Huang Weihua, Hao Lijing, Cai Xiayu, Xiao Chujie, Yin Huinan, Xu Weikang
Institute of Biological and Medical Engineering, Guangdong Academy of Sciences, Guangzhou, China.
Department of Orthopaedic Surgery, The Affiliated Qingyuan Hospital (Qingyuan People's Hospital), Guangzhou Medical University, Guangzhou, China.
Bone Joint Res. 2025 Apr 3;14(4):292-308. doi: 10.1302/2046-3758.144.BJR-2024-0080.R3.
This study aimed to examine the differences in bone induction and osseointegration performance of acellular extracellular matrix bone at different sites.
We decellularized bone from bovine epiphysis near the marrow cavity (NMC), the middle of the cancellous bone (MCB), and near the cartilage (NC). The characterization, physicochemical properties, and effectiveness of the decellularization process of decellularized extracellular matrix (dECM) were analyzed. The proliferation, adhesion, seeding efficiency, and osteogenic differentiation properties of bone marrow mesenchymal stem cells (BMSCs) on decellularized extracellular matrix were investigated. The osteogenicity and osteointegration of dECM from different sources were verified in vivo by animal experiments, and the compatibility of dECM in vivo was also verified.
The NC group had the most significant compressive properties, where the compressive strength was about 1.62 times higher than that of the MCB group (p = 0.022) and 1.34 times higher than that of the NMC group (p < 0.001). dECM scaffolds had good histocompatibility and supported the adhesion and proliferation of BMSCs. In vitro, compared with the remaining two groups, the MCB group significantly upregulated the expression of osteogenic genes (alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), osteopontin (OPN), collagen type 1 (COL1), and bone morphogenetic protein 2 (BMP2)) and marker proteins (ALP, BMP2), whereas the NC group showed the weakest osteoinductive properties. In vivo, we confirmed that the MCB group possessed the most significant osteogenic and osseointegrative properties, followed by the NMC group, and the NC group proved to be the weakest. In particular, the MCB group possessed the ability to endogenously immunomodulate macrophage M1 phenotype to M2 phenotype polarization, creating the most favourable immune microenvironment for osteogenesis.
Our data indicated that the xenogenic dECM scaffolds in MCB position possess the most significant biocompatibility and in vitro and in vivo induced osteogenesis and osseointegration properties. This study provides a more complete basis for the selection of dECM scaffolds in bone defect repair. In future studies of dECM composites applied to bone tissue engineering (BTE), utilizing the middle part of cancellous bone may be the best solution.
本研究旨在探讨不同部位的脱细胞细胞外基质骨在骨诱导和骨整合性能方面的差异。
我们从靠近骨髓腔（NMC）、松质骨中部（MCB）和靠近软骨（NC）的牛骨骺中提取脱细胞骨。分析了脱细胞细胞外基质（dECM）的表征、理化性质及脱细胞过程的有效性。研究了骨髓间充质干细胞（BMSCs）在脱细胞细胞外基质上的增殖、黏附、接种效率和成骨分化特性。通过动物实验在体内验证了不同来源dECM的成骨能力和骨整合能力，同时也验证了dECM在体内的相容性。
NC组的压缩性能最为显著，其抗压强度比MCB组高约1.62倍（p = 0.022），比NMC组高1.34倍（p < 0.001）。dECM支架具有良好的组织相容性，支持BMSCs的黏附和增殖。在体外，与其余两组相比，MCB组显著上调了成骨基因（碱性磷酸酶（ALP）、 runt相关转录因子2（RUNX2）、骨桥蛋白（OPN）、Ⅰ型胶原（COL1）和骨形态发生蛋白2（BMP2））和标记蛋白（ALP、BMP2）的表达，而NC组的骨诱导性能最弱。在体内，我们证实MCB组具有最显著的成骨和骨整合性能，其次是NMC组，而NC组被证明是最弱的。特别是，MCB组具有将巨噬细胞M1表型内源性免疫调节为M2表型极化的能力，为成骨创造了最有利的免疫微环境。
我们的数据表明，位于MCB位置的异种dECM支架具有最显著的生物相容性以及体外和体内诱导成骨和骨整合性能。本研究为骨缺损修复中dECM支架的选择提供了更完整的依据。在未来应用于骨组织工程（BTE）的dECM复合材料研究中，利用松质骨中部可能是最佳解决方案。
