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The proteins of synovial fluidA study of the ╨Ю┬▒1╨Ю┬▒2 globulin ratio.

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The Proteins of Synovial Fluid : A Study
of the “,/a2Globulin Ratio
By J. P. BINETTEAND K. SCHMID
Normal, postmortem, traumatic and
rheumatoid synovial fluids were studied
by paper electrophoresis, ultracentrifugation and starch gel electrophoresis. It
could be demonstrated that the change
in the a 1 / a 2 globulin ratio, particularly
prominent in normal and postmortem
synovial fluids as compared with normal
serum, is a consequence of the greatly
increased relative concentration of alacid glycoprotein and a significant decrease in the relative concentration of
the haptoglobins and the 19s a2-macroglobulin.
Specimens de liquido synovial-normal,
post morte, traumatic, e rheumatoideeesseva studiate per electrophorese a
papiro, ultracentrifugation, e electrophorese a gel de amylo. I1 esseva possibile demonstrar que le deviation del
proportion globulina a1 a globulina a2
ab le valor trovate in le sero-un deviation particularmente marcate in liquido
synovial normal e post morte-es un
consequentia del grandemente augmentate concentration relative de glycoproteina acide a1 e del significativemente reducite concentration relative
del haptoglobinas e del macroglobulina
“ 2 19s.
S
EVERAL INVESTIGATORS have demonstrated that the electrophoretic
distribution of the proteins of certain synovial fluids is unlike that of
serum and, further, that considerable differences in this distribution also exist
among the major groups of pathological joint fluids.*-l* Postmortem and
traumatic synovial fluids are known to exhibit a highly increased relative
concentration of the a,-globulins and a lower relative concentration of the
a2-globulins when compared with normal serum.3S6 Thus, the al/a2-globulin
ratio of these synovial fluids is significantly higher than that of normal
~ e r u m . ~ In
, ~ ,our
’ ~ earlier studies on the characterization of the synovial fluid
proteins, these constituents were fractionated by a modification of Cohn’s
Method 10.15 On the basis of the obtained results it was not possible, however, to explain the differences in the above mentioned ratio of these two
body fluids.
The present investigation was undertaken lto determine the proteins that
account for the differences in the al/a2-globulin ratio between certain synoFrom the Department of Medicine, State University of New York at Bu@o, Buffalo,
New York, and the Department of Biochemistry, Aoston University School of Medicine,
Boston University Medical Center, Boston, Mats.
Part of this work which was presented at the American Rheumatism Association
Meeting in Dallas, Texas, December 9, 1960 (Abstract, Arthritis (L7 Rheumatism, 4:104,
1961), was carried out at tlw Robert W. Lovett Mcmorial Unit, MassachusMs General
Hospital, Boston, Mass. Thus this is publication No. 383 of the Robert W. Lovett Memorial Unit. This study tms supported b y grants from the National lnstitutes cf Health,
U. S . Public Health Service (A-3564, A-5581, and GM-10374).
14
ARTHRITISAND RHEUMATISM,VOL. 8, No. 1 (FEBRUARY),
19135
15
PROTEINS OF SYNOVIAL FLUID
vial fluids and serum, and to compare the protein distribution of normal with
that of pathological joint fluids.
MATERIALAND METHODS
Normal synovial fluid was aspirated from the knee joints of six apparently healthy men
ranging in age from 20 to 40 years. The postmortem synovial specimens were obtained from the knee joints of nine patients shortly after their death. These patients
showed no clinical evidence of arthritis. The traumatic synovial fluids were aspirated
from each knee of nine apparently healthy men who had sustained joint injury and
whose age was between 18 and 22 years. Fourteen rheumatoid synovial fluids were taken
from patients afflicted with definite rheumatoid arthritis. Additional pathological specimens were obtained from knee joints of two patients with neuropathic joint disease and
from one patient with degenerative joint disease.
Treatment with hyaluroizidase (Wyeth, 1080 USP units per milligram) of all synovial
fluids was carried out as follows: Three mg. of enzyme powder were dissolved in 1 ml.
of 0.15 N NaCl, and 0.2 ml. of this solution was added to 1 ml. of synovial fluid. The
digestion was carried out at 37" C. for ninety minutes. The resulting digests were subsequently dialyzed against distilled water and lyophilized.
