Cytokine Profiling in Acute Anterior Cruciate Ligament Injury

doi:10.1016/j.arthro.2010.02.011

Arthroscopy: The Journal of Arthroscopic & Related Surgery

Volume 26, Issue 10, October 2010, Pages 1296-1301

https://doi.org/10.1016/j.arthro.2010.02.011 Get rights and content

Purpose

To evaluate the presence and relative concentrations of cytokines, known to be involved in the inflammatory cascade, in acute anterior cruciate ligament (ACL) injury.

Methods

We evaluated an extensive cytokine profile in synovial fluid from 12 patients with acute ACL injury undergoing arthroscopy compared with 15 control subjects using a BioPlex assay (Bio-Rad Laboratories, Hercules, CA) to measure the concentration of 17 inflammatory cytokines.

Results

In patients with acute ACL injury compared with asymptomatic control subjects, the following cytokines were identified at significantly increased concentrations (P < .001, Mann-Whitney U test) compared with control samples: interleukin 6 (105 ± 72 v 0 ± 0 pg/ml), interferon γ (1,544 ± 608 v 9 ± 7.5 pg/ml), macrophage inflammatory protein 1β (16 ± 3.8 v 0.3 ± 0.2 pg/ml), and monocyte chemotactic protein 1 (35 ± 13 v 0.5 ± 0.4 pg/ml). There was no case of a cytokine exhibiting increased levels in asymptomatic compared with symptomatic knee samples.

Conclusions

This investigation identified 4 specific cytokines (interleukin 6, interferon γ, monocyte chemotactic protein 1, and macrophage inflammatory protein 1β) out of a panel of 17 inflammatory molecules for which the levels were consistently elevated in the context of ACL injury compared with non-painful, non–acutely injured knees in a volunteer population.

Level of Evidence

Level IV, prognostic case series.

Section snippets

Inclusion and Exclusion Criteria

The inclusion criteria were patients aged greater than 18 years presenting with signs and symptoms consistent with acute ACL instability in 1 knee, occurring within 6 weeks of an acute event causing signs and symptoms including pain, instability, and knee effusion. In addition, patients had to have magnetic resonance imaging (MRI) results consistent with an isolated ACL complete or near-complete tear.

The exclusion criteria were patients aged less than 18 years and those with recent (within 3

Results

Cytokine analysis was completed for 12 patients undergoing unilateral knee arthroscopy for acute pain and/or disability consistent with ACL injury and 15 asymptomatic control subjects.

Discussion

The occurrence in this study of multiple inflammatory cytokines—IL-6, IFN-γ, MCP-1, and MIP-1β—underscores the inherent complexity of the inflammatory cascade in painful ligamentous injury. This observation may not only enhance our understanding of the inflammatory profile, but further investigation may possibly lead to prevention of the degenerative cascade leading to cartilage destruction and subsequent OA.

Several in vitro as well as in vivo studies in animal models have detected few

Conclusions

This investigation identified 4 specific cytokines (IL-6, IFN-γ, MCP-1, and MIP-1β) out of a panel of 17 inflammatory molecules for which the levels were consistently elevated in the context of acute, painful ACL injury compared with non-painful, non–acutely injured knees in a volunteer population.

References (36)

D.C. Fithian et al.
Fate of the anterior cruciate ligament-injured knee
Orthop Clin North Am
(2002)
H. Louboutin et al.
Osteoarthritis in patients with anterior cruciate ligament rupture: A review of risk factors
Knee
(2009)
M. Liden et al.
Osteoarthritic changes after anterior cruciate ligament reconstruction using bone-patellar tendon-bone or hamstring tendon autografts: A retrospective, 7-year radiographic and clinical follow-up study
Arthroscopy
(2008)
F. Nelson et al.
Early post-traumatic osteoarthritis-like changes in human articular cartilage following rupture of the anterior cruciate ligament
Osteoarthritis Cartilage
(2006)
K. Irie et al.
Intraarticular inflammatory cytokines in acute anterior cruciate ligament injured knee
Knee
(2003)
P.H. Marks et al.
Inflammatory cytokine profiles associated with chondral damage in the anterior cruciate ligament-deficient knee
Arthroscopy
(2005)
T. Sugiyama
Involvement of interleukin-6 and prostaglandin E2 in periarticular osteoporosis of postmenopausal women with rheumatoid arthritis
J Bone Miner Metab
(2001)
D. Bryan et al.
Cytokine gene expression in a murine wound healing model
Cytokine
(2005)
X.J. Xu et al.
Intrathecal interferon-gamma facilitates the spinal nociceptive flexor reflex in the rat
Neurosci Lett
(1994)
K.S. Vikman et al.
Interferon-gamma induces characteristics of central sensitization in spinal dorsal horn neurons in vitro
Pain
(2003)

