Distinctive Roles of Wnt Signaling in Chondrogenic Differentiation of BMSCs under Coupling of Pressure and Platelet-Rich Fibrin
Distinctive Roles of Wnt Signaling in Chondrogenic Differentiation of BMSCs under Coupling of Pressure and Platelet-Rich Fibrin
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DOI：
10.1007/s13770-022-00456-2
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发表时间：
2022-04-25
影响因子：
3.6
通讯作者：
Zhang, Min
中科院分区：
文献类型：
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作者：
Cheng, Baixiang;Feng, Fan;Zhang, Min
背景：虽然新形成的可行的压力预调节骨髓间充质干细胞（BMSC）和富血小板纤维蛋白（PRF）构建体显示出生物力学灵活性和卓越的软骨再生能力，但目前仍不清楚BMSC和种子细胞如何感受机械刺激并将其转化为生物信号，并且机械和化学信号背后的信号转导差异也不清楚。方法：为了确定机械刺激（静水压）和化学信号（富血小板纤维蛋白，PRF）是否激活 BMSC 中的经典或非经典 Wnt 信号传导，对与 PRF 共培养的 BMSC 进行静水压加载，并通过蛋白质印迹和聚合酶链反应（PCR）测定 Wnt 信号分子的激活以及软骨相关蛋白和基因的表达。采用经典或非经典Wnt信号传导抑制剂XVX-939或L690,330来研究Wnt信号分子在机械促进BMSCs软骨形成分化中的作用。结果：120 kPa的静水压同时激活Wnt/β-catenin信号和Wnt/Ca2+信号，且在60 min时促进作用最大。 PRF 对 Wnt/β-catenin 信号传导激活没有协同作用。然而，PRF释放的生长因子可能会逆转压力对Wnt/Ca2+信号传导的促进作用。实时PCR和Western blotting结果表明，压力可以激活与PRF共培养的BMSCs中Col-II、Sox9和聚集蛋白聚糖的表达。阻断实验发现Wnt/β-catenin信号传导在BMSCs的软骨分化中具有积极作用，而Wnt/Ca2+信号传导则具有消极作用。压力下 BMSC 中经典和非经典 Wnt 信号之间存在相互抑制。结论：Wnt信号参与了与PRF共培养的BMSCs的压力促进软骨形成过程，在此过程中典型和非典型信号通路发挥着不同的作用。
Background: Although newly formed constructs of feasible pressure-preadjusted bone marrow mesenchymal stem cells (BMSCs) and platelet-rich fibrin (PRF) showed biomechanical flexibility and superior capacity for cartilage regeneration, it is still not very clear how BMSCs and seed cells feel mechanical stimuli and convert them into biological signals, and the difference in signal transduction underlying mechanical and chemical cues is also unclear. Methods: To determine whether mechanical stimulation (hydrostatic pressure) and chemical cues (platelet-rich fibrin, PRF) activate canonical or noncanonical Wnt signaling in BMSCs, BMSCs cocultured with PRF were subjected to hydrostatic pressure loading, and the activation of the Wnt signaling molecules and expression of cartilage-associated proteins and genes were determined by western blotting and polymerase chain reaction (PCR). Inhibitors of canonical or noncanonical Wnt signaling, XVX-939 or L690,330, were adopted to investigate the role of Wnt signaling molecules in mechanically promoted chondrogenic differentiation of BMSCs. Results: Hydrostatic pressure of 120 kPa activated both Wnt/beta-catenin signaling and Wnt/Ca2+ signaling, with the the maximum promotion effect at 60 min. PRF exerted no synergistic effect on Wnt/beta-catenin signaling activation. However, the growth factors released by PRF might reverse the promotion effects of pressure on Wnt/Ca2+ signaling. Real-time PCR and Western blotting results showed that pressure could activate the expression of Col-II, Sox9, and aggrecan in BMSCs cocultured with PRF. Blocking experiment found a positive role of Wnt/beta-catenin signaling, and a negative role of Wnt/Ca2+ signaling in chondrogenic differentiation of the BMSCs. Mutual inhibition exists between canonical and noncanonical Wnt signaling in BMSCs under pressure. Conclusion: Wnt signaling participates in the pressure-promoted chondrogenesis of the BMSCs co-cultured with PRF, with canonical and noncanonical pathways playing distinct roles during the process.