Novel optical imaging technique to determine the 3-D orientation of collagen fibers in cartilage: variable-incidence angle polarization-sensitive optical coherence tomography.

OBJECTIVE To investigate a novel optical method to determine the three dimensional (3-D) structure of articular cartilage collagen non-destructively. METHODS Polarization-sensitive optical coherence tomography was used to determine the apparent optical birefringence of articular cartilage for a number of different illumination directions. A quantitative method based on the theory of light propagation in uniaxial crystalline materials was validated on equine flexor tendon. Qualitative maps of fiber polar and azimuthal orientation at sites on the posterior and anterior segments of the equine third metacarpophalangeal (fetlock) joint were produced, and the azimuthal orientations compared with data from a split-line experiment. RESULTS Polar and azimuthal angles of cut flexor tendon broadly agreed with the nominal values but suggested that the accuracy was limited by our method of determining the apparent birefringence. On intact equine fetlock joints we found a non-zero polar tilt that changed in direction at various points along the apex, moving from the sagittal ridge outwards. The azimuthal orientation changes from being parallel to the sagittal ridge in the posterior region to being inclined to the ridge in the anterior region. This broadly agrees with split-line data for the anterior region but differs in the posterior region, possibly reflecting depth-dependent orientation changes. CONCLUSION General quantitative agreement was found between our method and histology in validation experiments. Qualitative results for cartilage suggest a complicated 3-D structure that warrants further study. There is potential to develop this approach into a tool that can provide depth-resolved information on collagen orientation in near real-time, non-destructively and in vivo.