Biochemical, histochemical, and immunohistochemical characterization of distal tibial osteochondrosis in horses

J. D. Lillich From the Departments of Veterinary Clinical Sciences (Lillich, Bertone, Ruggles) and Veterinary Pathobiology (Weisbrode), The Ohio State University, 601 Vernon L Tharp St, Columbus, OH 43210-1089, and the Departments of Medicine (Malemud) and Orthopedics and Anatomy (Stevenson), Case Western Reserve University, Cleveland, OH 44106.

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Alicia L. Bertone From the Departments of Veterinary Clinical Sciences (Lillich, Bertone, Ruggles) and Veterinary Pathobiology (Weisbrode), The Ohio State University, 601 Vernon L Tharp St, Columbus, OH 43210-1089, and the Departments of Medicine (Malemud) and Orthopedics and Anatomy (Stevenson), Case Western Reserve University, Cleveland, OH 44106.

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Charles J. Malemud From the Departments of Veterinary Clinical Sciences (Lillich, Bertone, Ruggles) and Veterinary Pathobiology (Weisbrode), The Ohio State University, 601 Vernon L Tharp St, Columbus, OH 43210-1089, and the Departments of Medicine (Malemud) and Orthopedics and Anatomy (Stevenson), Case Western Reserve University, Cleveland, OH 44106.

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Steven E. Weisbrode From the Departments of Veterinary Clinical Sciences (Lillich, Bertone, Ruggles) and Veterinary Pathobiology (Weisbrode), The Ohio State University, 601 Vernon L Tharp St, Columbus, OH 43210-1089, and the Departments of Medicine (Malemud) and Orthopedics and Anatomy (Stevenson), Case Western Reserve University, Cleveland, OH 44106.

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Alan J. Ruggles From the Departments of Veterinary Clinical Sciences (Lillich, Bertone, Ruggles) and Veterinary Pathobiology (Weisbrode), The Ohio State University, 601 Vernon L Tharp St, Columbus, OH 43210-1089, and the Departments of Medicine (Malemud) and Orthopedics and Anatomy (Stevenson), Case Western Reserve University, Cleveland, OH 44106.

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Sharon Stevenson From the Departments of Veterinary Clinical Sciences (Lillich, Bertone, Ruggles) and Veterinary Pathobiology (Weisbrode), The Ohio State University, 601 Vernon L Tharp St, Columbus, OH 43210-1089, and the Departments of Medicine (Malemud) and Orthopedics and Anatomy (Stevenson), Case Western Reserve University, Cleveland, OH 44106.

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Abstract

Objective

To compare the biochemical, histochemical, and immunohistochemical profiles of articular cartilage from horses with naturally acquired distal tibial osteochondrosis (OC) with cartilage from a similar location in clinically normal horses.

Animals

9 affected horses (group 1, 16 OC lesions) and 4 control horses (group 2, 8 normal osteochondral specimens).

Procedure

OC specimens were collected during arthroscopic removal of the fragment, and control specimens were collected by aseptic osteotomy. Uronic acid, total protein, total glycosaminoglycan (GAG), chondroitin sulfate (CS), and keratan sulfate (KS) contents were determined. Histomorphologic, histochemical, and immunohistochemical examinations were performed on specimens after snap freezing at −80 C and cryosectioning. Monoclonal antibodies (MAB) 3B3 and 5D4 were applied for location of epitopes of CS and KS, respectively.

Results

OC lesions had significantly lower quantity of uronic acid, total GAG, and CS, compared with normal cartilage. OC cartilage had significantly less intense staining with toluidine blue, along with irregular cellularity and tidemark characteristics, compared with normal cartilage. Monoclonal antibodies 3B3 and 5D4 stained OC cartilage, whereas MAB 5D4 did not stain control cartilage. Additionally, MAB 3B3 and 5D4 stained the fibrous tissue that was found firmly attached to the OC lesion located between the parent distal portion of the tibia and OC fragment.

Conclusion

OC cartilage lesions of the distal intermediate ridge of the tibia in horses are biochemically, histochemically, and immunohistochemically distinct from normal cartilage from the same location. Results may reflect the inability of the chondrocyte of the developing joint to alter matrix components that would allow proper maturation and differentiation into bone. (Am J Vet Res 1997;58:89–98)

Abstract

Objective

To compare the biochemical, histochemical, and immunohistochemical profiles of articular cartilage from horses with naturally acquired distal tibial osteochondrosis (OC) with cartilage from a similar location in clinically normal horses.

Animals

9 affected horses (group 1, 16 OC lesions) and 4 control horses (group 2, 8 normal osteochondral specimens).

Procedure

OC specimens were collected during arthroscopic removal of the fragment, and control specimens were collected by aseptic osteotomy. Uronic acid, total protein, total glycosaminoglycan (GAG), chondroitin sulfate (CS), and keratan sulfate (KS) contents were determined. Histomorphologic, histochemical, and immunohistochemical examinations were performed on specimens after snap freezing at −80 C and cryosectioning. Monoclonal antibodies (MAB) 3B3 and 5D4 were applied for location of epitopes of CS and KS, respectively.

Results

OC lesions had significantly lower quantity of uronic acid, total GAG, and CS, compared with normal cartilage. OC cartilage had significantly less intense staining with toluidine blue, along with irregular cellularity and tidemark characteristics, compared with normal cartilage. Monoclonal antibodies 3B3 and 5D4 stained OC cartilage, whereas MAB 5D4 did not stain control cartilage. Additionally, MAB 3B3 and 5D4 stained the fibrous tissue that was found firmly attached to the OC lesion located between the parent distal portion of the tibia and OC fragment.

Conclusion

OC cartilage lesions of the distal intermediate ridge of the tibia in horses are biochemically, histochemically, and immunohistochemically distinct from normal cartilage from the same location. Results may reflect the inability of the chondrocyte of the developing joint to alter matrix components that would allow proper maturation and differentiation into bone. (Am J Vet Res 1997;58:89–98)

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