K.S. Vikman et al.

Increased responsiveness of rat dorsal horn neurons in vivo following prolonged intrathecal exposure to interferon-gamma

Neuroscience

(2005)

P. Conti et al.

Differential production of RANTES and MCP-1 in synovial fluid from the inflamed human knee

Immunol Lett

(2002)

R.M. Borzi et al.

Flow cytometric analysis of intracellular chemokines in chondrocytes in vivo: Constitutive expression and enhancement in osteoarthritis and rheumatoid arthritis

FEBS Lett

(1999)

G.R. Squires et al.

The pathobiology of focal lesion development in aging human articular cartilage and molecular matrix changes characteristic of osteoarthritis

Arthritis Rheum

(2003)

Z. Tang et al.

Contributions of different intraarticular tissues to the acute phase elevation of synovial fluid MMP-2 following rat ACL rupture

J Orthop Res

(2009)

H. Higuchi et al.

Changes in biochemical parameters after anterior cruciate ligament injury

Int Orthop

(2006)

P.E. Bijur et al.

Validation of a verbally administered numerical rating scale of acute pain for use in the emergency department

Acad Emerg Med

(2003)

W. de Jager et al.

Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells

Clin Diagn Lab Immunol

(2003)

Cited by (70)

Interferon-gamma modulates articular chondrocyte and osteoblast metabolism through protein kinase R-independent and dependent mechanisms
2022, Biochemistry and Biophysics Reports
Osteoarthritis (OA) affects multiple tissues of the synovial joint and is characterised by articular cartilage degeneration and bone remodelling. Interferon-γ (IFN-γ) is implicated in osteoarthritis pathology exerting its biological effects via various mechanisms including activation of protein kinase R (PKR), which has been implicated in inflammation and arthritis. This study investigated whether treatment of articular cartilage chondrocytes and osteoblasts with IFN-γ could induce a degradative phenotype that was mediated through the PKR signalling pathway. IFN-γ treatment of chondrocytes increased transcription of key inflammatory mediators (TNF-α, IL-6), matrix degrading enzymes (MMP-13), the transcription factor STAT1, and PKR. Activation of PKR was involved in the regulation of TNF-α, IL-6, and STAT1. In osteoblasts, IFN-γ increased human and mouse STAT1, and human IL-6 through a mechanism involving PKR. ALP, COL1A1 (human and mouse), RUNX2 (mouse), and PHOSPHO1 (mouse) were decreased by IFN-γ. The number of PKR positive cells were increased in post-traumatic OA (PTOA). This study has revealed that IFN-γ propagates inflammatory and degenerative events in articular chondrocytes and osteoblasts via PKR activation. Since IFN-γ and PKR signalling are both activated in early PTOA, these mechanisms are likely to contribute to joint degeneration after injury and might offer attractive targets for therapeutic intervention.
Assessing collagen alterations in enzymatic degradation models of osteoarthritis via second harmonic generation microscopy
2021, Osteoarthritis and Cartilage
Structural changes in the collagen II architecture of osteoarthritis (OA) are poorly understood, which is a large shortcoming in the early diagnosis of this disease. Though degradation can be simulated by enzymes including trypsin and bacterial collagenase, the specific structural features of each digestion and their relationship to naturally occurring OA remain unclear.
We used collagen sensitive/specific Second Harmonic Generation (SHG) microscopy in conjunction with optical scattering measurements to probe the resulting architecture changes in bovine knee cartilage upon trypsin and collagenase degradation. Image features extracted from SHG images were used to train a linear discriminant (LD) model capable of classifying enzymatic degradation, which was then applied to human cartilage with varied modified Mankin histological scores.
The treatment of cartilage with these enzymes resulted in more disorganized collagen structure, where this effect was greatest with collagenase treatment. Using the LD model, we classified the control and degraded tissues in the three zones with >92% accuracy, showing that these enzymes have distinct activity on the collagen assembly. Application of the LD model to human cartilage indicated that collagenase effects were more representative of in vivo degeneration and were also consistent with damage beginning at the articular surface and progressing into deeper zones.
SHG and optical scattering measurements successfully delineate trypsin and collagenase degradation and suggest that collagen alterations in human OA are better simulated by the latter mechanism. These results lay the groundwork for using high-resolution SHG and optical scattering as an earlier diagnostic tool than is currently available.
Identifying effector molecules, cells, and cytokines of innate immunity in OA
2020, Osteoarthritis and Cartilage
Inflammatory changes are observed in affected joints of osteoarthritis (OA) patients and are thought to be involved in the pathology that develops along OA progression. This narrative review provides an overview of the various cell types that are present in the joint during OA and which alarmins, cytokines, chemokines, growth factors, and other mediators they produce. Moreover, the involvement of more systemic processes like inflammaging and its associated cellular senescence in the context of OA are discussed.
Osteoarthritis-related biomarkers profile in chronic anterior cruciate ligament injured knee
2020, Knee
