Proteomics and N‐glycoproteomics analysis of an extracellular matrix‐based scaffold‐human treated dentin matrix

Abstract

Extracellular matrix (ECM)-based biomaterials developed from mammalian tissues have been successfully used in preclinical and clinical tissue engineering applications.We have previously reported about the applicability of dentin-based scaffold, treated dentin matrix (TDM), for tooth root regeneration. However, TDM protein composition has not been characterized. Here, we used a shotgun proteomic strategy to profile human TDM proteome. N-glycoproteins were enriched by lectin affinity chromatography and identified by mass spectrometry. The total human TDM proteome was compared with the previously published human dentin proteome, and bioinformatics analysis were performed accordingly. In total, 708 proteins were identified by mass spectrometry in human TDM, of which 208 were Nglycoproteins with 318 identified glycosylation sites. Collagens, proteoglycans, small integrin-binding ligand N-linked glycoproteins (SIBLINGs), and growth factors, such as COL1A1, biglycan, dentin sialoprotein, and transforming growth factor beta 1, were identified. Glycoproteins were enriched in "biological processes" Gene Ontology terms such as cellular process, biological regulation, response to stimulus, metabolic process, immune system process, and biological adhesion. Thus, our comprehensive study of the human TDM proteome revealed that dentin proteins are more heterogeneous than previously documented. Our findings provide clues for designing new biomaterials for tooth root regeneration and understanding dentin formation.

Functional Dental Pulp Regeneration: Basic Research and Clinical Translation

“…Similarly, the transcription factor SP1 could regulate KLF4 through a binding site lying in a CpG island in the promoter region of Klf4 [74]. The dentin matrix may be another promising natural biomaterial with which to regenerate the pulpodentinal complex due to its nonimmunogenicity and richness in dentinogenesis-related growth factors [87]. Some possible application forms have been proposed, such as nanostructure demineralised dentin matrix (DDM) treated with different demineralised agents, extracted dentin matrix components, and treated dentin matrix paste (TDMP) [88][89][90][91].…”

Adipose-derived stromal/stem cells are verified to be potential seed candidates for bio-root regeneration in three-dimensional culture

“…The current research results show that the 3D culture system constructed in this study can solve the problems of cell adhesion density and tightness, and promote cell proliferation and differentiation. For one thing, the protein composition of TDM contains cell adhesion-related proteins, such as fibronectin 1, vitronectin, and laminin, to ensure the tight adhesion of cells attached to its surface [ 8 ]. Ultra-low adhesion dishes allow more cells to grow on the surface of pTDM rather than traditional dishes, resulting in most cells adhering to the bottom of the dishes.…”

“…Treated dentin matrix (TDM) from natural tooth tissue has been used as a biological scaffold for bioengineering tooth organs owing to its biological property of providing an odontogenic microenvironment [ 8 , 9 ]. Several studies report the advantages of dental-derived stem cells combined with TDM in bio-root regeneration [ 10 – 14 ].…”

“…3D culture methods include scaffold-free and scaffold-based systems, in which scaffolds include biological scaffolds and synthetic scaffolds [ 19 – 21 ]. TDM, as a biological scaffold material for bio-root construction, provides the mechanical shape of natural root [ 22 ], presents good biocompatibility and bioactivity, and releases abundant important proteins including DSPP, DMP1, COL-1, TGF-β1, which play key roles in tooth development, providing a natural odontogenic 3D induction microenvironment [ 8 , 13 , 15 , 23 ].…”

Treated Dentin Matrix in Tissue Regeneration: Recent Advances

“…It also contains bone morphogenetic proteins (BMPs) [ 39 ]. The results of a shotgun proteomic strategy applied to profile human TDM proteome showed that collagens, proteoglycans, small integrin-binding ligand N-linked glycoproteins and growth factors were identified [ 40 ], which supplements the evidence showing that a TDM contains a similar protein composition to a native dentin matrix.…”