Paper electrophrresis was performed at pH 8.6 in l-/2 0.1 sodium citrate-diethylbarbiturate buffer. Solution containing 5 per cent of- lyophilized synovial fluid proteins in
0.15 N NaCl was utilized for these experiments. After the electrophoresis, the paper strips
were dried at 105" and then stained with amidoblack 10B. The relative protein concentrations were determined with an Analytrol (Spinco Model RB). The PAS technique of
McGuckin and McKenzielF was employed to stain the electropherograms for glycoproteins
in terms of their sialic acid content.
Ultracentrifugal analysis was performed with a Spinco Model E ultracentrifuge equipped
with an automatic temperature controller (R.T.I.C. type) at 56,100 rev/min. and 20" C.
The psthological joint fluids and sera were diluted before centrifugation with 0.15 N NaCl
so that a protein concentration of approximately 1.5 per cent was obtained. The enlarged
tracings of the patterns revealing three or four peaks were resolved in Gaussian curves.
The relative concentration of each boundary was calculated from the planimetric measurements of these curves.
Vertical starch gel electrophoresis was carried out in borate buffer according to
Smithies.17.ls For each analysis 6 mg. of lyophilized synovial fluid proteins were dissolved in 0.1 ml. of this buffer mixed with starch powder and transferred into an appropriate trough of the starch gel block. Concomitantly 0.1 ml. of serum or 6 mg. of
dialyzed and lyophilized serum proteins were analyzed. The electrophoresis was carried
out at 5.5 V/cm for 15 hours. The relative percentages of the protein zones of the
gels stained with amido-black 10B were estimated by visual comparison.
Normal human serum or the patient's serum when available served as control for
paper electrophoresis, starch gel electrophoresis and ultracentrifugation.
RESULTS
1. Normal Synovial Fluid
Because of the small volume of each normal synovial fluid specimen, only
starch gel electrophoresis could be carried out, except for one sample which
permitted performance of paper electrophoresis and determination of the
protein concentration (table 1). The fluid volumes ranged from 0.2 to 0.4
ml. per knee. The protein concentration was found to be 1.8 per cent agredng
with the range of 1.6 to 2.2 per cent reported by Bole.19 Paper electrophoresis
indicated a significant increase in the relative concentration of the albumin
5
4
10
10
19
10
17
10
7.0
2.4
2.3
3.0
2.2
3.5
2.8
2.6
2.8
2.5
1.1
2.2
2.5
1.4
1.0
1.1
1.4
1.8
1.8
0.2-0.4
2
5
10
4
6
8
9
12
8
7
%
Normal H u m n Serum
McCo
Sa
McCa
Am
Average
Ji
Le
At
Average
Th
Mr
Pb
Sr
Mh
Ma
Te
co
To
Case
Protein
Concen.
(Biuret)
Volume
of Fluid
per Knee
ml.
Table l.--dnalysis
49
55
57
58
60
50
58
47
55
59
53
59
54
49
42
40
37
46
50
63
Albumin
B
6
4
6
6
4
4
5
7
12
17
Control
Class I l l : Traumatic
11
9
10
11
7
14
10
6
8
11
12
7
10
8
8
11
16
18
22
18
18
24
18
31
21
0.5
0.7
0.6
1.0
0.8
0.6
0.5
0.8
-
1.2
0.8
1.0
1.0
1.1
1.2
2.1
1.1
1.3
6
6
Class 11: Postmortem
5
14
16
8
10
23
12
12
17
7
7
10
20
6
6
16
23
9
8
11
30
11
9
12
28
15
7
9
32
8
7
12
27
9
7
12
23
Ratio
aj/ai-Glohulin
1.0
Class I : Normal
7
9
Y
14
7
LYz-
(Relative Percentage)
1y1-
Paper Electrophoresis at pH 8.6
2
2
3
2
3
3
6
3
2
1
4
1
1
2
0
1
2
2
-
19s
12
7
3
6
9
9
17
8
14
10
12
15
6
6
12
16
11
12
-
86
89
93
91
88
88
77
88
82
88
84
80
90
86
84
82
86
83
-
7s
4s
(Relative Percentage)
Ultracentrifugal Analysis
of the Proteins of Normal, Postmortem and Traumatic Synouial Fluids
0
1
0
0
2
1
1
0
-
6
4
1
1
3
3
4
0
2
1
-
1s
EEi
%
a
z
39
34
E
c3
c-r
PROTEINS OF SYNOVIAL FLUID
17
while the corresponding values of the a,-globulins remained constant. The
relative concentration of the a2- and 8-globulins was greatly decreased. The
a1/a2-globulin ratio of 1.0 was twice as high as that of normal serum. The
starch gel electrophoretic patterns of normal synovial fluids (fig. 1 ) showed
a protein distribution which reveals more clearly the changes observed on
paper electrophoresis: a highly increased relative amount of al-acid glycoprotein which in this system is known to be identical with the slower moving
of the two prealbumins. The level of this protein is increased to such an
extent that the prealbumin area is almost completely covered. Further, the
albumin zone appears markedly enlarged. A decrease in the relative concentration of the haptoglobins and particularly of the 7-globulins is evident.
