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Depth-varying Density and Organization of Chondrocytes in Immature and Mature Bovine Articular Cartilage Assessed by 3D Imaging and Analysis

Kyle D. Jadin, Benjamin L. Wong, Won C. Bae, Kelvin W. Li, Amanda K. Williamson, Barbara L. Schumacher, Jeffrey H. Price and Robert L. Sah

Department of Bioengineering, University of California–San Diego, La Jolla, California

Correspondence to: Robert L. Sah, Department of Bioengineering, University of California–San Diego, 9500 Gilman Drive, MC 0412, La Jolla, CA 92093-0412. E-mail: rsah{at}ucsd.edu

Articular cartilage is a heterogeneous tissue, with cell density and organization varying with depth from the surface. The objectives of the present study were to establish a method for localizing individual cells in three-dimensional (3D) images of cartilage and quantifying depth-associated variation in cellularity and cell organization at different stages of growth. Accuracy of nucleus localization was high, with 99% sensitivity relative to manual localization. Cellularity (million cells per cm³) decreased from 290, 310, and 150 near the articular surface in fetal, calf, and adult samples, respectively, to 120, 110, and 50 at a depth of 1.0 mm. The distance/angle to the nearest neighboring cell was 7.9 µm/31°, 7.1 µm/31°, and 9.1 µm/31° for cells at the articular surface of fetal, calf, and adult samples, respectively, and increased/decreased to 11.6 µm/31°, 12.0 µm/30°, and 19.2 µm/25° at a depth of 0.7 mm. The methodologies described here may be useful for analyzing the 3D cellular organization of cartilage during growth, maturation, aging, degeneration, and regeneration. (J Histochem Cytochem 53:1109–1119, 2005)

Key Words: three-dimensional imaging • cartilage • cell organization • chondrocyte • development • growth • histology • nucleus

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