Abstract
Metallic chambers were implanted into the proximal tibiae of rabbits to permit microscopic examination of living bonein situ. The bone repair process secondary to the injury produced during installation of the chamber, was visualized. Six to 8 weeks after implantation, osteoid and/or bone could be seen.
The effects of various doses of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) on the repair and regeneration processes following chamber implantation were studied. Data from various techniques indicated that: (1) following low dose EHDP (0.25 mg/kg/day) chambers contained bone tissue morphologically and ultrastructurally indistinguishable from controls; and (2) with higher doses of EHDP (2.5 or 10 mg/kg/day) chamber contained spicules of normal osteoid, osteoblasts and osteocytes, but were devoid of osteoclasts.
The effects of the various regimes of EHDP also were assessed on regenerated, trabecular bone contained within the tibia chambers three months after implantation of the chambers.
Data from various methods of analysis supported the following conclusions: (1) the low dose of EHDP (0.25 mg/kg/day) had no toxic effects on the trabecular bone within the chambers but there appeared to be an increase in bone formation as compared to saline control; (2) higher doses of EHDP (2.5 or 10 mg/kg/day) were not toxic to bone cells but thick osteoid seams formed on the trabecular bone within the chambers. No osteoclasts were found associated with the bone apparently due to the coverage of bone surfaces by osteoid seams; and (3) osteoid which accumulated after EHDP treatment of 2.5 mg/kg/day for 2 months remained uncalcified for as long as 2 months following withdrawal of EHDP administration.
The results showed the value of tibial chamber for examining microscopically living bonein situ and demonstrated the inhibitory effect of EHDP on mineralization of newly formed osteoid and a lack of effect on bone cells.
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References
Baylink, D., Wergedal, J. Thompson, E.: Loss of protein polysaccharides at sited where bone mineralization is initiated. J. Histochem. Cytochem.20, 279–292 (1972)
Gasser, A. B., Morgan, D. B., Fleisch, H., Richelle, L. J.: The influence of two diphosphonates on calcium metabolism in the rat. Clin. Sci.43, 31–45 (1972)
Goldner, M. M.: A modification of the Masson trichrome technique for routine laboratory purposes. Amer. J. Path.14, 237–243 (1938)
Gray, J. A., Opdyke, D. L.: A device for thin sectioning of hard tissue. J. dent. Res.41, 172–181 (1962)
King, W. R., Francis, M. D., Michael, W. R.: Effect of disodium ethane-1-hydroxy-1, 1-diphosphonate on bone formation. Clin. Orthop.78, 251–270 (1971)
Luft, J. H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol.9, 409–414 (1961)
Mallory, F. B.: Pathological techniques, p. 144. New York: Hafner Publishing Co. 1962
McClugage, S. G., McCuskey, R. S.: Relationship of the microvascular system to bone resorption and growthin vivo. Microvas. Res.6, 132–134 (1973)
McCuskey, R. S., McClugage, S. G., Younker, W. J.: Microscopy of living bone marrowin situ. I. Design and use of a bone chamber for chronic microscopic examination of living bone marrowin situ. Blood38, 87–95 (1971)
Michael, W. R., King, W. R., Francis, M. D.: Effectiveness of diphosphonates in preventing osteoporosis of dissue in the rat. Clin. Orthop.78, 271–276 (1971)
Miller, S. C., Jee, W. S. S.: The effectiveness of diphosphonates in inhibiting bone resorption in rats. Abstract, 51st General Session of the International Association for Dental Research, 1973
Mühlbauer, R. C., Russell, R. G. G., Williams, D. A., Fleisch, H.: The effects of diphosphonates, polyphosphates and calcitonin on “immobilization osteoporosis” in rats. Europ. J. clin. Invest.1, 336–344 (1970)
Rosenblum, I. Y.: The effects of disodium ethane-1-hydroxy-1, 1-diphosphonate (EHDP) on mineral and organic components of rat bone. Calcif. Tiss. Res.16, 239–250 (1974a)
Rosenblum, I. Y.: The effects of disodium ethane-1-hydroxy-1, 1-diphosphonate (EHDP) on bone and serum chemistry in the rabbit. Calcif. Tiss. Res.16, 145–152 (1974b)
Russell, R. G. G., Kislig, A. M., Casey, P. A., Fleisch, H., Thornton, J., Schenk, R., Williams D. A.: Effect of diphosphonates and calcitonin on the chemistry and quantitative histology of rat bone. Calcif. Tiss. Res.11, 179–195 (1973)
Russell, R. G. G., Mühlbauer, R. C., Bisaz, S., Williams, D. A., Fleich, H.: The influence of pyrophosphate, condensed phosphates, phosphonates and other phosphate compounds on the dissolution of hydroxyapatitein vitro and on bone resorption induced by parathyroid hormaone in tissue culture and in thyroparathyroidectomized rats. Calcif. Tiss. Res.6, 183–196 (1970)
Schenk, R., Merz, W. A., Mühlbauer, R., Russell, R. G. G., Fleisch, H.: Effect of ethane-1-hydroxy-1, 1-diphosphonate (EHDP) and dichloromethylene diphosphonate (Cl2MDP) on the calcification and resorption of cartilage and bone in the tibial epiphysis and metaphysis of rats. Calcif. Tiss. Res.11, 209–217 (1973)
Spurr, A. R.: A low viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res.26, 31–43 (1969)
Townsend, S. F.: Regeneration of gastric mucosa in rats. Amer. J. Anat.109, 133–147 (1961)
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Rosenblum, I.Y., McCuskey, R.S., McNeal, N.C. et al. The effects of EHDP on regenerating trabecular bone usingin vivo microscopic, light and electron microscopic, and electron microprobe techniques. Calc. Tis Res. 20, 91–104 (1976). https://doi.org/10.1007/BF02546400
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DOI: https://doi.org/10.1007/BF02546400