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In vitro correlation of ultrastructural morphology and creatine phosphokinase release in L6 skeletal muscle cells after exposure to parenteral antibiotics

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Summary

Morphologic changes in a rat skeletal muscle cell line (L6) exposed for 1 h to the parenteral antibiotics amphotericin B (AMP), tetracycline-HCl (TET), erythromycin lactobionate (ERY), and cephaloridine (CEP) were characterized by transmission and scanning electron microscopy and compared to cellular release of creatine phosphokinase (CRK). AMP (0.05, 0.1, 0.5 mg/ml) caused a concentration-related swelling of nuclei, endoplasmic reticulum, and mitochondria. Loss of membrane integrity associated with AMP exposure was evident at the middle concentration and extensive at the high concentration, which correlated well with the 43 and 90% depletion of CPK from the muscle cells, respectively. TET (0.25, 1.0, 2.5 mg/ml) caused dilation of endoplasmic reticulum and cytoplasmic blebbing at the low concentration but had no effect on the cytoplasmic membrane or CPK. Cells exposed to the high concentration of TET had extensive damage to the cytoplasmic membrane, and CPK was completely depleted. ERY (2.5, 5.0, 25 mg/ml) caused a pattern of morphologic changes and CPK depletion similar to TET. CEP (4.0, 20, 50 mg/ml) had no effect on membrane integrity or CPK; however, membranous whorls were prominent in the cytoplasm. A good correlation between CPK release and cytoplasmic membrane integrity was evident and the ability of these agents to release CPK from muscle cells in culture correlated with the known irritancy potential of these parenteral antibiotics. Furthermore, CPK depletion seems to be a reliable indicator of muscle cell damage after cytoplasmic membrane perturbation and is therefore an appropriate index of toxicity in this in vitro muscle irritation model.

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References

  1. Aubert-Tulkens, G.; Von Hoof, F.; Tulkens, P. Gentamicin-induced lysosomal phospholipidosis in cultured rat fibroblasts. Lab. Invest. 40:(4)481–491:1979.

    PubMed  CAS  Google Scholar 

  2. Bagdon, R. D.; Prince, H. N. Assessment of primary irritation of parenteral drugs in vitro. (abstract). In: Frohberg, H., ed. Proceedings XXIVth congress of the European Society of Toxicology: Rome. Berlin: Springer International; 1983:93.

    Google Scholar 

  3. Benoit, P. W.; Belt, D. Destruction and regeneration of skeletal muscle after treatment with a local anaesthetic, bupivacaine (Marcaine). J. Anat. 107:(3)547–556; 1970.

    PubMed  CAS  Google Scholar 

  4. Cingolani, E.; Cingolani, G.; Mosconi, P., et al. Local tolerance evaluation of intramuscular drug preparations. Farmaco Edigione Pratica (Pavia) 41:89–97; 1985.

    Google Scholar 

  5. Comereski, C. R.; Williams, P. D.; Bregman, C. L., et al. Pain on injection and muscle irritation: a comparison of animal models for assessing parenteral antibiotics Fundam. Appl. Toxicol. 6:335–338; 1986.

    Article  PubMed  CAS  Google Scholar 

  6. Davies, W. A.; Stossel, T. P. Peripheral hyline blebs (podosomes) of macrophages. J. Cell Biol. 75:941–955; 1977.

    Article  PubMed  CAS  Google Scholar 

  7. Dukes, M. N. G. Index of side effects. In: Dukes, M. N. G., ed., Meylers side effect's of drugs. New York: Elsevier; 1984.

    Google Scholar 

  8. Ericsson, J. L. B.; Brunk, U. T. Alterations in lysosomal membranes as related to disease processes. In; Trump, B. J.; Arstila, A. U., eds., Pathobiology of cell membranes, vol. 1. New York: Academic Press; 1975:217–253.

    Google Scholar 

  9. Grim, M.; Rerabkova, L.; Carlson, B. M. A test for muscle lesions and their regeneration following intramuscular drug application. Toxicol. Pathol. 16:(4)432–442; 1988.

    Article  PubMed  CAS  Google Scholar 

  10. Hall-Craggs, E. C. B. Early ultrastructural changes in skeletal muscle exposed to the local anesthetic bupivacaine (Marcaine). Br. J. Exp. Pathol. 61:139–149; 1980.

    PubMed  CAS  Google Scholar 

  11. Hayat, M. A. In: Principals and techniques of electron microscopy biological applications, vol. 1; 2nd ed. Baltimore, MD: University Park Press; 1981:57.

    Google Scholar 

  12. Holohan, P. D.; Sokol, P. P.; Ross, C. R., et al. Gentamicin-induced increases in cytosolic calcium in pig kidney cells (LLC-PK1). J. Pharmacol. Exp. Ther. 247:(1)349–354; 1988.

    PubMed  CAS  Google Scholar 

  13. Houslay, M. D.; Stanley, K. K. Dynamic membrane-associated processes. In: Dynamics of biological membranes. New York: John Wiley & Sons; 1984.

    Google Scholar 

  14. Hruban, Z.; Slesers, A.; Hopkins, E. Drug induced and naturally occurring myeloid bodies. Lab. Invest. 27:(1)62–70; 1972.