In addition, and more important, the slow moving a,-globulin which is
designated as Sa2 and is identical with the 19s a2-globulin,* is significantly
decreased in its relative concentration. This finding is in agreement with the
observation by Schur and Sandson20 who demonstrated by ultracentrifugation
the presence in normal synovial fluid of a very low content of the total 19s
proteins ( 19Sa2- plus 19Sr2-globulins).
2. Postmo.l-tem Synovial Fluid
The average volume of this series of fluids varying from 2 to 12 ml. suggested that some of these specimens were derived from patients with
edema.1J3,21The range of the protein concentration and its average agrees
with the corresponding values reported by others.1J3J1,22,23The results of the
paper electrophoretic analyses of postmortem synovial fluids are given in
table 1. A duplicate electropherogram of a pattern with a highly increased
relative concentration of the al-globulin stained by the PAS-technique corroborates this high increase in relative concentration of a glycoprotein ( s ) ,
probably al-acid g l y ~ o p r o t e i n ,in
~ ~the al-globulin zone. The relatively low
albumin values and the high 7-globulin levels may reflect the serum protein
changes brought about by the disease of the patients. As indicated in table 1
the al/a2-globulin ratios of the investigated postmortem synovial fluids ranged
from 0.8 to 2.1 and averaged 1.3. Ultracentrifugation of postmortem synovial
fluid revealed an average concentration of the 19s globulins which was comparable to that of normal serum. The 7s and 4s components were found to
be present in the same ratio as in normal serum. It is of particular interest
that an additional component ( s ) with
~ a sedimentation coefficient of lS, not
detected in serum, was found in relatively high concentration, represented by
a sharp spike in pattern B of figure 2. The 1s peak is observed after digestion of
these fluids with hyaluronidase. Further investigations are needed to establish whether this peak represents a hitherto unknown constituent or a high
molecular weight degradation product of hyaluronic acid. An apparently
*Diagrammatic presentations of the starch gel electrophoretic pattern of normal human
serum has been published by many authors (P. Grabar and P. Burtin, Analyse ImmunoElectrophorktique, Masson & Cie, Paris, 1960, page 94). Such diagrams serve to identify
the relative location of the major serum proteins.
1s
BINETTE AND SCHMID
different component with a sedimentation coefficient lower than that of
albumin was noted by Schur and Sandson20 in normal synovial fluid without
digestion with hyaluronidase. The starch gel electrophoretic analysis (fig. 3 )
revealed striking differences between these fluids and normal serum. A slightly
increased concentration of al-acid glycoprotein and very low relative concentrations of the Sa2-globulin and haptoglobins were noted. The synovial fluid
and serum used for this starch gel electrophoretic analysis were taken from
a 17 year old male who had been in good health until approximately 24 hours
before death which was caused by an acute cerebrovascular accident. Although
similar observations were made regarding the nine postmortem synovial fluids
investigated, distinct differences in the relative concentration of the albumin
and globulins were observed (table 1). It was also noted that the synovial
fluid specimens with higher al/ap-globulin ratios tend to reveal a lower concentration of Sa2-globulin on starch gel electrophoresis. Moreover, it was found
that the higher the al/a2-globulin ratio, the larger the area of al-globulin ratio
and that of a,-acid glycoprotein appeased on paper electrophoresis and on
starch gel electrophoresis, respectively.
