Temporal dynamic changes in synthesis of chondroitin sulfate isomers in canine articular chondrocyte culture.
To investigate temporal dynamic changes in the synthesis of chondroitin 6-sulfate (CS6) and chondroitin 4-sulfate (CS4) in vitro, normal articular cartilage of femoral heads was harvested from three dogs. Chondrocytes were isolated and cultured in alginate microspheres for 21 days. On days 7, 14 and 21, DNA content was quantified by fluorometric assay using Hoechst 33258. On days 14 and 21, proteoglycans were extracted, and the amounts of CS6 and CS4 were quantified after chondroitinase ABC digestion using capillary electrophoresis. The DNA content and amounts of CS6 and CS4 increased during the culture period. The amounts of CS6 and CS4 divided by DNA content revealed that the synthesis of CS6 was more up-regulated than CS4.
Maeda
Sadatoshi
S
Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainsville, FL 32610-0126, USA.
Miyabayashi
Takayoshi
T
Yamamoto
Janet K
JK
Roberts
Gregory D
GD
Lepine
Allan J
AJ
Clemmons
Roger M
RM
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000096:biosynthesis*; D004247:DNA / Q000378:metabolism; D004285:Dogs; D007700:Kinetics; D013997:Time Factors
D002809:Chondroitin Sulfates; D004247:DNA