Three-dimensional culture environments enhance osteoblast differentiation.
OBJECTIVE
In previous work from our laboratory, we demonstrated that the three-dimensional (3D) cell cultures developed in simulated microgravity environments enhanced osseous-like aggregate formation and accelerated preosteoblast cell differentiation. Thus, as described here, we hypothesize that aggregate formation and mineralization would occur with fewer than 10 x 10(6) cells as previously described.


METHODS
Human preosteoblastic cells were cultured at different concentrations in a rotary wall vessel to simulate microgravity for 7 days. Aggregate size was assessed, and mineralization and collagen expression detected using Von Kossa and Masson Trichrome staining. Scanning electron microscopy was used for structural and elemental analysis. Immunohistochemistry was used to detect expression of the osteogenic markers BSPII and osteopontin (OP).


RESULTS
Size and calcium expression were dependent upon cultured cell number (p < 0.01). Calcium and collagen expression were detected throughout the aggregate, but organization was independent of cell number. Aggregates had similar microscopic structural patterns demonstrating organized development. Presence of BSPII and OP showed that the aggregates share common differentiation proteins with in vivo bone formation.


CONCLUSIONS
These results may lead to novel bone engineering techniques associated with dental treatment.
University of Iowa College of Dentistry, Iowa City, IA 52242-1001, USA.
Jessica
J
Boehrs
Rebecca S
RS
Zaharias
John
J
Laffoon
Y Joon
YJ
Ko
Galen B
GB
Schneider
D002113:Calcification, Physiologic; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D004577:Electron Probe Microanalysis; D006801:Humans; D007150:Immunohistochemistry; D058574:Integrin-Binding Sialoprotein; D059630:Mesenchymal Stem Cells / Q000166:cytology; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D053495:Osteopontin / Q000096:biosynthesis; D021362:Palate, Hard / Q000196:embryology; D012795:Sialoglycoproteins / Q000096:biosynthesis; D014893:Weightlessness
C546871:IBSP protein, human; D058574:Integrin-Binding Sialoprotein; D012795:Sialoglycoproteins; D053495:Osteopontin; D003094:Collagen