Laminin-Modified Dental Pulp Extracellular Matrix for Dental Pulp Regeneration

Abstract

Native dental pulp extracellular matrix (DPEM) has proven to be an effective biomaterial for dental pulp regeneration. However, as a significant extracellular matrix glycoprotein, partial laminins were lost during the decellularization process, which were essential for odontoblast differentiation. Thereby, this study investigated the feasibility of LN supplementation to improve the surface of DPEM for odontoblast layer regeneration. The influences of laminin on cell adhesion and odontogenic differentiation were evaluated in vitro. Then, we fabricated laminin-modified DPEM based on the physical coating strategy and observed the location and persistency of laminin coating by immunofluorescent staining. Finally, laminin-modified DPEM combined with treated dentin matrix (TDM) was transplanted in orthotopic jaw bone of beagles (n = 3) to assess the effect of LNs on dental pulp tissue regeneration. The in vitro results showed that laminins could improve the adhesion of dental pulp stem cells (DPSCs) and promoted DPSCs toward odontogenic differentiation. Continuous odontoblastic layer-like structure was observed in laminin-modified DPEM group, expressing the markers for odontoblastogenesis, dentine matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP). Overall, these studies demonstrate that the supplementation of laminins to DPEM contributes to the odontogenic differentiation of cells and to the formation of odontoblast layer in dental pulp regeneration.

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

“…Slices [ 61 , 69 – 71 , 82 ] were 8 µm thickness [ 61 , 69 – 71 ], chips were 1 mm thickness [ 75 ], and blocks [ 79 , 81 , 83 ] were 5 mm diameter and 2 mm thickness [ 79 ] or 5 mm diameter and 2 mm height [ 81 ] or 2–3 mm thickness and 4 mm diameter [ 83 ] whereas discs were 5 mm thickness [ 24 ]. The sizes of the root-shaped matrix [ 18 , 20 , 27 , 28 , 30 , 32 , 34 – 38 , 41 , 43 , 48 ] for human premolars were 10 mm in length and 1.0 mm in thickness [ 18 , 32 , 41 ] or 10 mm in length and 3-5 in mm diameter [ 38 ]. Regarding molars, the average length and width were 13.08 and 8.41 mm, respectively, and then perforated with uniformly distributed thirty pores measuring 1 mm in diameter [ 48 ].…”

“…The human-derived disc-shaped scaffolds were demineralized with 10% EDTA for 3 days, and then successively soaked in 17%, 10%, and 5% for 20 min, 20 min, and 20 min [ 24 ], respectively. Demineralization of the root-shaped matrix obtained from human sources was performed with 17%, 10%, and 5% EDTA for 5 min, 5 min, and 10 min [ 18 , 32 , 35 , 38 , 41 , 43 ] respectively, or for 12 min, 12 min, 20 min [ 27 ], for 30 min, 30 min, 30 min, respectively, or demineralized with 0.34 N HNO 3 for 30 min [ 48 ] whereas those obtained from porcine deciduous incisors were demineralized with 17%, 10%, and 5% EDTA for 20 min, 18 min, and 15 min [ 18 , 28 , 30 ] or for 20 min, 20 min, 10 min [ 34 ], or for 10 min, 10 min, 5 min [ 36 ], respectively, and those obtained from dogs were demineralized with 17%, 10%, and 5% for 12 min, 12 min, and 20 min [ 20 ], or for 8 min, 8 min, 12 min [ 37 ], respectively. Dentin tubes obtained from rat incisors were demineralized with 0.6 N HCl for 3 h [ 84 ].…”

“…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 ]. The matrices were preserved in α-MEM media containing 50 units/ml of penicillin and 50 μg/ml of streptomycin or in 0.1 x PBS and kept in refrigerator at 4 °C [ 18 , 20 , 24 , 27 , 28 , 30 – 32 , 34 , 36 , 39 – 42 , 44 , 47 ] cryopreserved at -196°C [ 38 ], −80 °C [ 26 ], −18 °C [ 83 ], and -20°C [ 25 ] or stored at room temperature [ 45 , 52 , 65 ]. Other methods of preservation included syringes for hydrogel [ 21 ], package for barrier membranes [ 46 ] and particles [ 58 – 60 ], or freshly prepared during operation [ 53 , 55 , 63 ].…”

“…The matrices were preserved in α-MEM media containing 50 units/ml of penicillin and 50 μg/ml of streptomycin or in 0.1 x PBS and kept in refrigerator at 4 °C [ 18 , 20 , 24 , 27 , 28 , 30 – 32 , 34 , 36 , 39 – 42 , 44 , 47 ] cryopreserved at -196°C [ 38 ], −80 °C [ 26 ], −18 °C [ 83 ], and -20°C [ 25 ] or stored at room temperature [ 45 , 52 , 65 ]. Other methods of preservation included syringes for hydrogel [ 21 ], package for barrier membranes [ 46 ] and particles [ 58 – 60 ], or freshly prepared during operation [ 53 , 55 , 63 ].…”

Efficient generation of functional pancreatic β cells from dental-derived stem cells via laminin-induced differentiation

“…There is compelling evidence that indigenous extracellular matrix (ECM) promotes stem cell survival and to regulate stem cell differentiation through maintaining its inherent structural, biochemical, and biomechanical properties [ 28 , 38 , 50 ]. Laminin specifically, which is a major constituent of the basement membrane, plays a role in supporting cell proliferation and differentiation [ 22 ]. During organ development, laminins interplay with several other ECM molecules and connect cells through cell surface receptors, which results in modulating cellular phenotypes and differentiation [ 24 , 51 , 53 ].…”