The influence of hydrocortisone on aggrecan metabolism in human articular chondrocyte cultures: comparison of two different matrices.
OBJECTIVE
Numerous reports of negative effects as well as protective effects of glucocorticoids on articular cartilage convinced us to study the influence of hydrocortisone on aggrecan synthesis of isolated phenotypically stable human articular chondrocytes cultured in two different matrices.


METHODS
Macroscopically normal human articular cartilage was obtained from femoral condyles within 24 hours postmortem. Chondrocytes were isolated and cultured in gelled agarose or in alginate. After 14 days in culture, hydrocortisone was added for 5 days at concentrations ranging from 0.005 microgram to 1 mg/ml for the agarose cultures and from 0.005 microgram to 1 microgram/ml for the alginate culture system. Aggrecan synthesis was measured by the incorporation of 35Sulphate, and the proportion of neosynthesized aggrecan that bound to hyaluronan to form aggrecan aggregates was analyzed by gel chromatography.


RESULTS
At concentrations from 0.005 to 1 microgram/ml, hydrocortisone was found to produce a similar dose-dependent stimulation of aggrecan synthesis in both matrices. The synthesis of aggrecans remained at the same level for concentrations of 1 microgram/ml up to 100 micrograms/ml of hydrocortisone. When supraphysiological concentrations of hydrocortisone were added the aggrecan synthesis rate plateau declined. Simultaneously with the increase in aggrecan synthesis, the proportion of low-molecular weight 35S-proteoglycans decreased in favour of 35S-aggrecan aggregates and monomers in the agarose system. The chondrocytes cultured in alginate showed this increase of aggrecan aggregates and monomeric aggrecans in both the cell-associated and the inter-territorial matrix.


CONCLUSIONS
Hydrocortisone is a stimulator of aggrecan synthesis by normal human articular chondrocytes cultured in vitro. The two culture systems (agarose and alginate) tested in this experiment showed a comparable aggrecan synthesis rate, increasing under the influence of hydrocortisone at concentrations up to 1 microgram/ml. The proportions of 35Sulphate incorporated in aggrecan aggregates and monomeric aggrecan were also higher under the influence of hydrocortisone.
Almqvist
K F
KF
Department of Rheumatology, Ghent University Hospital, Ghent, Belgium. fredrik.almqvist@rug.ac.be
Wang
L
L
Broddelez
C
C
Verdonk
R
R
Veys
E M
EM
Verbruggen
G
G
D053674:Aggrecans; D000464:Alginates; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism*; D016326:Extracellular Matrix Proteins; D005782:Gels; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D037181:Lectins, C-Type; D011509:Proteoglycans / Q000096:biosynthesis*; D012685:Sepharose
D053674:Aggrecans; D000464:Alginates; D000893:Anti-Inflammatory Agents; D016326:Extracellular Matrix Proteins; D005782:Gels; D006603:Hexuronic Acids; D037181:Lectins, C-Type; D011509:Proteoglycans; D020723:Glucuronic Acid; D012685:Sepharose; D006854:Hydrocortisone