Three-dimensional in situ zonal morphology of viable growth plate chondrocytes: a confocal microscopy study

Longitudinal growth, occurring in growth plates with structurally distinct zones, has clinical implications in the treatment of progressive skeletal deformities. This study documents the three-dimensional morphology of chondrocytes within histological zones of growth plate using confocal microscopy combined with fluorescent labeling techniques. Three-dimensional reconstruction of Calcein AM-labeled chondrocytes was made from stacks of confocal images recorded in situ from 4-week-old swine growth plates. Three-dimensional quantitative morphological measurements were further performed and compared at both tissue and cell levels. Chondrocyte volume and surface area increased about five- and threefold, respectively, approaching the chondro-osseous junction from the pool of reserve cells. Chondrocytes from the proliferative zone were the most discoidal cells (sphericity of 0.81 ± 0.06) among three histological zones. Minimum and maximum cell/matrix volume ratios were identified in the reserve (11.0 ± 2.2) and proliferative zones (16.8 ± 3.0), respectively. Evaluated parameters revealed the heterogeneous and zone-dependent morphological state of the growth plate. Tissue and cellular morphology may have noteworthy contribution to the growth plate behavior during growth process. The ability to obtain in situ cell morphometry and monitor the changes in the growth direction could improve our understanding of the mechanisms through which abnormal growth is triggered.