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Experimentally Induced Degenerative Joint Lesions Following Partial Meniscectomy in the Rabbit.

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Experimentally Induced Degenerative Joint Lesions
Following Partial Meniscectomy in the Rabbit
Roland W. Moskowitz, Wirt Davis, James Sammarco, Mark Martens,
Jack Baker, Michael Mayor, A. H. Burstein and V. H. Frankel
Degenerative lesions characterized by osteochondrophyte spur formation
and cartilage degeneration were induced by partial meniscectomy in the
rabbit knee. Cartilage degenerative lesions included ulceration, fissure and
cyst formation, and diminished concentration of matrix proteinpolysaccharide. Proliferation of chondrocytes represented efforts at repair.
Osteophytes increased in size with time. Changes resembled certain components of degenerative joint disease. The partial meniscectorny animal
may be useful in studies of osteoarthritis pathogenesis, pathology and
treatment.
Degenerative joint changes frequently follow
meniscus injury in the human (1,2). In the
present study degenerative lesions characterized
by osteochondrophyte spur formation and cartilage degeneration were induced by partial meniscectomy in the rabbit knee. Since the pathologic changes observed resemble certain
components of degenerative joint disease, the
partial meniscectomy animal may be useful in
studies of osteoarthritis pathogenesis, pathology
and treatment.
From the Departments of Medicine and Orthopedic Surgery, Case Western Reserve University School of Medicine,
Cleveland, Ohio.
This work was supported by Research Grant AM 12546
and Graduate Training Program in Orthopedics, 5T01AM 05437, The National Institutes of Health, US Public
Health Service, and Rainbow Research Fund.
ROLAND W. MOSKOWITZ, MD: Associate Professor of Medicine; WIRT DAVIS, MD: Formerly, Assistant Professor of Orthopedic Surgery; JAMES SAMMARCO, MD: The National Institutes of Health Research Fellow; MARK MARTENS, MD:
Research Fellow in Biomechanics; JACK BAKER, MI): The
National Institutes of Health Research Fellow; MICHAEL
MAYOR, MD: Formerly, Resident in Orthopedic Surgery and
The National Institutes of Health Research Fellow; A.H.
BURSTEIN, PHD: Associate Professor of Mechanics and Orthopedic Surgery; V.H. FRANKEL, MD, PHD: Professor of
Orthopedic Surgery and Biomechanical Engineering.
Reprint requests should be addressed to: Dr. Roland W.
Moskowitz, 2073 Abington Road, Cleveland, Ohio 44106.
Submitted for Publication Oct 2, 1972; accepted Dec 28,
1972.
MATERIALS AND METHODS
New Zealand white rabbits were divided into three
groups for study. The animals in the three groups were of
relatively similar characteristics with respect to range of
weight and to average and median weights (Table 1).
Group 1, 82 rabbits, comprised the experimental meniscus
surgery model. The rabbits were anesthetized with IV sodium pentothal (Diabutalm, E.L. Squibb and Sons, New
York, NY) after which, under sterile surgical conditions, an
anteromedial incision was made in the right knee. The soft
tissue and retinaculum were incised and reflected medially
and laterally. The joint capsule was then incised and the
medial compartment of the knee exposed. Using iris scissors, the peripheral attachment of the anterior one-half of
the medial meniscus was released and the anterior one-third
of the meniscus excised (Figure 1). The capsule and medial
retinaculum were closed with three 5-0 catgut sutures and
the skin closed with 4-0 silk suture. The operated extremity
was placed in a long-leg plaster cylinder cast with the knee
and ankle joints maintained at angles of approximately 9W.
After 1 week the cast and skin sutures were removed and
the animals allowed to move freely in their cages. Sacrifice
using intravenous injection of Diabutal was carried out at
periodic intervals of 2, 5, 8 and 12 weeks postoperatively
(Table 1). The largest number of animals in the series comprised those sacrificed at 12 weeks, since this became the
standard model being used in ongoing experimental studies.
