Targeting class A GPCRs for hard tissue regeneration.

G protein-coupled receptors (GPCRs) play important roles in various pathogeneses and physiological regulations. Owing to their functional diversity, GPCRs are considered one of the primary pharmaceutical targets. However, drugs targeting GPCRs have not been developed yet to regenerate hard tissues such as teeth and bones. Mesenchymal stromal cells (MSCs) have high proliferation and multi-lineage differentiation potential, which are essential for hard tissue regeneration. Here, we present a strategy for targeting class A GPCRs for hard tissue regeneration by promoting the differentiation of endogenous MSCs into osteogenic and odontogenic progenitor cells. Through in vitro screening targeted at class A GPCRs, we identified six target receptors (LPAR1, F2R, F2RL1, F2RL2, S1PR1, and ADORA2A) and candidate drugs with potent biomineralization effects. Through a combination of profiling whole transcriptome and accessible chromatin regions, we identified that p53 acts as a key transcriptional activator of genes that modulate the biomineralization process. Moreover, the therapeutic potential of class A GPCR-targeting drugs was demonstrated in tooth pulpotomy and calvarial defect models. The selected drugs revealed potent regenerative effects in both tooth and bone defects, represented by newly formed highly mineralized regions. Consequently, this study provides translational evidence for a new regenerative strategy for damaged hard tissue.

Functional expression of oxytocin receptors in pulp-dentin complex.

Dental pulp-derived stromal cells (DPSCs) are a crucial cell population for maintaining the tissue integrity of the pulp-dentin complex. The oxytocin receptor (OXTR), a member of the G protein-coupled receptor (GPCR) superfamily, plays versatile roles in diverse biological contexts. However, the role of OXTR in dental pulp has not yet been fully understood. Here, we demonstrate the biological functions and significance of OXTR in DPSCs through a multidisciplinary approach. Microarray data of 494 GPCR genes revealed high OXTR expression in human DPSCs (hDPSCs). Blocking OXTR activity increased the expression of osteogenic and odontogenic marker genes, promoting hDPSC differentiation. Additionally, we found that OXTR is involved in extracellular matrix (ECM) remodeling through the regulation of the gene expression related to ECM homeostasis. We further demonstrated that these genetic changes are mediated by trascriptional activity of Yes-associated protein (YAP). Based on the results, a preclinical experiment was performed using an animal model, demonstrating that the application of an OXTR inhibitor to damaged pulp induced significant hard tissue formation. These results provide new insight into the oxytocin-OXTR system in the regenerative process of pulp-dentin complex.

Low-intensity pulsed ultrasound attenuates replacement root resorption of avulsed teeth stored in dry condition in dogs.

This study aimed to investigate the effects of low-intensity pulsed ultrasound (LIPUS) on replacement root resorption after replantation of avulsed teeth stored in a dry condition in dogs. A total of 73 premolar roots from four male mongrel dogs were intentionally avulsed with forceps and divided into four groups-HN, HL, DN, and DL-according to storage conditions and whether or not they received LIPUS treatment. Thirty-eight roots were kept in Hanks' Balanced Salt Solution for 30 min (HN and HL groups), whereas the remaining 35 roots were left to dry in the air for an hour (DN and DL groups) prior to replantation. Following replantation, the roots in the HL and DL groups (21 and 18 roots, respectively) received a 20-min daily LIPUS treatment for 2 weeks. The animals were euthanized 4 weeks after the operation. Micro-computed tomography images were acquired for each root and the amount of replacement root resorption was measured three-dimensionally. Histological assessments were also carried out. There was significantly less replacement root resorption for the roots in the DL group compared to the DN group (p < 0.01). Histological findings in the DN group demonstrated evident replacement root resorption, whereas the DL group revealed less severe resorption compared to the DN group. Within the limitations, these results suggest that LIPUS could attenuate the replacement resorption of avulsed teeth stored in a dry condition, thereby improving their prognosis.

Color and Translucency Stability of Three-Dimensional Printable Dental Materials for Crown and Bridge Restorations.

