Fibrillogenesis in dense collagen solutions: a physicochemical study.
Fibrillogenesis, the formation of collagen fibrils, is a key factor in connective tissue morphogenesis. To understand to what extent cells influence this process, we systematically studied the physicochemistry of the self-assembly of type I collagen molecules into fibrils in vitro. We report that fibrillogenesis in solutions of type I collagen, in a high concentration range close to that of living tissues (40-300 mg/ml), yields strong gels over wide pH and ionic strength ranges. Structures of gels were described by combining microscopic observations (transmission electron microscopy) with small- and wide-angle X-ray scattering analysis, and the influence of concentration, pH, and ionic strength on the fibril size and organization was evaluated. The typical cross-striated pattern and the corresponding small-angle X-ray scattering 67-nm diffraction peaks were visible in all conditions in the pH 6 to pH 12 range. In reference conditions (pH 7.4, ionic strength=150 mM, 20 degrees C), collagen concentration greatly influences the overall macroscopic structure of the resultant fibrillar gels, as well as the morphology and structure of the fibrils themselves. At a given collagen concentration, increasing the ionic strength from 24 to 261 mM produces larger fibrils until the system becomes biphasic. We also show that fibrils can form in acidic medium (pH approximately 2.5) at very high collagen concentrations, beyond 150 mg/ml, which suggests a possible cholesteric-to-smectic phase transition. This set of data demonstrates how simple physicochemical parameters determine the molecular organization of collagen. Such an in vitro model allows us to study the intricate process of fibrillogenesis in conditions of molecular packing close to that which occurs in biological tissue morphogenesis.
Gobeaux
F
F
Chimie de la Matière Condensée, UMR 7574 CNRS-Université Pierre et Marie Curie, ENSCP-Ecole Pratique des Hautes Etudes, 12 rue Cuvier, 75005 Paris, France.
Mosser
G
G
Anglo
A
A
Panine
P
P
Davidson
P
P
Giraud-Guille
M-M
MM
Belamie
E
E
D000818:Animals; D024042:Collagen Type I / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D046529:Microscopy, Electron, Transmission; D051381:Rats; D017208:Rats, Wistar; D014961:X-Ray Diffraction
D024042:Collagen Type I