MMP-13 In Cartilage Research Articles

Yiqi Yangxue formula inhibits cartilage degeneration in knee osteoarthritis by regulating LncRNA-UFC1/miR-34a/MMP-13 axis

Ethnopharmacological relevanceKnee osteoarthritis (KOA) is a prevalent and disabling clinical condition affecting joint structures worldwide. The Yiqi Yangxue formula (YQYXF) is frequently prescribed in clinical settings for the treatment of KOA. Existing research has primarily focused on alterations in drug metabolism, with limited investigation into the epigenetic effects of YQYXF, particularly in relation to non-coding RNA. Aim of the studyExploring the effects of YQYXF on critical factors of long chain non-coding RNA UFC1/miR-34a/matrix metalloproteinase-13 (MMP-13) axis and their interrelationships. MethodsUHPLC-QE-MS technology was used to identify the YQYXF ingredients in rat serum. KEGG and GO analysis were performed on the targets of blood components acting on KOA using a database. Simultaneously, a protein interaction network was constructed using target proteins and metabolites to identify the core components and key pathways of YQYXF. The KOA rat model was established using an improved Hulth method. SPF SD rats were randomly divided into normal group, sham surgery group, model group, celecoxib capsules group (18 mg/kg), YQYXF low, medium and high dose groups (4.6g/kg, 9.2g/kg, 18.4g/kg). Observe the synovial and cartilage tissues of rats using pathological methods. RT-PCR was used to detect the levels of UFC1, miR-34a, and MMP-13 in cartilage. Immunohistochemistry was used to detect the levels of MMP-13 and ADAMTS-5 in cartilage. ELISA method was used to detect the levels of MMP-13 and ADAMTS-5 in serum. In addition, we further validated the regulation of crucial factor expression levels of UFC1/miR-34a/MMP-13 axis in rat chondrocytes and degenerative chondrocytes of KOA patients by YQYXF, providing a basis for its treatment of KOA. ResultsThe compounds that YQYXF enters the bloodstream mainly contain flavonoids and phenylpropanoid compounds. The core components that act on OA include quercetin, fisetin, and demethylweldelolactone. The main target pathways are the IL-17 signaling pathway, lipid and atherosclerosis, cellular sensitivity, inflammatory mediator regulation of TRP channels, TNF signaling pathway, relaxin signaling pathway and C-type lectin receptor signaling pathway. YQYXF inhibited the expression of miR-34a and MMP-13 mRNA, and reduced the protein levels of MMP-13 and ADAMTS-5. In vitro studies have confirmed that 20% YQYXF serum promoted UFC1 and reduce miR-34a levels. In addition, miR-34a in sh-UFC1+10% YQYXF serum and sh-UFC1+20% YQYXF serum groups significantly decreased compared to the sh-UFC1 group. ConclusionThe anti-KOA cartilage degeneration effect of YQYXF might be related to inhibiting cell apoptosis and promoting cell proliferation, which regulated the lncRNA-UFC1/miR-34a/MMP-13 axis.

Effects of Etanercept on Experimental Osteoarthritis in Rats: Role of Histone Deacetylases.

Mounting evidence suggests that histone deacetylases (HDAC) inhibitors reduce cartilage destruction in animal models of osteoarthritis (OA). Tumor necrosis factor (TNF)-α-blocking treatment for OA may provide effective joint protection by slowing joint damage. To investigate the effects of intraperitoneal administration of etanercept (a TNF-α inhibitor) on OA development in rats and changes in the nociceptive behavior of rats and expression of HDACs, RUNX2, and MMP13 in cartilage. Induction of OA in Wistar rats was accomplished through anterior cruciate ligament transection (ACLT). One or five milligrams (mg) of etanercept was administered intraperitoneally for 5 consecutive weeks after ACLT to the ACLT + etanercept (1 and 5 mg/kg) groups. Nociceptive behavior and changes in knee joint width were analyzed. Cartilage was evaluated histologically and immunohistochemically. ACLT + etanercept significantly improved mechanical allodynia and weight-bearing distribution compared to ACLT alone. In OA rats treated with etanercept, cartilage degeneration and synovitis were significantly less pronounced than those in ACLT rats. OA-affected cartilage also showed reduced expression of HDAC 6, 7, RUNX-2, and MMP-13 in response to etanercept but increased expression of HDAC4. Our study demonstrated that etanercept therapy (1) attenuated the development of OA and synovitis in rats, (2) reduced nociception, and (3) regulated chondrocyte metabolism, possibly by inhibiting cell HDAC6 and HDAC7, RUNX2, and MMP13 and increasing HDAC4 expression. Based on new evidence, etanercept may have therapeutic potential in OA.