The purpose of this study was to examine and compare color and translucency stability of three-dimensional (3D) printable dental materials for crown and bridge restorations. Five different materials were investigated, and twelve disc-shaped specimens of two different thicknesses (1 and 2 mm) were prepared using a digital light processing 3D printer. Color measurements were made according to the CIELAB color scale (L*, a*, and b*) using a spectrophotometer 1 h, 1 day, 1 week, one month, and six months after post-curing of the materials, and the translucency parameter (TP) was calculated. The L*, a*, b*, and TP values were compared among the different materials and storage periods using repeated measures analysis of variance. Color and translucency changes of the specimens after the different storage periods were compared with 1 h measurements to determine whether they exceeded clinically perceivable thresholds. The L*, a*, b*, and TP values showed significant differences according to the storage periods, as well as among the materials. Until one month, some materials demonstrated distinct color differences, while others showed small color differences below a clinically perceivable threshold. The translucency differences were not clinically perceivable for any specimen. After six months, all specimens demonstrated large color changes, whereas the changes in translucency were relatively small. In conclusion, the color of 3D printable dental materials changed with time, and the differences varied with the materials used. On the contrary, the changes in translucency were small. Overall, the materials became darker, more yellowish, and more opaque after six months of water storage.

Effects of Post-Curing Time on the Mechanical and Color Properties of Three-Dimensional Printed Crown and Bridge Materials.

Three-dimensional (3D) printing is increasingly being utilized in the dental field. After fabricating a prosthesis using a 3D printed resin, a post-curing process is required to improve its mechanical properties, but there has been insufficient research on the optimal post-curing conditions. We used various 3D printed crown and bridge materials in this study, and evaluated the changes in their properties according to post-curing time by evaluating the flexural strength, Weibull modulus, Vickers hardness, color change, degree of conversion, and biocompatibility. The obtained results confirmed that the strength of the 3D printed resin increased when it was post-cured for 60-90 min. The Vickers hardness, the degree of conversion, and biocompatibility of the 3D printed resins increased significantly around the beginning of the post-curing time, and then increased more gradually as the post-curing time increased further. It was observed that the color tone also changed as the post-curing time increased, with some groups showing a ΔE00 value of ≥ 2.25, which can be recognized clinically. This study has confirmed that, after the printing process of a 3D printed resin was completed, a sufficient post-curing time of at least 60 min is required to improve the overall clinical performance of the produced material.

Incorporation of zwitterionic materials into light-curable fluoride varnish for biofilm inhibition and caries prevention.

We incorporated zwitterionic materials into light-curable fluoride varnish (LCFV) in order to inhibit biofilm accumulation and prevent dental caries, and the properties of LCFV with three different zwitterionic materials, namely, 2-methacryloyloxyethyl phosphorylcholine (MPC), carboxybetaine methacrylate (CBMA), and sulfobetaine methacrylate (SBMA) polymers (each at a weight percentage of 3%), were compared; unmodified LCFV without any zwitterionic material was used as a control. Material properties including film thickness and degree of conversion (DC) of each type of LCFV were evaluated. In addition, protein-repellent effects and inhibitory effects on Streptococcus mutans adhesion and saliva-derived biofilm accumulation of LCFV were estimated. Finally, the preventive effect of LCFV on enamel demineralization was assessed in vitro on extracted human teeth specimens stored in S. mutans-containing medium. The film thickness of LCFV significantly decreased with the incorporation of zwitterionic materials compared to the control LCFV, whereas there were no significant differences in the DC among all of the LCFV groups. Furthermore, the amount of adsorbed protein, adherent S. mutans colony-forming unit (CFU) counts, and saliva-derived biofilm thickness and biomass were all significantly lower for LCFV with incorporated zwitterionic materials compared with the control. All LCFV groups including the control showed certain preventive effects against enamel demineralization during a 14-day immersion in the medium with S. mutans and sucrose, and the depth of demineralization was significantly lower in LCFV with zwitterionic materials than in the control. Thus, the incorporation of zwitterionic materials such as MPC, CBMA, and SBMA appears to confer superior antifouling effects to LCFV.