Appendix 2. Final inclusion list—study summaries by category
A. Diagnostic (n = 16)
Biomarker
identified
TIMP-1, HA
Tissue type, assays, and
techniques used
Serum; ELISA
45 symptomatic OA
versus 33 control
subjects
YKL-40 , CRP
Serum; immunoassay,
ultrasensitive
immunonephrotelemetry
Serum YKL-40 was significantly increased in
patients with hip OA
40 hip OA versus 75
control subjects
uHelix-II, uCTXII
Serum; ELISA
Increased uHelix-II levels are associated with
RDOA, independently of uCTX-II
40 hip OA versus 65
control subjects
CTX-II, free DPD
Urine; ELISA, LC
Patients with hip OA have increased CTX-II
degradation; increased uCTX-II levels are
associated with RDOA
55 advanced OA versus
30 control subjects
TNFα, TIMP-1,
IL-1α, IL-8, IL10, MMP-2,
MMP-3, MMP-9,
TIMP-1, CRP
CTX-II
Serum; ELISA, immunoassay;
OA pathology is proinflammatory condition
Urine; ELISA
Collagen type II derived fragments may serve as
markers for OA
58 hip OA versus 19
control subjects
RANKL, OPG,
OC, b-ALP
Serum; ELISA, immunoassay
None of the markers studied are adequate
surrogates for radiographic imaging in hip OA
7 OA versus 19 control
subjects
CRP,TGF-β1,
GM-CSF, INF-γ;
IL-8, IL-1β, IL-6,
IL-10, TNF-α, IL12p70; IL-2, IL-4,
IL-5, IL-10, TNFα
Serum; ELISA; cytometric
bead assay; BD cytomeric
human T-helper cell type 1/Thelper cell type 2 cytokine kit
CS-CC; assess
associations between
serum TGF-β1 and
radiographic hip OA
68 radiographic hip OA
versus 241 control
subjects
TGF-β1
Serum; ELISA
2013
CC; assess the activity of
CAT-D and alpha-1
antitrypsin
(AAT) in hip OA
46 hip OA (37
unilateral, 9 bilateral)
versus 54 control
subjects
CAT-D, AAT
Serum; Anson's method,
Eriksson's method
Reijman et
al [37]*
2004
123 OA versus 1112
control subjects
CTX-II
Urine; ELISA
Salvatierra
et al [38]
1999
CS-L; investigate the
association between
uCTX-II and the
prevalence and
progression
of radiographic OA of the
knee and hip
CS-CC; assess
associations between NO
concentration and hip OA
27 hip OA (16
ambulatory HOA; 11
surgical HOA) versus
12 control subjects
CRP, NO (as
Nitrile)
Serum; nephelometry, Greiss
reaction + OD550
Silvestri et
al [40]
2006
34 hip OA versus 38
control subjects
sIL-4R
Serum; ELISA
Stewart et al
[43]
1999
CS-CC; assess
associations between
serum sIL-4R and
different forms of OA
(hand, hip, knee) and
healthy control subjects
CS-CC; assess
associations among bone
mass, bone turnover, and
OA and control groups had comparable CRP as
well as proinflammatory and antiinflammatory
cytokine concentrations; the major exception
was TGF-b, which was higher in OA patient
plasma; the H and L subunits
of ferritin in the bone marrow macrophages were
both significantly higher in OA subjects as
compared with control subjects; a role for TGF-b
in the upregulation of the
expression of the H-subunit and possibly the Lsubunit of ferritin is postulated in osteoarthritis
Serum TGF-b1 varied by race and gender and
several rOA variables but did not exhibit
independently significant associations with
presence, laterality, or severity of knee or hip
rOA by KeL grade or IRFs, suggesting that
serum TGF-b1 is unlikely to be useful as
a standalone biomarker in OA studies
Preoperative CAT-D activity in hip OA pt is
significantly lower than CAT-D activity in
healthy patients (uni- and bilateral); preoperative
AAT activity in patients with hip OA is
significantly higher than ATT activity in healthy
patients (uni- and bilateral); the low CAT-D
activity in OA of big joints is associated with a
decrease of cartilage cells during the
degenerative process; the higher activity of acute
phase protein AAT in OA patients’ blood serum
confirms the inflammatory component in the
osteoarthritis process
This study shows that CTX-II is associated with
both the prevalence and the progression
of radiographic OA at the knee and hip;
importantly, this association is independent of
known clinical risk factors for OA and seems
stronger in subjects with joint
pain
Study shows increased NO levels in joint
