Absence of Glutamine Supplementation Prevents Differentiation of Murine Calvarial Osteoblasts to a Mineralizing Phenotype
Absence of Glutamine Supplementation Prevents Differentiation of Murine Calvarial Osteoblasts to a Mineralizing Phenotype
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DOI：
10.1007/s00223-011-9537-6
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发表时间：
2011-12-01
影响因子：
4.2
通讯作者：
Crockett, Julie C.
中科院分区：
文献类型：
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作者：
Brown, Philip M.;Hutchison, James D.;Crockett, Julie C.
体外成骨细胞在转移到富含β-甘油磷酸盐和抗坏血酸的培养基后从增殖分化为矿化表型。在这个分化过程的不同阶段，细胞的营养需求是未知的。在其他细胞类型中，手术期间的营养补充可以改善患者恢复速度和预后方面的结果。因此，有可能在骨折修复或骨移植部位补充关键的成骨细胞营养因子，以潜在地加速愈合。在这项研究中，我们调查所需的共同细胞营养素的增殖和矿化阶段的成骨细胞分化。含有5.5 mM葡萄糖的培养基足以实现原代颅骨成骨细胞和人成骨细胞系的最大增殖，并具有额外补充谷氨酰胺的一些额外益处。然而，当细胞被刺激矿化时，葡萄糖不足以支持它们的能量需求。只有当细胞补充葡萄糖和谷氨酰胺时，在培养物中观察到高水平的骨钙素表达和矿化结节，这表明这将是一种有用的组合，可用于评估原代人成骨细胞培养物，以确定其是否在骨折手术、骨移植和非骨水泥型关节成形术植入物固定过程中具有有益作用。
Osteoblasts in vitro differentiate from a proliferating to a mineralizing phenotype upon transfer to a medium rich in beta-glycerophosphate and ascorbic acid. The nutritional requirements of the cells at different stages of this differentiation process are not known. In other cell types, nutritional supplementation during surgery can improve the outcome in terms of speed of patient recovery and prognosis. There is therefore the potential for supplementation at the site of fracture repair or bone grafting with critical osteoblast nutritional factors to potentially accelerate healing. In this study we investigate which common cell nutrients are required for the proliferating and mineralizing stages of osteoblast differentiation. Medium containing 5.5 mM glucose was sufficient to achieve maximal proliferation of primary calvarial osteoblasts and human osteoblast cell lines, with some added benefit of additional glutamine supplementation. However, when cells were stimulated to mineralize, glucose was insufficient to support their energetic requirements. Only when cells were supplemented with glucose together with glutamine were high levels of osteocalcin expression observed together with mineralized nodules in culture, suggesting that this would be a useful combination to assess in cultures of primary human osteoblasts to determine whether it may have beneficial effects during fracture surgery, bone grafting, and fixation of uncemented arthroplasty implants.