PDF(Pubmed)
Abstract：
The osteogenic differentiation process, by which bone marrow mesenchymal stem cells and osteoprogenitors transform into osteoblasts, is regulated by several growth factors, cytokines, and hormones. Plasma Rich in Growth Factors (PRGF) is a blood-derived preparation consisting of a plethora of bioactive molecules, also susceptible to containing epigenetic factors such as ncRNAs and EVs, that stimulates tissue regeneration. The aim of this study was to investigate the effect of the PRGF clot formulation on osteogenic differentiation. Firstly, osteoblast cells were isolated and characterised. The proliferation of bone cells cultured onto PRGF clots or treated with PRGF supernatant was determined. Moreover, the gene expression of Runx2 (ID: 860), SP7 (ID: 121340), and ALPL (ID: 249) was analysed by one-step real-time quantitative polymerase chain reaction (RT-qPCR). Additionally, alkaline phosphatase (ALPL) activity determination was performed. The highest proliferative effect was achieved by the PRGF supernatant in all the study periods analysed. Concerning gene expression, the logRGE of Runx2 increased significantly in osteoblasts cultured with PRGF formulations compared with the control group, while that of SP7 increased significantly in osteoblasts grown on the PRGF clots. On the other hand, despite the fact that the PRGF supernatant induced ALPL up-regulation, significantly higher enzyme activity was detected for the PRGF clots in comparison with the supernatant formulation. According to our results, contact with the PRGF clot could promote a more advanced phase in the osteogenic process, associated to higher levels of ALPL activity. Furthermore, the PRGF clot releasate stimulated a higher proliferation rate in addition to reduced SP7 expression in the cells located at a distant ubication, leading to a less mature osteoblast stage. Thus, the spatial relationship between the PRGF clot and the osteoprogenitors cells could be a factor that influences regenerative outcomes.
摘要：
成骨分化过程，骨髓间充质干细胞和骨祖细胞转化为成骨细胞，受几个生长因子的调节，细胞因子,和荷尔蒙。富含生长因子的血浆（PRGF）是一种由多种生物活性分子组成的血液衍生制剂，也容易含有表观遗传因素，如ncRNAs和EV，刺激组织再生。这项研究的目的是研究PRGF凝块制剂对成骨分化的影响。首先,分离并鉴定成骨细胞。测定在PRGF凝块上培养或用PRGF上清液处理的骨细胞的增殖。此外，Runx2的基因表达（ID：860），SP7（ID：121340），通过一步实时定量聚合酶链反应（RT-qPCR）分析ALPL（ID：249）。此外,进行碱性磷酸酶(ALPL)活性测定。在所分析的所有研究阶段中,通过PRGF上清液实现了最高的增殖效果。关于基因表达，与对照组相比，用PRGF制剂培养的成骨细胞中Runx2的logRGE显著增加，而在PRGF凝块上生长的成骨细胞中，SP7的含量显着增加。另一方面,尽管PRGF上清液诱导ALPL上调，与上清液制剂相比,对于PRGF凝块检测到显著更高的酶活性。根据我们的结果,与PRGF凝块接触可以促进成骨过程的更晚期，与较高水平的ALPL活性相关。此外，PRGF凝块释放除了减少SP7表达外，还刺激了位于远处的细胞的更高的增殖率，导致成骨细胞不成熟。因此，PRGF凝块和骨祖细胞之间的空间关系可能是影响再生结果的一个因素.
