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Rotary Culture Enhances Pre-osteoblast Aggregation and Mineralization
S.R. Facer1,
R.S. Zaharias2,
M.E. Andracki3,
J. Lafoon2,
S.K. Hunter3 and
G.B. Schneider2,4,*
1 Department of Endodontics, University of Iowa, College of Dentistry;
2 Dows Institute of Dental Research, University of Iowa, College of Dentistry, Iowa City, IA 52242, USA;
3 Department of Maternal Fetal Medicine, University of Iowa Health Care and Clinics; and
4 Department of Prosthodontics, University of Iowa, College of Dentistry;
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Figure 1. Pre-osteoblast monolayer cell cultures on tissue culture plastic mineralized in 4 wks. Alizarin Red S staining for calcium was not evident in cells cultured in the absence (A) or presence (B) of β-glycerophosphate (β-GP) and ascorbate at 1 wk. At 4 wks, calcium deposition was greatly enhanced in the presence of β-GP and ascorbate (D), as compared with non-treated cultures (C). N = 3.
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Figure 2. At 1 wk, osteoblast cells cultured in 3D environments had aggregated. Gross and scanning electron microscopy revealed a mass of cells in untreated cultures (A,C) and microclusters within the aggregate in treated cultures (B,D). N = 3. Magnification, 300x. Bar = 50 µm.
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Figure 3. Enhanced mineralization was observed in cultures maintained in 3D environments for 1 wk. Uniform cell distribution and minimal calcium and phosphorus deposition were noted in non-treated cultures (A,C,E). When treated with β-GP and ascorbate, the cell distribution and enhanced mineralization pattern matched the pattern of microclusters within the aggregate (B,D,E). Magnification, 20x. Bar = 100 µm. Asterisks (*) indicate significant differences between conditions (N = 3) at P < 0.05.
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Figure 4. 3D pre-aggregated cells (A) or aggregates (B) when re-introduced into 2D tissue culture conditions had cellular outgrowth (B, arrow) mimicking that of normal tissue cultured cells. N = 3. Magnification, 10x. Bar = 10 µm.
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Journal of Dental Research, Vol. 84, No. 6,
542-547 (2005)
DOI: 10.1177/154405910508400611
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