The operated knees were carefully dissected and examined
for gross changes, Particular note was made of the presence
of osteophyte-like spur formation, pitting and ulcerative
erosions. Severity of spur formation was graded as follows:
mild-spur present and less than 2 mm in maximal width;
moderate-spur 2 mm in maximal width: and severespur exceeding 2 mm in maximal width. Ulcers were graded
Arthritis and Rheumatism, Vol. 16, No. 3 (Yay-June 1973)
397
MOSKOWITZ ET AL
Table 1.Comparison of Gross Pathologic Changes Observed in Tibias
Femur
Osteophyte formation
~~
Weeks
Initial weight (kg)
Present
Mild
Moderate
Severe
---after
No. of
surgery animals Range Average Median No. 70 No. 70 No. 9%
No. 9%
Partial meniscectomy model
Sharn-operated
controls
Nonoperated
controls
2
5
8
12
10
9
13
50
1.9-4.4
1.7-3.7
2.0-4.0
1.5-3.6
2.1
2.4
2.7
2.4
2.3
2.4
2.5
2.3
4
5
7
21
40
56
54
42
4
2
5
9
100
40
71
43
0
3
1
3
0
60
15
14
0
0
1
9
0
0
15
43
3to 20
28
2.2-4.6
2.8
2.7
1
4
0
0
0
0
1
100
-
24
1.5-3.6
2.4
2.2
1
4
1
100
0
0
0
0
'Ulcers graded 1 through 3 in severity according to depth of erosion (see Materials and Methods).
according to depth of erosion and size. Grade 1 ulcers extended in depth to involve only superficial layers of cartilage; Grade 2 ulcers extended into the mid-layers of cartilage; and Grade 3 ulcers extended down to subchondral
bone. Area size of ulcers was measured by multiplying
length and width, with the result given in square millimeters.
Distal femures and proximal tibias were prepared for
histologic examination by fixation in 10% formalin followed
by decalcification in a formic-citrate solution. Coronal sections of the distal femur and proximal tibia were embedded
in paraffin and cut in 8-r sections. Stains were made with
hematoxylin and eosin for routine study and Safranin 0
with fast green counterstain to assess loss of proteinpolysaccharide concentration in cartilage ground substance (3,4).
Tissues were mounted in synthetic mounting medium for
study. Group 2,28 rabbits, represented a sham-operated series. Surgical procedures and plaster immobilization as described in Group 1 were carried out, except that the meniscus and its attachments were left intact. Animal sacrifices
were carried out at 3 weeks (2 animals), 5 (1 animal), 7 (1
animal), 10 (1 animal), 11 (3 animals), 12 (11 animals), 16
(3 animals), 18 (1 animal) and 20 (5 animals) weeks postoperatively. Group 3,24 rabbits, represented a further control series comprising normal left knees from animals in
Groups 1 and 2. Gross and histologic examination were
carried out as described in the partial meniscectomy series.
RESULTS
Gross Examination
In the partial rneniscectomy series, os398
teophyte-like spur formation on both the tibia
and femur was a prominent finding (Table 1).
In most specimens tibial spurs involved the entire rim of the medial tibial plateau (Figure 2)
rather than being limited to the area of meniscus removal. T h e site from which the meniscus had been removed revealed replacement
with a proliferating pannus of connective tissue
which was adherent to the remainder of the
meniscus and adjacent structures. This substitute tissue increased in size and firmness in
parallel to duration of time till sacrifice. Femoral spurs were located primarily along the medial edge of the distal medial condyle. S,pur formation was seen as early as 2 weeks after surgery. Tibial spurs generally preceded femoral
spurs in development and were more frequent
at all time intervals studied. Tibial spurs severe
in grade were seen early, and the percentage of
animals with severe spurs increased with time.
Femoral spurs of severe grade similarly increased in frequency with time but generally
required a longer period to develop. Spurs in
late-sacrifice animals tended to be more firm
and almost like bone in consistency. Tibial osteophytes alone were noted in 39 animals; 36
animals had both tibial and femoral os-
Arthritis and Rheumatism, Vol. 16, No. 3 (May-June 1973)
DEGENERATIVE JOINT LESIONS
and Femurs of Partial Meniscectomy and Control Series of Animals
Tibia
Ulcer formation
Pitting
Present
No.
%
No.
%
1
6
10
22
10
67
77
44
3
2
6
32
30
22
46
64
1
4
0
0
0
0
Osteophyte formation
Average
size Average
(sq mm) grade*
Mild
Present
No.
%
No.
Moderate
Severe
%
No.
%
No.
%
6
2.3
5.5
11.0
1.8
1.7
2.0
1.5
9
9
10
47
90
100
77
94
2
1
1
7
22
11
10
15
6
4
3
11
66
44
30
23
1
4
6
29
11
44
60
62
0
-
-
3
11
0
0
1
33
2
67
0
-
-
1
4
1
0
0
1
0
0
0
0
teophytes. Osteophytes on the femur alone were
seen in only 1 animal in the series. Spur formation was infrequent in the sham-operated and
nonoperated control series (Table 1).