Quantitative evaluation of warm acupuncture-moxibustion in improvement of cartilage damage in rabbits with early knee osteoarthritis based on MR T2 mapping.

To observe the application value of MR T2 mapping for evaluating the effect of warm acupuncture-moxibustion on articular cartilage degeneration, and to observe the relationship between T2 value and expression of matrix metalloproteinases (MMP)-1 and MMP-13 of chondrocytes in rabbits with early knee osteoarthritis (KOA). Thirty male New Zealand rabbits were randomly divided into blank control, KOA model and warm acupuncture-moxibustion groups, with 10 rabbits in each group. The early KOA model was established by right hind limb tubular plaster extension fixation method for 2 weeks. The rabbits of the warm acupuncture-moxibustion group received warm acupuncture-moxibustion stimulation at "Heding"(EX-LE2), "Neixiyan"(EX-LE4), "Waixiyan" (EX-LE5) and"Zusanli"(ST36) on the right hind limb for 15 min, once a day for 2 weeks. After intervention, MR T2 mapping of the right knee joint was performed in each group. The H.E. staining was used to evaluate the histopathological changes of cartilage, followed by giving a score according to the standards of Mankin scoring. The TUNEL method was used to analyze the apoptosis state of chondrocytes, and the positive expressions of MMP-1 and MMP-13 in the articular cartilage were detected by immunohistochemical staining. Compared with the blank control group, the Mankin score, chondrocyte apoptosis index, T2 value and the positive expressions of MMP-1 and MMP-13 in the cartilage tissue were significantly increased in the model group (P<0.01). Compared with the model group, the Mankin score, chondrocyte apoptosis index, T2 value and the positive expressions of MMP-1 and MMP-13 in the cartilage tissue were markedly decreased in the warm acupuncture-moxibustion group (P<0.01). The T2 value was positively correlated with the expression levels of MMP-1 and MMP-13 (P<0.01). H.E. staining showed disordered arrangement of chondrocytes and thinner cartilage layer in the model group, and a clear and relative ordered arrangement of chondrocyte in the warm acupuncture-moxibustion group. Warm acupuncture-moxibustion can reduce the T2 value of articular cartilage in early KOA rabbits, which is positively correlated with the decreased expression of MMP-1 and MMP-13 in the extracellular matrix of cartilage. The MR T2 mapping has certain value in evaluating the effect of warm acupuncture-moxibustion on KOA rabbits with early cartilage degeneration.

CCL2 induces articular chondrocyte MMP expression through ERK and p38 signaling pathways

ObjectiveIn previous studies, we determined an association between increased serum and articular cartilage levels of CCL2 with osteoarthritis (OA) progression, cartilage damage and increased MMP13 in cartilage. Here we analyzed CCL2 downstream signaling mediators that lead to gene expression of cartilage catabolic markers, in healthy and OA human articular chondrocytes. DesignHuman articular chondrocytes obtained from healthy or OA subjects were treated with or without recombinant human CCL2; cell lysates or mRNA were collected for immunoblotting or qRT-PCR. For pathway analysis, chondrocytes were pre-incubated with an inhibitor of CCR2 (the unique CCL2 receptor), ERK inhibitor or p38 inhibitor prior to CCL2 treatment. ResultsCCL2 treatment of both healthy and OA chondrocytes activated ERK and p38 via CCR2. In healthy chondrocytes, short (6h) and prolonged (24–72h) CCL2 treatments led to Ccr2, Mmp-1, Mmp-3, Mmp-13 and Timp1 upregulation. In OA chondrocytes, CCL2 induced expression of Ccr2, Mmp-1 and Mmp-3, but not Mmp1 and Timp1, and only following longer treatments (72h). In both healthy and OA chondrocytes, the CCL2-mediated upregulation of Ccr2 and cartilage catabolic markers was mediated by ERK and p38 signaling. ConclusionsThe triggering of the CCL2/CCR2 axis in articular chondrocytes activates specific MAPK pathways leading to gene expression of cartilage degrading enzymes. However, some differences in the response to CCL2 stimulation are detected in healthy vs OA chondrocytes with respect to the number of activated genes and to the time of exposure to CCL2, suggesting that CCL2 action in articular cartilage may be dependent on OA stage and severity.

