Clegg, P.D.

Innes, J.

Redmond, C.

Rhodes, N.

Vaughan-Thomas, A.

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Osteoarthritis and Cartilage Vol. 14, Supplement B S45
Fig. 2. Knee joint, articular cartilage, normal control, ank+/ank+ mouse. Figure
width = 100 μm.
capsule and periosteum occurs, followed by osseous metaplasia
and ultimately ankylosis. With age, the sequence of histologic
changes proceeds progressively to involve the proximal joints as
well.
Conclusions: The ank-/ank- mouse demonstrates chondrocyte
hypertrophy, joint space calcification and excessive peri-articular
ossification, all features that occur to more or less extent in
osteoarthritis. As these features develop and progress over 18
weeks of life, we conclude that the ank-/ank- mouse is a useful
model for investigating the pathogenesis and regulation of these
articular morphologic features of osteoarthritis.
A57
CHONDROGENESIS OF INFRAPATELLAR FAT PAD AND
SYNOVIAL MEMBRANE CELLS IN COMPARISON TO
ARTICULAR CARTILAGE CHONDROCYTES
P.D. Clegg, A. Vaughan-Thomas, C. Redmond, N. Rhodes,
J. Innes
The University of Liverpool, Neston, United Kingdom
Purpose: Cartilage is frequently damaged and only has a limited
capacity for repair. There is growing interest in the use of cell
based therapies for enhancing cartilage repair. Autologous chon-
drocytes have been used in the repair of focal articular cartilage
defects, although there are issues with donor site morbidities.
There has been recent interest in the use of undifferentiated
adult progenitor cells from other sources, which under appro-
priate culture conditions can be differentiated into chondrocytes.
Such cells can be sourced from a variety of tissues, includ-
ing the synovial membrane (SM) and the infrapatellar fat pad
(IPFP). In this study we identified the chondrogenic potential of
canine cells sourced from the SM and the IPFP, and compared
their potential with articular cartilage chondrocytes (ACC). We
hypothesised that IPFP and SM derived cells would have compa-
rable chondrogenic potential in comparison to ACC in an in-vitro
chondrogenesis model.
Methods: IPFP, SM and articular cartilage were harvested from
the femoropatellar joint of 4 skeletally mature dogs (aged 3-5
years) which had no signs of joint disease. Cells were iso-
lated by collagenase digestion and proliferated for two passages
in monolayer culture. 3D cell aggregates were formed from
500,000 cells by centrifugation. The cell aggregates were placed
in chondrogenic media and cultured for 2 weeks. Chondrogene-
sis was assessed by: Wet weight, Histologically (H&E, safranin
O staining and collagen-II immunohistochemistry), Gene expres-
sion (real-time PCR quantification of collagen-I, -II, aggrecan),
and Biochemically (Total glycosaminoglycan (GAG) and DNA
content).
Results: ACC derived cell aggrecates were significantly bigger
than aggrecates derived from either IPFP or SM cells (p<0.05),
with the mean wet weight of the aggrecates being 5.2mg for
the ACC cells, 1.2mg for the IPFP cells and 1.1mg for the SM
cells. Histologically the IPFP and SM cell derived aggrecates
showed significantly lower matrix accumulation in comparison to
the aggrecates from ACC cells, and stained much more poorly
for both GAG and Collagen-II. Similarly, both total GAG and DNA
content of the aggrecates were significantly lower in both the
SM and IPFP cell derived aggrecates in comparison to the ACC
cell derived aggrecates. Average GAG content per cell aggrecate
was over five fold lower in aggrecates derived from SM cells, and
three fold lower in aggrecates derived from IPFP, in comparison to
those derived from ACC. ACC derived cell aggrecates expressed
high levels of both collagen-II and aggrecan genes, although
collagen-I gene expression was of a similar magnitude to that of
collagen-II expression. Average ratio of collagen-II to collagen-I
gene expression per cell aggrecate was over 500 fold lower in
aggrecates derived from SM cells, and nearly a 100 fold lower
in aggrecates derived from IPFP, in comparison to those derived
from ACC.
Conclusions: Under the culture conditions used in this study, cell
aggrecates derived from ACC were able to produce a cartilage
like matrix with the cells showing gene expression indicative of a
chondrogenic phenotype. However, the ACC cells showed similar
levels of collagen-I gene expression in comparison to collagen-
II gene expression indicating a failure to fully recapitulate the
phenotype of cells found within hyaline articular cartilage. Using
the cell extraction protocol and culture conditions used in this
study, neither SM nor IPFP derived cells demonstrated ability
to produce a cartilage like matrix, nor did these cells demon-
strate a gene expression profile consistent with chondrogenic
differentiation.
A58
PREVALENCE OF RADIOGRAPHIC OSTEOARTHRITIS
OF KNEE AND LUMBAR SPINE, AND ITS ASSOCIATION
WITH PAIN: THE RESEARCH ON OSTEOARTHRITIS
AGAINST DISABILITY (ROAD) STUDY
H. Kawaguchi1, N. Yoshimura1, S. Muraki1, H. Oka1,
A. Mabuchi1, Y. En-yo2, M. Yoshida2, A. Saiga2, T. Suzuki3,
H. Yoshida3, H. Ishibashi3, S. Yamamoto3, K. Nakamura1
1University of Tokyo, Tokyo, Japan, 2Wakayama Medical
University, Wakayama, Japan, 3Tokyo Metropolitan Institute of
Gerontology, Tokyo, Japan
Purpose: Although osteoarthritis (OA) of knee and lumbar spine
is a major cause of disability in the elderly, few epidemiologic
studies have been performed. We established a large-scale na-
tionwide clinical study called ROAD (research on osteoarthritis
against disability) in 2005 to clarify the environmental and genetic
backgrounds. We began the study by creating a comprehensive
and systemic database including clinical and genomic informa-
tion in two cohorts of urban and rural areas. From their baseline
data, we investigated the prevalence of radiographic OA of knee
and lumbar spine, as well as its association with the respective
local pain.
Methods: We recruited 1,885 inhabitants older than 50 years
(mean age=73.5 yrs.): 1,030 (355 men & 675 women; mean
age=76.9 yrs.) from the urban and 855 (317 men & 538 women;
69.3 yrs.) from the rural areas. The radiographic severity of OA
was determined according to the Kellgren/Lawrence (KL) grade
(0-4) at femoral-tibial joints of bilateral knees and at intervertebral
spaces from L1/2 to L5/S1 of the lumbar spine by a blinded
orthopaedic surgeon. Logistic regression analysis was performed
after adjustment for age to determine the association.
Results: Prevalence of radiographic OA (KL> or = 2) in either


A57 CHONDROGENESIS OF INFRAPATELLAR FAT PAD AND SYNOVIAL MEMBRANE CELLS IN COMPARISON TO ARTICULAR CARTILAGE CHONDROCYTES 

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A57 CHONDROGENESIS OF INFRAPATELLAR FAT PAD AND SYNOVIAL MEMBRANE CELLS IN COMPARISON TO ARTICULAR CARTILAGE CHONDROCYTES

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