Liu Jiani, Liu Rui, Wang Hong, Zhang Zijie, Wang Jixiao, Wei Fulan
Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, No. 44-1 Wenhua Road West, Jinan, Shandong, 250012, China.
J Orthop Surg Res. 2024 Apr 22;19(1):257. doi: 10.1186/s13018-024-04727-7.
The mechanotransduction mechanisms by which cells regulate tissue remodeling are not fully deciphered. Circular RNAs (circRNAs) are crucial to various physiological processes, including cell cycle, differentiation, and polarization. However, the effects of mechanical force on circRNAs and the role of circRNAs in the mechanobiology of differentiation and remodeling in stretched periodontal ligament stem cells (PDLSCs) remain unclear. This article aims to explore the osteogenic function of mechanically sensitive circular RNA protein kinase D3 (circPRKD3) and elucidate its underlying mechanotransduction mechanism.
PDLSCs were elongated with 8% stretch at 0.5 Hz for 24 h using the Flexcell® FX-6000™ Tension System. CircPRKD3 was knockdown or overexpressed with lentiviral constructs or plasmids. The downstream molecules of circPRKD3 were predicted by bioinformatics analysis. The osteogenic effect of related molecules was evaluated by quantitative real-time PCR (qRT-PCR) and western blot.
Mechanical force enhanced the osteogenesis of PDLSCs and increased the expression of circPRKD3. Knockdown of circPRKD3 hindered PDLSCs from osteogenesis under mechanical force, while overexpression of circPRKD3 promoted the early osteogenesis process of PDLSCs. With bioinformatics analysis and multiple software predictions, we identified hsa-miR-6783-3p could act as the sponge of circPRKD3 to indirectly regulate osteogenic differentiation of mechanically stimulated PDLSCs.
Our results first suggested that both circPRKD3 and hsa-miR-6783-3p could enhance osteogenesis of stretched PDLSCs. Furthermore, hsa-miR-6783-3p could sponge circPRKD3 to indirectly regulate RUNX2 during the periodontal tissue remodeling process in orthodontic treatment.
细胞调节组织重塑的机械转导机制尚未完全阐明。环状RNA（circRNA）对包括细胞周期、分化和极化在内的各种生理过程至关重要。然而，机械力对circRNA的影响以及circRNA在拉伸的牙周膜干细胞（PDLSCs）分化和重塑的力学生物学中的作用仍不清楚。本文旨在探讨机械敏感的环状RNA蛋白激酶D3（circPRKD3）的成骨功能，并阐明其潜在的机械转导机制。
使用Flexcell® FX - 6000™张力系统以0.5Hz的频率对PDLSCs进行8%的拉伸处理24小时。通过慢病毒构建体或质粒敲低或过表达circPRKD3。通过生物信息学分析预测circPRKD3的下游分子。通过定量实时PCR（qRT-PCR）和蛋白质印迹评估相关分子的成骨作用。
机械力增强了PDLSCs的成骨作用，并增加了circPRKD3的表达。敲低circPRKD3阻碍了机械力作用下PDLSCs的成骨，而过表达circPRKD3促进了PDLSCs的早期成骨过程。通过生物信息学分析和多种软件预测，我们确定hsa-miR-6783-3p可作为circPRKD3的海绵，间接调节机械刺激的PDLSCs的成骨分化。
我们的结果首次表明，circPRKD3和hsa-miR-6783-3p均可增强拉伸的PDLSCs的成骨作用。此外，在正畸治疗的牙周组织重塑过程中，hsa-miR-6783-3p可作为circPRKD3的海绵间接调节RUNX2。