Relationship Between Magnetic Resonance T2-Mapping and Matrix Metalloproteinase 1,3 in Knee Osteoarthritis

ObjectiveTo investigate the relationship between quantitative analysis of MRI (T2-mapping) and the expression of matrix metalloproteinase (MMP-1, MMP-3) in osteoarthritis of the knee joint and the role of MMP-1,3 in the pathogenesis of osteoarthritis.MethodsThirty cases of knee osteoarthritis (KOA) patients with total knee arthroplasty (TKA) (lesion group) and 30 healthy adult volunteers (control group) were scanned with 1.5 T routine MR and T2-mapping, and their T2 values were measured and statistically analyzed. The pathological examination of the knee cartilage that was replaced during the operation and the immunohistochemical assay were used to measure the expression of MMP-1,3. The correlation between the T2 value of magnetic resonance imaging and the expression of MMP-1,3 was analyzed.Results(1) According to the Recht grading standard for magnetic resonance, the T2 value of magnetic resonance increased significantly with the increase of cartilage degeneration. The differences in T2 values between each level and the normal group were statistically significant (P < 0.05). (2) The T2 value of magnetic resonance imaging increased with the severity of the cartilage degeneration pathological Mankin grading, and the difference was statistically significant (P < 0.05). (3) The expression of MMP-1,3 increased with cartilage degeneration. (4) The T2 value and the expression of MMP-1 in cartilage showed a linear trend. The result of Spearman correlation analysis showed that the expression of MMP-1,3 increased as the cartilage T2 value increased. There was a positive linear correlation between the two.ConclusionThe T2 value of magnetic resonance increased with the degeneration of KOA cartilage. The expression of MMP-1,3 increased with the severity of articular cartilage destruction. The T2 value of KOA magnetic resonance was positively correlated with the expression of MMP-1,3.

LncRNA-GAS5 Inhibits Expression of miR 103 and Ameliorates the Articular Cartilage in Adjuvant-Induced Arthritis in Obese Mice.

We explored whether long noncoding RNA growth arrest-specific transcript 5 (LncRNA-GAS5) small interfering RNA (siRNA) reduced cartilage destruction in obese mice with adjuvant-induced arthritis. We studied the effects of LncRNA-GAS5 siRNA on the polyarthritis index; hind paw swelling; and the serum levels of certain biochemicals, cytokines, and oxidative stress parameters. We measured the expression levels of matrix metalloproteinases (MMP)-13, NF-κB, fibroblast growth factor (FGF) 21, p38, Akt, and PI3K in cartilage via Western blotting and quantitative reverse transcription PCR. Long noncoding RNA-GAS5 siRNA reduced joint swelling; the serum levels of arthritis-associated biochemicals, cytokines, and oxidative stress markers; and cartilage MMP-13, NF-κB, FGF21, p38, Akt, and PI3K levels. Cartilage miR-103 expression was reduced. Histopathologically, LncRNA-GAS5 siRNA ameliorated the pathological changes of cartilage. Long noncoding RNA-GAS5 siRNA prevented cartilage destruction by inhibiting miR-103 expression.

CircTMBIM6 promotes osteoarthritis-induced chondrocyte extracellular matrix degradation via miR-27a/MMP13 axis.

Osteoarthritis is a degenerative disease characterized by degeneration of articular cartilage, but the current mechanism is unclear. Circular RNA (circRNA) plays a significant role in a series of biological processes related to osteoarthritis, but its mechanism remains unclear. The purpose of this study was to investigate the role of the circTMBIM6/miR-27a/MMP13 axis in osteoarthritis. The expression levels of circTMBIM6, miR-27a and MMP13 in cartilage of osteoarthritis patients and normal human cartilage were detected by reverse transcription polymerase chain reaction (RT-PCR). Osteoarthritis cell model was induced by IL-1β and TNF-α, and the expression changes of circTMBIM6, miR-27a and MMP13 in the in vitro model were detected. In addition, the in vitro regulation of circTMBIM6 and miR-27a in osteoarthritis was verified by transfection of circTMBIM6 and miR-27a plasmids, and the regulation of miR-27a on MMP13 was also verified. The dimethylmethylene blue (DMMB) method was used to analyze the secretion and formation of soluble glycosaminoglycan sulfate (sGAG), and the effects of circTMBIM6 and miR-27a chondrocytes were evaluated. The expression levels of circTMBIM6 and MMP13 in cartilage tissue of patients with osteoarthritis were higher than that of normal group, while the expression level of miR-195 in cartilage tissue of patients with osteoarthritis was lower. After IL-1β and TNF-α treatment, the expression of circTMBIM6 and MMP13 in chondrocytes increased, while the expression of miR-27a decreased. CircTMBIM6 overexpression reduced miR-27a expression but increased MMP13 expression. The circTMBIM6 gene knockout showed the opposite effect. CircTMBIM6 promotes osteoarthritis-induced chondrocyte extracellular matrix degradation via miR-27a/MMP13 axis.

