Microrheological characterization of collagen systems: from molecular solutions to fibrillar gels.
Collagen is the most abundant protein in the extracellular matrix (ECM), where its structural organization conveys mechanical information to cells. Using optical-tweezers-based microrheology, we investigated mechanical properties both of collagen molecules at a range of concentrations in acidic solution where fibrils cannot form and of gels of collagen fibrils formed at neutral pH, as well as the development of microscale mechanical heterogeneity during the self-assembly process. The frequency scaling of the complex shear modulus even at frequencies of ∼10 kHz was not able to resolve the flexibility of collagen molecules in acidic solution. In these solutions, molecular interactions cause significant transient elasticity, as we observed for 5 mg/ml solutions at frequencies above ∼200 Hz. We found the viscoelasticity of solutions of collagen molecules to be spatially homogeneous, in sharp contrast to the heterogeneity of self-assembled fibrillar collagen systems, whose elasticity varied by more than an order of magnitude and in power-law behavior at different locations within the sample. By probing changes in the complex shear modulus over 100-minute timescales as collagen self-assembled into fibrils, we conclude that microscale heterogeneity appears during early phases of fibrillar growth and continues to develop further during this growth phase. Experiments in which growing fibrils dislodge microspheres from an optical trap suggest that fibril growth is a force-generating process. These data contribute to understanding how heterogeneities develop during self-assembly, which in turn can help synthesis of new materials for cellular engineering.
Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada.
Marjan
M
Shayegan
Nancy R
NR
Forde
D000143:Acids / Q000737:chemistry; D001696:Biomechanical Phenomena; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism; D004548:Elasticity; D005109:Extracellular Matrix / Q000737:chemistry* / Q000378:metabolism; D024022:Fibrillar Collagens / Q000737:chemistry* / Q000378:metabolism; D005782:Gels / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D052898:Optical Tweezers; D012212:Rheology; D014783:Viscosity
D000143:Acids; D024022:Fibrillar Collagens; D005782:Gels; D003094:Collagen