Biochemical and Biophysical Research Communications
Physical stress by magnetic force accelerates differentiation of human osteoblasts☆
Section snippets
Materials and methods
Cell culture. A normal human osteoblast (NHOst) cell system was purchased from BioWhittaker (Walkersville, MD). The cells were seeded in 60-mm culture dishes (Nalge Nunc International, Roskilde, Denmark). The cell concentration was 2 × 106 cells/ml and maintained in an osteoblast basal medium (OBM, CC-3208: BioWhittaker, Walkersville, MD) supplemented with 10% fetal bovine serum (FBS: CC-4102), 200 nM ascorbic acid (CC-4398), and 200 nM gentamicin/amphotericin-B (CC-4381).
An osteoblast
Uptake of trypan blue and BrdU
There was no difference in the percentages of viable cells (trypan blue-negative cells) or in the percentages of BrdU-uptake cells among group C, group MP, group MF, and group MP–MF 0.01, 0.03, 0.05 T (Fig. 1).
Morphology of cultured osteoblasts in the magnetic force
Differentiation of human osteoblasts in group C was compared with that in group MP, group MF, and group MP–MF (0.01, 0.03, and 0.05 T). In the group C the osteoblasts were thin spindle-shaped. The cells are oriented randomly on day 1 (Fig. 2A). They began to form colonies by day 3. The
Discussion
We originated a physical stimulation method, in which magnetic microparticles were introduced into cultured cells and the culture was placed in a magnetic field [5]. This physical stimulation accelerated differentiation of myoblasts [5], as well as osteoblasts shown in the present study in which the appearance of AlPase activity, formation of bone nodules, and calcification were seen earlier. In the present study we also found that this osteoblast differentiation acceleration is mainly
Acknowledgements
We are grateful to Mr. Toshiaki Mouri of Himeji Denshi Co. for providing technical information. We also thank Mrs. N.J. Johnson for comments on the manuscript. This study was supported by the “Ground Research Announcement for Space Utilization” promoted by the National Space Development Agency of Japan and the Japan Space Forum (to L.Y.), “Space Utilization Research” from ISAS (to L.Y.), and by Grants-in-Aid for scientific research (C), No. 10838021 and No. 12832032 (to L.Y.), from the Ministry
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Abbreviations: MAPK, mitogen-activated protein kinase; ERK, extracellular signal-regulated kinases; SAPK/JNK, stress-activated protein kinase/jun N-terminal kinase; MAPKK, mitogen-activated protein kinase kinase; AlPase, alkaline phosphatase; Cbfa 1, core-binding factor A1.