Tran Y Nhu, Chansaenroj Ajjima, Jivaphetthai Araya, Osathanon Thanaphum, Arunmanee Wanatchaporn
Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
Center of Excellence for Dental Stem Cell Biology, Faculty of Dentistry, Chulalongkorn University, Bangkok 10330, Thailand; Department of Animal Husbandry, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
Arch Oral Biol. 2025 Feb;170:106138. doi: 10.1016/j.archoralbio.2024.106138. Epub 2024 Nov 28.
Jagged1, a Notch ligand, is essential for osteogenic differentiation in human periodontal ligament cells (hPDLs) by interacting with Notch2 to induce osteogenic markers, alkaline phosphatase activity, and mineral deposition. However, its large size hampers absorption and distribution of biomaterials. This study aimed to identify the critical region of Jagged1 necessary for its interaction with Notch2 to create a truncated version that retains osteogenic activity but with improved delivery characteristics.
Truncated versions of Jagged1 were designed by deleting C-terminal regions, focusing on the importance of the N-terminal domain. Both truncated and full-length Jagged1 were fused with human IgG1 Fc (Jagged1-Fc) and expressed in Chinese hamster ovary cells. hPDLs treated with these constructs were analyzed for Notch target gene expression using real-time PCR. Mineral deposition was assessed using alizarin red S staining.
Both truncated and full-length Jagged1-Fc increased the expression of Notch target genes (Hes1, Hey1, and ALP) in hPDLs, indicating successful activation of Notch signaling. However, only the full-length Jagged1-Fc enhanced mineral deposition, while the truncated version did not.
Full-length Jagged1-Fc is required for mineral deposition and complete osteogenic differentiation in hPDLs. The truncated versions, while capable of activating Notch signaling, are ineffective in promoting mineralization, underscoring the importance of the entire protein for clinical applications in bone regeneration.
锯齿蛋白1（Jagged1）是一种Notch配体，通过与Notch2相互作用诱导成骨标志物、碱性磷酸酶活性和矿物质沉积，对人牙周膜细胞（hPDL）的成骨分化至关重要。然而，其较大的尺寸阻碍了生物材料的吸收和分布。本研究旨在确定Jagged1与Notch2相互作用所必需的关键区域，以创建一个保留成骨活性但具有更好递送特性的截短版本。
通过删除C端区域设计Jagged1的截短版本，重点关注N端结构域的重要性。将截短的和全长的Jagged1与人IgG1 Fc融合（Jagged1-Fc），并在中国仓鼠卵巢细胞中表达。使用实时PCR分析用这些构建体处理的hPDL的Notch靶基因表达。使用茜素红S染色评估矿物质沉积。
截短的和全长的Jagged1-Fc均增加了hPDL中Notch靶基因（Hes1、Hey1和ALP）的表达，表明Notch信号成功激活。然而，只有全长的Jagged1-Fc增强了矿物质沉积，而截短版本则没有。
全长Jagged1-Fc是hPDL中矿物质沉积和完全成骨分化所必需的。截短版本虽然能够激活Notch信号，但在促进矿化方面无效，这突出了整个蛋白质在骨再生临床应用中的重要性。