cartilage of patients with hip OA; higher NO
levels found in macroscopically deteriorated
areas; high serum NO levels in patients with hp
OA may be the result of joint cartilage
destruction
Increased levels of sIL-4R observed in patients
with OA compared with healthy individuals; this
may reduce availability of IL-4 and its effects on
chondrocytes
30 hip OA versus 30
control subjects
OC, Pyd, DPD
Serum; in-house assay, HPLC,
HPLC
Study
Year
Study design/objectives
Definition population
Chevalier et
al [9]*
2001
29 hip OA versus 225
control subjects
Conrozier et
al [11]
2000
Garnero et
al [18]*
2006
Garnero et
al [19]*
2003
Hulejová et
al [21]
2007
Jung et al
[22]
2004
Kenanidis et
al [25]
2011
Koorts et al
[27]
2012
Prosp CC; determine
predictive value of TIMP1 and HA in a 1-year
prospective study of hip
OA
CS-CC; study serum
levels of YKL-40 in hip
OA and its relation with
CRP
CS-CC; investigate the
association of rapidly
destructive hip OA with
urinary Helix-II and
urinary CTX-II
CS-CC; compare type II
collagen degradation in
patients with RDOA and
SPOA
CS-CC; investigate the
expression of several
cytokines, MMPs and
TIMP-1 in OA and control
sera
CS-CC; evaluate
degradation of articular
cartilage using fragments
of collagen type II
CS-CC; determine serum
biomarker levels in
patients with OA of
varying radiographic
severity and healthy
control subjects
CC; investigate the
expression of the H- and
L-subunits of ferritin in
bone marrow
macrophages of patients
with OA
Nelson et al
[34]
2009
OlszewskaSlonina et al
[36]
51 hip OA versus 48
control subjects
Main outcome of the study
TIMP-1 serum level may serve to predict the
evolution of patients with hip OA
Increased bone turnover was restricted to the OA
group
Sweet et al
[44]
1988
Vos et al
[46]
2010
radiological OA in OA,
OP, and control groups
CS-CC; assess the
associations between KS
concentration and hip OA
before and after THA
CS; study the role of
cartilage AGE levels in
OA by analyzing skin and
urine samples
31 OA versus 41
control subjects
KS
Serum; ELISA
67 hip OA (grade ≥ 2)
versus 138 control
subjects
Pentosidine,
CTX-II
Skin, urine; HPLC, ELISA
Tissue type, assays, and
techniques used
Serum, HTRF
Patients with hypertrophic OA may have a
generalized imbalance of cartilage proteoglycan
metabolism; measurements of serum KS are
likely to prove most useful in studying this
particular subset of patients with generalized OA
The higher skin and urinary pentosidine levels in
those with mild compared with no radiographic
joint damage and low versus high cartilage
breakdown, respectively, suggest that AGEs may
contribute to disease susceptibility and/or
progression; however, relations are weak and
cannot be used as surrogate markers of severity
of OA
B. Disease staging (n = 15)
Author
Year
Study design/objectives
Definition population
Abe et al [1]
2014
136 hips (33 RDOA, 57
OA, 10 RA, 36 ON)
Berger et al
[5]
2005
38 OA (18 RDOA, 20
SPOA) subjects
ICTP, bALP, OC,
PINP
Serum; ELISA, immunoassay
RDOA is associated with elevated serum ICTP
levels
Catterall et
al [6]
2012
Serum; ELISA
D-COMP was associated with radiographic hip
OA; total COMP associated with knee OA
1998
450 subjects from
population-based
sampling
35 OA (10 RDOA, 20
SPOA) subjects
D-COMP, total
COMP
Conrozier et
al [12]
CRP
Serum;
immunonephrotelemetry
RDOA may be associated with some degree of
inflammation
Conrozier et
al [15]*
1998
48 hip OA subjects
COMP, BSP,
CRP
Serum; ELISA,
immunonephrotelemetry
COMP seems to be a surrogate marker of OA
and may be of interest for the detection of
patients at risk of rapid progressing disease
Conrozier et
al [13]
2007
59 OA (17 atrophic
OA, 42 hypertrophic
OA) subjects
CS846, CPII,
C2C and C1/C2
Serum; ELISA
Atrophic hip OA is associated with reduced
collagen synthetic activity
Conrozier et
al [14]
2004
50 OA (25 atrophic, 25
hypertrophic) subjects
CRP, MMP1,
TIMP-1, HA,
COMP, CP-I,
CTX-I
Serum; ELISA, immunoassay,
immunonephrotelemetry,
peptide digestion
Serum biomarkers able to demonstrate
