Construction of vascularized tissue-engineered bone with a double-cell sheet complex.
Construction of vascularized tissue-engineered bone with a double-cell sheet complex.
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具有双细胞片复合物的血管化组织工程骨的构建。
DOI：
10.1016/j.actbio.2018.07.024
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发表时间：
2018
影响因子：
9.7
通讯作者：
Deng Yan
中科院分区：
文献类型：
--
作者：
Zhang Hualin;Zhou Yueli;Zhang Wen;Wang Kairong;Xu Lihua;Ma Hairong;Deng Yan
本研究开发了一种由成骨细胞片和血管内皮细胞片组成的具有成骨和血管形成潜能的双细胞片（DCS）复合物。诱导培养兔脂肪间充质干细胞，获得成骨和血管内皮细胞片。成骨细胞片呈茜素红、von Kossa和碱性磷酸酶（ALP）染色阳性。血管内皮细胞片层中可见W-P小体，CD 31表达阳性，在Matrigel基质中自发形成血管网状结构。4组DCS、DCS-珊瑚羟基磷灰石（CHA）复合物及CHA支架组裸鼠皮下移植结果显示，12周时各组均有胶原纤维矿化和血管化，但各组间矿化和血管化程度存在差异。B组的模式包括内皮细胞片被成骨细胞片覆盖，表现出最好的结果。此外，DCS-CHA复合物的矿化程度比同类DCS复合物和CHA支架更成熟，毛细血管数比同类DCS复合物和CHA支架更大。因此，CHA支架增强了DCS复合物的成骨和血管形成潜力。同时，DCS复合物也促进了CHA支架的成骨和血管形成潜力。本研究将为构建血管化组织工程骨治疗骨缺损提供基础。意义本研究开发了一种由成骨细胞片和血管内皮细胞片组成的具有成骨和血管形成潜能的双细胞片复合体。诱导培养兔脂肪间充质干细胞，获得成骨细胞和血管内皮细胞。DCS复合物和DCS-CHA复合物在体内表现出成骨和血管形成潜力。CHA可增强DCS复合物体内成骨和血管形成能力。同时，DCS复合物也促进了CHA支架的成骨和血管形成潜力。B组的DCS复合物和DCS-CHA复合物具有最好的成骨和血管形成能力。
A double-cell sheet (DCS) complex composed of an osteogenic cell sheet and a vascular endothelial cell sheet with osteogenesis and blood vessel formation potential was developed in this study. The osteogenic and vascular endothelial cell sheets were obtained after induced culture of rabbit adipose-derived mesenchymal stem cells. The osteogenic cell sheet showed positive alizarin red, von Kossa, and alkaline phosphatase (ALP) staining. The vascular endothelial cell sheet exhibited visible W-P bodies in the cells, the expression of CD31 was positive, and a vascular mesh structure was spontaneously formed in a Matrigel matrix. The subcutaneous transplantation results for four groups of DCS and DCS-coral hydroxyapatite (CHA) complexes, and the CHA scaffold group in nude mice revealed mineralization of collagen fibers and vascularization in each group at 12 weeks, but the degrees of mineralization and vascularization showed differences among groups. The pattern involving endothelial cell sheets covered with osteogenic cell sheets, group B, exhibited the best results. In addition, the degree of mineralization of the DCS-CHA complexes was more mature than those of the same group of DCS complexes and the CHA scaffold, and the capillary number was greater than those of the same group of DCS complexes and the CHA scaffold. Therefore, the CHA scaffold strengthened the osteogenesis and blood vessel formation potential of the DCS complexes. Meanwhile, the DCS complexes also promoted the osteogenesis and blood vessel formation potential of the CHA scaffold. This study will provide a basis for building vascularized tissue-engineered bone for bone defect therapy.Statement of SignificanceThis study developed a double-cell sheet (DCS) complex composed of an osteogenic cell sheet and a vascular endothelial cell sheet with osteogenesis and blood vessel formation potential. Osteogenic and vascular endothelial cell sheets were obtained after induced culture of rabbit adipose-derived mesenchymal stem cells. The DCS complex and DCS-CHA complex exhibited osteogenic and blood vessel formation potential in vivo. CHA enhanced the osteogenesis and blood vessel formation abilities of the DCS complexes in vivo. Meanwhile, the DCS complexes also promoted the osteogenesis and blood vessel formation potential of the CHA scaffold. Group B of the DCS complexes and DCS-CHA complexes exhibited the best osteogenesis and blood vessel formation abilities.