eliminate
strain hardening is consistent with this model for visco-
elasticity. Applying tension to strands within a network
would serve to pull out the slack within network
strands, and thereby increase their stiffness, rather than
increasing the number of elastically active junctions.
This picture is consistent both with the images in
Fig. 3, which show no increase in the number of obser-
vable junction points, and with previous results that the
increase in shear modulus of plasma clots caused by
platelets could by mimicked by application of an elon-
gational strain to a platelet poor plasma clot (Jen and
McIntire, 1984).
Summaryand conclusions
Much of the elastic resistance to deformation of platelet
rich plasma clots, and therefore presumably of clots
formed in vivo, depends on the presence of platelets. In
part, the contribution of platelets may arise from an in-
creased number of contacts between fibrin strands, but
the relatively small increase in interfilament contacts
within the inherently branched and covalently ligated fi-
brin network is too small to explain the nearly ten fold
increase in shear modulus observed. An alternative ex-
planation suggested by the data in this report is that the
contractile function of platelets pulls out the slack in fi-
brin strands between crosslinks, and thereby mimics the
large increase in shear modulus of pure fibrin gels at in-
creasing shear strains. The strain hardening observed in
fibrin networks may be a general feature of networks
formed by highly elongated protein networks with mesh
sizes on the order of a micron.
The rheological analysis of fibrin gels over the years
has yielded many insights into the physics and chemis-
try of polymeric systems as well as shed light on the
molecular origins of pathologic states due to defects in
a number of clotting factors. Since many of the visco-
elastic properties of fibrin and other biopolymers gels,
such as strain hardening, are radically different from
those of common rubber-like materials, continued study
of such materials is likely to provide data valuable to
testing theories of polymer dynamics as well as under-
standing basic physiology and disease.
Acknowledgments The authors gratefully acknowledge the technical
assistance of Louise Z. Wang. This work was supported by grants
G957 from the Cystic Fibrosis Foundation and NS29031 from the
National Institutes of Health. E A.J. expresses his deepest gratitude to
John Ferry for the introduction to rheology.
268
References
Bale MD, Ferry JD (1988) Strain enhance-
ment of elastic modulus in fine fibrin
clots. Thromb Res 52:565-572
Bale MD, Janmey PA, Ferry JD, Lorand L
(1984) Kinetics of formation of fibrin oli-
gomers. III. Ligation kinetics concurrent
with and subsequent to oligomer assem-
bly. Biopolymers 23:127-138
Doi M, Kuzuu NY (1980) Nonlinear elastic-
ity of rodlike macromolecules in con-
densed state. J Polym Sci Polym Phys Ed
18:409-419
Ferry J, Morrison P (1947) Preparation and
properties of serum and plasma proteins.
VIII. The conversion of human fibrino-
gen to fibrin under various conditions. J
Am Chem Soc 69:388-400
Ferry JD (1983) The conversion of fibrino-
gen to fibrin: events and recollections
from 1942 to 1982. Ann NY Acad Sci
408:1-10
Galanakis DK, Lane BP, Simon SR (1987)
Albumin modulates lateral assembly of fi-
brin polymers: evidence of enhanced fine
fibril formation and of unique synergism
with fibrinogen. Biochem 26:2389-2400
Gittes F, Mickey B, Nettleton J, Howard J
(1993) Flexural rigidity of microtubules
and actin filaments measured from ther-
mal fluctuations in shape. J Cell Biol
120:923-934
Handen MS (1984) Fibronectin and coagula-
tion factor XIII increases blood platelet ad-
hesion to fibrin. Thromb Res 34:551-556
Hartwig JH, Bokoch GM, Carpenter CL, Jan-
mey PA, Taylor LA, Toker A, Stossel TP
(1995) Thrombin receptor ligation and ac-
tivated Rac uncap actin filament barbed
ends through phosphoinositide synthesis
in permeabilized human platelets. Cell
82:643-653
Rheologica Acta, Vol. 36, No. 3 (1997)
© Steinkopff Verlag 1997
Hermans J, McDonagh J (1982) Fibrin: smac-
ture and interactions. Sere Thrombos He-
mostas 8:11-24
Janmey P, Amis E, Ferry J (1983) Rheology
of fibrin clots. VI. Stress relaxation,
creep, and differentiai dynamic modulus
of fine clots in large shearing deforma-
tions. J Rheol 27:135-153
Janmey PA, Euteneuer U, Tranb P, Schliwa
M (1991) Viscoelastic properties of vi-
mentin compared with other filamentous
biopolymer networks. J Bell Biol
113:155-160
Janmey PA, Hvidt S, Peetermans J, Lamb J,
Ferry JD, Stossel TP (1988) Viscoelastic-
ity of F-actin and F-actin/gelsolin com-
plexes. Biochem 27:8218-8227
Jen CJ, McIntire LV (1984) The structural
properties and contractile force of a clot.
Cell Motil 2:445-455
Kamykowski GW, Mosher DF, Lorand L,
Ferry JD (1981) Modification of shear
modulus and creep compliance of fibrin
clots by fibronectin. Biophys Chem
13:25-28
Kfis J, Strey H, Sackmann E (1994) Direct
imaging of reptation for semiflexible ac-
tin filaments. Nature 368:226-229
Kishino A, Yanagida T (1988) Force mea-
sttrements by micromanipulation of a sin-
gle actin filament by glass needles. Na-
ture 334:74-76
Leterrier IF, Kas J, Hartwig J, Vegners R,
Janmey PA (1996) Mechanical effects of
neurofilament cross-bridges. Modulation
by phosphorylation, lipids, and interac-
tions with F-actin. J Biol Chem
271:15 687-15 694
MacKintosh F, Kiis J, Janmey P (1995) Elas-
ticity of semiflexible biopolymer net-
works. Phys Rev Lett 75:4425-4428
Mosesson M (1990) Fibrin polymerization
and its regulatory role in hemostasis. J
Lab Clin Med 116:8-17
Mosher DF, Blout ER (1973) Heterogeneity
of bovine fibrinogen and fibrin. J Biol
Chem 248:6896--6903
Perkins TT, Smith DE, Chu S (1994) Direct
observation of tube-like motion of a sin-
gle polymer chain. Science 264:819-822
Roberts WW, Lorand L, Mockros LF (1973)
Viscoelastic properites of fibrin clots.
Biorheol 10:29-42
Ross-Murphy SB, Shatwell KP (1993) Poly-
saccharide strong and weak gels. Bio-
rheol 30:217-227
Schreiber WE, Schmer G (1984) Application
of mechanical impedance measurements
to the study of dysfibrinogens. J Lab Clin
Med 104:494-500
Shen L, Lorand L (1983) Contribution of fi-
brin stabilization to clot strength. Supple-
mentation of factor XIlI-deficient plasma
with the purified zymogen. J Clin Invest
71:1336-1341
Torbet J (1986) Fibrin assembly in human
plasma and fibrinogerdalbumin mixtures.
Biochem 25:5309-5314
Wagreich H, Tarlov IM (1945) Studies on the
strength of fibrinogen-tin'ombin clots. Ar-
chives of Biochem 7:345-352
Wang LX, Bloomfield VA (1991) Small-an-
gle x-ray scattering of semidilute rodlike
DNA solutions-polyelectrolyte behavior.
Macromol 24:5791-5795
Yanagida T, Nakase M, Nishiyama K, Oosa-
wa F (1984) Direct observation of motion
of single F-actin filaments in the presence
of myosin. Nature 307:58-60