differences between the atrophic and
hypertrophic patterns of OA are lacking
Garnero et
al [19]*
2003
40 hip OA (12 RDOA;
28 SPOA) subjects
CTX-II, free DPD
Urine; ELISA, LC
Patients with hip OA have increased CTX-II
degradation; increased uCTX-II levels are
associated with RDOA
Garnero et
al [20]
2005
376 hip OA subjects
PINP, PIIINP,
COMP, YKL-40,
HA, MMP1,
MM3, CRP,
uCTX-I, uCTX-II
Serum, urine; ELISA, RIA,
immunonephrotelemetry
CTX-II showed the most consistent association
with symptoms and joint damage of OA
Garnero et
al [18]*
2006
40 hip OA subjects
uHelix-II, uCTXII
Serum; ELISA
Increased uHelix-II levels are associated with
RDOA, independently of uCTX-II
Kawasaki et
al [23]
2001
CS; evaluate cytokine
level characteristics in the
hip fluid
CS; compare ICTP in
patients with RDOA and
SPOA
CS; investigate the use of
deaminated COMP to
monitor OA
CS; compare serum CRP
levels in patients with
RDOA versus SPOA
L; evaluate COMP and
BSP as predictors of
disease progression in hip
OA
CS; determine if serum
levels of biomarkers can
distinguish between
atrophic and hypertrophic
hip OA
CS; determine the
epidemiological,
radiological, and
biological differences
between hypertrophic and
atrophic hip OA
CS-CC; compare type II
collagen degradation in
patients with RDOA and
SPOA
CS; investigate the
associations of molecular
markers and joint tissue
turnover with clinical and
radiological variables of
hip OA
CS-CC; investigate the
association of rapidly
destructive hip OA with
urinary Helix-II and
urinary CTX-II
CS; evaluate
concentrations of YKL-40
in hip diseases
Biomarker
identified
IL-1β, IL-6, IL-8,
TNFα
YKL-40
Synovial fluid; ELISA
YKL-40 reflects the degree of inflammation
rather than cartilage metabolism
Stannus et
al [42]
2010
CS; describe the
associations among leptin,
IL-6, and hip radiographic
OA in older adults
45 OA (19 OA
secondary to DDH, 21
OA secondary to
ONFH, 5 failed THA)
subjects
193 subjects
Leptin, IL-6
Serum; RIA
Yamada et
al [47]
1999
50 hip OA (0 pre, 17
early, 6 advanced, 27
terminal; 8 primary OA,
42 secondary OA to
DDH) subjects
C4S, C6S
Synovial fluid; HPLC
Yamada et
al [48]
2000
50 hip OA (6 early, 12
advanced, 32 terminal;
8 primary OA, 42
secondary OA to DDH)
subjects
L2, L4 (fragments
of digested KS)
Synovial fluid; digestion +
HPLC
The levels of KS-related disaccharide isomer
vary with severity of disease in hip OA; analysis
of these KS-related disaccharide isomers by
HPLC provides information on both the content
and sulfation pattern of KS in SF, reflecting the
metabolism of cartilage aggrecan
Yamaguchi
et al [49]
2014
CS; investigate
significance of chondroitin
sulfate isomers as markers
reflecting extracellular
matrix metabolism in joint
tissues
CS-CS; determine content
and sulfation pattern of
KS in synovial fluid in
patients with hip OA and
investigate its significance
as a marker of cartilage
matrix metabolism
CS; compare the levels of
bone and cartilage
metabolism markers in the
synovial fluid of the hip
between patients with
Metabolic and inflammatory mechanisms may
play a role in the etiology of hip
OA; the associations between body composition
and hip JSN may be mediated by leptin,
particularly in women
The ratio of CS isomers in synovial fluid in hip
OA varies with the severity of disease; these
molecules in synovial fluid may serve as a useful
marker reflecting extracellular matrix
metabolism in OA
41 OA (21 RDA, 20
DDH [7 early OA, 13
advanced OA]) subjects
BAP, TRACP-5b,
MMP-3, KS
Synovial fluid; ELISA
The patients with ONFH showed a relatively
bone formative condition before the
osteoarthritic
stage and maintained a higher rate of cartilage
turnover throughout several stages compared
Main outcome of the study
IL-8 in RDOA was higher than in other hip
diseases
secondary OA to ONFH,
RDA, or DDH
with the patients with RDA and DDH; patients
with RDA were characterized by a significantly
high osteoclast activity
C. Prognostic (n = 11)
677 white women
Biomarker
identified
COMP, NTX
Tissue type, assays, and
techniques used
Serum; ELISA
