Evidence for multiple bone resorption-stimulating factors produced by normal human keratinocytes in culture

R. M. Fried, E. F. Voelkel, R. H. Rice, L. Levine, A. H. Tashjian

Research output: Contribution to journalArticle

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Abstract

Conditioned medium from cultured normal human foreskin keratinocytes enhanced the release of calcium from neonatal mouse calvaria in organ culture. Unfractionated keratinocyte-conditioned medium (KCM) stimulated bone resorption in a dose-dependent manner, but it did not increase the concentration of prostaglandin E2 (PGE2) in the bone culture medium until a maximal dose of KCM for resorption was used. Furthermore, inhibitors of PGE2 synthesis, indomethacin, ibuprofen, and piroxicam, did not inhibit KCM-induced calcium release. High concentrations of KCM increased cAMP production by calvaria in the presence of isobutylmethylxanthine, but the increase was small compared with that produced by a dose of bovine PTH that caused a similar level of bone resorption. The bone resorption-stimulating activity of KCM was not lost after incubation at 56 C for 60 min, but it was lost after heating at 100 C for 10 min. Fractionation of KCM by gel filtration chromatography revealed two distinct peaks of bone resorption-stimulating activity. One peak, KCM(I), caused a significant increase in bone resorption at 2 μg protein/ml. KCM(I) did not increase medium PGE2, and inhibition of PGE2 synthesis in bone had no effect on KCM(I)-induced bone resorption. KCM(I) failed to increase cAMP production by human osteosarcoma SaOS-2 cells. Another peak, KCM(II), caused a dose-dependent increase in bone resorption, and a significant increase in medium calcium was noted at a 20-fold lower concentration (0.1 μg protein/ml) than with KCM(I). In contrast to KCM(I), the increase in bone resorption stimulated by KCM(II) was accompanied by a parallel increase in the production of PGE2, and inhibition of PGE2 synthesis completely inhibited the bone resorption-stimulating activity of KCM(II). KCM(II) also caused an increase in cAMP production by SaOS-2 cells. We conclude that KCM contains at least two distinct bone resorption-stimulating factors, one of which acts via a PG-mediated mechanism and the other by a PG-independent pathway.

Original languageEnglish (US)
Pages (from-to)2467-2475
Number of pages9
JournalEndocrinology
Volume122
Issue number6
StatePublished - 1988
Externally publishedYes

Fingerprint

Conditioned Culture Medium
Keratinocytes
Bone Resorption
Dinoprostone
bone resorption factor
Calcium
Skull
Foreskin
Piroxicam
Bone and Bones
Ibuprofen
Organ Culture Techniques
Osteosarcoma
Indomethacin
Heating

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Fried, R. M., Voelkel, E. F., Rice, R. H., Levine, L., & Tashjian, A. H. (1988). Evidence for multiple bone resorption-stimulating factors produced by normal human keratinocytes in culture. Endocrinology, 122(6), 2467-2475.

Evidence for multiple bone resorption-stimulating factors produced by normal human keratinocytes in culture. / Fried, R. M.; Voelkel, E. F.; Rice, R. H.; Levine, L.; Tashjian, A. H.

In: Endocrinology, Vol. 122, No. 6, 1988, p. 2467-2475.

Research output: Contribution to journalArticle

Fried, RM, Voelkel, EF, Rice, RH, Levine, L & Tashjian, AH 1988, 'Evidence for multiple bone resorption-stimulating factors produced by normal human keratinocytes in culture', Endocrinology, vol. 122, no. 6, pp. 2467-2475.
Fried RM, Voelkel EF, Rice RH, Levine L, Tashjian AH. Evidence for multiple bone resorption-stimulating factors produced by normal human keratinocytes in culture. Endocrinology. 1988;122(6):2467-2475.
Fried, R. M. ; Voelkel, E. F. ; Rice, R. H. ; Levine, L. ; Tashjian, A. H. / Evidence for multiple bone resorption-stimulating factors produced by normal human keratinocytes in culture. In: Endocrinology. 1988 ; Vol. 122, No. 6. pp. 2467-2475.
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abstract = "Conditioned medium from cultured normal human foreskin keratinocytes enhanced the release of calcium from neonatal mouse calvaria in organ culture. Unfractionated keratinocyte-conditioned medium (KCM) stimulated bone resorption in a dose-dependent manner, but it did not increase the concentration of prostaglandin E2 (PGE2) in the bone culture medium until a maximal dose of KCM for resorption was used. Furthermore, inhibitors of PGE2 synthesis, indomethacin, ibuprofen, and piroxicam, did not inhibit KCM-induced calcium release. High concentrations of KCM increased cAMP production by calvaria in the presence of isobutylmethylxanthine, but the increase was small compared with that produced by a dose of bovine PTH that caused a similar level of bone resorption. The bone resorption-stimulating activity of KCM was not lost after incubation at 56 C for 60 min, but it was lost after heating at 100 C for 10 min. Fractionation of KCM by gel filtration chromatography revealed two distinct peaks of bone resorption-stimulating activity. One peak, KCM(I), caused a significant increase in bone resorption at 2 μg protein/ml. KCM(I) did not increase medium PGE2, and inhibition of PGE2 synthesis in bone had no effect on KCM(I)-induced bone resorption. KCM(I) failed to increase cAMP production by human osteosarcoma SaOS-2 cells. Another peak, KCM(II), caused a dose-dependent increase in bone resorption, and a significant increase in medium calcium was noted at a 20-fold lower concentration (0.1 μg protein/ml) than with KCM(I). In contrast to KCM(I), the increase in bone resorption stimulated by KCM(II) was accompanied by a parallel increase in the production of PGE2, and inhibition of PGE2 synthesis completely inhibited the bone resorption-stimulating activity of KCM(II). KCM(II) also caused an increase in cAMP production by SaOS-2 cells. We conclude that KCM contains at least two distinct bone resorption-stimulating factors, one of which acts via a PG-mediated mechanism and the other by a PG-independent pathway.",
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