The effects of oxygen level and glucose concentration on the metabolism of porcine TMJ disc cells.
The effects of oxygen level and glucose concentration on the metabolism of porcine TMJ disc cells.
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DOI：
10.1016/j.joca.2015.05.021
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发表时间：
2015-10
影响因子：
7
通讯作者：
Yao H
中科院分区：
文献类型：
--
作者：
Cisewski SE;Zhang L;Kuo J;Wright GJ;Wu Y;Kern MJ;Yao H
目的：探讨氧浓度和葡萄糖浓度对颞下颌关节盘细胞活力、ATP生成和基质合成的影响。从6-8月龄的猪中分离TMJ椎间盘细胞，并以单层培养。细胞培养物分别用0、1.5、5或25 mM葡萄糖DMEM在1%、5%、10%或21%O2水平下预处理48小时。使用WST-1测定法测量细胞活力。使用荧光素-荧光素酶测定法测定ATP产生。胶原和蛋白多糖的合成分别通过测量[2，3- 3 H]脯氨酸和[35 S]硫酸盐掺入细胞来确定。无葡萄糖时，TMJ椎间盘细胞活力显著降低（P<0.0001）。在葡萄糖存在的情况下，降低的氧水平显著增加了存活率（P<0.0001），而葡萄糖浓度的降低显著降低了存活率（P<0.0001）。在葡萄糖存在的情况下，降低氧水平显著降低ATP产生（P<0.0001）和基质合成（P<0.0001）。降低葡萄糖浓度显著降低胶原合成（P<0.0001）。葡萄糖和氧之间的相互作用在细胞活力（P<0.0001）、ATP产生（P=0.00015）、胶原蛋白（P=0.0002）和蛋白聚糖合成（P<0.0001）方面是显著的。虽然葡萄糖和氧气都很重要，但葡萄糖是TMJ椎间盘细胞存活的限制性营养素。在低氧水平下，ATP、胶原蛋白和蛋白聚糖的产生受到严重抑制。这些结果表明，陡峭的营养梯度可能存在于颞下颌关节盘，它可能容易受到病理事件，阻碍营养供应。
To determine the combined effect of oxygen level and glucose concentration on cell viability, ATP production, and matrix synthesis of temporomandibular joint (TMJ) disc cells. TMJ disc cells were isolated from pigs aged 6-8 months and cultured in a monolayer. Cell cultures were preconditioned for 48 hours with 0, 1.5, 5, or 25mM glucose DMEM under 1%, 5%, 10%, or 21% O2 level, respectively. The cell viability was measured using the WST-1 assay. ATP production was determined using the Luciferin-Luciferase assay. Collagen and proteoglycan synthesis were determined by measuring the incorporation of [2, 3-3H]proline and [35S]sulfate into the cells, respectively. TMJ disc cell viability significantly decreased (P<0.0001) without glucose. With glucose present, decreased oxygen levels significantly increased viability (P<0.0001), while a decrease in glucose concentration significantly decreased viability (P<0.0001). With glucose present, decreasing oxygen levels significantly reduced ATP production (P<0.0001) and matrix synthesis (P<0.0001). A decreased glucose concentration significantly decreased collagen synthesis (P<0.0001). The interaction between glucose and oxygen was significant in regards to cell viability (P<0.0001), ATP production (P=0.00015), and collagen (P=0.0002) and proteoglycan synthesis (P<0.0001). Although both glucose and oxygen are important, glucose is the limiting nutrient for TMJ disc cell survival. At low oxygen levels, the production of ATP, collagen, and proteoglycan are severely inhibited. These results suggest that steeper nutrient gradients may exist in the TMJ disc and it may be vulnerable to pathological events that impede nutrient supply.