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Journal of Dental Research
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Phenytoin at Micromolar Concentrations is an Osteogenic Agent for Human-mandible-derived Bone Cells in vitro

O. Nakade

Departments of Medicine and Biochemistry, Loma Linda University, and Mineral Metabolism Unit (151), Jerry L. Pettis Memorial V.A. Medical Center, 11201 Benton Street, Loma Linda, California 92357, USA, Department of Oral Pathology, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido 061-02, Japan

D.J. Baylink

Departments of Medicine and Biochemistry, Loma Linda University, and Mineral Metabolism Unit (151), Jerry L. Pettis Memorial V.A. Medical Center, 11201 Benton Street, Loma Linda, California 92357, USA

K.-H.W. Lau

Departments of Medicine and Biochemistry, Loma Linda University, and Mineral Metabolism Unit (151), Jerry L. Pettis Memorial V.A. Medical Center, 11201 Benton Street, Loma Linda, California 92357, USA

The present study sought to test the hypothesis that phenytoin acts on normal human-mandible-derived bone cells to induce osteogenic effects. To test the effects of phenytoin on bone cell proliferation, we measured [ 3H]-thymidine incorporation into cell DNA during the final four hr of a 24-hour incubation with phenytoin. Phenytoin at micromolar concentrations significantly stimulated the [3H]-thymidine incorporation in a dose-dependent, biphasic, manner with a maximal effect at from 10 to 50 uM. We confirmed the proliferative effect of phenytoin by counting cell number. To evaluate the effects of phenytoin on osteoblastic differentiation, we determined alkaline-phosphatase specific activity and found that phenytoin at micromolar concentrations significantly increased that activity in a dose-dependent manner, with maximal stimulation at approximately 1 µM. To investigate the effects of phenytoin on mature osteoblastic activities, we measured de novo collagen synthesis and osteocalcin secretion. Mitogenic doses of phenytoin significantly increased collagen synthesis and osteocalcin secretion in a dose-dependent, biphasic, manner, with the maximal stimulatory dose at from 5 to 10 uM. In summary, phenytoin at micromolar ranges increased (a) [3H]-thymidine incorporation and cell number, (b) alkaline-phosphatase specific activity, (c) collagen synthesis, and (d) osteocalcin secretion in monolayer cultures of normal human-mandible-derived bone cells. These observations are consistent with the premise that low doses of phenytoin act on human craniofacial bone cells to stimulate cell proliferation, differentiation, and mature osteoblastic activities to stimulate bone formation.

Key Words: phenytoin • mandibular bone cells • osteogenesis • proliferation • differentiation.