3. Traumatic Synovial Fluid
The volume of this series of joint fluids varied from 4 to 19 ml. per knee.
The protein concentration, found to be significantly higher than that of
postmortem synovial fluids, ranged from 2.2 to 3.5 per cent, i.e. from that of
the postmortem to {thatof the majority of fluids from patients with rheumatoid
arthritis (see below). Paper electrophorstic analysis of the traumatic joint
fluids showed that the relative concentration of albumin was distinctly higher
than that of normal serum. The relative level of the a,-globulin is essentially
unchanged whereas that of the a,-globulins slightly decreased when compared
with the corresponding values of normal serum. The a,/a,-globulin ratios
varied between 0.5 and 1.0. The ultracentrifugal analysis revealed an average
composition which differed from that of normal serum: The majority of these
fluids showed an increase in the 4s peak and a corresponding decrease in the
7s peak.
On starch gel electrophoresis, the relative concentration of a,-acid glycoprotein was approximately the same as that of normal serum (fig. 4). This
confirms the earlier reports on electrophoretic analysis of traumatic joint
fl~ids.l,~
The
s ~ average of the Sa2-globulin and the r-globulin levels appeared
significantly decreased. It should be pointed out that considerable variation
in the relative concentrations of the a,-acid glycoprotein, Sa2-globulin and
haptoglobins was noted in this group of synovial fluids, so that some patterns
resembled those of postmortem synovial fluid while others were somewhat
similar to those of normal serum.
4 . Synovial Fluid of Patients with Rheumatoid Arthritis
The protein concentration of the synovial fluids of this group of patients
ranged from 1.1to 6.0 per cent.
19
PROTEINSOF SYNOVIAL FLUID
3
I
Y
T
A
O1
A
B
B
A
Fig. 1.-Starch gel electrophoretic patterns of normal synovial fluid ( A ) and
normal serum (B) of the same individual. The location of the following proteins is
indicated: 7s 7-globulins ( y ) , 19a,-macroglobulin (Sa,) , transferrin (PI),Albumin
( A ) , and a,-acid glycoprotein or orosomucoid (a1).The position of the trough is
indicated by T.
Fig. 3.-Starch gel electrophoretic pattern of the postmortem synovial fluid (A)
and serum (B) of a patient without apparent joint disease and without edema.
Paper electrophoresis of this limited series of fluids revealed the following
findings. The relative concentration in the synovial fluid of albumin was often
increased, that of the a2- and P-globulins mostly decreased, and that of the
al- and r-globulins unchanged, although exceptionally highly elevated Yglobulin values were noted (2,4) when compared with the corresponding
values of the paired serum. Significant differences in the distribution of the
protein were noted from pair to pair so that no correlation could be found
between the relative changes of the different groups of the electrophoretically
separated proteins. As expected, these patterns also differed to varying degrees
from those of normal serum because of the changes effected by the disease
20
BINETTE AND SCHMID
19s
1 4 7s
A
B
Fig. 2.-Ultracentrifugal pattern of a postmortem synovial fluid. Pattern A was
obtained after centrifugation at 56,000 rev./min. and 20 C. for 12 minutes and pattern B after 100 minutes. Pattern A shows the low relative concentration of 19s
proteins. Pattern B demonstrates the component sedimenting with a coefficient of
1s.The direction of the centrifugal movement is from left to right.
states.l-14 The al/a2-globulin ratio of the rheumatoid fluids varied from 0.3
to 1.1and averaged 0.5. The corresponding values of the rheumatoid sera were
0.2 to 0.5, averaging 0.3. On ultracentrifugation, essentially all synovial fluids
and sera revealed a greatly increased relative content of the 19s globulins
which was more pronounced with the sera where a several fold increase over
the normal value was observed. The relative percentage of the 7S-globulin
was almost always elevated significantly, in some cases over 300 per cent of
the normal value. In certain pairs of fluids this value was higher in the joint
fluid whereas in others it was higher in the serum. The area of the albumin
gradient was correspondingly decreased. The 1s-peak was not detected in this
class of joint fluids with the exception of one specimen. No apparent correlation could be noted between the changes observed on ultracentrifugation and
electrophoresis. In table 2 the analysis of only four pairs of fluids is given.
