The integration of 3D bioprinting and stem cells is of great promise in facilitating the reconstruction of cranial defects. However, the effectiveness of the scaffolds has been hampered by the limited cell behavior and functions. Herein, a therapeutic cell-laden hydrogel for bone regeneration is therefore developed through the design of a void-forming hydrogel. This hydrogel is prepared by digital light processing (DLP)-based bioprinting of the bone marrow stem cells (BMSCs) mixed with gelatin methacrylate (GelMA)/dextran emulsion. The 3D-bioprinted hydrogel can not only promote the proliferation, migration, and spreading of the encapsulated BMSCs, but also stimulate the YAP signal pathway, thus leading to the enhanced osteogenic differentiation of BMSCs. In addition, the in vivo therapeutic assessments reveal that the void-forming hydrogel shows great potential for BMSCs delivery and can significantly promote bone regeneration. These findings suggest that the unique 3D-bioprinted void-forming hydrogels are promising candidates for applications in bone regeneration.

Infections to dental pulp commonly result in pulpitis and pulp necrosis, and surgical removal of the infected tissues is the only therapeutic approach. Dental pulp injury remains a challenging medical issue due to the limited regenerative capability of dental pulp. In this work, a dental pulp guidance construct (DPGC) with the instructive niche was bioprinted to mimic native teeth for dentin and neovascular-like structure reconstruction. GelMA-Dextran aqueous emulsion was used as an ink for in situ printing of porous DPGC to induce predominant nuclear localization of Yes-associated protein (YAP) in the encapsulated dental pulp stem cells (DPSCs) and enhance their stemness properties. Furthermore, the DPSCs encapsulated in DPGC with microporous structures exhibited enhanced viability, migration, and spreading. Meanwhile, we found that DPGC could promote capillary tube formation and induce neurogenesis. In a mouse subcutaneous implant model, the DPGC consisted of porous structures, such as odontoblasts and newly formed vascular structures, that mimic dental pulp characteristics. This study demonstrated a new strategy to design DPGC with instructive niche for dental pulp regeneration, presenting a potential treatment alternative to root canal therapy.

Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is limited in hypoxia, and HIF-1α is key to the response to hypoxia. However, its mechanisms remain largely unknown. This study discovered an osteogenesis-related gene sensitive to hypoxia in PDLSCs, and investigated the molecular mechanisms between HIF-1α and the gene. NOG, a gene that negatively regulates osteogenesis, was discovered by RNA-seq. Under normoxic conditions, HIF-1α overexpression led to enhanced expression of NOG/Noggin and inhibited the expression of osteogenesis-related genes, while inhibition of HIF-1α reversed this effect. The expression of HIF-1α, NOG/Noggin and the osteogenesis-related genes were detected by qRT-PCR or Western blot. Mechanistically, we verified that HIF-1α binds to the hypoxia response element (-1505 to -1502) in the promotor of NOG to enhance secretion of Noggin by chromatin immunoprecipitation and a dual-luciferase reporter assay. IHC staining findings in an animal model verified that Noggin-associated osteogenic differentiation was inhibited in hypoxia. NOG displayed a concordant relationship with HIF-1α, and secreted more with increasing of HIF-1α. Hypoxia stabilized HIF-1α, which bound to the HRE (-1505 to -1502) of the NOG promotor to enhance NOG transcription resulted in inhibiting osteogenic differentiation of PDLSCs. This study offers a promising therapy for periodontitis.

Repairing defects in alveolar bone is essential for regenerating periodontal tissue, but it is a formidable challenge. One promising therapeutic approach involves using a strategy that specifically recruits periodontal ligament cells (PDLCs) with high regenerative potential to achieve in situ regeneration of alveolar bone. In this study, we have created a new type of microsphere conjugated with an antibody to target p75 neurotrophin receptor (p75NTR), which is made of nano-hydroxyapatite (nHA) and chitosan (CS). The goal of this design is to attract p75NTR⁺hPDLCs selectively and promote osteogenesis. In vitro experiments demonstrated that the antibody-conjugated microspheres attracted significantly more PDLCs compared to non-conjugated microspheres. Incorporating nHA not only enhances cell adhesion and proliferation on the surface of the microsphere but also augments its osteoinductive properties. Microspheres effectively recruited p75NTR⁺ cells at bone defect sites in SD rats, as observed through immunofluorescent staining of p75NTR antibodies. This p75NTR antibody-conjugated nHA/CS microsphere presents a promising approach for selectively recruiting cells and repairing bone defects.

Correct torque of the incisors is beneficial in the assessment of the effects of orthodontic treatment. However, evaluating this process effectively remains a challenge. Improper anterior teeth torque angle can cause bone fenestrations and exposure of the root surface. A three-dimensional finite element model of the maxillary incisor torque controlled by a homemade four-curvature auxiliary arch was established. The four-curvature auxiliary arch placed on the maxillary incisors was divided into four different state groups, among which 2 groups had tooth extraction space retracted traction force set to 1.15 N. Initial displacements and pressure stresses of the periodontal tissue in the maxillary incisors and molars were calculated after torque forces (0.5, 1, 1.5, and 2 N) were applied to the teeth at different stable states. The effect of using the four-curvature auxiliary arch on the incisors was significant but did not affect the position of the molars. Given the absence of tooth extraction space, when the four-curvature auxiliary arch was used in conjunction with absolute anchorage, the recommended force value was < 1.5 N. In the other 3 groups (i.e., molar ligation, molar retraction, and microimplant retraction groups), the recommended force value was < 1 N. The application of a four-curvature auxiliary arch did not influence the molar periodontal and displacement. A four-curvature auxiliary arch may treat severely upright anterior teeth and correct cortical fenestrations of the bone and root surface exposure.