Demineralized Dentin Matrix Particle-Based Bio-Ink for Patient-Specific Shaped 3D Dental Tissue Regeneration.

Demineralized dentin matrix (DDM)-based materials have been actively developed and are well-known for their excellent performance in dental tissue regeneration. However, DDM-based bio-ink suitable for fabrication of engineered dental tissues that are patient-specific in terms of shape and size, has not yet been developed. In this study, we developed a DDM particle-based bio-ink (DDMp bio-ink) with enhanced three-dimensional (3D) printability. The bio-ink was prepared by mixing DDM particles and a fibrinogen-gelatin mixture homogeneously. The effects of DDMp concentration on the 3D printability of the bio-ink and dental cell compatibility were investigated. As the DDMp concentration increased, the viscosity and shear thinning behavior of the bio-ink improved gradually, which led to the improvement of the ink's 3D printability. The higher the DDMp content, the better were the printing resolution and stacking ability of the 3D printing. The printable minimum line width of 10% w/v DDMp bio-ink was approximately 252 µm, whereas the fibrinogen-gelatin mixture was approximately 363 µm. The ink's cytocompatibility test with dental pulp stem cells (DPSCs) exhibited greater than 95% cell viability. In addition, as the DDMp concentration increased, odontogenic differentiation of DPSCs was significantly enhanced. Finally, we demonstrated that cellular constructs with 3D patient-specific shapes and clinically relevant sizes could be fabricated through co-printing of polycaprolactone and DPSC-laden DDMp bio-ink.

In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and ß-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells.

3D printed biomaterials have been extensively investigated and developed in the field of bone regeneration related to clinical issues. However, specific applications of 3D printed biomaterials in different dental areas have seldom been reported. In this study, we aimed to and successfully fabricated 3D poly (lactic-co-glycolic acid)/ß-tricalcium phosphate (3D-PLGA/TCP) and 3D ß-tricalcium phosphate (3D-TCP) scaffolds using two relatively distinct 3D printing (3DP) technologies. Conjunctively, we compared and investigated mechanical and biological responses on human dental pulp stem cells (hDPSCs). Physicochemical properties of the scaffolds, including pore structure, chemical elements, and compression modulus, were characterized. hDPSCs were cultured on scaffolds for subsequent investigations of biocompatibility and osteoconductivity. Our findings indicate that 3D printed PLGA/TCP and ß-tricalcium phosphate (ß-TCP) scaffolds possessed a highly interconnected and porous structure. 3D-TCP scaffolds exhibited better compressive strength than 3D-PLGA/TCP scaffolds, while the 3D-PLGA/TCP scaffolds revealed a flexible mechanical performance. The introduction of 3D structure and ß-TCP components increased the adhesion and proliferation of hDPSCs and promoted osteogenic differentiation. In conclusion, 3D-PLGA/TCP and 3D-TCP scaffolds, with the incorporation of hDPSCs as a personalized restoration approach, has a prospective potential to repair minor and critical bone defects in oral and maxillofacial surgery, respectively.

Bioprinting of three-dimensional dentin-pulp complex with local differentiation of human dental pulp stem cells.

Numerous approaches have been introduced to regenerate artificial dental tissues. However, conventional approaches are limited when producing a construct with three-dimensional patient-specific shapes and compositions of heterogeneous dental tissue. In this research, bioprinting technology was applied to produce a three-dimensional dentin-pulp complex with patient-specific shapes by inducing localized differentiation of human dental pulp stem cells within a single structure. A fibrin-based bio-ink was designed for bioprinting with the human dental pulp stem cells. The effects of fibrinogen concentration within the bio-ink were investigated in terms of printability, human dental pulp stem cell compatibility, and differentiation. The results show that micro-patterns with human dental pulp stem cells could be achieved with more than 88% viability. Its odontogenic differentiation was also regulated according to the fibrinogen concentration. Based on these results, a dentin-pulp complex having patient-specific shape was produced by co-printing the human dental pulp stem cell-laden bio-inks with polycaprolactone, which is a bio-thermoplastic used for producing the overall shape. After culturing with differentiation medium for 15 days, localized differentiation of human dental pulp stem cells in the outer region of the three-dimensional cellular construct was successfully achieved with localized mineralization. This result demonstrates the possibility to produce patient-specific composite tissues for tooth tissue engineering using three-dimensional bioprinting technology.