Starch gel electrophoresis (fig. 5) of the synovial fluid and serum of each
patient revealed similar patterns. In this series of analyses the relative concentration of al-acid glycoprotein of both the synovial fluids and sera was
greater than that of normal serum. Haptoglobins and Sa2-globulin often appeared slightly decreased in their levels in the synovial fluids as compared with
the corresponding sera. The synovial fluid of a patient with neuropathic joint
disease and of a patient with osteoarthritis revealed similar starch gel patterns.
A resolution of the r-globulins, observed earlier by the same techniques in
multiple myeloma and other diseases,25126was seen in the serum and synovial
fluid of a young patient with severe rheumatoid arthritis of long duration. It
was further noted that the resolution of the r-globulin of this patient was
more apparent in the synovial fluid than in the plasma (fig. 6 ) .
21
PROTEINS OF SYNOVIAL FLUID
5
4
Y
T
Sag
A
B
A
B
Fig. 4.-Starch gel electrophoretic pattern of the synovial fluid (A) and serum
(B) of a patient with traumatic effusion.
Fig. 5.--Starch gel electrophoretic pattern of the synovial fluid (A) and serum
(B) of a patient with definite rheumatoid arthritis.
5. Control Experimmts
The following control experiments were carried out to study whether the
distribution of the synovial fluid proteins was affected (1) by the digestion
with hyaluronidase by proteolytic enzymes possibly present in the hyaluronidase preparation or ( 2 ) by direct contribution of the hyaluronidase proteins
to one or more bands of the synovial fluid proteins or ( 3 ) by binding of
the hyaluronic acid breakdown products to the synovial fluid proteins and,
thus, leading to a change in the apparent electrophoretic mobility and consequently in the relative distribution of the synovial fluid proteins. One ml.
of normal serum was incubated with 0.6 mg. of the enzyme for 90 minutes
and subsequently subjected to starch gel electrophoresis in parallel with
untreated serum. No appreciable proteolytic activity could be demonstrated
in this experiment, i.e. the protein patterns were identical. Further, two
43
30
4.8
6.3
Syn. fluid
Serum
Ma
44
35
Syn. fluid
Serum
Br
3.5
7.7
Syn. fluid
Serum
c1
B
Y
10
13
5
5
4
7
10
15
9
13
4
4
13
17
12
13
15
19
0.5
0.5
0.4
0.3
30
33
0.4
0.3
0.4
0.2
at/nz-Globulin
Ratio
29
32
32
28
Class IV: Rheumatoid Arthritis
12
16
11
56
5
58
3
17
12
11
39
36
4.2
6.6
Syn. fluid
Serum
Ha
a2-
(Relative Percentage)
(YI-
Paper Electrophoresis at pH 8.6
Albumin
5.6
8.8
%
Protein
Cone.
(Biuret)
5
2
3
5
4
5
6
6
19s
27
41
28
19
25
22
17
18
71
54
68
77
74
70
77
76
7s
4s
(Relative Percentage)
Ultracentrifugal Analysis
of the Proteins of Synovial Fluids and Sera from Patients with Rheumatoid Arthritis
Case
Table 2.-Analysis
0
0
0
1
0
0
0
0
IS
23
PROTEINS OF SYNOVIAL FLUID
Y
T
A
‘I1
A
Fig. 6.-Starch
3
gel electrophoretic pattern of the synovial fluid (A) and serum
(B) of a patient with juvenile rheumatoid arthritis.
hyaluronidase samples, 0.6 and 2.0 mg. respectively, were subjected to starch
gel electrophoresis. The 0.6 mg.-specimen which is equal to the amount of
enzyme used for the digestion of each joint fluid showed a single weak zone
in the albumin area and its contribution to the synovial fluid albumin zone
appeared to be negligible. The 2 mg.-pattern displayed seven bands, the largest
migrating with the albumin and the next with the haptoglobins. A small amount
of protein migrated with the mobilities of the 7-globulins.
