Effect of epithelial debridement on human cornea proteoglycans.
Corneal transparency is attributed to the regular spacing and diameter of collagen fibrils, and proteoglycans may play a role in fibrillogenesis and matrix assembly. Corneal scar tissue is opaque and this opacity is explained by decreased ultrastructural order that may be related to proteoglycan composition. Thus, the objectives of the present study were to characterize the proteoglycans synthesized by human corneal explants and to investigate the effect of mechanical epithelial debridement. Human corneas unsuitable for transplants were immersed in F-12 culture medium and maintained under tissue culture conditions. The proteoglycans synthesized in 24 h were labeled metabolically by the addition of (35)S-sulfate to the medium. These compounds were extracted by 4 M GuHCl and identified by a combination of agarose gel electrophoresis, enzymatic degradation with protease and mucopolysaccharidases, and immunoblotting. Decorin was identified as the main dermatan sulfate proteoglycan and keratan sulfate proteoglycans were also prominent components. When the glycosaminoglycan side chains were analyzed, only keratan sulfate and dermatan sulfate were detected (approximately 50% each). Nevertheless, when these compounds were (35)S-labeled metabolically, the label in dermatan sulfate was greater than in keratan sulfate, suggesting a lower synthesis rate for keratan sulfate. (35)S-Heparan sulfate also appeared. The removal of the epithelial layer caused a decrease in heparan sulfate labeling and induced the synthesis of dermatan sulfate by the stroma. The increased deposit of dermatan sulfate proteoglycans in the stroma suggests a functional relationship between epithelium and stroma that could be related to the corneal opacity that may appear after epithelial cell debridement.
Departamento de Bioquímica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brasil.
E S
ES
Soriano
M S
MS
Campos
J A
JA
Aguiar
Y M
YM
Michelacci
D003315:Cornea / Q000378:metabolism*; D065306:Corneal Injuries; D003319:Corneal Stroma / Q000378:metabolism; D003646:Debridement / Q000009:adverse effects; D003871:Dermatan Sulfate / Q000096:biosynthesis; D004587:Electrophoresis, Agar Gel; D005109:Extracellular Matrix; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000302:isolation & purification; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D007632:Keratan Sulfate / Q000378:metabolism; D011509:Proteoglycans / Q000096:biosynthesis* / Q000302:isolation & purification; D017154:Stromal Cells / Q000378:metabolism
D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D007632:Keratan Sulfate; D011509:Proteoglycans