Treated dentin matrix induces odontogenic differentiation of dental pulp stem cells via regulation of Wnt/β-catenin signaling

Abstract

Treated dentin matrix (TDM) is an ideal scaffold material containing multiple extracellular matrix factors. The canonical Wnt signaling pathway is necessary for tooth regeneration. Thus, this study investigated whether the TDM can promote the odontogenic differentiation of human dental pulp stem cells (hDPSCs) and determined the potential role of Wnt/β-catenin signaling in this process. Different concentrations of TDM promoted the dental differentiation of the hDPSCs and meanwhile, the expression of GSK3β was decreased. Of note, the expression of the Wnt/β-catenin pathway-related genes changed significantly in the context of TDM induction, as per RNA sequencing (RNA seq) data. In addition, the experiment showed that new dentin was visible in rat mandible cultured with TDM, and the thickness was significantly thicker than that of the control group. In addition, immunohistochemical staining showed lower GSK3β expression in new dentin. Consistently, the GSK3β knockdown hDPSCs performed enhanced odotogenesis compared with the control groups. However, GSK3β overexpressing could decrease odotogenesis of TDM-induced hDPSCs. These results were confirmed in immunodeficient mice and Wistar rats. These suggest that TDM promotes odontogenic differentiation of hDPSCs by directly targeting GSK3β and activating the canonical Wnt/β-catenin signaling pathway and provide a theoretical basis for tooth regeneration engineering.

Demineralized Dentin Matrix for Dental and Alveolar Bone Tissues Regeneration: An Innovative Scope Review

“… Forms and sizes The TDM/DDM were produced in different forms likely as particles, liquid extract, hydrogel, and paste, and in different shapes such as sheets, slices, chips, blocks, discs, root-shaped, barrier membranes, and press-fitted according to the defect shape. Dentin grinder and ball mill machines were used for preparing particulate dentin [ 17 , 19 , 22 , 25 , 26 , 45 , 49 – 52 , 55 , 56 – 60 , 62 , 65 , 68 , 73 , 74 , 76 , 77 , 80 ]. The diameter of the particle size was measured in either µm or mm.…”

“…There are several demineralizing protocols tested with different study designs. Excluding the size parameter of the dentin matrix, the particles obtained from human permanent teeth were demineralized with consecutive gradient concentrations of 17%, 10%, and 5% EDTA [ 17 , 26 , 49 ] with different time frames. Correspondingly they were 10 min, 5 min, and 10 min [ 17 ], 10 min, 10 min, and 10 min [ 26 ], and 5 min, 5 min, and 10 min [ 49 ].…”

“…Excluding the size parameter of the dentin matrix, the particles obtained from human permanent teeth were demineralized with consecutive gradient concentrations of 17%, 10%, and 5% EDTA [ 17 , 26 , 49 ] with different time frames. Correspondingly they were 10 min, 5 min, and 10 min [ 17 ], 10 min, 10 min, and 10 min [ 26 ], and 5 min, 5 min, and 10 min [ 49 ]. In addition, 0.6 N HCl [ 45 , 50 , 52 , 57 , 65 , 68 , 73 , 77 , 80 ] was used for demineralization and the exposure time was considered once and varied between 15 min [ 77 ], 30 min [ 52 , 73 ] and 12 h [ 50 ].…”

“…Figure 6 presents DDM with opened dentinal tubules after demineralization. Sterilization was performed by maintaining the matrices in sterile phosphate-buffered saline (PBS) with 100 units/ml of penicillin and 100 μg/ml of streptomycin at 37 °C, for 72 h and then washed in sterilized deionized water for 5 min [ 17 , 21 , 22 , 24 , 28 , 30 – 36 , 38 – 44 ], socking into penicillin and streptomycin [ 72 ] only, preservation in 5% peracetic acid and 75% ethanol for 10 min [ 53 , 80 , 81 ], immersion in 5 ml alcohol/2 ml of gentamicin [ 61 , 69 – 71 ], rinsing with sterile saline for 10 min [ 75 ], washing in sterilized deionized water for 10 min [ 82 ], rinsing twice in 0.1 M Tris-HCl (pH 7.4) for 10 min [ 55 , 62 ] and using gradient ethanol concentrations [ 47 ]. Other methods of sterilization were gamma irradiation processing using Cobalt 60 radiation with a dose of 5 kGy [ 66 ], ethylene oxide gas sterilization at low temperatures [ 19 , 45 , 49 , 52 , 57 , 58 , 60 , 67 , 68 , 76 ], steam sterilization at 121°C and a pressure of 1 bar for 15 min [ 23 ], lyophilization and freeze-drying [ 46 ], or mixing was accomplished in a sterile container [ 65 ].…”

“…In addition, slices were stored at 2°C until implantation [ 61 , 69 – 71 ]. Outcome The TDM/DDM were considered for osteogenic differentiation [ 39 , 49 , 50 ] and bone/guided bone regeneration [ 45 , 46 , 49 , 51 , 53 , 55 – 62 , 66 , 69 – 71 , 80 – 82 , 84 ], odontogenic differentiation [ 17 , 40 ], and dentin-pulp tissue regeneration [ 25 , 38 ] (Fig. 7 ), bio-root regeneration [ 18 , 24 , 26 , 27 , 34 ], tooth tissue remodeling and regeneration [ 30 ], socket preservation and alveolar ridge augmentation [ 52 , 55 , 73 , 76 , 82 ], periodontal tissue regeneration [ 32 , 64 , 78 ], furcation perforation repair [ 31 ] and alveolar bone defect regeneration [ 57 ], osteoclastogenesis and osteoclastic resorption [ 28 ], and maxillary sinus augmentation [ 55 , 77 ] (Fig.…”