REFERENCES

  • Beresford JN, Gallagher JA, Russell RGG (1986). 1,25-Dihydroxyvitamin D3 and human bone-derived cells in vitro: effects on alkaline phosphatase, type I collagen and proliferation. Endocrinology 119:1776-1785.[Abstract/Free Full Text]
  • Brown JP, Malaval L., Chapuy MC, Delmas PD, Edouard C., Meunier PJ (1984). Serum bone Gla-protein: a specific marker for bone formation in postmenopausal osteoporosis. Lancet 1:1091-1093.[Medline] [Order article via Infotrieve]
  • Caine DB (1973). The drug treatment of epilepsy. Br J Hosp Med 9:171-175.
  • Caspary WF (1972). Inhibition of intestinal calcium transport by diphenylhydantoin in rat duodenum. Naunyn-Schmiedeberg's Arch F Pharmakol 274:146-153.
  • Centrella M., McCarthy TL, Canalis E. (1991). Current concepts review: transforming growth factor-beta and remodeling of bone. J Bone Joint Surg 73(A):1418-1428.[Free Full Text]
  • Christiansen C., Rodbro P., Lund M. ( 1973a). Incidence of anticonvulsant osteomalacia and effect of vitamin D: controlled therapeutic trial. Br Med J 4:695-698.[Abstract/Free Full Text]
  • Christiansen C., Rodbro P., Lund M. ( 1973b). Effect of vitamin D on bone mineral mass in normal subjects and in epileptic patients on anticonvulsants: a controlled therapeutic trial. Br Med J 2:208-209.[Abstract/Free Full Text]
  • De Lorenzo RJ, Glaser GH (1976). Effect of diphenylhydantoin on the endogenous phosphorylation of brain proteins. Brain Res 105:381-386.[CrossRef][Medline] [Order article via Infotrieve]
  • Delgado-Escueta AV, Horan MP (1980). Phenytoin: biochemical membrane studies. Adv Neurol 27:377-398.[Medline] [Order article via Infotrieve]
  • Dietrich JW, Duffield R. (1980). Effects of diphenylhydantoin on synthesis of collagen and noncollagen protein in tissue culture. Endocrinology 106:606-610.[Abstract/Free Full Text]
  • Dill RE, Miller EK, Weil T., Lesley S., Farmer GR, Iacopino AM (1993). Phenytoin increases gene expression for platelet-derived growth factor B chain in macrophages and monocytes. J Periodontol 64:169-173.[Medline] [Order article via Infotrieve]
  • Dziak R., Vernillo A., Rifkin B. (1988). The effects of phenytoin on calcium uptake in osteoblastic cells. J Bone Min Res 3:415-420.[Medline] [Order article via Infotrieve]
  • Ebadi MS, Scott PM (1971). Increase in collagen level by diphenylhydantoin as possible mechanism of drug-induced gingival hyperplasia. Clin Toxicol 4:39-46.[Medline] [Order article via Infotrieve]
  • Farley JR, Baylink DJ (1986). Skeletal alkaline phosphatase activity as a bone formation index in vitro. Metabolism 35:563-571.[Medline] [Order article via Infotrieve]
  • Farley JR, Jorch UM (1983). Differential effects of phospholipids on skeletal alkaline phosphatase activity in extracts, in situ, and in circulation. Arch Biochem Biophys 221:477-488.[CrossRef][Medline] [Order article via Infotrieve]
  • Ferrendelli JA (1980). Phenytoin: cyclic nucleotide regulation in the brain. Adv Neurol 27:429-433.[Medline] [Order article via Infotrieve]
  • Goddard GA, Robinson JD (1976). Uptake and release of calcium by rat brain synaptosomes. Brain Res 110:331-350.[Medline] [Order article via Infotrieve]
  • Goldberg MA (1977). Phenytoin, phospholipids and calcium. Neurology (Minneap) 27:827-833.[Abstract/Free Full Text]
  • Hahn TJ, Avioli LV (1975). Anticonvulsant osteomalacia. Arch Intern Med 135:997-1000.[Abstract/Free Full Text]
  • Hahn TJ, Birge SJ, Scharp CR, Avioli LV (1972). Phenobarbital-induced alterations in vitamin D metabolism. J Clin Invest 51:741-748.[Medline] [Order article via Infotrieve]
  • Hahn TJ, Hendin BA, Scharp CR, Boisseau VC, Haddad JG (1975). Serum 25-hydroxycalciferol levels and bone mass in children on chronic anticonvulsant therapy. N Engl J Med 292:550-554.
  • Harris M., Jenkins MV, Wills MR (1974). Phenytoin inhibition of parathyroid hormone induced bone resorption in vitro. J Pharmacol ( Paris) 50:405-408.
  • Hassell TM, Hefti AF (1991). Drug-induced gingival overgrowth: old problem, new problem. Crit Rev Oral Biol Med 2:103-137.[Free Full Text]
  • Jenkins MV, Harris M., Wills MR (1974). The effect of phenytoin on parathyroid extract and 25-hydroxycholecalciferol- induced bone resorption: adenosine 3',5' cyclic monophosphate production. Calcif Tissue Res 16:163-167.[CrossRef][Medline] [Order article via Infotrieve]
  • Kattan KR (1970). Calvarial thickening after dilantin medication. Am J Roentgenol Radium Ther Nucl Med 110:102-106.[Medline] [Order article via Infotrieve]
