Three-dimensional culture of canine articular chondrocytes on multiple transplantable substrates

James L. Cook From the Departments of Veterinary Medicine and Surgery (Cook, Payne, Tomlinson) and Pathobiology (Kreeger), College of Veterinary Medicine, University of Missouri-Columbia, 379 F. Campus Dr, Columbia, MO 65211.

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John M. Kreeger From the Departments of Veterinary Medicine and Surgery (Cook, Payne, Tomlinson) and Pathobiology (Kreeger), College of Veterinary Medicine, University of Missouri-Columbia, 379 F. Campus Dr, Columbia, MO 65211.

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John T. Payne From the Departments of Veterinary Medicine and Surgery (Cook, Payne, Tomlinson) and Pathobiology (Kreeger), College of Veterinary Medicine, University of Missouri-Columbia, 379 F. Campus Dr, Columbia, MO 65211.

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James L. Tomlinson From the Departments of Veterinary Medicine and Surgery (Cook, Payne, Tomlinson) and Pathobiology (Kreeger), College of Veterinary Medicine, University of Missouri-Columbia, 379 F. Campus Dr, Columbia, MO 65211.

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 DVM, MVSc

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Abstract

Objective

To determine the effects of transplantable substrates on canine chondrocytes grown in three-dimensional culture.

Animals

3 canine cadavers.

Procedure

Articular cartilage harvested from canine cadavers was used to obtain chondrocytes for primary culture. Subcultured chondrocytes were grown in agarose alone (AG), or in agarose on canine cancellous bone (CB), polypropylene mesh, or oxidized regenerated cellulose substrate. Cell proliferation, proteoglycan and glycosaminoglycan (GAG) production, and collagen production were assessed on days 3, 6, 10, 15 and 20.

Results

Chondrocytes from groups AG and CB proliferated and produced matrix over the entire 20-day study period. Group-CB chondrocytes had significantly more GAG than did chondrocytes of all other groups on days 6 (P = 0.0297) and 15 (P = 0.00272). Those of groups AG and CB contained significantly (P = 0.0235) more GAG on day 20. Chondrocytes of the polypropylene mesh group proliferated and produced matrix through day 10 in culture, but were no longer viable and had no matrix production on days 15 and 20. Regenerated cellulose appeared to be toxic to canine chondrocytes during all stages of in vitro three-dimensional culture.

Conclusions

Three-dimensional culture of canine chondrocytes in agarose appears to produce favorable results with respect to chondrocyte proliferation and matrix production. Canine CB appears to have beneficial effects with regard to early GAG synthesis. Polypropylene mesh and oxidized regenerated cellulose had detrimental effects on cellular proliferation and matrix production. (Am J Vet Res 1997;58:419–424)

Abstract

Objective

To determine the effects of transplantable substrates on canine chondrocytes grown in three-dimensional culture.

Animals

3 canine cadavers.

Procedure

Articular cartilage harvested from canine cadavers was used to obtain chondrocytes for primary culture. Subcultured chondrocytes were grown in agarose alone (AG), or in agarose on canine cancellous bone (CB), polypropylene mesh, or oxidized regenerated cellulose substrate. Cell proliferation, proteoglycan and glycosaminoglycan (GAG) production, and collagen production were assessed on days 3, 6, 10, 15 and 20.

Results

Chondrocytes from groups AG and CB proliferated and produced matrix over the entire 20-day study period. Group-CB chondrocytes had significantly more GAG than did chondrocytes of all other groups on days 6 (P = 0.0297) and 15 (P = 0.00272). Those of groups AG and CB contained significantly (P = 0.0235) more GAG on day 20. Chondrocytes of the polypropylene mesh group proliferated and produced matrix through day 10 in culture, but were no longer viable and had no matrix production on days 15 and 20. Regenerated cellulose appeared to be toxic to canine chondrocytes during all stages of in vitro three-dimensional culture.

Conclusions

Three-dimensional culture of canine chondrocytes in agarose appears to produce favorable results with respect to chondrocyte proliferation and matrix production. Canine CB appears to have beneficial effects with regard to early GAG synthesis. Polypropylene mesh and oxidized regenerated cellulose had detrimental effects on cellular proliferation and matrix production. (Am J Vet Res 1997;58:419–424)

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