Increasing evidence has shown the involvement of the innate immune system and inflammatory response in osteoarthritis (OA) pathogenesis; however, anterior cruciate ligament (ACL) tears are recognized risk factors for development of post-traumatic OA. We investigated (1) whether inflammatory mediators involved in OA pathogenesis are also present at significant concentrations in the knee joint sometime after ACL complete tear and may be considered as prognostic biomarkers of progression to secondary OA; and (2) whether quantification in serum may surrogate synovial fluid (SF) measurements in both cases.
Thirty-seven end-stage OA patients and 33 patients with ACL complete tear that were included on the waiting list for knee surgery were consecutively recruited. Serum and SF samples were taken before surgery, and tumor necrosis factor-alpha, (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), matrix metalloproteinase-1 (MMP1), matrix metalloproteinase-3 (MMP3), tissue inhibitor of metalloproteinase-1 (TIMP1), bone morphogenetic protein-7 (BMP7), regulated upon activation normal t-cell expressed and secreted (RANTES), cytokine interferon-γ-induced protein 10 (IP-10) and heat shock protein family A (Hsp70) member 1A (HSPA1A) were quantified by enzyme-linked immunosorbent assay (ELISA.) Normally distributed data were compared using a one-way analysis of variance (ANOVA) test. Data not normally distributed were analyzed using a nonparametric Mann–Whitney rank sum test. Differences in means were compared using a Student's t-test. Correlations were analyzed using Pearson's coefficient of variation.
Eighty-seven percent of patients with OA and 86% of those with ACL tear had quantifiable levels of biomarkers in SF. SF levels of IL-6, IL-8, MMP1, MMP3, RANTES, IP-10, BMP7 and HSPA1A were significantly lower in ACL injury knees compared with those with OA, but much higher than those reported in control subjects. Serum levels of IL-6, IP-10, and MMP1 were also lower in patients with ACL tears, who had, however, significantly higher TNF-α, HSPA1A, and TIMP1 levels when compared with OA patients. Levels of biomarkers tested in serum and SF samples were significantly different.
Our data propose that cytokines IL-6 and IL-8 and the chemokines RANTES, IP-10, MMP1, MMP3, and HSPA1A may be involved in the inflammatory process leading to synovitis, the central lesion in OA onset and development; persistent high levels of these substances sometime after ACL injury suggest that they could play an etiopathogenic role in the maintenance of the inflammatory environment leading to post-traumatic OA. Serum biomarker levels do not appear to faithfully reflect what occurs inside the joint. Thus, most biomarkers cannot yet be considered as useful inflammatory biomarkers of knee joint diseases.
ACL reconstruction with lateral plasty reduces translational and rotatory laxity compared to anatomical single bundle and non-anatomical double bundle surgery: An in vivo kinematic evaluation with navigation system
2019, Clinical Biomechanics
Citation Excerpt :
Moreover, recent in vivo studies have underlined how SB reconstructions were not able to restore normal rotatory stability during daily life activities (Georgoulis et al., 2007; Hemmerich et al., 2011; Kim et al., 2015; Ristanis et al., 2003; Ristanis et al., 2005; Tashman et al., 2007). Residual laxities are acknowledged risk factors for long-term functional impairment and development of knee OA (Bigoni et al., 2013; Cuellar et al., 2010; Øiestad et al., 2009; Potter et al., 2012; Swärd et al., 2013). Extra-articular procedures in addition to SB ACL reconstruction have recently resurged interest among surgeons, partially because of the discovery of the importance of the anterolateral ligament (ALL) (Claes et al., 2013; Lubowitz et al., 2014; Vincent et al., 2012).
Significantly better stability may be achieved with a Single Bundle Lateral Plasty (SBLP) reconstruction compared with the Single Bundle (SB) and Double Bundle (DB) procedures.
The study included 42 patients who underwent ACL reconstruction. Patients were randomly selected for one of the following surgical procedure defining three study groups: Single-Bundle-Lateral-Plasty, Single-Bundle and Double-Bundle procedures. Laxity evaluation was performed with an intraoperative navigation system. Lachman test (AP30), Drawer test (AP90), Varus-Valgus stress test at 0° and 30° knee flexion (VV0, VV30), Internal-External rotation (IE30, IE90), and pivot shift (PS) test are the clinical test executed for the laxity evaluation.
Laxity reduction was defined as the difference between laxity before the fixation of the graft used for the reconstruction and the laxity just after its fixation.
For all the analyzed surgical techniques, the pre-reconstruction laxity values were statistically higher (P < 0.05) than the post-reconstruction values for all the analyzed tests. The analysis of the Drawer test and Internal-External rotation at 30° and 90° of knee flexion, highlighted a significant difference at time zero after surgery among the three study groups. The results showed that the SBLP technique had the highest reduction values compared to SB (P_IE90 = 0.001) and DB (P_AP90 = 0.012; P_IE30 = 0.021; P_IE90 = 0.003) techniques.
SBLP technique showed significantly superior results in terms of antero-posterior and internal-external laxity reduction at time-zero after ACL reconstruction.
Two Amnion-Derived Mesenchymal Stem-Cells Injections to Osteoarthritic Elbows in Dogs—Pilot Study
2023, Animals

