Abstract
Fluorescent components in bone and dentine were separated from alkaline hydrolysates of their matrices on Sephadex C25 CM cationic exchange columns. The fluorescence levels, and the excitation (λ max 330 nm) and emission (λ max 395 nm) spectra, were the same as those observed in the intact and gelatinised matrices. The fluorescence parameters were unaltered by the hydrolysis procedure. Gel filtration on Sephadex G. 10 columns further resolved the isolated material into two components with the same fluorescence and UV absorption properties. The fluorescence was independent of pH over the range 3.5–9.5. Dialysis and gel filtration studies on the gelatinised matrices indicated a firmly-bonded association of the fluorescent material with the collagen polypeptide chains.
Résumé
Des composants fluorescents de l'os et la dentine sont séparés des hydrolysats alcalins de leur marice sur des colonnes Sephadex C25 CM d'échange cationique. Les concentrations en fluorescence et le spectre d'excitation (λ max 330 nm) et d'émission (λ max 395 nm) sont les mêmes que ceux observés au niveau des matrices intactes et gélatinisées. Les paramètres de fluorescence ne sont pas altérés par hydrolyse. La filtration sur gel à l'aide de colonnes Sephadex G 10 perment de différencier le matériel isolé en deux composants, ayant la même fluorescence et la même absorption UV. La fluorescence est indépendante de pH de 3.5–9.5. Des études de dialyse et de filtration sur gel de matrices gélatinisées indiquent une association étroite du matériel fluorescent avec les chaines polypeptidiques de collagène.
Zusammenfassung
Fluorescierende Bestandteile aus Knochen und Dentin wurden in Sephadex C25 CM Kationen-Austauschersäulen von alkalischen Hydrolysaten ihrer Matrices getrennt. Die Fluorescenzintensitäten sowie die Erregungs- (λ max 330 nm) und Emissions- (λ max 395 nm) Spektren waren dieselben wie bei intakten und gelatinisierten Matrices. Die Fluorescenzparameter wurden durch die Hydrolyse nicht verändert. Eine Gelfiltration über Sephadex-G10-Säulen trennte das isolierte Material in 2 Komponenten auf, welche gleiche Fluorescenz- und UV-Absorptionseigenschaften zeigten. Im pH-Bereich zwischen 3,5 und 9,5 war die Fluorescenz unabhängig vom pH. Dialysierversuche sowie Gelfiltrationsexperimente mitden gelatinisierten Matrices zeigten eine starkgefügte Bindung des fluorescierenden Materials mit den Polypeptidketten des Kollagens.
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References
Armstrong, W. G.: Further studies on the action of collagenase on sound and carious human dentine. J. dent. Res.37, 1001–1011 (1958).
—: Ultra-violet spectrometric estimation of dentine protein in solution. Arch. oral Biol.7, 771–772 (1962).
—: Fluorescent characteristics of sound and carious human dentine preparations. Arch. oral Biol.8, 79–90 (1963a).
—: The presence of ultra violet absorbing material and its relation to fluorescence. “Quenching” effects in carious dentine. Arch. oral Biol.8, 223–231 (1963b).
—: Modification of the properties and composition of the dentine matrix caused by dental caries. Adv. oral biol.1, 309–332 (1964).
—, Horsley, H. J.: Isolation of fluorescent components from ox-bone human dentine and gelatine. Nature (Lond.)211, 981 (1966).
Bailey, A. J., Peach, C. M., Fowler, L. J.: Chemistry of collagen cross-links. Biochem. J.117 819–831 (1970).
Hartles, R. L., Leaver, A.: The fluorescence of teeth under ultraviolet irradiation. Biochem. J.54, 632–638 (1953).
Hoerman, K. C., Balekjian, A. Y.: On the probability that the cross-link of collagen is its prime luminescent centre. J. dent. Res. (Suppl.) I.A.D.R. Abstract No 86, p. 59 1966a.
—: Some quantum aspects of collagen. Fed. Proc.25, 1016–1021 (1966b).
—, Mancewicz, A.: Phosphorescence of tissues. Arch. oral Biol.9, 517–534 (1964).
Horsley, H. J.: An investigation into the isolation and properties of fluorescent material in calcified tissues. M. Phil. thesis (University of London) 1967.
LaBella, F. S.: Analysis of fluorescence in elastin, collagen and procollagens. 2nd Canad. Conf. on Research in Rheumatic Diseases. Publ., p. 221–224 Can. Arthritis and Rheumatism Soc. Toronto 1961.
—: Separation of fluorescent and pigmented components of elastin. Possible relation to age pigments and certain invertebrate pigments. J. Geront.17, 8–13 (1962).
—, Thornhill, D. P.: Effects of ultraviolet irradiation on human and bovine collagen and elastin relationship of tyrosine to native fluorescence. Third Canad. Conf. in Res. in Rheumatic Diseases, p. 246–258. Toronto: Univ. Toronto Press 1966.
Laurila, U. R., Mancewicz, S. A., Forziati, A. F.: Isolation and partial fractionation of fluorescent material from human teeth. J. dent. Res.39, 714 (1960).
Jacobs, S.: An improved method for the quantitative determination of amino acids. Analyst81, 502–507 (1956).
Mancewicz, S., Hoerman, K. C.: Characteristics of Insoluble protein of tooth and bone I. Arch. oral Biol.9, 505–549 (1964).
McDevitt, C. A.: Isolation and characterisation studies on the fluorescent compounds in calcified tissues. M. Phil. Thesis (University of London) 1970.
Mechanic, G.: Stable intermolecular crosslinks in collagen after reduction. In: “Chemistry and molecular biology of the intercellular matrix”, vol. 1, p. 305–311. ed. E. A. Balaz. London-New York: Acad. Press 1970.
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Armstrong, W.G., Joan Horsley, H. Isolation and properties of fluorescent components associated with calcified tissue collagen. Calc. Tis Res. 8, 197–210 (1971). https://doi.org/10.1007/BF02010138
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DOI: https://doi.org/10.1007/BF02010138