Effect of Varied Ionic Calcium on Human Adipose-Derived Stem Cell Mineralization
Effect of Varied Ionic Calcium on Human Adipose-Derived Stem Cell Mineralization
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DOI：
10.1089/ten.tea.2009.0691
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发表时间：
2010-06-01
影响因子：
4.1
通讯作者：
Loboa, Elizabeth G.
中科院分区：
文献类型：
--
作者：
McCullen, Seth D.;Zhan, Jackie;Loboa, Elizabeth G.
人脂肪干细胞（hASC）是一种相对丰富且易于获取的干细胞来源，具有多向分化能力，在骨组织工程应用中具有巨大潜力。骨组织工程的成功与模拟天然骨中存在的天然矿物质的矿化基质的生产密切相关。在这项研究中，我们研究了 1.8（细胞培养基中的正常浓度）、8 和 16mM 离子钙水平对接种在二维单层和三维电纺支架中并在完全生长培养基 (CGM) 或成骨分化培养基 (ODM) 中培养的 hASC 的影响。确定了补钙对 hASC 活力、增殖和矿物质沉积的影响。 hASC 在所有实验治疗中均保持活力。添加8mM Ca2+ CGM 后hASC 增殖增加，但16mM Ca2+ CGM 处理后hASC 增殖减少。使用四种技术对 hASC 沉积的材料进行分析：(1) 茜素红 S 组织学染色，(2) 钙定量，(3) 傅里叶变换红外光谱，以及 (4) 广角 X 射线衍射。在生长和成骨培养基条件下，通过增加细胞外Ca2+，矿物质沉积显着增强。最大的矿物质沉积发生在 ODM 8mM Ca2+ 处理组。傅里叶变换红外光谱分析表明，8mM Ca2+ 的钙浓度升高显着增加了 ODM 的 PO4 量和 PO4 与蛋白质的比率。 X射线衍射表明，CGM和ODM中Ca2+含量升高所产生的矿物都具有羟基磷灰石的晶体结构特征。离子钙应被认为是 hASC 矿化的有效调节剂，并且可以作为一种潜在的治疗方法，在植入前诱导 hASC 接种的骨组织工程支架迅速骨化。
Human adipose-derived stem cells (hASCs) are a relatively abundant and accessible stem cell source with multilineage differentiation capability and have great potential for bone tissue engineering applications. The success of bone tissue engineering is intimately linked with the production of a mineralized matrix that mimics the natural mineral present within native bone. In this study, we examined the effects of ionic calcium levels of 1.8 (normal concentration in cell culture medium), 8, and 16mM on hASCs seeded in both two-dimensional monolayer and three-dimensional electrospun scaffolds and cultured in either complete growth medium (CGM) or osteogenic differentiation medium (ODM). The impact of calcium supplementation on hASC viability, proliferation, and mineral deposition was determined. hASCs remained viable for all experimental treatments. hASC proliferation increased with the addition of 8mM Ca2+ CGM, but decreased for the 16mM Ca2+ CGM treatment. Materials deposited by hASCs were analyzed using four techniques: (1) histological staining with Alizarin Red S, (2) calcium quantification, (3) Fourier transform infrared spectroscopy, and (4) wide-angle X-ray diffraction. Mineral deposition was significantly enhanced under both growth and osteogenic medium conditions by increasing extracellular Ca2+. The greatest mineral deposition occurred in the ODM 8mM Ca2+ treatment group. Fourier transform infrared spectroscopy analysis indicated that elevated calcium concentrations of 8mM Ca2+ significantly increased both PO4 amount and PO4 to protein ratio for ODM. X-ray diffraction indicated that mineral produced with elevated Ca2+ in both CGM and ODM had a crystalline structure characteristic of hydroxyapatite. Ionic calcium should be considered a potent regulator in hASC mineralization and could serve as a potential treatment for inducing prompt ossification of hASC-seeded scaffolds for bone tissue engineering prior to implantation.