Effects of strontium-doped calcium polyphosphate on behavior and angiogenic growth factors expression of co-cultured osteoblasts and endothelial cells

doi:10.3969/j.issn.2095-4344.2014.03.006

Abstract

Abstract:

BACKGROUND: Our previous studies have shown that strontium-doped calcium polyphosphate containing low-dose strontium appears to have a significant effect on angiogenesis-related behaviors of monocultured umbilical vein endothelial cells and osteoblasts.
OBJECTIVE: To investigate the effect of strontium-doped calcium polyphosphate on angiogenesis-related behaviors of umbilical vein endothelial cells and osteoblasts co-cultured, including cell adhesion, spreading, proliferation, as well as the protein secretion of vascular endothelial growth factor and basic fibroblast growth factor from co-culture system in vitro.
METHODS: Human umbilical vein endothelial cells and osteoblastic cells (MG63) were utilized in this study. Cells from passage 3 were used for preparation of the cell-scaffold constructs. After placed in 24-well plate at a ratio of 2:1, human umbilical vein endothelial cells and MG63 cells were seeded onto strontium-doped calcium polyphosphate, calcium polyphosphate and hydroxyapatite scaffolds and co-cultured for 7 days. The vascular endothelial growth factor and basic fibroblast growth factor protein levels were determined through a double ligand enzyme-linked immunosorbent assay. The colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide assay was performed to quantify the effect of scaffolds on cell proliferation.
RESULTS AND CONCLUSION: Compared with those on calcium polyphosphate and hydroxyapatite scaffolds, cells on strontium-doped calcium polyphosphate scaffolds attached and spread better with a significantly improved cell proliferation. More importantly, the vascular endothelial growth factor and basic fibroblast growth factor expressions were significantly higher in the strontium-doped calcium polyphosphate group than the other two groups (P < 0.05), indicating strontium-doped calcium polyphosphate can up-regulate levels of vascular endothelial growth factor and basic fibroblast growth factor proteins.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: biocompatible materials, calcium phosphates, strontium, endothelial cells, osteoblasts

CLC Number:

R318

Peng Hong, Gu Zhi-peng, Huang Cheng-cheng, Xu Yuan-ting, Yu Xi-xun. Effects of strontium-doped calcium polyphosphate on behavior and angiogenic growth factors expression of co-cultured osteoblasts and endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(3): 365-370.

Figures/Tables 3

2.3 共培养的细胞与支架材料复合培养的生长曲线图3A为共培养的成骨肉瘤细胞和内皮细胞与3种支架材料复合培养不同时间的细胞增殖情况。随着培养时间的延长，各组细胞增殖都呈上升趋势，峰值出现在第10天，到第14天，由于培养液中细胞分泌物的过多累积，细胞活性整体回落。在14 d的复合培养时间段内，掺锶聚磷酸钙组的细胞数量一直明显高于聚磷酸钙组和羟基磷灰石组，细胞增殖速率也一直明显高于聚磷酸钙组和羟基磷灰石组。另外，聚磷酸钙组细胞增殖速率也一直明显高于羟基磷灰石组。相比于聚磷酸钙和羟基磷灰石，在掺锶聚磷酸钙表面共培养细胞可更好地黏附和生长，可能是因为培养液中同时有释放出的锶元素能进一步刺激促进细胞的生长行为，使得掺锶聚磷酸钙能显著促进细胞的增殖。 2.4 不同支架材料上细胞血管内皮生长因子和碱性成纤维细胞生长因子的蛋白分泌量图3B显示了共培养的成骨细胞和内皮细胞与不同支架材料复合培养7 d后，细胞培养上清液中血管内皮生长因子和碱性成纤维细胞生长因子蛋白分泌量的检测情况。由图可见，掺锶聚磷酸钙组细胞分泌的血管内皮生长因子和碱性成纤维细胞生长因子明显高于聚磷酸钙组和羟基磷灰石组(P < 0.05)。另外，聚磷酸钙组细胞分泌的血管内皮生长因子和碱性成纤维细胞生长因子也要高于羟基磷灰石组(P < 0.05)。ELISA实验结果表明掺锶聚磷酸钙明显上调了血管内皮生长因子和碱性成纤维细胞生长因子的蛋白表达，原因可能是掺锶聚磷酸钙降解释放出锶元素，同时材料优良的三维结构和表面形貌为细胞生长提供了良好的微环境，有利于细胞功能的表达。"

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