Cartilage pitting and ulcerative erosions
were observed on the femur only (Table 1). Pitting was characterized by multiple punctate
erosions distributed over various-sized areas of
cartilage. Changes were localized primarily to
the distal aspect of the medial femoral cartilage
in contact with the medial tibial plateau. Pitting
was observed in 1 animal as early as 2 weeks after surgery. A diminished frequency of pitting
was noted in the 12-week animals. This most
likely reflects an increased severity of cartilage
degeneration, with pitting areas coalescing to
form ulcers, seen with increased frequency at
this ,time period. Ulcerative lesions consisted of
larger erosions of various size and depth. In 24
animals, single ulcerations were. seen; 17 animals had 2 ulcers and 2 animals had 3 ulcers
Fig 1. Right tibia, partial meniscectomy model.
The peripheral attachment of the anterior one-half
of the medial meniscus is released and the anterior one-third of the meniscus excised.
Fig2. Right and left tibias, 12 weeks after partial
rneniscectomy. Osteophyte-like formation (arrow)
is seen along the entire r i m of the medial tibial
plateau of the right tibia. Left tibia is normal.
Arthritis and Rheumatism, V d . 16, No. 3 (MayJune 1973)
399
MOSKOWITZ E l AL
Fig 3. Chondrophyte, medial tibial plateau, 8 weeks after partial meniscectomy.
The spur is composed mainly of young hyaline and fibrocartilage(H&E. x 80).
young hyaline cartilage at the base with fibrocartilage toward the surface (Figure 3). Uptake of Safranin 0 stain in the spur was minimal or absent (Figure 4). Late firm spurs (osteochondrophytes) consisted of hyaline and
fibrocartilage overlying a proliferating projection of bone (Figures 5 and 6). The junction between bone and overlying cartilage was often
indistinct. Clones of proliferating chondrocytes
were at times seen at the margin of the spur and
normal cartilage. Spur formation in the shamoperated and nonoperated control knees was
characterized in 2 cases by small irregular projections of mature bone covered with normal
appearing cartilage. Configurational changes
associated with sectioning of tissues could not be
Histologic Examination
excluded. In the 4 cases in which gross spur forThe tibial and femoral spurs observed grossly mation was moderate or severe the histologic
in the partial meniscectomy animals were histo- appearance of the spurs was similar to that delogically similar in nature and varied with their scribed in the partial meniscectomy animals.
stage of development. Early soft spurs
The ulcerations noted grossly in the femurs
(chondrophytes) were composed mainly of in the partial meniscectomy model were charac-
each. As observed with pitting lesions, ulcers
were noted most frequently at the distal aspect
of the medial femoral condyle, being seen in this
location in 37 of the 43 animals with ulcers. U1cer formation was next most common at the
posterior aspect of the medial femoral condyle,
occurring in 24 animals. Three animals had ulcers at the distal aspect of the lateral femoral
condyle. Ulcer size was greatest in the 12-week
series, with depth of erosion somewhat less at
this time interval. As noted in Table 1, gross
pitting or ulceration were absent in all but 1 animal in the sham-operated and nonoperated
control series.
400
Arthritis and Rheumatism, Vol. 16. No. 3 (May-June 1973)
DEGENERATIVE JOINT LESIONS
Fig 4. Chondrophyte, medial tibial plateau, 8 weeks after partial meniscectomy. The
spur (arrow 1) stains minimally with Safranin 0 in contrast to normal cartilage (arrow
2) (Safranin 0 with fast green counterstain. x 45).
terized histologically by degeneration of cells,
prominent fissure formation, fibrillation and
loss of cartilage layers to various degree (Figure
7). Matrix proteinpolysaccharide concentration
in these regions was often markedly diminished
as evidenced by decreased Safranin 0 stain.
Proliferation of chondrocytes in clones (Figure
7), similar in appearance to Weichselbaum’s lacunae (S), was seen. In several specimens cyst
formation was seen in close proximity to the ulcers (Figure 7). Early cyst formation was characterized by degeneration of cells lying at the
center of proliferating clones of chondrocytes.
Mature cysts were filled with nonsp&ific detritus. Fissure formation was often striking, particularly in areas adjacent to the ulcers. Histologic study of the tibias in the partial
meniscectomy series revealed only infrequent
evidence of cartilage ulceration. Such ulceration, when seen, consisted of loss of superficial
layers of cartilage over a small area. Fissure formation was seen primarily on the medial tibial
plateau and did not exceed that described pre-
viously in the normal rabbit knee (6).
