Kaysinger K K, Ramp W K
Baxter Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina 28232, USA.
J Cell Biochem. 1998 Jan 1;68(1):83-9. doi: 10.1002/(sici)1097-4644(19980101)68:1<83::aid-jcb8>3.0.co;2-s.
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.
本研究调查了培养基pH值对培养的人成骨细胞活性的影响。从人松质骨外植体获得的成骨细胞生长至汇合后进行传代培养。第一代细胞在初始pH值从7.0调节至7.8的Hepes缓冲培养基中孵育。通过测量乳酸生成、碱性磷酸酶活性、脯氨酸羟化、DNA含量和胸苷掺入来评估成骨细胞功能。根据最终48小时孵育期开始和结束时记录的培养基pH值确定培养基pH值的变化。随着培养基pH值升高至7.6,胶原蛋白合成、碱性磷酸酶活性和胸苷掺入增加。DNA含量从pH 7.0增加至7.2,在pH 7.2至7.6保持稳定,然后从pH 7.6再次增加至7.8。培养基pH值在pH 7.0和7.8时变化最大,在pH 7.4和7.6时变化适中,在7.2时不变,表明pH值正朝着pH 7.2迁移。乳酸生成在pH 7.0时增加,但从7.2至7.8保持恒定。这些结果表明,在7.0 - 7.6的pH范围内,人成骨细胞的活性随pH值升高而增加,这种活性增加不需要糖酵解活性增加,并且pH 7.2可能是这些细胞的最佳pH值。