  • Koch H.-U, Kraft D., Von Herrath D., Schaefer K. (1972). Influence of diphenylhydantoin and phenobarbital on intestinal calcium transport in the rat. Epilepsia 13:829-834.[CrossRef][Medline] [Order article via Infotrieve]
  • Kutt H., Solomon GE (1980). Phenytoin: relevant side effects. Adv Neurol 27:435-445.[Medline] [Order article via Infotrieve]
  • Lau K.-HW, Lee MY, Linkhart TA, Mohan S., Vermeiden J., Liu CC, et al. (1985). A mouse tumor-derived osteolytic factor stimulates bone resorption by a mechanism involving local prostaglandins production in bone. Biochim Biophys Acta 840:56-68.[Medline] [Order article via Infotrieve]
  • Lau K-HW, Nakade O., Taylor AK, Houchin K., Baylink DJ (1994). Low dose phenytoin is osteogenic for the human species in vitro and in vivo (abstract). J Bone Miner Res 9(Suppl 1): S152.
  • Lefebvre EB, Haining RG, Labbe RF (1972). Coarse facies, calvarial thickening and hyperphosphatasia associated with long-term anticonvulsant therapy. N Engl J Med 286:1301-1302.[Medline] [Order article via Infotrieve]
  • Levy RH ( 1980). Phenytoin: biopharmacology. Adv Neurol 27:315-321.[Medline] [Order article via Infotrieve]
  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951). Protein measurement with the folin phenol reagent. J Biol Chem 198:265-275.
  • Narayanan AS, Meyers DF, Page RC ( 1988). Regulation of collagen production in fibroblasts cultured from normal and phenytoin-induced hyperplastic human gingiva. J Periodont Res 23:118-121.[Medline] [Order article via Infotrieve]
  • Ohta T., Wergedal JE, Gruber HE, Baylink DJ, Lau K.-HW (1995). Low dose phenytoin is an osteogenic agent in the rat. Calcif Tissue Int 56:42-48[Medline] [Order article via Infotrieve]
  • Peterkofsky B. (1972). The effect of ascorbic acid on collagen polypeptide synthesis and proline hydroxylation during the growth of cultured fibroblasts. Arch Biochem Biophys 152:318-328.[CrossRef][Medline] [Order article via Infotrieve]
  • Pincus JH, Grove I., Marino BB, Glaser GH (1970). Studies on the mechanism of action of diphenylhydantoin. Arch Neurol 22:566-571.[Abstract/Free Full Text]
  • Reynolds EH (1975). Chronic antiepileptic toxicity: a review. Epilepsia 16:319-352.[CrossRef][Medline] [Order article via Infotrieve]
  • Richens A., Rowe DJF (1970). Disturbance of calcium metabolism by anticonvulsant drugs. Br Med J 4:73-76.[Abstract/Free Full Text]
  • Seymour RA, Smith DG, Turnbull DN (1985). The effects of phenytoin and sodium valproate on periodontal health of adult epileptic patients. J Clin Periodontol 12:413-419.[CrossRef][Medline] [Order article via Infotrieve]
  • Silver J., Neale G., Thompson GR (1974). Effect of phenobarbitone treatment on vitamin D metabolism in mammals. Clin Sci Molec Med 46:433-448.
  • Sklans S., Taylor RG, Shklar G. (1967). Effect of diphenylhydantoin sodium on healing of experimentally produced fractures in rabbit mandibles. J Oral Surg 25:310-319.[Medline] [Order article via Infotrieve]
  • Taylor AK, Linkhart SG, Mohan S., Baylink DJ (1988). Development of a new radioimmunoassay for human osteocalcin: evidence for a mid-molecule epitope. Metabolism 37:872-877.[CrossRef][Medline] [Order article via Infotrieve]
  • Valimaki MJ, Tiihonen M., Laitinen K., Tahtela R., Karkkainen M., Lamberg-Allardt C., et al. (1994). Bone mineral density measured by dual-energy x-ray absorptiometry and novel markers of bone formation and resorption in patients on antiepileptic drugs. J Bone Miner Res 9:631-637.[Medline] [Order article via Infotrieve]
  • Vernillo AT, Schwartz NB (1987). The effects of phenytoin (5,5-diphenylhydantoin) on human gingival fibroblasts in culture. J Periodont Res 22:307-312.[Medline] [Order article via Infotrieve]
  • Vemillo AT, Rifkin BR, Hauschka PV (1990a). Phenytoin affects osteocalcin secretion from osteoblastic rat osteosarcoma 17/2.8 cells in culture. Bone 11:309-312.
  • Vernillo AT, Ramamurthy NS, Lee HM, Rifkin BR (1990b). The effect of phenytoin on collagenase and gelatinase activities in UMR 106-01 rat osteoblastic osteosarcoma cells. Matrix 10:27-32.[Medline] [Order article via Infotrieve]
  • Wergedal JE, Baylink DJ (1984). Characterization of cells isolated and cultured from human bone. Proc Soc Exp Biol Med 176:60-69.[CrossRef][Medline] [Order article via Infotrieve]
  • Woodbury DM (1958). Effects of diphenylhydantoin on electrolytes and radio-sodium turnover in brain and other tissues of normal, hyponatremic and postictal rats. J Pharmacol Exp Ther 115:74-95.

Journal of Dental Research, Vol. 74, No. 1, 331-337 (1995)
DOI: 10.1177/00220345950740010801
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