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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Ahrens PBA, Solursh M, Reiter RS (1977) Stage-related capacity for limb chondrogenesis in cell culture. Dev Biol 60:69–82
Binderman I, Green RM, Pennypacker JP (1979) Calcification of differentiating skeletal mesenchyme in vitro. Science 206:222–225
Solursh M (1987) Use of tissue culture in the analysis of limb chondrogenesis. Mechanistic approaches to developmental toxicology. In: McLachlin JM, Pratt RM, Marker CL (ed) Developmental toxicology: mechanisms and risk, Banbury Report no 26. Cold Spring Harbor Laboratory, New York
Boskey AL, Stiner D, Doty S, Binderman I (1991) Requirement of vitamin C for cartilage calcification in a mesenchymal cell culture. Bone 12:277–282
Boskey AL, Stiner D, Leboy P, Doty S, Binderman I (1992) Optimal conditions for cartilage calcification in differentiating chick limb-bud mesenchymal cells. Bone Miner 16:11–37
Mendelsohn R, Hassankhani A, DiCarlo E, Boskey A (1989) FT-IR microscopy of endochondral ossification a 20 μ spatial resolution. Calcif Tissue Int 44:20–24
Boskey AL (1990) Bone mineral and matrix: are they altered in osteoporosis? Orthop Clin North Am 21:19–29
Pleshko NL (1991) Applications of FTIR microscopy to biomineralization. PhD Thesis, Rutgers University, Newark NJ
Pleshko NL, Boskey AL, Mendelsohn R (in press) An FTIR investigation of the effect of tissue preservation on bone. Calcif Tissue Int 51:72–77
Bailey RT, Holt C (1989) Fourier transform infrared spectroscopy and characterization of biological calcium phosphates. In: Hukins DWL (ed) Calcified tissue. Macmillan Press Ltd, Houndmills, Basingstoke, Hampshire, pp 93–120
Rey C, Lian JB, Grynpas M, Shapiro F, Zylerberg L, Glimcher MJ (1989) Non-apatitic environments in bone mineral. FT-IR detection, biological properties, and changes in several disease states. Connect Tissue Res 21:267–273
Sauer GR, Wuthier RE (1988) Fourier transform infrared characterization of mineral phases formed during induction of mineralization by collagenase-released matrix vesicles. J Biol Chem 263:13718
Termine JD, Lundy DR (1973) Hydroxide and carbonate in rat bone mineral and its synthetic analogues. Calcif Tissue Res 13:73–82
Termine JD, Lundy DR (1974) Vibrational spectra of some phosphate salts amorphous to x-ray diffraction. Calcif Tissue Res 15:55–70
Pleshko NL, Mendelsohn R, Boskey A (1991) A novel IR spectroscopic method for the determination of crystallinity of hydroxyapatite minerals. Biophys J 60:786–793
Willis JB (1960) Determination of metals in blood serum by atomic absorption spectroscopy. I. Calcium. Spectrochim Acta 16:259–271
Heinonen JK, Lahti RJ (1981) A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphates. Anal Biochem 113:313–317
Termine JD, Posner AS (1966) Amorphous/crystalline interrelationship in bone mineral. Calcif Tissue Res 1:8–23
Roufosse AH, Landis WJ, Sabine WK, Glimcher MJ (1979) Identification of brushite in newly deposited bone mineral from embryonic chicks. J Ultrastruct Res 68:235–255
Bellows CG, Aubin JE, Heersche JNM (1991) Initiation and progression of mineralization of bone nodules formed in vitro: the role of alkaline phosphatase and organic phosphate. Bone Miner 14:27–40
Satomura K, Hiraiwa K, Nagasayama M (1991) Mineralized nodule formation in rat bone marrow stromal cell culture without β-glycerophosphate. Bone Miner 14:41–54
Khouja HI, Bevington A, Kamp GJ, Russell RGG (1990) Calcium and orthophosphate deposits in vitro do not imply osteoblast-mediated mineralization: mineralization by beta glycerophosphate in the absence of osteoblasts. Bone 11:385–391
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Boskey, A.L., Camacho, N.P., Mendelsohn, R. et al. FT-IR microscopic mappings of early mineralization in chick limb bud mesenchymal cell cultures. Calcif Tissue Int 51, 443–448 (1992). https://doi.org/10.1007/BF00296678
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00296678