Extracellular pH modulates the activity of cultured human osteoblasts.
The effect of medium pH on the activity of cultured human osteoblasts was investigated in this study. Osteoblasts derived from explants of human trabecular bone were grown to confluence and subcultured. The first-pass cells were incubated in Hepes-buffered media at initial pHs adjusted from 7.0 to 7.8. Osteoblast function was evaluated by measuring lactate production, alkaline phosphatase activity, proline hydroxylation, DNA content, and thymidine incorporation. Changes in medium pH were determined from media pHs recorded at the beginning and end of the final 48 h incubation period. As medium pH increased through pH 7.6, collagen synthesis, alkaline phosphatase activity, and thymidine incorporation increased. DNA content increased from pH 7.0 to 7.2, plateaued from pH 7.2 to 7.6, and increased again from pH 7.6 to 7.8. The changes in the medium pH were greatest at pHs 7.0 and 7.8, modest at pHs 7.4 and 7.6, and did not change at 7.2, suggesting that the pHs are migrating towards pH 7.2. Lactate production increased at pH 7.0 but remained constant from 7.2 to 7.8. These results suggest that in the pH range from 7.0-7.6 the activity of human osteoblasts increases with increasing pH, that this increase in activity does not require an increase in glycolytic activity, and that pH 7.2 may be the optimal pH for these cells.
Kaysinger
K K
KK
Baxter Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina 28232, USA. kkaysinger@carolinas.org
Ramp
W K
WK
D000469:Alkaline Phosphatase / Q000378:metabolism; D002452:Cell Count; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D003470:Culture Media; D017077:Culture Media, Conditioned; D004247:DNA / Q000378:metabolism; D005110:Extracellular Space / Q000737:chemistry*; D006019:Glycolysis / Q000502:physiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019344:Lactic Acid / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D013997:Time Factors
D003470:Culture Media; D017077:Culture Media, Conditioned; D019344:Lactic Acid; D003094:Collagen; D004247:DNA; D000469:Alkaline Phosphatase