Summary
Simple and rapid methods for the determination and detection of autofluorescence and oxytetracycline (OTC) induced fluorescence in powdered and intact bone (without previous chemical extraction of the antibiotic) were used to estimate age and sex related differences in the rat femur. The autofluorescence always had collagen characteristics; it increased with age and was not sex dependent. The intensity of OTC induced fluorescence decreased with age, being higher in males than in females. In the oldest animals, however a slight increase in the fluorescence intensity was observed, and sex differences disappeared.
The spectrofluorometric measurements of intact femora indicate a higher OTC retention on the bone surface of young animals and adult males but not in adult females. The results indicate that the determination of OTC retention in the bone using our quantitative fluorometric method and the detection of OTC deposition at bone surface using a spectrofluorometric technique can be a sensitive and inexpensive tool for experimental studies of bone changes in physiological and pathological conditions.
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Argauer, R. J., White, C. E.: Fluorescent compounds for calibration of excitation and emission units of spectrofluorometer. Anal. Chem.36, 368–371 (1964)
Armstrong, W. G., Horsley, H. J.: Isolation and porperties of fluorescent components associated with calcified tissue collagen. Calcif. Tiss. Res.8, 197–210 (1972)
Armstrong, W. G., Erholtz, L., Prentice, A. I. D.: Age-related accumulation in calcified connective tissues of UV absorbing material masking an increased collagen-bound fluorescence found in older tissues. IVth International Osteological Symposium, Abstract 4, Prague 1972
Bachman, C. H., Ellis, E. H.: Fluorescence of bone. Nature206, 1328–1331 (1965)
Blanuša, M.: Utjecaj nekih faktora na kinetiku metabolizma kalcija (PhD Thesis), University Zagreb 1974
Buyske, D. A., Eisner, H. J., Kelly, R. G.: Concentration and persistence of tetracycline and chlortetracycline in bone. J. Pharmacol. Exp. Ther.130, 150–156 (1960)
Dekanić, D.: Odredjivanje tetraciklinskih antibiotika u kostima (MSc Thesis), University of Zagreb 1971
Dekanić, D., Blanuša, M., Kostial, K., Weber, K.: Odredjivanje tetraciklinskih antibiotika u koštanom tkivu i zubima mjerenjem fluorescencije. In: Antibiotici i antibiotska terapija (J. Fališevac et al., eds.), pp. 364–366. Zagreb: Ognjen Prica 1972
Dekanić, D.: Primjena tetraciklinskih antibiotika za studij metabolizma kalcija (PhD Thesis), University of Zagreb 1975
Epker, B. N., Frost, H. M.: Periosteal appositional bone growth from age two to age seventhy in man. A tetracycline evaluation. Anat. Rec.154, 573–577 (1966)
Frost, H. M.: Tetracycline-based histological analysis of bone remodeling. Calcif. Tiss. Res.3, 211–237 (1969)
Harris, W. H., Jackson, R. H., Jowsey, J.: The in vivo distribution of tetracyclines in canine bone. J. Bone Joint Surg. Am.44, 1308–1320 (1962)
Hawkins, K. I.: Fluorometric determination of demethylchlortetracycline and tetracycline in mammalian bone. Anal. Biochem.45, 128–136 (1972)
Himel, C. M., Mayer, R. T.: 5-Dimethylaminonaphtalene-1-sulfonic acid (DANS acid) as standard for quantum yield of fluorescence. Anal. Chem.42, 130–132 (1970)
Jowsey, J.: Age changes in human bone. Clin. Orthop.17, 210–218 (1960)
LaCroix, P., Coutelier, L.: Microscopie de fluorescence et microradiographie dans l'étude du vieillissement de l'os. In: Orthopaedic Surgery and Traumatology (J. Delchef, et al., eds.), pp. 361–363. Amsterdam: Excerpta Medica 1973
Lippert, E., Nägele, W., Seibold-Blankenstein, I., Staiger, U., Voss, W.: Messung von Fluorescenzspektren mit Hilfe von Spectralphotometern und Vergleichsstandards. Z. anal Chem.170, 1–18 (1959)
Milch, R. A., Rall, D. P., Tobie, J. E.: Fluorescence of tetracycline antibiotics in bone. J. Bone Joint Surg. Am.40, 897–910 (1958)
Prentice, A. I. D.: Bone autofluorescence and mineral content. Nature206, 1167 (1965)
Spasić, P., Weber, K.: Über die natürliche Fluoreszenz der Knochen. Acta Pharm. Jug.21, 151–158 (1971)
Steendijk, R.: Studies on the mechanism of the fixation of the tetracyclines to bone. In: Bone and Tooth (H. J. J. Blackwood, ed.), pp. 49–63. Oxford: Pergamon Press 1964
Urist, M. R., Ibsen, K. H.: Chemical reactivity of mineralized tissue with oxytetracycline. Arch. Pathol.76, 484–496 (1963)
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This study was supported in part by the “Pliva”, Pharmaceutical Works, Zagreb, Yugoslavia and US Department of Agriculture, Beltsville, Maryland
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Dekanić, D., Weber, K. & Kostial, K. Determination of oxytetracycline induced fluorescence and autofluorescence as an indicator of age and sex differences in rat femur. Pflugers Arch. 370, 77–80 (1977). https://doi.org/10.1007/BF00707949
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DOI: https://doi.org/10.1007/BF00707949