title: Effect of different cell sheet ECM microenvironment on the formation of vascular network.
abstract: The repair and reconstruction of large bone defects remains as a significant clinical challenge mainly due to the insufficient vascularization. The prefabrication of vascular network based on cell sheet technique brings a promising potential for sufficient vascularization due to rich extracellular matrix (ECM) of cell sheets. However, the effect of different cell sheet ECM micro-environment on the formation of a vascular network has not been well understood. Here our goal is to study the effect of different cell sheets on the formation of a vascular network. First we cultured human bone marrow mesenchymal stem cells (hBMSCs) under two culture conditions to obtain osteogenic differentiated cell sheet (ODCS) and undifferentiated cell sheet (UDCS), respectively. Then the human umbilical vein endothelial cells (HUVECs) were seeded onto the surface of the two sheets at different seeding densities to fabricate pre-vascularized cell sheets. Our results indicated that the two sheets facilitated the alignment of HUVECs and promoted the formation of vascular networks. Quantitative analysis showed that the number of networks in ODCS was higher than that in the UDCS. The ECM of the two sheets was remodeled and rearranged during the tubulogenesis process. Furthermore, results showed that the optimal seeding density of HUVECs was 5×10(4)cell/cm(2). In summary, these results suggest that the vascularized ODCS has a promising potential to construct pre-vascularized tissue for bone repair.
authors: School of Stomatology, Lanzhou University, Lanzhou, Gansu, 730000, China; Hospital of Stomatology, General Hospital of Ningxia Medical University, Yingchuan, Ningxia, 750004, China.
authors: Jia
authors: J
authors: Chen
authors: School of Stomatology, Lanzhou University, Lanzhou, Gansu, 730000, China.
authors: Dan
authors: D
authors: Zhang
authors: School of Stomatology, Lanzhou University, Lanzhou, Gansu, 730000, China.
authors: Qin
authors: Q
authors: Li
authors: Department of Stomatology, The First people's Hospital of Jiayuguan, Jiayuguan, Gansu, 735100, China.
authors: Dan
authors: D
authors: Yang
authors: School of Stomatology, Lanzhou University, Lanzhou, Gansu, 730000, China.
authors: Zengjie
authors: Z
authors: Fan
authors: Department of Oral and Maxillofacial Surgery, Lanzhou General Hospital, Lanzhou Command of PLA, Gansu 730050, China.
authors: Dongyang
authors: D
authors: Ma
authors: School of Stomatology, Lanzhou University, Lanzhou, Gansu, 730000, China. Electronic address: renlil@lzu.edu.cn.
authors: Liling
authors: L
authors: Ren
mesh_terms: D001808:Blood Vessels / Q000166:cytology / Q000254:growth & development*; D001846:Bone Development / Q000235:genetics; D001854:Bone Marrow Cells / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D018919:Neovascularization, Physiologic / Q000235:genetics*; D010012:Osteogenesis / Q000235:genetics
keywords: Cell sheet; Extracellular matrix; Human bone marrow mesenchymal stem cells; Human umbilical vein endothelial cells; Vascularization