In vitro remineralization by various ion-releasing materials of artificially demineralized dentin: A micro-CT study.

OBJECTIVE: The aim of this study was to evaluate the remineralizing properties of ion-releasing restorative materials on pH cycling-induced carious dentin. METHODS: Fifty sound molars were freshly extracted. The occlusal surfaces were abraded using water-cooled sandpaper (800 grit). The residual crowns were embedded in self-cured acrylic resin with the flat dentin surface exposed. A mesio-distal trench was created using a calibrated 0.5 mm deep occlusal reduction burr, and artificial dentin caries were generated by pH cycling. Then, teeth were randomly assigned to five groups according to the ion-releasing material used. For each sample, micro-CT acquisitions were performed at various intervals. Remineralization was assessed by mean gray value (MGV) measurements after registration and segmentation of the region of interest with 3D Slicer software. One-way repeated-measures ANOVA followed by Tukey's post hoc test was used to investigate the difference in MGVs among the various groups. RESULTS: Only Cention Forte showed significantly increased MGVs after 4 weeks compared to demineralized dentin. MGVs were higher, but not significantly, after placement of the restorative materials, including in the resin composite control group. These results can be explained by the radiopacity of the materials. SIGNIFICANCE: Cention Forte, the material with the highest radiopacity, showed a significant increase in the MGVs of artificially carious dentin after 4 weeks. However, the study of dentin remineralization by micro-CT could be impacted by the radiopacity of the restorative materials used. The relevance of this examination for the study of dentinal remineralization should be investigated.

Evaluation of the Impact of Calcium Silicate-Based Sealer Insertion Technique on Root Canal Obturation Quality: A Micro-Computed Tomography Study.

BACKGROUND: Calcium silicate-based sealers have gained in popularity over time due to their physicochemical/biological properties and their possible use with single-cone obturation. The single cone technique is a sealer-based obturation and there is still a knowledge gap regarding the potential impact of the sealer insertion method on the root canal-filling quality. Therefore, the aim of this micro-CT study was to assess the impact of the calcium silicate-based sealer insertion technique on void occurrence and on the sealer extrusion following single-cone obturation. METHODS: Thirty-six single-rooted mandibular premolars with one canal were shaped with Reciproc® R25 (VDW, Munich, Germany) then divided randomly into four groups of nine canals, each depending on the TotalFill® BC Sealer insertion technique used with single cone obturation: injection in the coronal two-thirds (group A); injection in the coronal two-thirds followed by direct sonic activation (group B); injection in the coronal two-thirds followed by indirect ultrasonic activation on tweezers (group C); sealer applied only on the master-cone (control group D). Samples were then scanned using micro-CT for voids and sealer extrusion calculation. Data were statistically analyzed using v.26 IBM; Results: No statistically significant differences were noted between the four groups in terms of voids; nevertheless, sonic activation (group B) followed by ultrasonic activation on the tweezers (group C) showed the best results (p = 0.066). Group D showed significantly less sealer extrusion when compared with group C (p = 0.044), with no statistically significant differences between groups D, A and B (p > 0.05). CONCLUSIONS: Despite no significant differences observed between the different sealer placement techniques, the use of sonic and ultrasonic activation might be promising to reduce void occurrence. Further investigations are needed to demonstrate the potential benefit of calcium silicate-based sealer activation especially in wide and oval root canals in order to improve the quality of the single-cone obturation.

NLRP3 Is Involved in Neutrophil Mobilization in Experimental Periodontitis.