DISCUSSION
In the present study a paper electrophoretic, ultracentrifugal and starch gel
electrophoretic investigation of four main groups of synovial fluids, normal,
postmortem, traumatic and rheumatoid joint fluids, are described. Although
certain data obtained from each of these three investigations represent confirmations of earlier observations, the increased aJaz-globulin ratio noted with
most joint fluids can now be explained on the basis of the information inte-
24
BINETTE AND SCHMID
grated from this study. Moreover, since this investigation includes the analysis
of a series of synovial fluids of each group, except that of the normal, the
relative rangc of variation of certain parameters of the different classes of
joint fluids can be indicated. As such parameters, protein concentration,
relative concentration of the 19s or Sa2-glycoprotein, a,-acid glycoprotein
and haptoglobins and the aJaZ-globulin ratio were considered (table 3 ) .
The decrease in the relative concentration of the 19s aa-macroglobulin and
haptoglobins which accounts for 45 and 15 per cent, respectively, of the total
a2-globulins of normal serum, is particularly striking for normal and postmortem synovial fluids. This decrease was observed to a lesser extent in many
pathological, including traumatic joint fluids of the present series and is in
agreement with reports of others who employed immunoele~trophoresis~~
and
electrophoresis in polyacrylamid.28 The relative concentration of a,-acid
glycoprotein accounting far about 20 per cent of the al-globulin fraction of
normal serum, was particularly increased in postmortem and normal synovial
fluids.
The difference in the al/a2-globulin ratio between the four groups of
synovial fluids studied and normal serum is thus primarily due to the change
in the relative concentration of three proteins: The ( 3S)a1-acid glycoprotein
is greatly increased and the 19s a,-glycoprotein and the (4s) haptoglobins
are significantly decreased in certain joint fluids as compared with the corresponding values of normal serum. The low relative level of the haptoglobins
and other plasma proteins has been reported previously.ll In an earlier study
on the characterization of the synovial fluid proteins6 using Cohn’s Method
1015 it was possible to demonstrate the highly increased level of cul-acid
glycoprotein, but the 19s az-glycoprotein escaped detection. This is probably
the major reason why the changed al/a2-globuiin ratios could not be explained
at that time.
The mechanism responsible for partial exclusion of the high molecular
weight 19s proteins and the increase of the low molecular weight proteins
with sedimentation coefficients of 4s and 3s (and perhaps that of IS),
respectively, is due in large part to the partial permeability of the plasma
proteins through the normal synovial membrane.1~3J1~2g
Comparable protein
distributions were reported for other extravascular fluids. In cerebrospinal
fluid the relative concentration of the 19s aa-globulins was found to be
significantly lower and that of the a,-globulin higher than that of normal
serum.3o Further, Villa and Rugarli31 who carried out pressure filtration experiments through the iliac artery wall, naked that the resulting filtrate contained a lower total protein concentration with a higher relative concentration
of albumin and al-globulin and a lower relative concentration of the other
globulins. Additional mechanisms must exist at least in the synovial membrane
to exclude almost completely specific proteins such as haptoglobinsll and
those of the clotting mechanism.29Suter and Majnd2 detected two kinds of
capillaries in the joint capsule. Permeability f a ~ t o r s might
~ ~ , ~regulate
~
the
permeability of these capillaries at different rates. In addition, the hyaluronic
acid, particularly that of the perisynovial connective tissue, may play an
25
I’ROTEINS OF SYNOVIAL FLUID
Table 3.--Comparison of Certain Properties of Normal Synovial
Fluid with Those of Nmml Serum
Ultracentrifugation
Technique
Biuret
Parameter:
Protein
coneentration
Body Fluid
9%
Normal
synovial
fluid
Normal
serum
2.0
7
Starch Gel Electrophoresis
Paper
electrophoresis
a~/al-
Globulin
ratio
1s1%Components Components
(Relative Percentage)
%
%
2
4*
1
0.5
ot
4
a:-Acid
19s-azglycopro- glycoprokin
tein
mg. %
mg. %
-250s
60r
-EOB
240Z
Haptoglobins
mg.
%
-608
loo$
*Highest value of postmortem synovial fluids.
?Not detectable.
$These values are taken from “The Plasma Proteins” by F. R . Putnam, ed., Academic Press, New
York, 1960.
