Dong Rui, Bai Yun, Dai Jingjin, Deng Moyuan, Zhao Chunrong, Tian Zhansong, Zeng Fanchun, Liang Wanyuan, Liu Lanyi, Dong Shiwu
Department of Biomedical Materials Science, School of Biomedical Engineering, Third Military Medical University, Chongqing, China.
Department of Orthopedics, Southwest Hospital, Third Military Medical University, Chongqing, China.
J Tissue Eng. 2020 Jun 7;11:2041731420926918. doi: 10.1177/2041731420926918. eCollection 2020 Jan-Dec.
Recently, extracellular matrix-based tissue-engineered bone is a promising approach to repairing bone defects, and the seed cells are mostly mesenchymal stem cells. However, bone remodelling is a complex biological process, in which osteoclasts perform bone resorption and osteoblasts dominate bone formation. The interaction and coupling of these two kinds of cells is the key to bone repair. Therefore, the extracellular matrix secreted by the mesenchymal stem cells alone cannot mimic a complex bone regeneration microenvironment, and the addition of extracellular matrix by preosteoclasts may contribute as an effective strategy for bone regeneration. Here, we established the mesenchymal stem cell/preosteoclast extracellular matrix -based tissue-engineered bones and demonstrated that engineered-scaffolds based on mesenchymal stem cell/ preosteoclast extracellular matrix significantly enhanced osteogenesis in a 3 mm rat femur defect model compared with mesenchymal stem cell alone. The bioactive proteins released from the mesenchymal stem cell/ preosteoclast extracellular matrix based tissue-engineered bones also promoted the migration, adhesion, and osteogenic differentiation of mesenchymal stem cells in vitro. As for the mechanisms, the iTRAQ-labeled mass spectrometry was performed, and 608 differentially expressed proteins were found, including the IGFBP5 and CXCL12. Through in vitro studies, we proved that CXCL12 and IGFBP5 proteins, mainly released from the preosteoclasts, contributed to mesenchymal stem cells migration and osteogenic differentiation, respectively. Overall, our research, for the first time, introduce pre-osteoclast into the tissue engineering of bone and optimize the strategy of constructing extracellular matrix-based tissue-engineered bone using different cells to simulate the natural bone regeneration environment, which provides new sight for bone tissue engineering.
近年来，基于细胞外基质的组织工程骨是修复骨缺损的一种有前景的方法，种子细胞大多为间充质干细胞。然而，骨重塑是一个复杂的生物学过程，其中破骨细胞进行骨吸收，成骨细胞主导骨形成。这两种细胞的相互作用和耦合是骨修复的关键。因此，仅由间充质干细胞分泌的细胞外基质无法模拟复杂的骨再生微环境，而破骨前体细胞添加细胞外基质可能是一种有效的骨再生策略。在此，我们构建了基于间充质干细胞/破骨前体细胞外基质的组织工程骨，并证明与单独使用间充质干细胞相比，基于间充质干细胞/破骨前体细胞外基质的工程支架在3毫米大鼠股骨缺损模型中显著增强了成骨作用。基于间充质干细胞/破骨前体细胞外基质的组织工程骨释放的生物活性蛋白在体外也促进了间充质干细胞的迁移、黏附和成骨分化。至于机制，进行了iTRAQ标记的质谱分析，发现了608种差异表达蛋白，包括IGFBP5和CXCL12。通过体外研究，我们证明主要由破骨前体细胞释放的CXCL12和IGFBP5蛋白分别促进了间充质干细胞的迁移和成骨分化。总体而言，我们的研究首次将破骨前体细胞引入骨组织工程，并优化了使用不同细胞构建基于细胞外基质的组织工程骨以模拟天然骨再生环境的策略，为骨组织工程提供了新的视角。