Changes in cartilage and subchondral bone in a growing rabbit experimental model of developmental trochlear dysplasia of the knee

ABSTRACT Purpose: Trochlear dysplasia is one of the most frequent lower extremities deformities. Aim of this research was to investigate the changes in cartilage and subchondral bone of trochlea after patellar dislocation in growing rabbits. Materials and Methods: Ninety-six knees from 48 one-month-old rabbits were divided into two groups (experimental, control). Lateral patellar dislocation was established in the experimental group and distal femurs were collected at 4, 8, 12 and 24-week time points, respectively. General examination and histological observations were conducted to research the anatomical structure of the trochlear cartilage and subchondral bone. Structural parameters of trochlear subchondral bone were measured by MicroCT. Subsequently, the expression of TRPV4, collagen II and MMP-13 in cartilage were detected by western blot and RT-PCR analysis, respectively. Results: Subchondral bone loss was found in experimental group from 4 weeks after patellar dislocation, accompanied by increased TRAP-positive osteoclasts in subchondral bone. The trochlear dysplasia model was well established from 8 weeks after patellar dislocation. In addition, degeneration of cartilage was found from 8 weeks, accompanied by decreased expression of mechanically sensitive TRPV4 and collagen II, and increased expression of MMP-13. Conclusions: This study proved that trochlear dysplasia can be caused by patellar dislocation in growing rabbits, accompanied by significant subchondral bone loss. What is more, this study also shows that degenerative cartilage changes occur in the patellar dislocation model and become aggravated with time, accompanied by decreased TRPV4 and collagen II, but increased MMP-13

T140 blocks the SDF-1/CXCR4 signaling pathway and prevents cartilage degeneration in an osteoarthritis disease model.

Osteoarthritis (OA) is one of the most common diseases affecting older people; however, there remains no effective targeted drug to combat OA. The aims of this study were (1) to explore the effect of T140 in regulating degeneration of articular cartilage in vivo by targeted blocking of the SDF-1/CXCR4 signaling pathway, and (2) to provide experimental evidence for the development of a novel OA-targeted pharmacotherapy. Thirty-six healthy Hartley guinea pigs were randomly divided into three groups: a T140-treated group (n = 12), a phosphate buffer saline control group (n = 12) and an untreated control group (n = 12). At 2, 4, 6, 8, 10 and 12 weeks of treatment, SDF-1 in serum was quantified by enzyme-linked immunosorbent assay. At 12 weeks of treatment, the cartilage from knee tibial plateau in the knee joint was collected for H&E, Safranin-O staining and Mankin grading; measurement for mRNA levels of matrix metalloproteinases (MMP-3, MMP-9 and MMP-13), aggrecan (ACAN) and collagen II (Col II) using RT-PCR; and measurement for Col II protein levels by western blot. Results showed that SDF-1 in serum increased in the T140 group and increased in the control groups. H&E and Safranin-O staining revealed less cartilage loss in T140-treated animals compared to controls. The mRNA levels of MMP-3, MMP-9 and MMP-13 in cartilage were much lower in the T140 group than other groups, but mRNA levels of ACAN and Col II in cartilage were higher in the T140-treated group. Col II protein levels in the T140 group and control groups were different. T140 can downregulate the expression of matrix-degrading enzyme and lessen the degeneration of cartilage by blocking the SDF-1/CRCR4 signaling pathway in vivo. This mechanism may present a pharmacological target for the treatment of OA.

Astaxanthin ameliorates cartilage damage in experimental osteoarthritis

Purpose. Astaxanthin is a red-pigment carotenoid found in certain marine animals and plants. Astaxanthin has been shown to inhibit matrix metalloproteinases (MMPs) expression in vitro. However, the effect of astaxanthin on cartilage is still unclear. The aim of this study was to investigate the effects of astaxanthin on cartilage in experimental osteoarthritis (OA).Methods. New Zealand rabbits underwent anterior cruciate ligament transection to induce OA in right knee. Rabbits received intra-articular injection containing 0.3 ml of vehicle (dimethyl sulfoxide) or astaxanthin (50 μM). Injection was started on the day of operation, and the injection were performed once weekly for six consecutive weeks. Then, rabbits were sacrificed and the right knees were harvested for study.Results. Cartilage degradation was reduced by astaxanthin, as assessed by morphological and histological examination. Astaxanthin inhibited the gene expression of MMP-1, MMP-3, and MMP-13 in cartilage as compared with the vehicle group.Conclusions. The results suggest that astaxanthin may be considered as pharmaceutical agent in OA treatment.

