Department of Bioengineering, Clemson University, Clemson, SC.
Department of Orthopedics, Medical University of South Carolina (MUSC), Charleston, SC.
Spine (Phila Pa 1976). 2018 Jan 15;43(2):E60-E67. doi: 10.1097/BRS.0000000000002252.
In vitro measurements of the oxygen consumption rates (OCR) of human intervertebral disc (IVD) cells.
The aim of this study was to determine the differences in the OCR of nondegenerate and degenerate human annulus fibrosus (AF), nucleus pulposus (NP), and cartilage endplate (CEP) cells at different glucose concentrations.
The avascular nature of the IVD creates a delicate balance between rate of nutrient transport through the matrix and rate of disc cell consumption necessary to maintain tissue health. Previous studies have shown a dependence of OCR for animal (e.g., bovine and porcine) IVD cells on oxygen level and glucose concentration. However, the OCR of nondegenerate human IVD cells compared to degenerate human IVD cells at different glucose concentrations has not been investigated.
IVD cells were isolated from the AF, NP, and CEP regions of human cadaver spines and surgical samples. The changes in oxygen concentration were recorded when cells were sealed in a metabolic chamber. The OCR of cells was determined by curve fitting using the Michaelis-Menton equation.
Under identical cell culture conditions, the OCR of degenerate human IVD cells was three to five times greater than that of nondegenerate human IVD cells. The degenerate IVD cells cultured in low-glucose medium (1 mmol/L) exhibited the highest OCR compared to degenerate cells cultured at higher glucose levels (i.e., 5 mmol/L, 25 mmol/L), whereas no significant differences in OCR were found among the nondegenerate IVD cells for all glucose levels.
Considering the significantly higher OCR and unique response to glucose of degenerate human IVD cells, the degeneration of the IVD is associated with a cell phenotypic change related to OCR. The OCR of IVD cells reported in this study will be valuable for understanding human IVD cellular behavior and tissue nutrition in response to disc degeneration.
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体外测量人类椎间盘（IVD）细胞的耗氧量（OCR）。
本研究旨在确定不同葡萄糖浓度下非退变和退变人纤维环（AF）、髓核（NP）和软骨终板（CEP）细胞的 OCR 差异。
IVD 的无血管特性在营养物质通过基质的运输速度和维持组织健康所需的椎间盘细胞消耗速度之间创造了微妙的平衡。先前的研究表明，动物（如牛和猪）IVD 细胞的 OCR 依赖于氧水平和葡萄糖浓度。然而，尚未研究不同葡萄糖浓度下非退变与退变人 IVD 细胞的 OCR。
从人尸体脊柱和手术样本的 AF、NP 和 CEP 区域分离 IVD 细胞。当细胞在代谢室中密封时，记录氧浓度的变化。使用米氏方程进行曲线拟合来确定 OCR。
在相同的细胞培养条件下，退变人 IVD 细胞的 OCR 比非退变人 IVD 细胞高 3 到 5 倍。与在较高葡萄糖水平（即 5 mmol/L、25 mmol/L）下培养的退变 IVD 细胞相比，在低葡萄糖培养基（1 mmol/L）中培养的退变 IVD 细胞的 OCR 最高，而所有葡萄糖水平下的非退变 IVD 细胞的 OCR 均无显著差异。
考虑到退变人 IVD 细胞的 OCR 明显较高且对葡萄糖的反应独特，IVD 的退变与与 OCR 相关的细胞表型变化有关。本研究报告的 IVD 细胞的 OCR 将有助于理解人类 IVD 细胞行为和组织营养对椎间盘退变的反应。
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