The NLRP3 inflammasome is overexpressed in gingiva of periodontitis patients but its role remains unclear. In our study, we use a periodontitis mouse model of ligature, impregnated or not with Porphyromonas gingivalis, in WT or NLRP3 KO mice. After 28 days of induction, ligature alone provoked exacerbated periodontal destruction in KO mice, compared to WT mice, with an increase in activated osteoclasts. No difference was observed at 14 days, suggesting that NLRP3 is involved in regulatory pathways that limit periodontitis. In contrast, in the presence of P. gingivalis, this protective effect of NLRP3 was not observed. Overexpression of NLRP3 in connective tissue of WT mice increased the local production of mature IL-1ß, together with a dramatic mobilization of neutrophils, bipartitely distributed between the site of periodontitis induction and the alveolar bone crest. P. gingivalis enhanced the targeting of NLRP3-positive neutrophils to the alveolar bone crest, suggesting a role for this subpopulation in bone loss. Conversely, in NLRP3 KO mice, mature IL-1ß expression was lower and almost no neutrophils were mobilized. Our study sheds new light on the role of NLRP3 in periodontitis by highlighting the ambiguous role of neutrophils, and P. gingivalis which affects NLRP3 functions.

Combining sclerostin neutralization with tissue engineering: An improved strategy for craniofacial bone repair.

Scaffolds associated with different types of mesenchymal stromal stem cells (MSC) are extensively studied for the development of novel therapies for large bone defects. Moreover, monoclonal antibodies have been recently introduced for the treatment of cancer-associated bone loss and other skeletal pathologies. In particular, antibodies against sclerostin, a key player in bone remodeling regulation, have demonstrated a real benefit for treating osteoporosis but their contribution to bone tissue-engineering remains uncharted. Here, we show that combining implantation of dense collagen hydrogels hosting wild-type (WT) murine dental pulp stem cells (mDPSC) with weekly systemic injections of a sclerostin antibody (Scl-Ab) leads to increased bone regeneration within critical size calvarial defects performed in WT mice. Furthermore, we show that bone formation is equivalent in calvarial defects in WT mice implanted with Sost knock-out (KO) mDPSC and in Sost KO mice, suggesting that the implantation of sclerostin-deficient MSC similarly promotes new bone formation than complete sclerostin deficiency. Altogether, our data demonstrate that an antibody-based therapy can potentialize tissue-engineering strategies for large craniofacial bone defects and urges the need to conduct research for antibody-enabled local inhibition of sclerostin. STATEMENT OF SIGNIFICANCE: The use of monoclonal antibodies is nowadays broadly spread for the treatment of several conditions including skeletal bone diseases. However, their use to potentialize tissue engineering constructs for bone repair remains unmet. Here, we demonstrate that the neutralization of sclerostin, through either a systemic inhibition by a monoclonal antibody or the implantation of sclerostin-deficient mesenchymal stromal stem cells (MSC) directly within the defect, improves the outcome of a tissue engineering approach, combining dense collagen hydrogels and MSC derived from the dental pulp, for the treatment of large craniofacial bone defects.

Morphological variation of the deciduous second molars in the Baka Pygmies.

The Baka Pygmies are known for their short stature resulting from a reduced growth rate during infancy. They are peculiar also for their teeth erupt earlier than in any other African population, and their posterior dentition is larger than in non-Pygmy populations. However, the Baka's dental morphology, like several other aspects of their biology, is still understudied. Here, we explore the variation of the Baka's deciduous upper and lower second molars (dm2s) in comparison to a geographically heterogeneous human sample by means of 3D geometric morphometrics and analysis of dental traits. Our results show that the different populations largely overlap based on the shape of their dm2s, especially the lower ones. Their distal region and the height of the dentinal crown differ the most, with the Baka showing the most extreme range of variation. Upper and lower dm2s covary to a great extent (RV = 0.82). The Baka's and South Americans' dm2s were confirmed among the largest in our sample. Despite the Baka's unique growth pattern, long-lasting isolation, and extreme dental variation, it is not possible to distinguish them from other populations based on their dm2s' morphology only.

Priming Dental Pulp Stem Cells from Human Exfoliated Deciduous Teeth with Fibroblast Growth Factor-2 Enhances Mineralization Within Tissue-Engineered Constructs Implanted in Craniofacial Bone Defects.