Effect of semen sojae germinatum on experimental osteoarthritis in the rabbit knee

As a complementary therapeutic approach, soy ingredients, such as soy protein and soybean unsaponifiables, recently attract more attention for relieving symptoms of osteoarthritis (OA). Semen sojae germinatum (SSG) processed from bean-sprout coming from germinating Glycine max L. Merr. is a traditional Chinese herbal medicine treating knee OA. We evaluated the effect of SSG as feed on the development of anterior cruciate ligament transaction (ACLT) -induced knee OA in rabbits. The results showed that SSG reduced the severity of cartilage degeneration in macroscopic grading scale, and delayed the degradation of the cartilage matrix. The contents of prostaglandin E2 (PGE2), nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin-6(IL-6) and IL-1β in synovial fluid, mRNA and protein expressions of matrix metalloproteinases (MMP)-3, MMP-13 in cartilage were effectively decreased in SSG group. These results demonstrate that SSG had a protective effect against the development of knee OA.

Activation of Indian hedgehog promotes chondrocyte hypertrophy and upregulation of MMP-13 in human osteoarthritic cartilage

The objectives of this study were to (1) determine the correlation between osteoarthritis (OA) and Indian hedgehog (Ihh) expression, and (2) establish the effects of Ihh on expression of markers of chondrocyte hypertrophy and matrix metalloprotease (MMP)-13 in human OA cartilage. OA cartilage and synovial fluid samples were obtained during total knee arthroplasty. Normal cartilage samples were obtained from intra-articular tumor resections, and normal synovial fluid samples were obtained from healthy volunteers and the contralateral uninjured knee of patients undergoing anterior cruciate ligament reconstruction. OA was graded using the Mankin score. Expression of Ihh in synovial fluid was determined by Western blot. Ihh, type X collagen and MMP-13 mRNA were determined by real time PCR. Protein expression of type X collagen and MMP-13 in cartilage samples was analyzed with immunohistochemistry. Chondrocyte size was measured using image analysis. Ihh expression was increased 2.6 fold in OA cartilage and 37% in OA synovial fluid when compared to normal control samples. Increased expression of Ihh was associated with the severity of OA and expression of markers of chondrocyte hypertrophy: type X collagen and MMP-13, and chondocyte size. Chondrocytes were more spherical with increasing severity of OA. There was a significant correlation between Mankin score and cell size (r(2) = 0.80) and Ihh intensity (r(2) = 0.89). Exogenous Ihh induced a 6.8 fold increase of type X collagen and 2.8 fold increase of MMP-13 mRNA expression in cultured chondrocytes. Conversely, knockdown of Ihh by siRNA and Hh inhibitor cyclopamine had the opposite effect. Ihh expression correlates with OA progression and changes in chondrocyte morphology and gene expression consistent with chondrocyte hypertrophy and cartilage degradation seen in OA cartilage. Thus, Ihh may be a potential therapeutic target to prevent OA progression.

Expression of MMP-1 in cartilage and synovium of experimentally induced rabbit ACLT traumatic osteoarthritis: immunohistochemical study

To observe the level and the distribution of MMP-1 in cartilage and synovium over the progression of OA in rabbit ACLT model, and to explore the relationship between the amount of MMP-1 expression and the degree of OA cartilage degradation. OA was induced in 20 rabbits by anterior cruciate ligament transection (ACLT) and ten rabbits were killed at 4 and 8 weeks after the operation, respectively. A further ten rabbits received unilateral knee arthrotomy as controls. Cartilage degradation was observed under dissecting microscope, immunohistochemical, and morphometric analysis were adopted to record the tissue level and distribution of MMP-1 in cartilage and synovium. Cartilage degradation in both ACLT groups was more severe than that in control group, and the degradation in 8-week ACLT group was more severe than that in 4-week ACLT group. MMP-1 was expressed predominantly in the superficial and upper intermediate layers of cartilage and in the lining layer of synovium. Its expression was increased steadily over the progression of OA both in cartilage and in synovium, but there was a little difference in the increase pattern between them: increase of MMP-1 in synovium lagged behind that in cartilage in early stage of OA. MMP-1 was involved in OA development in rabbit ACLT model and the amount of its expression was related with the degree of cartilage degradation. The increase of MMP-1 expression in synovium lagged behind that in cartilage, suggesting OA pathology was originated from cartilage, but synovitis may also paticipate in cartilage degradation, especially in middle and late stage of OA.