In the sham-operated and normal control series histologic evidence of cartilage ulceration
was rare and, when seen, was characterized by
superficial loss of cartilage over a small area. As
in the partial meniscectomy series, changes
were noted in femoral cartilage only. Fissure
formation on both the femoral and tibial surface
did not exceed that seen in normal rabbit
knees (6). Diminished Safranin 0 stain was observed on the medial femoral condyle in 2 animals.
DISCUSSION
Degenerative joint disease is the most commonly occurring form of arthritis. In many
cases it is benign and asymptomatic. Involvement of certain joints, such as the hip, knee
or cervical spine may lead, however, to disabling disease in a significant number of persons. Studies related to disease pathogenesis
and treatment would be greatly aided by the
Arthritis and Rheumatism, Vol. 16, No. 3 (May-June 1973)
401
MOSKOWITZ El AL
Fig 5. Osteochondrophyte, medial tibia1 plateau, 16 weeks after partial meniscectomy. The spur consists of hyaline and fibrocartilage (arrow) overlying a proliferating
projection of bone (H&E, x 45).
Fig 6. Osteochondrophyte.
medial femoral condyle, 12
weeks after partial meniscectorny. A large spur is seen,
consisting of hyaline and fibrocartilage overlying a large projection of bone at its core.
(H&E, x 20).
402
Arthritis and Rheumatism, Vol. 16, No. 3 (May-June 1973)
DEGENERATIVE JOINT LESIONS
Fig 7. Degenerative changes, medial femoral condyle, 12 weeks after partial
rneniscectorny. Fissure-formation, cell degeneration, proliferation of chondrocytes in
clones and cyst formation are seen (H&E. x 140).
availability of experimental models which simulated the naturally occuring disease in whole,
or in part, in pathogenesis and pathology. A
number of experimental technics which produce
various aspects of the lesions seen in osteoarthritis have been described (7-1 3). These technics utilize various procedures of induction, including surgical alteration of normal joint
structures (8, 9), compression (ll), intraarticular injection of various agents (7, 13) and
breeding of animal strains genetically predisposed to develop osteoarthritis (12). Partial
meniscectomy in the rabbit produces certain
pathologic changes which are similar to lesions
seen in osteoarthritis as it occurs in the human.
Chondrophyte and osteochondrophyte spur formation were seen on the medial tibia1 plateau
and the medial femoral condyle. Ulceration, fissure formation, cyst formation and diminished
concentration of matrix proteinpolysaccharide
were observed on the medial femoral condyles.
Attempts at repair, characterized by proliferation of chondrocytes in clones, were also noted.
Severity of osteophyte formation tended to increase with time. These more severe spurs were
of larger size, more firm and, on histologic examination, revealed increased bone proliferation. Changes of similar nature were seen only
infrequently in control animals.
In addition to its definition on the basis of a
characteristic set of histologic features, osteoarthritis has been defined on the basis of certain
observed biochemical changes. Studies utilizing
%O4 and 3H-thymidine have demonstrated an
increased synthesis of polysaccharides and
DNA, respectively, in human osteoarthritic cartilage (14). Similarly, increased synthesis of
DNA was noted in degenerating cartilage in
rabbit knee joints following experimentally induced degenerative joint disease (15). The validity of the partial mensicectomy procedure as
a model of degenerative joint disease would be
Arthritis and Rheumatism, Vol. 16, No. 3 (May-June 1973)
403
MOSKOWITZ ET AL
strengthened if similar biochemical changes pathologic components characteristically dewere seen. Preliminary analysis of studies by us scribed in osteoarthritis. T h e development of
in this model do reveal an increased intake of degenerative lesions simulates a naturally ocboth 35S0, and 3H-thymidine by cartilage com- curring sequence in the human in which deponents in the areas of ulceration and os- generative changes follow meniscus injury (1,
teophyte formation. Uptake of 3H-thymidine 2). It is suggested that this experimental techwas maximal at 5 weeks, whereas maximal up- nic may be of value in studies of degenerative
take of 35S0, occurred at 12 weeks.
joint disease in the human.
No detailed analysis has been made at this
time to relate the pathologic changes noted in
ACKNOWLEDGMENTS
this study to rabbit age and weight. T h e relaThe technical assistance of Mary Lee Berman and Linda
tively si,milar composition of the groups studied Fritz is gratefully acknowledged.
with respect to these factors allowed valid comparison for the purposes of this study without
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