View all citing articles on Scopus

: V.G.C., J.M.C., S.R.G., D.C.Y., and G.J.S. received support exceeding US $500 from Cytonics, Jupiter, Florida, related to this research.

View full text

Original ArticleCytokine Profiling in Acute Anterior Cruciate Ligament Injury

Purpose

Methods

Results

Conclusions

Level of Evidence

Section snippets

Inclusion and Exclusion Criteria

Results

Discussion

Conclusions

Orthop Clin North Am

Knee

Arthroscopy

Osteoarthritis Cartilage

Knee

Arthroscopy

J Bone Miner Metab

Cytokine

Neurosci Lett

Pain

Neuroscience

Immunol Lett

FEBS Lett

The pathobiology of focal lesion development in aging human articular cartilage and molecular matrix changes characteristic of osteoarthritis

Arthritis Rheum

Contributions of different intraarticular tissues to the acute phase elevation of synovial fluid MMP-2 following rat ACL rupture

J Orthop Res

Changes in biochemical parameters after anterior cruciate ligament injury

Int Orthop

Validation of a verbally administered numerical rating scale of acute pain for use in the emergency department

Acad Emerg Med

Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells

Clin Diagn Lab Immunol

Original Article
Cytokine Profiling in Acute Anterior Cruciate Ligament Injury