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. 1998 Jul;57(7):408–413. doi: 10.1136/ard.57.7.408

Local removal of phagocytic synovial lining cells by clodronate-liposomes decreases cartilage destruction during collagen type II arthritis

P L E M Van Lent 1, A Holthuysen 1, N Van Rooijen 1, L B A V De Putte 1, W B V Den Berg 1
PMCID: PMC1752671  PMID: 9797567

Abstract

OBJECTIVE—To investigate whether local removal of phagocytic synovial lining cells (SLCs) from the knee joint before onset of collagen type II arthritis has an effect on development of cartilage destruction.
METHODS—Phagocytic SLCs were selectively depleted by a single injection of clodronate laden liposomes in the knee joint seven days before induction of collagen type II arthritis (CIA). Clodronate laden liposomes were given in one knee joint either alone or in combination with a short-term oral treatment of dexamethasone. Cartilage damage including proteoglycan depletion and chondrocyte death was measured in total knee joints sections stained with safranin-o or haematoxylin.
RESULTS—Local removal of phagocytic SLCs, seven days before arthritis onset, prevented cell influx for the larger part. Chondrocyte death was significantly decreased in the SLC depleted arthritic joint both at an early (6 days) and late (12 days) time point after CIA induction. However, depletion of proteoglycans from femoral and patellar cartilage layers was not prevented. If the mild acute inflammation caused by a single clodronate laden liposome injection in the left knee joint, was blocked by a short-term (on consecutive days 9, 8, 7, 6, 5 before CIA onset) oral treatment with dexamethasone, cell influx, but also proteoglycan depletion was almost completely blocked. In the contralateral control right knee joint prominent cell influx and severe cartilage damage was observed, indicating that there was no effect of dexamethasone anymore at the onset of CIA.
CONCLUSIONS—This study shows that removal of phagocytic lining cells before CIA induction, particularly in the presence of a short-term treatment with dexamethasone, decreases cartilage destruction.

 Keywords: experimental arthritis; phagocytic lining cell; inflammation; chondrocyte

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Figure 1  .

Figure 1  

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Total knee joint sections stained with safranin-o, six days after accelerated collagen type II arthritis. Note the significantly decreased cell influx in the arthritic knee joint in which the lining was selectively removed seven days before CIA induction (B) compared with controls (A). Comparable PG depletion was found as seen by loss of red staining from the cartilage surface. Original magnification × 200. F=femur, P=patella, S=synovium.

Figure 2  .

Figure 2  

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Depletion of proteoglycan (PG) from cartilage layer of patella and femur. PG loss was scored using an arbritrary scale from 0-3 (0: no PG loss; 1:minor; 2:moderate; 3:marked). Clodronate (CL2MDP) laden liposomes, PBS laden liposomes or PBS alone was injected into the knee joint before arthritis induction. Knee joints were injected seven days before arthritis onset. PG loss was scored at either day 6 (A) or day 12 (B) after CIA onset. Note that the PG loss in the low inflamed, clodronate treated knee joints was not significantly different from high inflamed control arthritic joints. Values represent two experiments with 10 mice per group.

Figure 3  .

Figure 3  

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Chondrocyte death in patellar and femural cartilage layer of SLC depleted and control knee joints. Chondrocyte death was measured by determining the percentage chondrocyte death (empty lacunae) and total chondrocytes. Clodronate (CL2MDP) laden liposomes, PBS-laden liposomes or PBS alone was injected into the knee joint before arthritis induction. Knee joints were injected seven days before arthritis onset. Percentage chondrocyte death was measured at either day 6 (A) or day 12 (B) after CIA onset. Note that chondrocyte death in the low inflamed SLC depleted arthritic knee joints is significantly lower than that from highly inflamed control arthritic joints if clodronate laden liposomes were given seven days before CIA onset. Values represent two experiments with 10 mice per group. p<0.05 compared with PBS or PBS containing liposomes, by Mann-Whitney U test.

Figure 4  .

Figure 4  

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Total knee joint sections stained with haematoxylin and eosin, six days after accelerated collagen type II arthritis. Note the significantly decreased cell influx in the arthritic knee joint in which the lining was selectively removed seven days before CIA induction (B) compared with controls (A). Significantly less chondrocyte death is found particularly in the femur in lining depleted arthritic knee joints. Original magnification × 200. F=femur, P=patella, S=synovium.

Figure 5  .

Figure 5  

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Clinical severity of arthritis in the hindpaws of DBA/1 mice with accelerated type II collagen induced arthritis (CIA). CIA was accelerated by giving 40 µg of lipopolysaccharide (LPS) on day 28. Clodronate laden liposomes were injected in the left knee joint whereas PBS was injected in the right knee joint on day 21. One group of seven mice were treated orally with dexamethasone (1 mg/kg/day) on day 19, 20, 21, 22, 23. The control group was given the vehicle at similar time points. Clinical severity of the hind paw was scored using an arbritary scale from 0-2 per paw. Note that the clinical severity of the hind paws was not different between dexamethasone and vehicle treated animals.

Figure 6  .

Figure 6  

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Cell influx, proteoglycan depletion, and chondrocyte death in knee joints of DBA/1 mice with accelerated type II collagen induced arthritis (CIA). Total knee joint sections were stained with either haematoxylin and eosin (cell influx) or safranin-o (proteoglycan loss). Cell influx and proteoglycan loss was scored on a 0-3 scale. The percentage chondrocyte death was expressed as ratio of number of empty lacunae/number of total lacunae. Clodronate laden liposomes or PBS alone was injected in the knee joint before arthritis induction. One group of seven mice was treated orally with dexamethasone (1 mg/kg/day) on days 19, 20,21, 22, 23. Note the absence of cell infiltration and chondrocyte death and the minor proteoglycan loss in SLC depleted but not control arthritic knee joints of mice given dexamethasone. The control group was given the vehicle at similar time points. Clinical severity of the hind paws was not different between dexamethasone and vehicle treated animals (data not shown).

Figure 7  .

Figure 7  

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Total knee joint sections stained with safranin-o, six days after accelerated collagen type II arthritis that started at day 28 after immunisation. Mice were given dexamethasone (1 mg/kg/day) orally on day 19, 20, 21, 22, 23, around the intra-articular injection of clodronate laden liposomes at day 21 after immunisation. Note the significantly decreased cell influx and minor proteoglycan depletion in the arthritic knee joint in which the lining was selectively removed (B) compared with the contralateral control joint with intact lining (A). Original magnification × 100. F=femur, P=patella, S=synovium space.

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