The craniofacial area is prone to trauma or pathologies often resulting in large bone damages. One potential treatment option is the grafting of a tissue-engineered construct seeded with adult mesenchymal stem cells (MSCs). The dental pulp appears as a relevant source of MSCs, as dental pulp stem cells display strong osteogenic properties and are efficient at bone formation and repair. Fibroblast growth factor-2 (FGF-2) and/or hypoxia primings were shown to boost the angiogenesis potential of dental pulp stem cells from human exfoliated deciduous teeth (SHED). Based on these findings, we hypothesized here that these primings would also improve bone formation in the context of craniofacial bone repair. We found that both hypoxic and FGF-2 primings enhanced SHED proliferation and osteogenic differentiation into plastically compressed collagen hydrogels, with a much stronger effect observed with the FGF-2 priming. After implantation in immunodeficient mice, the tissue-engineered constructs seeded with FGF-2 primed SHED mediated faster intramembranous bone formation into critical size calvarial defects than the other groups (no priming and hypoxia priming). The results of this study highlight the interest of FGF-2 priming in tissue engineering for craniofacial bone repair. Stem Cells Translational Medicine 2019;8:844&857.

Characterization of two rat models of cystic fibrosis-KO and F508del CFTR-Generated by Crispr-Cas9.

BACKGROUND: Genetically engineered animals are essential for gaining a proper understanding of the disease mechanisms of cystic fibrosis (CF). The rat is a relevant laboratory model for CF because of its zootechnical capacity, size, and airway characteristics, including the presence of submucosal glands. METHODS: We describe the generation of a CF rat model (F508del) homozygous for the p.Phe508del mutation in the transmembrane conductance regulator (Cftr) gene. This model was compared to new Cftr -/- rats (CFTR KO). Target organs in CF were examined by histological staining of tissue sections and tooth enamel was quantified by micro-computed tomography. The activity of CFTR was evaluated by nasal potential difference (NPD) and short-circuit current measurements. The effect of VX-809 and VX-770 was analyzed on nasal epithelial primary cell cultures from F508del rats. RESULTS: Both newborn F508del and Knock out (KO) animals developed intestinal obstruction that could be partly compensated by special diet combined with an osmotic laxative. The two rat models exhibited CF phenotypic anomalies such as vas deferens agenesis and tooth enamel defects. Histology of the intestine, pancreas, liver, and lungs was normal. Absence of CFTR function in KO rats was confirmed ex vivo by short-circuit current measurements on colon mucosae and in vivo by NPD, whereas residual CFTR activity was observed in F508del rats. Exposure of F508del CFTR nasal primary cultures to a combination of VX-809 and VX-770 improved CFTR-mediated Cl- transport. CONCLUSIONS: The F508del rats reproduce the phenotypes observed in CFTR KO animals and represent a novel resource to advance the development of CF therapeutics.

Tissue-specific mineralization defects in the periodontium of the Hyp mouse model of X-linked hypophosphatemia.

X-linked hypophosphatemia (XLH) is a dento-osseous disorder caused by inactivating mutations in the PHEX gene, leading to renal phosphate wasting and hypophosphatemia, and impaired mineralization of bones and teeth. In the oral cavity, recent reports suggest a higher susceptibility of XLH patients to periodontitis, where patients present with impaired tooth cementum - a bone-like tissue involved in tooth attachment to the jaw bones and post-eruption tooth positioning - and a higher frequency of intrabony defects. In the present study, the pathobiology of alveolar bone and tooth cementum was investigated in the Hyp mouse, the murine analog of XLH. PHEX deficiency in XLH/Hyp dramatically alters the periodontal phenotype, with hypoplasia of tooth root cementum associated with a lack of periodontal ligament attachment and the presence of an immature apatitic mineral phase of all periodontal mineralized tissues. Challenging the Hyp periodontium in two surgical experimental models - ligature-induced periodontal breakdown and repair, and a model of tooth movement adaptation inducing cementum formation - we show that bone and cementum formation, and their healing, are altered. Bone and cementum mineralization appear similarly disturbed, where hypomineralized pericellular matrix surrounds cells, and where the protein osteopontin (OPN, a mineralization inhibitor) accumulates in a tissue-specific manner, most notably in the perilacunar matrix surrounding osteocytes. Although the pathobiology is different between XLH/Hyp bone and cementum, our results show a major XLH phenotype in oral mineralized tissues consistent with variations in patient susceptibility to periodontal disorders.