FT-IR microscopic mappings of early mineralization in chick limb bud mesenchymal cell cultures

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Abstract
Summary Chick limb bud mesenchymal cells differentiate into chondrocytes and form a cartilaginous matrix in culture. In this study, the mineral formed in different areas within cultures supplemented with 4 mM inorganic phosphate, or 2.5, 5.0, and 10 mM β-glycerophosphate (βGP), was characterized by Fourier-transform infrared (FT-IR) microscopy. The relative mineral-to-matrix ratios, and distribution of crystal sizes at specific locations throughout the matrix were measured from day 14 to day 30. The only mineral phase detected was a poorly crystalline apatite. Cultures receiving 4 mM inorganic phosphate had smaller crystals which were less randomly distributed around the cartilage nodules than those in the βGP-treated cultures. βGP-induced mineral consisted of larger, more perfect apatite crystals. In cultures receiving 5 or 10 mM βGP, the relative mineral-to-matrix ratios (calculated from the integrated intensities of the phosphate and amide I bands, respectively) were higher than in the cultures with 4mM inorganic phosphate or in the in vivo calcified chick cartilage.