J Korean Orthop Assoc.  2008 Jun;43(3):269-275. 10.4055/jkoa.2008.43.3.269.

The Effect of Repeated Intraarticular Bleeding in the Synovium and Articular Cartilage: An Animal Model

• Yoo MC
• Cho YJ
• Kim KI
• Chun SW
• So DH
• Cho HJ
• Yang HI
• Lee SH
• Lee YA

• KMID: 2186441
• DOI: http://doi.org/10.4055/jkoa.2008.43.3.269

Abstract

PURPOSE: We designed this study to demonstrate the pathophysiology of hemophilic arthropathy (HA) by creating an animal model for determining the effect of repeated intraarticular bleeding in the synovium and articular cartilage.
MATERIALS AND METHODS
20 normal male New Zealand white rabbits were used for this study. We injected 1 ml of autologous blood from the ear vein of the rabbits into the right knee joint three timeds a week for 18 weeks, and we injected 1 ml of normal saline into the left knee joint three times a week for 18 weeks as a control group. We examined the pathologic changes by microscopy and plain X-ray, and we determined the mRNA expression of proinflammatory cytokines in the synovium of the HA by performing real time RT-PCR at the 11th week and 18th week after starting blood-injection. We also examined the GAG and the PGE2 production in cultured chondrocytes that were extracted from the HA knees.
RESULTS
At the 11th week, after blood injection there were no remarkable gross changes in the HA knees and the control knee joints. At the 18th weeks, the experimental knee joints (HA knees) showed grossly swelling and degenerative changes by X-ray. The infiltration of inflammatory cells and the synovial proliferation in the HA knee joints were compared with that in the control knee joints by microscopic examination. The expressions of the mRNA of TNF-alpha, IL-1, MMP-1 and MMP-3 in the HA synovium were increased, as determined by real time RT- PCR, as compared with that in the control knee. In the cultured chondrocytes, the GAG production was decreased and the PGE2 was increased, but the MMP-1 and MMP-3 were not changed, as determined by ELISA.
CONCLUSION
Our results showed that the GAG production of chondrocytes of the HA knees was decreased and there was increased PGE2, so that the cartilage degeneration by intra-articular bleeding was caused by the decreased metabolism of chondrocytes rather than by increased catabolism of the chondrocytes. We suggest that HA was associated with synovitis and cartilage degeneration, but decreased cartilage metabolism was the major mechanism of HA.

Keyword

Synovium; Hemopohilia; Hemophilic arthropathy; Animal; Proinflammatory cytokine

MeSH Terms

Animals
Cartilage
Chondrocytes
Cytokines
Dinoprostone
Ear
Hemorrhage
Humans
Interleukin-1
Knee
Knee Joint
Male
Microscopy
Models, Animal
Polymerase Chain Reaction
Rabbits
RNA, Messenger
Synovial Membrane
Synovitis
Tumor Necrosis Factor-alpha
Veins
Cytokines
Dinoprostone
Interleukin-1
RNA, Messenger
Tumor Necrosis Factor-alpha

Figure

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  Fig. 1 The blood injected knee of a rabbit (right) was more swollen compared with the other knee.

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  Fig. 2 The blood injected right knee shows degenerative change by X-ray

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  Fig. 3 The synovium of the blood injected knee shows proliferation and the fat tissue was increased under the synovium. (A) The normal saline injected knee synovium and (B) the blood injected knee synovium (×400, H&E stain at the 11th week after of intra articular blood injection). (C) The normal saline injected knee synovium and (D) the blood injected knee synovium(×400, H&E stain at the 18th week after intra articular blood injection).

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  Fig. 4 The mRNA expression in the rabbit's synovium, as determined by RT-PCR, in an animal model of recurrent hemarthrosis. The control was a saline-injected knee. The mRNAs of TNF-α, IL-1, MMP-1 and MMP-3 were increased in the hemarthrosis animals. *GAPDH: Glyceraldehyde-3-phosphate degydrogenease.

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  Fig. 5 The results of quantative analysis of TNF-alpha, IL-1, MMP-1 and MMP-3 by real-time RT-PCR.

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  Fig. 6 The production of GAG, PGE2, MMP-1 and MMP-3 from chondrocyte culture by ELISA.

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