    PubMed  CAS  Google Scholar 

  15. Josepovitz, C.; Levine, R.; Lane, B., et al. Contrasting effects of gentamicin and mercuric chloride on urinary excretion of enzymes and phospholipids in the rat. Lab. Invest. 52:(4)375–386; 1985.

    PubMed  CAS  Google Scholar 

  16. Maunsbach, A. B. Functions of lysosomes in kidney cells. In: Dingle, J. T.; Fell, H. B., eds., Lysosomes in biology and pathology, vol. 2. Amsterdam: North-Holland; 1969.

    Google Scholar 

  17. Nakahara, M. Effects of antibiotics on lysosomes of skeletal muscle. J. Med. 5:234–243; 1974.

    PubMed  CAS  Google Scholar 

  18. Nelson, A. A.; Price, C. W.; Welch, H. Muscle irritation following the injection of various penicillin preparations in rabbits. J. Am. Pharm. Assoc. 38:237–239; 1949.

    CAS  Google Scholar 

  19. Nicolson, G. L.; Smith, J. R.; Poste, G. Effects of local anesthetics on cell morphology and membrane-associated cytoskeletal organization in BALB/3T3 cells. J. Cell Biol. 68:(2)395–402; 1976.

    Article  PubMed  CAS  Google Scholar 

  20. Oshida, S.; Degawa, K.; Takahashi, Y., et al. Physico-chemical properties and local toxic effects of injectables. Tohoku J. Exp. Med. 127:301–316; 1979.

    Article  PubMed  CAS  Google Scholar 

  21. Oradell, NJ: Physician's Desk Reference. Medical Economics Co.; 1986.

  22. Rosalki, S. B. An improved procedure for serum creatine phosphokinase determination. J. Lab. Clin. Chem. 69:696–705; 1967.

    CAS  Google Scholar 

  23. Svendsen, O.; Rasmussen, F.; Neilsen, P., et al. The loss of creatine phosphokinase (CK) from intramuscular injection sites in rabbits. A predictive tool for local toxicity. Acta Pharmacol. Toxicol. 44:324–328; 1979.

    Article  CAS  Google Scholar 

  24. Svendsen, O.; Hjelse, F.; Bagdon, R. E. Tests for local toxicity of intramuscular drug preparations: comparison of in vivo and in vitro models. Acta Pharmacol. Toxicol. 56:183–190; 1985.

    Article  CAS  Google Scholar 

  25. Svendsen, O. Histologic changes after intramuscular injection with antibiotic preparations. Nord. Vet. Med. 24:181–185; 1972.

    Google Scholar 

  26. Thelestam, M.; Molley, R. Cultured human fibroblasts as a model for evaluation of potential in-vivo toxicity of membrane damaging antibotics. Chem. Biol Interact. 29(3):315–326; 1980.

    Article  PubMed  CAS  Google Scholar 

  27. Trifillis, A. L.; Regec, A. L.; Trump, B. F. Isolation, culture and characterization of human renal tubular cells. J. Urol. 133:324–329; 1985.

    PubMed  CAS  Google Scholar 

  28. Trump, B. F.; Berezesky, I. K. Mechanisms of cell injury: role of ion regulation. In Vitro Cell. Dev. Biol. 22:49A; 1986.

    Google Scholar 

  29. Weinberg, J. M.; Hunt, D.; Humes, D. Distribution of gentamicin among subcellular fractions from rat renal cortex. Biochem. Pharm. 34:(10)1779–1787; 1985.

    Article  PubMed  CAS  Google Scholar 

  30. Weissmann, G.; Hirschhorn, R.; Pras, M., et al. Studies of lysosomes-VIII: the effects of polyene antibiotics on lysosomes. Biochem. Pharm. 16:1057–1069; 1967.

    Article  PubMed  CAS  Google Scholar 

  31. Weissmann, G. The role of lysosomes in inflammation and disease. Annu. Rev. Med. 18:97–112; 1967.

    Article  PubMed  CAS  Google Scholar 

  32. Wilkinson, J. H. Clinical significance of enzyme activity measurements. Clin. Chem. 16:882–890; 1970.

    PubMed  CAS  Google Scholar 

  33. Williams, P. D.; Masters, B. G.; Evans, L. D., et al. An in vitro model for assessing muscle irritation due to parenteral antibiotics. Fundam. Appl. Toxicol. 9:10–17; 1987.

    Article  PubMed  CAS  Google Scholar 

  34. Zollinger, H. H. Cytological studies with the phase microscope I. The formation of “blisters” on cells in suspension (potocytosis), with observations on the nature of the cellular membrane. Am. J. Pathol. 24:545–558; 1948.

    CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Lilly Research Laboratories, Toxicology Division, A Division of Eli Lilly and Company, 46140, Greenfield, Indiana

    Dennis A. Laska, Patricia D. Williams, Sandy L. White, Carol A. Thompson & Dennis M. Hoover

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  1. Dennis A. Laska
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  2. Patricia D. Williams
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Laska, D.A., Williams, P.D., White, S.L. et al. In vitro correlation of ultrastructural morphology and creatine phosphokinase release in L6 skeletal muscle cells after exposure to parenteral antibiotics. In Vitro Cell Dev Biol 26, 393–398 (1990). https://doi.org/10.1007/BF02623831

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  • DOI: https://doi.org/10.1007/BF02623831

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