1104 men
25(OH)D
Serum; mass spectrometry
Men with Vitamin D deficiencies are twice as
likely to have prevalent radiographic hip OA
29 hip OA subjects
TIMP-1, HA
Serum; ELISA
TIMP-1 serum level may serve to predict the
evolution of patients with hip OA
48 hip OA subjects
COMP, BSP,
CRP
Serum; ELISA,
immunonephrotelemetry
COMP seems to be a surrogate marker of OA
and may be of interest for the detection of
patients at risk of rapid progressing disease
5164 population cohort
CRP
Serum; latex high-sensitivity
assay
CRP was not associated with incidence of knee
or hip OA when confounding factors were taken
into account
186 incident OA
subjects (COMP); 198
incident OA subjects
(NTX); progressive OA
subjects (COMP); 197
progressive OA
subjects (NTX)
172 women
COMP, NTX
Serum, ELISA
Serum levels of COMP and NTX are modest risk
markers for the development of RHOA in a
community-based cohort of elderly white women
25-OH Vit D,
1,25-OH Vit D
Serum; RIA
Low serum levels of 25-Vitamin D may be
associated with incident changes of radiographic
hip OA characterized by joint space narrowing
179 incident hip OA
subjects; 167
progressive hip OA
subjects
FRP, Dkk-1
Serum; ELISA
618 subjects
TGF-β1
Serum; ELISA
Elevated circulating levels of Dkk-1 appeared to
be associated with reduced progression of
RHOA in elderly women, whereas the highest
quartile of serum FRP levels tended to be
associated with a modest reduction in risk of
incident RHOA
Levels of TGF-β1 do not predict incident or
progressive rOA, OST, or JSN at the hip or knee;
it is unlikely that TGF-b1 will be a robust
biomarker for rOA in future studies
1235 population cohort
CTX-II
Urine; ELISA
912 subjects
VCAM-1
Serum; ELISA
Biomarker
identified
FAC, INFɣ, IL-6,
IL-1, IL-RA,
MCP-1, eotaxin,
MIP-1β, IL-10,
PDGF-BB,
RANTES, TNFα,
VEGF
CRP, COMP
Tissue type, assays, and
techniques used
Synovial fluid; human
inflammatory multiplex panel,
ELISA
Serum; ELISA
Athletes with FAI demonstrate increased
cartilage turnover and systemic inflammation
COMP
Serum; ELISA
Serum COMP may be useful as a biomarker of
preradiographic hip pathology
Author
Year
Study design/objectives
Definition population
Chaganti et
al [7]
2008
Chaganti et
al [8]
2010
Chevalier et
al [9]*
2001
Conrozier et
al [15]*
1998
Engström et
al [17]
2009
Kelman et
al [24]
2006
L; determine the
association between
changes in serum COMP
and NTX over 6 years
with development and
progression of
radiographically apparent
hip OA
L; examine the crosssectional association of
25-OH Vitamin D with
prevalent radiographic hip
OA in elderly men
Prosp CC; determine
predictive value of TIMP1 and HA in a 1-year
prospective study of hip
OA
L; evaluate COMP and
BSP as predictors of
disease progression in hip
OA
L; explore relationships
among CRP, metabolic
syndrome, and incidence
of severe knee or hip OA
Retro CS-CC; assess the
association of baseline
serum COMP and NTX
and the development and
progression of
radiographic hip OA
Lane et al
[29]
1999
Lane et al
[30]
2007
Nelson et al
[35]
2010
Reijman et
al [37]*
2004
Schett et al
[39]
2009
L; determine relationship
of serum 25-OH Vitamin
D and 1,25-OH Vitamin D
to incident changes of
radiographic
hip OA
CS-CC; assess
associations between
serum FRP and Dkk-1 and
the development and
progression of
radiographic hip OA
Longitudinal; test whether
serum TGF-b1 predicts
incident and progressive
hip or knee radiographic
OA
CS-L; investigate the
association between
uCTX-II and the
prevalence and
progression
of radiographic OA of the
knee and hip
CS-L; determine
predictors
of the development of
severe OA, apart from age
and overweight
Main outcome of the study
Measurement of serum COMP at two distinct
time points may be a method of identifying
patients at risk for developing incident RHOA
This study shows that CTX-II is associated with
both the prevalence and the progression
of radiographic OA at the knee and hip;
importantly, this association is independent of
known clinical risk factors for OA and seems
stronger in subjects with joint
