Imposed state of deformation determines local collagen fibre orientation but not apparent mechanical properties

In a previous study, we have shown that the observed biaxial material behaviour of planar connective tissues is influenced by the sample gripping method. Commonly used suture attachments produced an apparently more compliant and extensible material compared to the same sample with clamped edges. We hypothesized that these differences were due to the imposed collagen fibre constraint under each method. In this study, we have directly compared the collagen fibre orientations which result under both gripping schemes. Small angle light scattering (SALS) was used to determine collagen fibre orientations in square bovine pericardial samples, before and after a 10% equibiaxial stretch. Local fibre distributions were determined at the sample centre and at the grip-sample interface. Resulting scattering patterns were statistically compared using repeated measures ANOVA. After stretch, collagen fibre distributions were identical at the sample centre where deformation is measured - but not at the sample boundaries. Therefore, the central fibre orientation distribution appears to be exclusively determined by the imposed deformation state. However, it is important to note that the loads necessary to achieve a given deformation depended strongly on gripping method. The resulting apparent differences in mechanical properties must be due to the method of load transmission. Indeed, fibres were observed to arc around suture attachment points, suggesting a discontinuous load transfer to the specimen which produced an apparent increase in extensibility and compliance. By contrast, only smooth transitions were observed at the clamped edges. Direct transmission of load from grip-to-grip in clamped samples (away from the sample centre) increased apparent stiffness.