Three-dimensional morphology of the pericellular matrix of intervertebral disc cells in the rat

Intervertebral disc cells are surrounded by a pericellular matrix that is biochemically and morphologically distinct from other extracellular matrix regions. Although the function of the pericellular matrix is not fully understood, prior studies of pericellular matrix-chondrocyte regions in articular cartilage (termed 'chondrons') suggest that the size, shape, and mechanical properties of the pericellular matrix significantly influence the micromechanical environment of the contained cells. A first step in understanding the role of the pericellular matrix in the intervertebral disc is to quantify the three-dimensional morphology and zonal variations of these regions across the disc. In this study, three-dimensional reconstructions and morphometric measurements of pericellular matrix-cell regions were obtained in situ using fluorescence confocal microscopy of en bloc sections of nucleus pulposus and anulus fibrosus of the rat disc immunolabeled for type VI collagen. The morphology of the pericellular matrix and cells varied significantly across regions, with distinct pericellular matrix aspect ratios (largest/smallest diameter) showing shapes that were generally large and rounded in the nucleus pulposus (average of 1.9), and ellipsoidal and discoidal in the inner (2.4) and outer anulus fibrosus (2.8). The average pericellular matrix volume per cell was found to be significantly larger in the nucleus (6424 microm(3)) than that of inner (1903 microm(3)) and outer (1433 microm(3)) anulus. Pericellular matrix regions containing 1 or 2 cells were the dominant subgroup in the rat intervertebral disc at both 1 and 12 months of age. Multicellular pericellular matrix regions were present more often in the younger nucleus pulposus and outer anulus fibrosus. The orientation of the pericellular matrix regions further varied significantly across the disc, reflecting local collagen matrix architecture. These studies provide new information on the organization and shape of intervertebral disc cells and their surrounding pericellular matrix, which may provide new insights into the mechanisms that regulate cell-matrix interactions.