$These values were estimated by comparing these bands with the corresponding bands of normal
serum.
important role in excluding certain plasma protein^.^^,^^,^^ Resorption of
specific proteins has also been proposedJ8 as a mechanism for achieving a
relative gradient of the proteins of these two fluids.
Wide variations in the above mentioned parameters of the synovial fluids
investigated in this and earlier studies6 as well as those by others1V6 were
noted particularly with traumatic joint fluids and synovial fluids from patients
with rheumatoid arthritis. The extreme values of these parameters (table 3 )
are those of normal synovial fluid on the one side and those of normal serum
on the other which form a clusteT on position A and B, respectively, as
a result of their constant composition (fig. 7). The postmortem synovial
fluids form a group of which some have properties similar to that of normal
joint and others cover a broad range toward normal serum. This presentation
clearly demonstrates that postmortem synovial fluid is not a substitute for
normal joint fluid. The group of the traumatic synovial fluids also exhibits a
wide range with regard to these parameters, but the position of the maximum
frequency of occurrence lies about half way between A and B, and forms
an intermediate range between postmortem and pathological fluids. The distribution of the joint fluids derived from patients with rheumatoid arthritis
differs significantly from that of the other groups of fluids. Most of these specimens are characterized by values which lie between those of traumatic joint
fluids and normal serum.
SUMMARY
Normal, postmortem and traumatic joint fluids and synovial fluids from patients with rheumatoid arthritis have been studied by paper electrophoresis,
ultracentrifugation and starch gel electrophoresis. This investigation confirms
certain results of earlier reported studies by these and other authors. Normal
and postmortem synovial fluids showed in comparison with normal serum a
significant increase in the relative concentration of albumin and especially of
-acid glycoprotein and a decrease in relative concentration of the hapto-
26
BINETTE AND SCHMID
8
I
PM
T
I
I
I
I
I
I
I
I
1
I
I
I
I
I
BI
I
2
3
4
5
6
7
A
Fig. 7.-Schematic representation of the distribution of four main groups of
synovial fluids. On the abscissa certain parameters (total protein concentration,
relative concentration of a,-acid glycoprotein, 19s a,-glycoprotein and al/a2-globulin ratio) are plotted in arbitrary units without regard to sign. Position H indicates
the corresponding values of normal synovial fluid ( F ) and position ( B ) that of
normal serum ( S ) . The range of distribution of the different synovial fluid groups
(postmortem without edema, PM; traumatic, T; rheumatoid arthritic, R.A. ) is indicated by the extension of the corresponding areas in the direction of the abscissa.
The width of these figures in the direction of the ordinate indicates a rough measure
of the relative frequency of occurrence of these fluids at a given position.
globins and particularly of the 19s a,-globulin. Further, the traumatic joint
fluids exhibited a 1s component which might represent a nevv finding.
Synovial fluids from patients with arthritis displayed patterns that, depending
on the joint involvement, were often similar to those of the patient’s serum.
Traumatic synovial fluids were found to represent a group of joint fluids which
affords patterns intermediate between normal and rheumatoid synovial fluids.
The present investigation demonstrates that the change in the al/cuz-globulin
ratio which is particularly prominent in normal and postmortem
synovial fluids
as compared with normal serum, is a consequence especially of the greatly
increased relative concentration of al-acid glycoprotein and a significant decrease in the relative concentration of the haptoglobins and the 19s a2globulin.
A schematic distribution of postmortem, traumatic and rheumatic joint
fluids between normal synovial fluid and normal serum is presented with
regard to certain parameters.
ACKNOWLEDGMENT
E. Schultze. Marburg, for the gift of tryptophanrich prealbumin, to Dr. Frank E. Hamerslag, Wyeth Laboratories of Medical Research,
for the generous gift of hyaluronidase, and to Drs. E. Calkins, M. Ropes, J. Heyl, J.
Bolzan, R. deAndrade, and J. D. Kenney for providing many synovial fluid specimens.
The expert technical assistance of Miss Alise Polis is acknowledged.
T h e authors are grateful to Prof. H.
27
PROTEINS OF SYNOVIAL FLUID
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K. Schmid, Dipl. rm. nsct., M.A., Dr. Phil., Associate Professor,
Department of Biochemistry, Boston University School of
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