Influence of exogenous hyaluronan on synthesis of hyaluronan and collagenase by equine synoviocytes

Timothy M. Lynch From the Departments of Large Animal Clinical Sciences (Lynch, Caron, Lloyd, Stick) and Pathology (Render) and the Laboratory for Comparative Orthopaedic Research (Arnoczky), College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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John P. Caron From the Departments of Large Animal Clinical Sciences (Lynch, Caron, Lloyd, Stick) and Pathology (Render) and the Laboratory for Comparative Orthopaedic Research (Arnoczky), College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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Steven P. Arnoczky From the Departments of Large Animal Clinical Sciences (Lynch, Caron, Lloyd, Stick) and Pathology (Render) and the Laboratory for Comparative Orthopaedic Research (Arnoczky), College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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James W. Lloyd From the Departments of Large Animal Clinical Sciences (Lynch, Caron, Lloyd, Stick) and Pathology (Render) and the Laboratory for Comparative Orthopaedic Research (Arnoczky), College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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John A. Stick From the Departments of Large Animal Clinical Sciences (Lynch, Caron, Lloyd, Stick) and Pathology (Render) and the Laboratory for Comparative Orthopaedic Research (Arnoczky), College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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James A. Render From the Departments of Large Animal Clinical Sciences (Lynch, Caron, Lloyd, Stick) and Pathology (Render) and the Laboratory for Comparative Orthopaedic Research (Arnoczky), College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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SUMMARY

Objective

To evaluate the influence of exogenous hyaluronan (HA) on in vitro synthesis of HA and collagenase by equine synoviocytes from normal and inflamed joints.

Animals

9 adult horses.

Procedure

Synoviocytes for culture were taken from the middle carpal joint of 3 horses with normal joints (control) and 6 horses with osteochondral fractures (principal). Synoviocytes were propagated in monolayer cultures and were incubated with 3 commercial HA products at concentrations of 0, 200, 400, and 1,500 μg/ml. Newly synthesized HA was radiolabeled with [3H]glucosamine and quantified by cetylpyridinium chloride precipitation and liquid scintillation counting. The hydrodynamic size of radioactive HA was determined by high-performance liquid chromatography, and collagenase activity was evaluated by use of a quantitative radioactive collagen film assay.

Results

Exogenous HA influenced neither the rate of synthesis nor the hydrodynamic size of the newly produced HA by control or principal cell cultures. Culture supernatants from abnormal synovium, exposed to 400 and 1,500 μg of exogenous HA/ml, contained significantly more collagenase activity than did those exposed to lower concentrations.

Conclusion

Although HA is thought to have beneficial effects in equine arthropathies, the principal mechanisms of action of HA do not appear to be stimulation of synthesis of HA of augmented molecular weight or marked inhibition of collagenase synthesis. (Am J Vet Res 1998;59:888–892)

SUMMARY

Objective

To evaluate the influence of exogenous hyaluronan (HA) on in vitro synthesis of HA and collagenase by equine synoviocytes from normal and inflamed joints.

Animals

9 adult horses.

Procedure

Synoviocytes for culture were taken from the middle carpal joint of 3 horses with normal joints (control) and 6 horses with osteochondral fractures (principal). Synoviocytes were propagated in monolayer cultures and were incubated with 3 commercial HA products at concentrations of 0, 200, 400, and 1,500 μg/ml. Newly synthesized HA was radiolabeled with [3H]glucosamine and quantified by cetylpyridinium chloride precipitation and liquid scintillation counting. The hydrodynamic size of radioactive HA was determined by high-performance liquid chromatography, and collagenase activity was evaluated by use of a quantitative radioactive collagen film assay.

Results

Exogenous HA influenced neither the rate of synthesis nor the hydrodynamic size of the newly produced HA by control or principal cell cultures. Culture supernatants from abnormal synovium, exposed to 400 and 1,500 μg of exogenous HA/ml, contained significantly more collagenase activity than did those exposed to lower concentrations.

Conclusion

Although HA is thought to have beneficial effects in equine arthropathies, the principal mechanisms of action of HA do not appear to be stimulation of synthesis of HA of augmented molecular weight or marked inhibition of collagenase synthesis. (Am J Vet Res 1998;59:888–892)

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