Abstract:
Bone injury is often associated with tears in the periosteum and changes in the internal stress microenvironment of the periosteum. In this study, we investigated the biological effects of periosteal prestress release on periosteum-derived cells (PDCs) and the potential mechanisms of endogenous stem cell recruitment. Decellularized periosteum with natural extracellular matrix (ECM) components was obtained by a combination of physical, chemical, and enzymatic decellularization. The decellularized periosteum removed immunogenicity while retaining the natural network structure and composition of the ECM. The Young\'s modulus has no significant difference between the periosteum before and after decellularization. The extracted PDCs were further composited with the decellularized periosteum and subjected to 20% stress release. It was found that the proliferative capacity of PDCs seeded on decellularized periosteum was significantly enhanced 6 h after stress release of the periosteum. The cell culture supernatant obtained after periosteal prestress release was able to significantly promote the migration ability of PDCs within 24 h. Enzyme-linked immunosorbnent assay (ELISA) experiments showed that the expression of stroma-derived factor-1α (SDF-1α) and vascular endothelial growth factor (VEGF) in the supernatant increased significantly after 3 h and 12 h of stress release, respectively. Furthermore, periosteal stress release promoted the high expression of osteogenic markers osteocalcin (OCN), osteopontin (OPN), and collagen type I of PDCs. The change in stress environment caused by the release of periosteal prestress was sensed by integrin β1, a mechanoreceptor on the membrane of PDCs, which further stimulated the expression of YAP in the nucleus. These investigations provided a novel method to evaluate the importance of mechanical stimulation in periosteum, which is also of great significance for the design and fabrication of artificial periosteum with mechanical regulation function.
摘要:
骨损伤通常与骨膜撕裂和骨膜内部应力微环境的变化有关。在这项研究中,我们研究了骨膜预应力释放对骨膜来源细胞(PDC)的生物学效应以及内源性干细胞募集的潜在机制。脱细胞骨膜与天然细胞外基质(ECM)成分是通过物理,化学,和酶解细胞化。脱细胞骨膜去除了免疫原性,同时保留ECM的天然网络结构和组成。脱细胞前后骨膜杨氏模量无显著差异。将提取的PDC进一步与脱细胞骨膜复合并进行20%的应力释放。发现在骨膜应力释放后6小时,接种在脱细胞骨膜上的PDC的增殖能力显着增强。骨膜预应力释放后获得的细胞培养上清液能在24h内显著促进PDCs的迁移能力。酶联免疫吸附试验(ELISA)实验表明,压力释放3h和12h后,上清液中基质衍生因子-1α(SDF-1α)和血管内皮生长因子(VEGF)的表达明显增加,分别。此外,骨膜应力释放促进成骨标志物骨钙蛋白(OCN)的高表达,骨桥蛋白(OPN),和PDC的I型胶原蛋白。由骨膜预应力释放引起的应力环境变化由整合素β1感知,整合素β1是PDC膜上的机械受体,这进一步刺激了YAP在细胞核中的表达。这些研究提供了一种新的方法来评估机械刺激在骨膜中的重要性。这对具有机械调节功能的人工骨膜的设计和制作也具有重要意义。