Effects of chondroitin sulfate-C on articular cartilage destruction in murine collagen-induced arthritis.

The effects of chondroitin sulfate-C (CAS 25322-46-7, Chondroitin ZS Tab) on type II collagen (CII)-induced arthritis (CIA) in mice were evaluated. DBA/1J mice were immunized with bovine CII emulsified in Freund's complete adjuvant, followed by a booster injection 21 days later. Chondroitin sulfate-C at doses of 100, 300 and 1000 mg/kg was administered orally once daily beginning 14 days before initial immunization. An arthritis index and hind paw edema were examined from day 0 to day 49, when the mice were killed by ether anesthesia for histopathological examination. The delayed-type hypersensitivity (DTH) reaction, serum anti-CII antibody titer, and histopathologic characteristics of both synovitis and destruction of articular cartilage were analyzed. Both the arthritis index and the serum anti-CII antibody titer were reduced by treatment with chondroitin sulfate-C in a dose-dependent manner. Chondroitin sulfate-C (1000 mg/kg) significantly inhibited hind paw edema, synovitis and destruction of the articular cartilage, but not DTH reaction.

Effects of chondroitin sulfate-C on bradykinin-induced proteoglycan depletion in rats.

Depletion of the proteoglycan content of articular cartilage was induced by injecting bradykinin (30-300 mumol/l, 50 microliters/knee) into the left knee articular cavities of rats 3 times a day for 2 days. The degree of the reduction in the intensity of histopathological safranin O staining was used as an index of proteoglycan depletion. Bradykinin reduced the cartilage proteoglycan contents of the knee joints of non-injected limbs in a dose-dependent manner and at 300 mumol/l markedly reduced these contents, but evoked no inflammatory changes. The extent of the reduction of the cartilage proteoglycan contents induced by bradykinin injection depended on the dose and injection frequency. Chondroitin sulfate-C (CAS 25322-46-7, Chondroitin ZS Tab) (30-1,000 mg/kg/day) administered orally to rats for 14 days inhibited the bradykinin-induced proteoglycan depletion of the articular cartilage in a dose-dependent manner. These results suggest that a reduction of the proteoglycan content of cartilage, like that associated with osteoarthritis, was induced by injecting bradykinin into the knee articular cavities of rats and chondroitin sulfate-C protected against this effect.

Age-depending effects of methotrexate treatment on systemic bone turnover in experimental adjuvant arthritis.

Adjuvant arthritis was induced in rats in the growth stage (aged 6 weeks) and those in the mature stage (aged 4 months), and changes in the systemic bone turnover and the effects of methotrexate (MTX, CAS 133073-73-1) were compared. After induction of adjuvant arthritis, the paw edema ratio and the urinary deoxypyridinoline (u-Dpy) level increased in both age groups. No marked changes were observed in the serum osteocalcin (s-OC) level in either group. In the 6-week-old rats, arthritis completely inhibited the bone mass, and strength of the femur and lumbar vertebral body. The 4-month-old rats showed more marked changes than the 6-week-old rats in the bone mass and strength of the lumbar, vertebral body. MTX administration (0.05, 0.1 and 0.2 mg/kg/day) resulted in significant dose-dependent inhibition of arthritis-induced changes, and the effects of MTX were similar between the two age groups. MTX was useful at each age. These results suggest that 4-month-old rats with arthritis are more appropriate as a model for evaluation of drugs for bone metabolic turnover in human chronic rheumatoid arthritis.

Methotrexate maintains bone mass by preventing both a decrease in bone formation and an increase in bone resorption in adjuvant-induced arthritic rats.

We examined the effects of low doses methotrexate (MTX) and indomethacin (IND) on bone mass and turnover in normal male Sprague-Dawley rats and those with adjuvant-induced arthritis. Normal and the adjuvant (heat-killed mycobacterium)-injected rats, 6 weeks of age, were given MTX at daily doses of 0.05, 0.1, or 0.2 mg/kg body weight (BW) or IND at a daily dose of 1.0 mg/kg BW. Rats were killed at the start, or at 14 and 28 days. In normal rats, the administration of these agents did not change the lumbar and femoral BMD values, nor did the serum osteocalcin or urinary deoxypyridinoline (D-Pyr) levels. Lumbar trabecular osteoclast number (Oc.N/BS) and osteoclast surface (Oc.S/BS) were decreased in the rats given IND. In the arthritic rats, the administration of MTX did not prevent an early increase of paw edema in the adjuvant-injected limb, but late inflammatory edema was alleviated in the non-injected limb. However, MTX administration at a dose of 0.1-0.2 mg/kg BW maintained an age-dependent increase in the lumbar and femoral BMD values. While serum osteocalcin levels were decreased and urinary D-Pyr values were increased in the arthritic control rats, these bone markers remained at the levels of the normal rats. Decreases in mineral apposition rate (MAR) and bone formation rate (BFR/BS) and increases in the trabecular Oc.N/BS and Oc.S/BS values were prevented by MTX. While IND almost completely prevented inflammatory paw edema, it did not improve the parameters of bone formation. An increase in osteoclasts was prevented and the osteopenia in the lumbar and the femoral bone was only partially prevented by IND. These data suggest that MTX improves bone mass and turnover in the arthritic rat, in which several cytokines that affect bone cells are involved. An increase in bone resorption may be due to prostaglandins, but bone formation defect was suggested to be due to other cytokines such as IL-1, IL-6, and TNF-alpha in this model.

Electron microscopical studies on the effect of lapsed time on the nerve endings of the outer hair cells in acoustically exposed rabbits.

Rabbits were exposed to pure tone (2 kHz, 100 dB for 2 h) and the changes in the nerve endings of the outer hair cells over a period of 1 month were studied using the electron microscope. In the afferent nerve endings mitochondria dilated and small vesicles decreased in number immediately after acoustic exposure. However, the afferent nerve endings recovered during the period of 1 month. In the efferent nerve endings mitochondria dilated and were distributed throughout the nerve endings, and small vesicles decreased in number immediately after acoustic exposure. The efferent nerve endings recovered more slowly than afferent nerve endings. In addition, the changes in the agglomerations of synaptic vesicles along the synaptic membrane and the gap-structures in the synaptic cleft in the efferent nerve endings after acoustic exposure are briefly discussed.

Electron microscopical studies on the nerve endings of the outer hair cells in acoustically exposed rabbits.

The afferent and efferent nerve endings of the outer hair cells of the rabbit after acoustic exposure were investigated under the electron microscope. After acoustic stimulation, the afferent and efferent nerve endings of normal range were respectively 57 per cent and 64 per cent. This result suggests that half of the afferent and efferent nerve endings maintain their function under the conditions of this experiment. An abnormal inclusion body forming an oblong dense body and cistern in addition to the postsynaptic cisterna was observed. The active zone and the vesicular gap-structure of the synapse are discussed.