Enhanced integrin-mediated human osteoblastic adhesion to porous amorphous calcium phosphate/poly(L-lactic acid) composite
Enhanced integrin-mediated human osteoblastic adhesion to porous amorphous calcium phosphate/poly(L-lactic acid) composite
复制标题
DOI：
10.3760/cma.j.issn.0366-6999.20140691
复制
发表时间：
2014-10-05
影响因子：
6.1
通讯作者：
He Rongxin
中科院分区：
文献类型：
--
作者：
Huang Xin;Qi Yiying;He Rongxin
背景 成骨细胞对材料的初始粘附是细胞-材料相互作用第一阶段的特征，并影响导致新骨形成的所有事件。在之前的工作中，我们开发了一种新型无定形磷酸钙（ACP）/聚（L-乳酸）（PLLA）材料，该材料表现出其微观结构的形态变化。本研究的目的是研究该材料与成骨细胞之间的初始相互作用。研究细胞附着和相应的信号转导途径。方法将多孔ACP/PLLA复合材料和PLLA支架（作为对照）在含有磷酸盐缓冲盐水（PBS）的胎牛血清（FBS）中孵育，并测定蛋白质吸附。将成骨细胞 MG63 细胞接种在材料上并培养 1、4、8 或 24 小时。使用MTS方法评估细胞附着。使用扫描电子显微镜（SEM）检查细胞形态。使用实时逆转录聚合酶链式反应（RT-PCR）测定编码整合素亚基α1、α5、αv、β1、粘着斑激酶（FAK）和3-磷酸甘油醛脱氢酶（GAPDH）的基因的表达水平。结果与纯材料相比，ACP/PLLA材料在1小时时的蛋白质吸附显着增加6.4倍，在24小时时显着增加2.4倍。 PLLA支架。成骨细胞在 ACP/PLLA 上的附着明显高于在 PLLA 支架上的附着。 SEM观察显示ACP/PLLA上细胞呈多边形展开形状，丝状伪足粘附于支架表面。相比之下，PLLA 支架上的细胞呈现球形或多边形形态。此外，实时RT-PCR显示编码整合素亚基α1、αv、β1和FAK的基因在ACP/PLLA复合材料上有较高水平的表达。结论ACP/PLLA复合材料促进了蛋白质吸附和成骨细胞粘附。增强的细胞粘附可能是通过整合素亚基α1、αv和β1的结合介导的，并且随后可以通过FAK信号转导途径进行调节。
Background The initial osteoblastic adhesion to materials characterizes the first phase of cell-material interactions and influences all the events leading to the formation of new bone. In a previous work, we developed a novel amorphous calcium phosphate (ACP)/poly(L-lactic acid) (PLLA) material that demonstrated morphologic variations in its microstructure. The aim of this study was to investigate the initial interaction between this material and osteoblastic cells. Cellular attachment and the corresponding signal transduction pathways were investigated.Methods A porous ACP/PLLA composite and PLLA scaffold (as a control) were incubated in fetal bovine serum (FBS) containing phosphate-buffered saline (PBS), and the protein adsorption was determined. Osteoblastic MG63 cells were seeded on the materials and cultured for 1, 4, 8, or 24 hours. Cell attachment was evaluated using the MTS method. Cell morphology was examined using scanning electron microscopy (SEM). The expression levels of the genes encoding integrin subunits alpha 1, alpha 5, alpha v, beta 1, focal adhesion kinase (FAK), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined using real-time reverse transcription polymerase chain reaction (RT-PCR).Results The ACP/PLLA material significantly increased the protein adsorption by 6.4-fold at 1 hour and 2.4-fold at 24 hours, compared with the pure PLLA scaffold. The attachment of osteoblastic cells to the ACP/PLLA was significantly higher than that on the PLLA scaffold. The SEM observation revealed a polygonal spread shape of cells on the ACP/PLLA, with the filopodia adhered to the scaffold surface. In contrast, the cells on the PLLA scaffold exhibited a spherical or polygonal morphology. Additionally, real-time RT-PCR showed that the genes encoding the integrin subunits alpha 1, alpha v, beta 1, and FAK were expressed at higher levels on the ACP/PLLA composite.Conclusions The ACP/PLLA composite promoted protein adsorption and osteoblastic adhesion. The enhanced cell adhesion may be mediated by the binding of integrin subunits alpha 1, alpha v, and beta 1, and subsequently may be regulated through the FAK signal transduction pathways.