The modulation of collagen fibril assembly and its structure by decorin: an electron microscopic study.
The present study was carried out to determine the effect of decorin in the process of collagen assembly. Collagen fibrils were obtained in vitro by aggregation from commercialized acid-soluble type I collagen with the addition of different concentrations of decorin (0-25 microg/ml). All specimens were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The distribution of collagen fibril diameters was also analyzed by TEM. In samples without or with low concentrations of decorin, highly porous collagen fiber networks were formed. On the other hand, dense networks were observed in samples treated with high concentrations of decorin. The influence of decorin secreted by cells on collagen fibrils was observed by SEM, and the fiber network elasticity was measured using a rheometer. SEM images showed that collagen fiber networks without fibroblasts were much looser than those cultured with normal fibroblasts. The networks cultured with the fibroblasts were composed of straight fibers with large diameters. On the other hand, collagen fiber networks cultured with siRNA-decorin-transfected (siDT) fibroblasts had loose, meandering fibers with small diameters. The elasticity of collagen fiber networks cultured with untransfected fibroblasts showed no significant difference over the 7-day incubation period. However, significantly lower elastic values were obtained for collagen fiber networks treated with siDT cells on days 3 and 7. In addition, after treatment with 5.0 or 25 microg/ml decorin, the l collagen fiber networks cultured with siDT cells exhibited an altered structure that showed a dense structure similar to that of the fiber networks cultured with untransfected fibroblasts. In conclusion, this in vitro study showed that decorin is a regulatory and architecturally small leucine-rich repeat proteoglycan in the process of collagen fibril assembly.
Iwasaki
Shunsuke
S
Department of Veterinary Anatomy, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Japan.
Hosaka
Yoshinao
Y
Iwasaki
Tomohito
T
Yamamoto
Katsuhiro
K
Nagayasu
Aya
A
Ueda
Hiromi
H
Kokai
Yasuo
Y
Takehana
Kazushige
K
D058575:Decorin; D016326:Extracellular Matrix Proteins / Q000378:metabolism / Q000494:pharmacology*; D024022:Fibrillar Collagens / Q000378:metabolism / Q000648:ultrastructure*; D020894:Microfibrils / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism / Q000494:pharmacology*; D034622:RNA Interference; D014162:Transfection
D058575:Decorin; D016326:Extracellular Matrix Proteins; D024022:Fibrillar Collagens; D011509:Proteoglycans