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Rotary Culture Enhances Pre-osteoblast Aggregation and Mineralization

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;

View larger version (91K): [in a new window]   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 ß-glycerophosphate (ß-GP) and ascorbate at 1 wk. At 4 wks, calcium deposition was greatly enhanced in the presence of ß-GP and ascorbate (D), as compared with non-treated cultures (C). N = 3.

View larger version (126K): [in a new window]   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.

View larger version (98K): [in a new window]   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 ß-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.

View larger version (67K): [in a new window]   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.