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Article
Effects of ascorbic acid, calcitriol, and retinoic acid on the differentiation of preosteoblasts
Dr. Peter F. M. Choong 1 *, T. John Martin 2, Kong Wah Ng 2
1Department of Orthopaedics, St. Vincent's Institute of Medical Research, St. Vincent's Hospital, Melbourne, Australia
2Department of Medicine, St. Vincent's Institute of Medical Research, St. Vincent's Hospital, Melbourne, Australia
*Correspondence to Peter F. M. Choong, St. Vincent's Institute of Medical Research, St. Vincent's Hospital, 41 Victoria Parade, Melbourne, Victoria 3065,
Australia

ABSTRACT
The responses of the immortalized rat preosteoblast UMR-201-10B to ascorbic acid (AA), 1,25(OH)2D3 (calcitriol), and retinoic acid (RA) were examined.
UMR-201-10B cells have an undetectable basal alkaline phosphatase (ALP) activity that is induced after 24 h of treatment with 10-6 M RA (4.64 ± 0.06
mol/h/mg of protein). The addition of 10-8 M calcitriol resulted in a slight induction of ALP activity after 72 h (0.43 ± 0.07 mol/h/mg of protein). When
calcitriol was added to RA, however, over the same period ALP activity was enhanced significantly compared with treatment with RA alone (RA and
calcitriol, 12.29 ± 0.86 mol/h/mg of protein). Treatment with AA (50 g/ml) alone had no effect on ALP activity but increased RA-induced ALP activity to
6.78 ± 0.28 mol/h/mg of protein at 24 h. In contrast, AA inhibited calcitriol-induced ALP activity after 7 days of combined treatment with calcitriol
(calcitriol, 7.73 ± 0.16 mol/h/mg of protein; AA and calcitriol, 1.44 ± 0.06 mol/h/mg of protein). Individually, RA and calcitriol induced mRNA expression
for ALP, matrix-gla protein (MGP), and osteopontin (OP). The steady state level of pro- 1(I) collagen mRNA also was increased significantly by treatment
with RA and AA individually. The combination of RA and calcitriol had a synergistic effect on ALP, OP, and especially MGP mRNA expression but
significantly reduced the expression of pro- 1(I) collagen mRNA. AA enhanced the effect of RA on the expression of pro- 1(I) collagen, MGP, and ALP
mRNAs as well as the effect of calcitriol on OP and MGP. The addition of AA to RA resulted in a decrease in the steady state level of OP, whereas its
cotreatment with calcitriol caused a decrease in pro- 1(I) collagen and ALP mRNA. In conclusion, these studies identify RA, calcitriol, and AA as
regulators of differentiated osteoblast function.

Received: 7 February 1992; Accepted: 27 January 1993

DIGITAL OBJECT IDENTIFIER (DOI)

10.1002/jor.1100110505 About DOI

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