Indirect Immobilised Jagged-1 Enhances Matrisome Proteins Associated with Osteogenic Differentiation of Human Dental Pulp Stem Cells: A Proteomic Study

Abstract

The indirect immobilisation of Jagged-1 (Jagged-1) promoted osteogenic differentiation of human dental pulp cells (hDPs). Furthermore, the analysis of the Reactome pathway of RNA sequencing data indicates the upregulated genes involved with the extracellular matrix (ECM). Hence, our objective was to investigate the effects of Jagged-1 on proteomic profiles of human dental pulp stem cells (hDPSC). hDPSCs were cultured on the surface coated with human IgG Fc fragment (hFc) and the surface coated with rhJagged1/Fc recombinant protein-coated surface. Cells were differentiated to the osteogenic lineage using an osteogenic differentiation medium (OM) for 14 days, and cells cultured in a growth medium were used as a control. The protein component of the cultured cells was extracted into the cytosol, membrane, nucleus, and cytoskeletal compartment. Subsequently, the proteomic analysis was performed using liquid chromatography–tandem mass spectrometry (LC-MS). Metascape gene list analysis reported that Jagged-1 stimulated the expression of the membrane trafficking protein (DOP1B), which can indirectly improve osteogenic differentiation. hDPSCs cultured on Jagged-1 surface under OM condition expressed COL27A1, MXRA5, COL7A1, and MMP16, which played an important role in osteogenic differentiation. Furthermore, common matrisome proteins of all cellular components were related to osteogenesis/osteogenic differentiation. Additionally, the gene ontology categorised by the biological process of cytosol, membrane, and cytoskeleton compartments was associated with the biomineralisation process. The gene ontology of different culture conditions in each cellular component showed several unique gene ontologies. Remarkably, the Jagged-1_OM culture condition showed the biological process related to odontogenesis in the membrane compartment. In conclusion, the Jagged-1 induces osteogenic differentiation could, mainly through the regulation of protein in the membrane compartment.

Biomedical engineering approaches for the delivery of JAGGED1 as a potential tissue regenerative therapy

“…Indirect affinity immobilization of JAG1 is a common method for JAG1 delivery. For example, protein G has been used by many studies for indirect immobilization and they have demonstrated the functionality of JAG1 in activating the canonical NOTCH gene expression pathway ( Osathanon et al, 2013 ; Manokawinchoke et al, 2017 ; Nowwarote et al, 2018 ; Chansaenroj et al, 2022 ). A 2009 study used self-assembled monolayers for indirect affinity immobilization and it also established the ability of SAMs to allow JAG1-induced NOTCH signaling activation ( Goncalves et al, 2009 ).…”

“…Moreover, this was the first study to demonstrate that JAG1 delivery transiently activates the NOTCH signaling pathway. Chansaenroj et al (2022) was the first study to perform a proteomic study on hDPSCs that were cultured with indirectly immobilized JAG1. Upon extracting the protein component of cells, it was found that JAG1 stimulated the expression of proteins associated with osteogenic differentiation and matrisome proteins played a role in osteogenic differentiation.…”

“…The results demonstrated that indirect affinity immobilized JAG1 on the membranes upregulated hPDL NOTCH target gene expression. Recently, Chansaenroj et al (2022) also used this immobilization method to understand the effect of JAG1 and the extracellular matrix (ECM) on the osteogenic differentiation of hDPSC. In this study, surfaces were incubated with protein G, human IgG Fc fragment, and rhJagged1/Fc recombinant protein.…”