Surface‐bound orientated Jagged‐1 enhances osteogenic differentiation of human periodontal ligament‐derived mesenchymal stem cells

Abstract

Notch signaling plays critical roles in various cell types by regulating cell fate determination and differentiation. Here, we investigated the ability to control differentiation of human periodontal ligament derived mesenchymal stem cells using modified surfaces containing the affinity immobilized Notch ligand, Jagged-1. After seeding human periodontal ligament derived mesenchymal stem cells (HPDLs) on Jagged-1 modified surfaces, expression of Notch signaling target genes, Hes-1 and Hey-1, was higher than those exposed to soluble Jagged-1 or control surfaces. Upregulation of Notch signaling target genes was attenuated after treatment with the γ secretase inhibitor. Upon seeding the cells on Jagged-1 immobilized surface and maintained in osteogenic medium, alkaline phosphatase enzymatic activity and mineralization as well as mRNA expression of alkaline phosphatase (ALP), collagen type I (COL I) and osteopontin (OPN) were significantly increased compared to those of controls. However, osteocalcin (OCN) mRNA expression level was decreased when cells were exposed to Jagged-1 modified surfaces. HPDLs on Jagged-1 modified surfaces expressed lower TWIST2 mRNA levels than the control, suggesting that the mechanism whereby Jagged-1 enhances osteogenic differentiation of HPDLs may occur through Notch signaling and TWIST regulation. In summary, an alteration of biomaterial interface using Notch ligands illustrates a promising system to control HPDLs differentiation toward osteogenic lineage.

Notch Signaling Is Involved in Neurogenic Commitment of Human Periodontal Ligament-Derived Mesenchymal Stem Cells

“…Regarding neuronal differentiation, our group and other investigators have illustrated that hPDLSCs were able to differentiate toward neurogenic lineage under specific conditions [4][5][6][7]. Neurite-like, long cellular processes and upregulation of neurogenic markers [b3-tubulin, nestin, and neurofilament (NF)] were noted after neurogenic induction [4,8].…”

“…Surface-bound Notch ligand, Jagged-1, was fabricated by an indirect affinity immobilization method previously reported by our group [4,20,21]. Briefly, recombinant protein G (50 mg/mL; Zymed) was incubated with the tissue culture plate surface for 16 h and subsequently, incubated with bovine serum albumin (10 mg/mL; Sigma) for 2 h. The surface was further incubated with recombinant human Jagged-1 (10 mM; R&D Systems) for 2 h. The surface was washed 3 times with sterile phosphate-buffered saline (PBS) between each step.…”

“…The periodontal ligament mainly consists of type I collagen fibers and several types of cells, for example, periodontal ligament fibroblast cells, vascular endothelial cells, and smooth muscle cells [1]. Moreover, several studies have shown that mesenchymal stem cells can be isolated from periodontal ligament tissues of both primary and permanent teeth [2][3][4]. Our previous study showed that cells isolated from human periodontal ligament tissues expressed embryonic and mesenchymal stem cell markers: Oct4, Rex-1, Nanog, CD44, CD73, and CD105 [5].…”

“…Neurite-like, long cellular processes and upregulation of neurogenic markers [b3-tubulin, nestin, and neurofilament (NF)] were noted after neurogenic induction [4,8]. Moreover, it has been demonstrated that hPDLSCs were able to integrate into brain tissues in vitro, evaluated by hippocampal organ culture model [9].…”

“…Subsequently, the cleavage of Notch protein occurs intracellularly, resulting in the translocation of Notch intracellular domain (NICD) into the nucleus and further regulating Notch target gene expression [14]. In periodontal ligament cells, Notch signaling was shown to participate in osteogenic differentiation process and induction of osteoclastogenesis [4,15,16]. However, evidence of Notch signaling in hPDLSCs differentiation toward neuronal lineage is yet unknown.…”