Modulation of proteoglycan metabolism by human fibroblasts maintained in an endogenous three-dimensional matrix.
This report describes synthesis and degradation of proteoglycans by human gingival fibroblasts growing in an endogenous three-dimensional matrix. Cells grown in the matrix cultures demonstrated a high rate of proteoglycan synthesis, varying between 2 and 4 times that of cells maintained in monolayer cultures. In addition, the relative amount deposited into the cell layer was increased in the matrix cultures, constituting 70% to 90% of the synthesized material during the first 24 h. Comparable levels for the monolayer cultures were 30% to 60%. The majority of the 35S-sulfate-labeled material in both matrix (80%) and monolayer (62%) cultures was susceptible to chondroitin ABC-lyase digestion. The major product was a low Mr (120,000) proteoglycan which could be immunoprecipitated by an antibody against PGII (decorin). In addition, the cells synthesized two chondroitin ABC-lyase-sensitive proteoglycans, one with Mr greater than 400,000, one with an apparent Mr of 250,000, as well as two heparan sulfate proteoglycans with Mr greater than 250,000. The low Mr dermatan sulfate, decorin, was also the major component deposited in the three-dimensional matrix, constituting about 60% of the total sulfate incorporation. In contrast, fibroblasts in monolayer cultures deposited only a small amount (13%) of decorin (PGII) in the cell layer, and the major proteoglycan in this compartment was heparin sulfate. The rate of release of the newly deposited proteoglycans was the same in the two culture conditions, although material released from the three-dimensional matrix cultures contained small Mr components indicating a higher degree of degradation. These studies show differences in proteoglycan metabolism by gingival fibroblasts grown in an endogenous matrix and in monolayer cultures.(ABSTRACT TRUNCATED AT 250 WORDS)
Qwarnström
E E
EE
Department of Pathology, University of Washington, Seattle.
Kinsella
M G
MG
MacFarlane
S A
SA
Page
R C
RC
Wight
T N
TN
D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000378:metabolism; D046508:Culture Techniques / Q000379:methods; D058575:Decorin; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins; D005347:Fibroblasts / Q000378:metabolism*; D005881:Gingiva / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D011509:Proteoglycans / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism
D002809:Chondroitin Sulfates; D058575:Decorin; D016326:Extracellular Matrix Proteins; D006497:Heparan Sulfate; D011509:Proteoglycans; D013431:Sulfates; C546878:DCN protein, human