Is 1, 25-dihydroxyvitamin D3 an ideal substitute for dexamethasone for inducing osteogenic differentiation of human adipose tissue-derived stromal cells in vitro?
BACKGROUND
Human adipose tissue-derived stromal cells (hADSCs) can be induced to differentiate along an osteoblastic lineage under stimulation of dexamethasone (DEX). Recent studies, however, have questioned the efficacy of glucocorticoids such as DEX in mediating the osteogenesis process of skeletal progenitor cells and processed lipoaspirate cells. Is it possible to find a substitute for DEX? Therefore, this study was designed to investigate osteogenic capacity and regulating mechanisms for osteoblastic differentiation of hADSCs by comparing osteogenic media (OM) containing either 1, 25-dihydroxyvitamin D(3) (VD) or DEX and determine if VD was an ideal substitute for DEX as an induction agent for the osteogenesis of hADSCs.


METHODS
Osteogenic differentiation of hADSCs was induced by osteogenic medium (OM) containing either 10 nmol/L VD or 100 nmol/L DEX. Differentiation of hADSCs into osteoblastic lineage was identified by alkaline phosphatase (ALP) staining, von Kossa staining, and reverse transcription-polymerase chain reaction assays for mRNA expression of osteogenesis-related genes such as type I collagen (COL I), bone sialoprotein (BSP), osteocalcin (OC), bone morphogenetic protein (BMP)-2, BMP-4, BMP-6, BMP-7, runt-related transcription factor 2/core binding factor alpha1 (Runx2/Cbfa1), osterix (Osx), and LIM mineralization protein-1 (LMP-1).


RESULTS
von Kossa staining revealed that the differentiated cells induced by both VD and DEX were mineralized in vitro. They also expressed osteoblast-related markers, such as ALP, COL I, BSP, and OC. Runx2/Cbfa1, Osx, BMP-6, and LMP-1 were upregulated during VD and DEX-induced hADSC osteoblastic differentiation, but BMP-4, BMP-7 were not. BMP-2 was only expressed in VD-induced differentiated cells.


CONCLUSIONS
VD or DEX-induced hADSCs differentiate toward the osteoblastic lineage in vitro. Runx2/Cbfa1, Osx, BMP-2, BMP-6, and LMP-1 are involved in regulating osteoblastic differentiation of hADSCs, but BMP-4, BMP-7 are not. VD, but not DEX, induces expression of BMP-2 during osteogenic induction of hADSCs. VD is an ideal substitute for DEX for osteogenic induction of hADSCs.
Zhou
Yong-sheng
YS
Department of Prosthodontics, School and Hospital of Stomatology, Peking University, Beijing 100081, China. kqzhouysh@bjmu.edu.cn
Liu
Yun-song
YS
Tan
Jian-guo
JG
D048868:Adaptor Proteins, Signal Transducing; D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D002117:Calcitriol / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics; D003598:Cytoskeletal Proteins; D003907:Dexamethasone / Q000494:pharmacology*; D006801:Humans; D058574:Integrin-Binding Sialoprotein; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D060588:LIM Domain Proteins; D008875:Middle Aged; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D015675:Osteocalcin / Q000235:genetics; D010012:Osteogenesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000032:analysis; D012795:Sialoglycoproteins / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects
D048868:Adaptor Proteins, Signal Transducing; D024042:Collagen Type I; D003598:Cytoskeletal Proteins; C546871:IBSP protein, human; D058574:Integrin-Binding Sialoprotein; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; C484380:PDLIM7 protein, human; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D015675:Osteocalcin; D003907:Dexamethasone; D002117:Calcitriol