P173 THE EFFECTS OF INTRA-ARTICULAR INJECTION OF P38 MAPK INHIBITOR ON MATRIX METALLOPROTEINASE IN CARTILAGE OF EXPERIMENTAL OSTEOARTHRITIS MODEL

Purpose: The primary aim of this study was to investigate, using an experimental rat model of osteoarthritis(OA), the effect of a selective p38 mitogen activated protein kinase inhibitor,SB203580, on the development of structural changes.Additional aims were to assess the effects of the inhibitor on levels of matrixmetalloproteinase 3 (MMP-3) and MMP-13(collagenase 3) in OA cartilage and to explore the relation between the MMP-3,13 expression and the severity of OA. Methods: OA was induced in 40 SD rats by anterior cruciate ligament transection (ACLT).After surgical, rats with OA were randomly divided into A_D groups: Rats of group A received 0.1 ml intra-articular injection of SB203580 at high concentration of 100um/L. Each treatment started immediately after surgery, once a week;those in group B were treated under the same condition using SB203580 with low concentration of 10um/L and those in group C received 0.1ml of intra-articular 0.9% Sodium Chloride injection,animals of group D were not injected as controls after ACLT. The animals were killed 8 weeks after surgery. Macroscopicand histologic studies were performed on the cartilage. The levels of MMP-3,13 in OA cartilage chondrocytes were evaluated by immunohistochemistry and western-blotting. Results: All ACLT knees demonstrated osteoarthritic changes. Cartilage degradation in the control group was significantly more severe than that in the experimental group both on the macroscopic grading scale and on Mankin’s grading scale(P <0.05). Immunohistochemical study showed that in the experimental group MMP-3,13 was predominantly expressed in the superficial and upper intermediate layers of cartilage, and the amount of MMP-3,13 in the experimental group was also lower than that in control group(P <0.05). In western-blotting the amount of MMP3,13 was reduced by the treatment of the inhibitor. The protein levels of MMP-3 and MMP-13 in cartilage of inhibitor injection groups were significantly lower than those of Sodium Chloride group and untreated group. There was no significant difference in MMP-3 and MMP-13 expression between the different concentration inhibitor injection groups. No significant difference in MMP-3 and MMP-13 expression in cartilage was found between Sodium Chloride group and control group. Conclusions: This study demonstrates that, in vivo,SB203580, a selective inhibitor of p38MAPK, can partially decrease the development of some of the structural changes in the early phases of experimental OA and significantly reduces the severity of cartilage degradation. This effect was associated with a reduction in the level of MMP-3,13 in OA cartilage, which probably explains the action of the drug and thus may be a potential drug for the treatment of OA.

Cloning, expression, and type II collagenolytic activity of matrix metalloproteinase-13 from human osteoarthritic cartilage.

Proteolysis of triple-helical collagen is an important step in the progression toward irreversible tissue damage in osteoarthritis. Earlier work on the expression of enzymes in cartilage suggested that collagenase-1 (MMP-1) contributes to the process. Degenerate reverse transcription polymerase chain reaction experiments, Northern blot analysis, and direct immunodetection have now provided evidence that collagenase-3 (MMP-13), an enzyme recently cloned from human breast carcinoma, is expressed by chondrocytes in human osteoarthritic cartilage. Variable levels of MMP-13 and MMP-1 in cartilage was significantly induced at both the message and protein levels by interleukin-1 alpha. Recombinant MMP-13 cleaved type II collagen to give characteristic 3/4 and 1/4 fragments; however, MMP-13 turned over type II collagen at least 10 times faster than MMP-1. Experiments with intact type II collagen as well as a synthetic peptide suggested that MMP-13 cleaved type II collagen at the same bond as MMP-1, but this was then followed by a secondary cleavage that removed three amino acids from the 1/4 fragment amino terminus. The expression of MMP-13 in osteoarthritic cartilage and its activity against type II collagen suggest that the enzyme plays a significant role in cartilage collagen degradation, and must consequently form part of a complex target for proposed therapeutic interventions based on collagenase inhibition.