Three-Dimensional Engineered Bone from Bone Marrow Stromal Cells and Their Autogenous Extracellular Matrix
Three-Dimensional Engineered Bone from Bone Marrow Stromal Cells and Their Autogenous Extracellular Matrix
复制标题
DOI：
10.1089/ten.tea.2007.0140
复制
发表时间：
2009-01-01
影响因子：
4.1
通讯作者：
Arruda, Ellen M.
中科院分区：
文献类型：
--
作者：
Syed-Picard, Fatima N.;Larkin, Lisa M.;Arruda, Ellen M.
大多数骨组织工程研究使用多孔三维（3D）支架进行细胞接种。本研究利用大鼠骨髓基质细胞（BMSCs）及其自体细胞外基质（ECM）构建无支架三维类骨组织。将BMSCs在诱导成骨分化的培养基中的2D基质上培养。在达到汇合并产生足够量的其自身ECM后，细胞围绕两个约束点收缩其组织单层，形成无支架的圆柱形工程化骨样构建体（EBC）。茜素红染色显示矿化和碱性磷酸酶活性，并含有I型胶原。EBC形成骨膜，其特征在于在构建体的外周上的成纤维细胞和未矿化的胶原。拉伸试验显示，培养物中的EBC在3D构建体形成后7天具有7.5 +/-0.5MPa的切线模量，在构建体形成后6周具有29 +/-9MPa的切线模量。在构建体形成后7天将EBC植入大鼠中导致进一步的骨发育和血管化。在4周时收集的组织外植体含有在天然骨中发现的所有三种细胞类型：成骨细胞、骨细胞和破骨细胞。由此产生的工程组织是第一个在不使用外源性支架的情况下开发的3D骨组织。
Most bone tissue-engineering research uses porous three-dimensional (3D) scaffolds for cell seeding. In this work, scaffold-less 3D bone-like tissues were engineered from rat bone marrow stromal cells (BMSCs) and their autogenous extracellular matrix (ECM). The BMSCs were cultured on a 2D substrate in medium that induced osteogenic differentiation. After reaching confluence and producing a sufficient amount of their own ECM, the cells contracted their tissue monolayer around two constraint points, forming scaffold-less cylindrical engineered bone-like constructs (EBCs). The EBCs exhibited alizarin red staining for mineralization and alkaline phosphatase activity and contained type I collagen. The EBCs developed a periosteum characterized by fibroblasts and unmineralized collagen on the periphery of the construct. Tensile tests revealed that the EBCs in culture had a tangent modulus of 7.5 +/- 0.5 MPa at 7 days post-3D construct formation and 29 +/- 9 MPa at 6 weeks after construct formation. Implantation of the EBCs into rats 7 days after construct formation resulted in further bone development and vascularization. Tissue explants collected at 4 weeks contained all three cell types found in native bone: osteoblasts, osteocytes, and osteoclasts. The resulting engineered tissues are the first 3D bone tissues developed without the use of exogenous scaffolding.