Experimental Trauma Surgery, Faculty of Medicine, Justus-Liebig University of Giessen, Aulweg 128, 35392 Giessen, Germany.
Institute for Systems Medicine, Faculty of Human Medicine, MSH Medical School Hamburg, 20457 Hamburg, Germany.
Int J Mol Sci. 2023 Feb 10;24(4):3569. doi: 10.3390/ijms24043569.
The success of fracture healing relies on overlapping but coordinated cellular and molecular events. Characterizing an outline of differential gene regulation throughout successful healing is essential for identifying crucial phase-specific markers and may serve as the basis for engineering these in challenging healing situations. This study analyzed the healing progression of a standard closed femoral fracture model in C57BL/6N (age = 8 weeks) wild-type male mice. The fracture callus was assessed across various days post fracture (D = days 0, 3, 7, 10, 14, 21, and 28) by microarray, with D0 serving as a control. Histological analyses were carried out on samples from D7 until D28 to support the molecular findings. Microarray analysis revealed a differential regulation of immune response, angiogenesis, ossification, extracellular matrix regulation, mitochondrial and ribosomal genes during healing. In-depth analysis showed differential regulation of mitochondrial and ribosomal genes during the initial phase of healing. Furthermore, the differential gene expression showed an essential role of Serpin Family F Member 1 over the well-known Vascular Endothelial Growth Factor in angiogenesis, especially during the inflammatory phase. The significant upregulation of matrix metalloproteinase 13 and bone sialoprotein from D3 until D21 asserts their importance in bone mineralization. The study also shows type I collagen around osteocytes located in the ossified region at the periosteal surface during the first week of healing. Histological analysis of matrix extracellular phosphoglycoprotein and extracellular signal-regulated kinase stressed their roles in bone homeostasis and the physiological bone-healing process. This study reveals previously unknown and novel candidates, that could serve as a target for specific time points in healing and to remedy cases of impaired healing.
骨折愈合的成功依赖于重叠但协调的细胞和分子事件。描述成功愈合过程中差异基因调控的概况对于鉴定关键的时相特异性标志物至关重要，并且可能为在具有挑战性的愈合情况下对这些标志物进行工程设计提供基础。本研究分析了 C57BL/6N（年龄= 8 周）野生型雄性小鼠标准闭合股骨骨折模型的愈合进展。通过微阵列分析在骨折后不同天数（D = 0、3、7、10、14、21 和 28）评估骨折痂，D0 作为对照。在 D7 至 D28 期间对样本进行组织学分析，以支持分子发现。微阵列分析显示，在愈合过程中，免疫反应、血管生成、成骨、细胞外基质调节、线粒体和核糖体基因的调节存在差异。深入分析显示，在愈合的初始阶段，线粒体和核糖体基因的调节存在差异。此外，差异基因表达显示丝氨酸蛋白酶家族 F 成员 1在血管内皮生长因子中具有重要作用，特别是在炎症阶段。基质金属蛋白酶 13 和骨涎蛋白从 D3 至 D21 的显著上调表明它们在骨矿化中的重要性。该研究还表明，在愈合的第一周，在骨细胞周围的骨化区的骨膜表面存在Ⅰ型胶原。基质细胞外磷糖蛋白和细胞外信号调节激酶的组织学分析强调了它们在骨稳态和生理骨愈合过程中的作用。本研究揭示了以前未知的和新的候选物，这些候选物可能作为愈合特定时间点的靶标，并弥补愈合受损的情况。