pain
The level of soluble VCAM-1 emerged as a
strong and independent predictor of the risk of
hip and knee replacement as a result of severe
OA
D. Prearthritic (n = 3)
Author
Year
Study design/objectives
Definition population
Abrams et
al [2]
2014
CS-CC; report hip
synovial fluid cytokine
concentration in hips with
and without radiographic
arthritis
17 OA subjects
(imaging evidence of
Tönnis grade > 2 OA)
undergoing hip
arthroplasty or
arthroscopy
Bedi et al
[4]
2013
10 male athletes with
radiographically
confirmed FAI
Dragomir et
al [16]
2002
CS-CC; comparison of
CRP and COMP in
athletes with and without
FAI
CS; examine crosssectional relationship
between serum COMP and
hip and knee clinical signs
or symptoms in adults
145 subjects without
radiographic hip or
knee OA but with
clinical symptoms or
signs
Main outcome of the study
Fibronectin-aggrecan complex (FAC) was
significantly higher in patients without
radiographic OA
without radiographic OA
Study included in more than one category; 1,25-OH Vit D = 1,25-hydroxy Vitamin D; 25(OH) Vitamin D = 25-hydroxy Vitamin D; AAT = alpha-1 antitrypsin; b-ALP = bone-specific
alkaline phosphatase; BAP = bone alkaline phosphatase; BSP = bone sialoprotein; C1,2C = types 1 and 2 collagens; C2C = collagenase of type II; C4S = chondroitin-4-sulphate; C6S =
chondroitin-6-sulphate; CAT-D = cathepsin D; COMP = cartilage oligomeric protein; CP-I = type 1 procollagen; CPII = C-propeptide; Cr = creatinine; CRP = C-reactive protein; CS 846 =
epitope on the chondroitin sulphate chains of aggrecan molecules; CS = cross-sectional study; CS-CC = cross-sectional case-control study; CTX-I = C-terminal crosslinking telopeptide of
type I collagen; CTX-II = type II collagen telopeptide; DCOMP = deaminated cartilage oligomeric protein; DDH = developmental dysplasia of the hip; DPD = deoxypyridinoline; ELISA =
enzyme-linked immunosorbent assay; FAC = fibronectin-aggrecan complex; FAI = femoroacetabular impingement; FRP = frizzled-related protein; GM-CSF = granulocyte macrophage
colony-stimulating factor; HA = hyaluronate; Helix-II = type II collagen helical telopeptide; HPLC = high-performance liquid chromatography; HTRF = homogeneous time resolved
fluorescence; ICTP = crosslinking C-terminal telopeptide; IL = interleukin; IL-RA = IL-1 receptor agonist; INFɣ = interferon gamma; JS = joint space; JSN = joint space narrowing; JSW =
joint space width; K-L = Kellgren-Lawrence; KS = keratin sulphate; L = longitudinal; L2+L4-KS = L2 +L4 fragments of digested keratan sulfate [L2 = B-galactosyl-(1-4)-6-0-sulfo-Nacetylglucosamine, L2-KS=B-galactosyl-(1-4)-6-0-sulfo-N-acetylglucosamine (L2 fragment of keratan sulfate), L4 = B-6-0-sulfo-galactosyl-(1-4)-6-0-sulfo-N-acetylglucosamine], L4/L2
ratio = [L4-KS fragment]/[L2-KS fragment], L4-KS = L4 = B-6-0-sulfo-galactosyl-(1-4)-6-0-sulfo-N-acetylglucosamine (fragment of digested keratan sulfate; LC = liquid chromatography;
MCP-1 = monocyte chemotactic protein-1; MIP-1β = macrophage inflammatory protein-1 beta; MMP1 = matrix metalloproteinase 1; MMP3 = matrix metalloproteinase 3; NO = nitrous
oxide; NTX = N-terminal telopeptide; OA = osteoarthritis; OC = osteocalcin; OPG = osteoprotegerin; PDGF-BB = platelet-derived growth factor-BB; PIIINP = N-terminal propeptide of
collagen type 3; PINP = N-terminal propeptide of collagen type 1; Prosp = prospective; PYD = pyridinoline; RANKL = receptor activator of nuclear factor kappa-K ligand; RANTES =
regulated on activation/normal T cell expressed; RDOA = rapidly progressive OA; Retro = retrospective; RHOA = radiographic hip osteoarthritis; RIA = radioimmunoassay; sIL-4R =
soluble IL-4 receptor; SPOA = slowly progressive OA; TGF-β1= transforming growth factor β1, TIMP-1 = TIMP metalloproteinase inhibitor 1; TNFα = tumor necrosis factor-alpha;
TRACP-5b = tartrate-resistant acid phosphatase 5b; VCAM-1 = vascular cell adhesion molecule 1; VEGF = vascular endothelial growth factor; YKL-40 = tyrosine, lysine, leucine-40 kD
molecular weight protein.
*