Wei Song-Guan, Chen Hui-Hong, Xie Liu-Rong, Qin Yuan, Mai Yu-Ying, Huang Lin-Hui, Liao Hong-Bing
Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction, College & Hospital of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
Department of Stomatology, The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou 545005, Guangxi Zhuang Autonomous Region, China.
World J Stem Cells. 2025 Apr 26;17(4):101290. doi: 10.4252/wjsc.v17.i4.101290.
degradation of bone scaffolds is significantly influenced by osteoclast (OC) activity, which is orchestrated by the interplay between receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG). The ratio of RANKL/OPG is a crucial determinant of OC-mediated bone resorption, which plays an integral role in bone remodeling and scaffold degradation. Elevated levels of RANKL relative to OPG enhance osteoclastogenesis, thereby accelerating the degradation process essential for integrating bone scaffolds into the host tissue.
To elucidate the effects of gene silencing on osteoclastogenesis within rat bone marrow-derived mesenchymal stem cells (BMSCs). By investigating these effects, the study aimed to provide deeper insights into the regulatory mechanisms that influence bone scaffold degradation, potentially leading to improved bone repair and regeneration strategies.
We employed recombinant lentiviral plasmids to silence the gene in rat BMSCs to achieve the aims. The efficacy of gene silencing was assessed using quantitative reverse transcription polymerase chain reaction and western blot analysis to measure the expression levels of OPG and RANKL. Tartrate-resistant acid phosphatase staining was utilized to evaluate the formation of OCs. Additionally, co-immunoprecipitation assays were conducted to explore the interactions between RANKL and OPG proteins, further assessing the biochemical pathways involved in osteoclastogenesis.
The silencing of the gene in BMSCs resulted in a significant increase in the RANKL/OPG ratio, evidenced by decreased expression levels of OPG and increased levels of RANKL. Enhanced osteoclastogenesis was observed through tartrate-resistant acid phosphatase staining, which indicated a substantial rise in OC formation in response to the altered RANKL/OPG balance. The co-immunoprecipitation assays provided concrete evidence of the direct interaction between RANKL and OPG proteins, substantiating their pivotal roles in regulating OC activity.
The findings from this study underscore the critical role of the RANKL/OPG axis in osteoclastogenesis. Silencing of the gene in BMSCs effectively increases the RANKL/OPG ratio, promoting OC activity and potentially enhancing bone scaffold degradation. This regulatory mechanism offers a promising avenue for modulating bone remodeling processes, which is essential for effective bone repair and the successful integration of bone scaffolds into damaged sites. Future research might focus on optimizing the control of this axis to better facilitate bone tissue engineering and regenerative therapies.
破骨细胞（OC）活性对骨支架的降解有显著影响，而OC活性是由核因子κB受体激活剂配体（RANKL）和骨保护素（OPG）之间的相互作用所调控的。RANKL/OPG的比值是OC介导的骨吸收的关键决定因素，在骨重塑和支架降解中起着不可或缺的作用。相对于OPG，RANKL水平升高会增强破骨细胞生成，从而加速骨支架与宿主组织整合所必需的降解过程。
阐明基因沉默对大鼠骨髓间充质干细胞（BMSCs）中破骨细胞生成的影响。通过研究这些影响，该研究旨在更深入地了解影响骨支架降解的调控机制，可能会带来改进的骨修复和再生策略。
我们使用重组慢病毒质粒沉默大鼠BMSCs中的基因以实现研究目的。通过定量逆转录聚合酶链反应和蛋白质印迹分析来评估基因沉默的效果，以测量OPG和RANKL的表达水平。利用抗酒石酸酸性磷酸酶染色来评估破骨细胞的形成。此外，进行免疫共沉淀试验以探索RANKL和OPG蛋白之间的相互作用，进一步评估破骨细胞生成所涉及的生化途径。
BMSCs中基因的沉默导致RANKL/OPG比值显著增加，表现为OPG表达水平降低和RANKL水平升高。通过抗酒石酸酸性磷酸酶染色观察到破骨细胞生成增强，这表明随着RANKL/OPG平衡的改变，破骨细胞形成大幅增加。免疫共沉淀试验提供了RANKL和OPG蛋白直接相互作用的确切证据，证实了它们在调节破骨细胞活性中的关键作用。
本研究结果强调了RANKL/OPG轴在破骨细胞生成中的关键作用。BMSCs中基因的沉默有效地增加了RANKL/OPG比值，促进了破骨细胞活性，并可能增强骨支架降解。这种调控机制为调节骨重塑过程提供了一条有前景的途径，这对于有效的骨修复以及骨支架成功整合到受损部位至关重要。未来的研究可能集中在优化对该轴的控制上，以更好地促进骨组织工程和再生治疗。
