关节软骨细胞三维复合体立体空间氧分压的测量  

In vitro measurement of local oxygen partial pressure in three-dimensional chondrocyte-agarose constructs

作　　者：李薇[1] 金丹[2] 倪国新[2] 陈滨[2] 胡振富[1] 

机构地区：[1]南方医科大学南方医院整形外科,广州510515 [2]南方医科大学南方医院创作骨科,广州510515

出　　处：《中华创伤骨科杂志》2010年第12期1169-1172,共4页Chinese Journal of Orthopaedic Trauma

基　　金：广东省自然科学基金（7300297） 国家重点基础研究发展（973）计划子课题（2009CB930003）

摘　　要：目的 探讨关节软骨细胞-琼脂糖复合体三维培养模型内最利于各部分细胞生长的氧气和葡萄糖条件,为体外工程化软骨组织的构建提供合适的能量代谢环境. 方法根据极谱电极法原则,自行制作并应用针形氧电极测定1×107、2×107、4×107个/mL 3种不同细胞密度牛关节软骨细胞-琼脂糖体外三维复合体模型内立体空间1～5 mm各个深度位置的氧分压值,每种细胞密度模型6个,重复测量3次,分析立体空间氧分压的分布情况.在相同细胞密度模型中按是否加入葡萄糖分为无糖和低糖两组,测定培养液内葡萄糖浓度对模型内立体空间氧分压的影响. 结果在3种细胞密度的关节软骨细胞-琼脂糖复合体体外三维培养模型1～3 mm深度位置范围内,氧分压梯度变化最明显,氧分压值的差值高达50 mm Hg（1mm Hg=0.13 kPa）.软骨细胞密度为2×107个/mL和4×107个/mL的模型中,氧分压梯度存在,但没有1×107个/mL模型中氧分压梯度变化明显.在相同细胞密度的情况下,低糖组氧分压高于无糖组. 结论关节软骨细胞-琼脂糖复合体模型内软骨细胞的氧耗量取决于培养基中的细胞密度和葡萄糖浓度环境2个方面,最佳细胞密度在1×107～2×107个/mL之间.当氧气和葡萄糖同时存在的条件下,复合体内的软骨细胞体现出消耗葡萄糖的倾向性.Objective To investigate spatial distribution of the oxygen partial pressure （PO2） within different levels of chondrocyte-agarose constructs with various articular chondrocyte densities to find out the optimal best oxygen and glucose conditions for cell growth. Methods The local levels of PO2 and the spatial gradient of PO2 within articular chondrocyte-agarose constructs with different cell densities （1×107,2 × 107, 4 × 107 cells/mL） were measured at different depths （1 to 5 mm） . The measurements were conducted using a self-developed oxygen-sensitive electrode in 6 samples of each cell density and each measurement was repeated twice. The effects of different concentrations of glucose on the PO2 spatial distribution within the articular chondrocyte-seeded constructs with various chondrocyte densities were determined. Results The PO2 gradients were particularly evident in the top layers （1 to 3 mm） of the constructs for each of the cell densities and the difference reached up to 50 mmHg, while no obvious difference was observed in the constructs between the densities of 2 × 107 cells/mL and 4 × 107 cells/mL. The PO2 in the low glucose group was higher than in the glucose-free one at the same cell density. Conclusions Oxygen depletion is dependent on both cell density and glucose concentration in the incubating medium. The optimal cell seeding density may range from 1 × 107 to 2 × 107 cells/mL. Chondrocytes within articular chondrocyte-agarose constructs have a preference for glucose metabolism when oxygen and glucose supplies are equally abundant.

关 键 词：软骨细胞 细胞骨架 葡萄糖 氧分压 

分 类 号：R686[医药卫生—骨科学]