Cervical wear pathobiology by robot-simulated 3-year toothbrushing - New methodological approach.

OBJECTIVES: An ex-vivo study was aimed at (i) programming clinically validated robot three-year random toothbrushing, (ii) evaluating cervical macro- and microwear patterns on all tooth groups of different functional age, (iii) documenting and codificating wear related morphological features at the cemento-enamel junction in young teeth and on roots in older teeth. DESIGN: Following ethical approval random toothbrushing (44 strokes per tooth horizontally, rotating, vertically; 2x/d) with manual toothbrushes and low-abrasive dentifrice was performed in an artificial oral cavity with brushing-force 3.5 N on 14 extracted human teeth. Morphological features were examined by SEM at baseline and after simulated 3 years using the replication technique. 3D-SEM analyses were carried out with a four-quadrant back scattered electron detector. Wilcoxon-Mann-Whitney-test was used for statistical analyses. RESULTS: 3-year random toothbrushing with horizontal, rotating and vertical brushing movements revealed morphological features classified as four enamel patterns, one dentin pattern and three cervical patterns. Negative impacts were enamel, cementum and dentin loss. Positive impact on oral health was removing dental calculus and straightening cervical traumatic and iatrogenic damages. The volume loss varied from x̅=34.25nl to x̅=87.75nl. Wear extended apically from 100 to 1500 micrometres. CONCLUSION: Robot simulated toothbrushing in an artificial oral cavity, with subsequent SEM and 3D-SEM assessment, elucidated both negative and oral health-contributing micromorphology patterns of cervical wear after simulated 3-year random toothbrushing. Cervical macro- and microwear of cementum revealed, for the first time, what we describe as overhanging enamel peninsulas and enamel islands on roots in young teeth, but no enamel islands on roots from older teeth after root cementum loss. In contrast, many older teeth exhibited enamel peninsulas.

Lead exposure across the life course and age at death.

BACKGROUND: Lead (Pb) exposure has been associated with an increased risk of all-cause mortality, even at low levels. Little is known about how the timing of Pb exposure throughout life may influence these relationships. Quantifying the amount of Pb present in various tissues of the body provides measurements of exposure from different periods of life. These include bone, tooth enamel, which is the hard outer layer of the crown, and tooth cementum, which is the calcified connective tissue covering the tooth root. The purpose of the study was to examine Pb exposure at multiple periods throughout life, including childhood (enamel), adulthood (cementum), and later life (bone), and to estimate their associations with age at death. METHODS: 208 skeleton donors (born 1910-1960) from an ongoing case-control study were included in this study. Pb was measured in tibia (shin), bone using X-Ray Florescence and in teeth using Laser-Ablation Inductively Coupled Plasma Mass Spectroscopy. After excluding unusually high measurements (>2sd), this resulted in a final sample of 111 with all exposure measures. Correlations across measures were determined using partial Spearman correlations. Associations between Pb exposure and age at death were estimated using Multivariable Linear Regression. RESULTS: Pb measures across exposure periods were all significantly correlated, with the highest correlation between cementum and tibia measures (r = 0.61). Donors were largely female (63.0 %), White (97.3 %), and attended some college (49.5 %). Single exposure models found that higher tooth cementum Pb (-1.27; 95 % CI: -2.48, -0.06) and tibia bone Pb (-0.91; 95 % CI: -1.67, -0.15) were significantly associated with an earlier age at death. When considered simultaneously, only cementum Pb remained significant (-1.51; 95 % CI: -2.92, -0.11). Secondary analyses suggest that the outer cementum Pb may be especially associated with an earlier age at death. CONCLUSION: Results suggest that higher Pb exposure is associated with an earlier age at death, with adulthood as the life period of most relevance. Additional studies using Pb exposure measures from different life stages should be conducted.

Biological response to endodontic treatment in one versus two-visit: a systematic review and meta-analysis of animal studies.

OBJECTIVES: To perform a systematic review of animal studies that compared the histopathological characteristics between teeth with apical periodontitis after endodontic treatment in one or two visits. MATERIALS AND METHODS: This systematic review was registered on the International Prospective Register of Systematic Reviews (PROSPERO) - CRD42022340849. Studies were collected from PubMed/MEDLINE, LILACS, EMBASE, Livivo, SciELO, Web of Science, Scopus, and Cochrane Library and manual and gray literature searches. Animal studies that evaluated histological characteristics after endodontic treatment of teeth with apical periodontitis in one or two visits were included. Risk of bias analysis of the included studies was performed using the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE) tool. Data synthesis of the included studies with quantitative data was performed, and meta-analysis was conducted with the Comprehensive Meta-Analysis software, using the random effects model and odds ratio (OR). RESULTS: Eighteen studies met the inclusion criteria (Kappa = 0.891). Meta-analyses indicated values in inflammatory infiltrate intensity with effect size of 5.5% (95% CI: 0.020-0.148; p < 0.001), periodontal ligament thickness: 25.6% (95% CI: 0.134-0.487; p < 0.001), dentin resorption: 13% (95% CI: 0.015-1.141; p = 0.066), cementum resorption: 7.1% (95% CI: 0.015-0.325; p = 0.001), bone resorption: 1.4% (95% CI: 0.002-0.130; p < 0.001), mineralized tissue resorption: 42.8% (95% CI: 0.110-1.671; p = 0.222), biological apical sealing: 13.1% (95% CI: 0.055-0.314; p < 0.001), and presence of microorganisms: 10.3% (95% CI: 0.014-0.747; p = 0.025). CONCLUSIONS: When considering animal studies, the two-visit endodontic treatment, using calcium hydroxide-based intracanal medication, resulted in better biological repair characteristics. CLINICAL RELEVANCE: A two-visit endodontic treatment with calcium hydroxide-based intracanal medication yields superior histopathological outcomes.

Lithiated porous silicon nanowires stimulate periodontal regeneration.

Periodontal disease is a significant burden for oral health, causing progressive and irreversible damage to the support structure of the tooth. This complex structure, the periodontium, is composed of interconnected soft and mineralised tissues, posing a challenge for regenerative approaches. Materials combining silicon and lithium are widely studied in periodontal regeneration, as they stimulate bone repair via silicic acid release while providing regenerative stimuli through lithium activation of the Wnt/ß-catenin pathway. Yet, existing materials for combined lithium and silicon release have limited control over ion release amounts and kinetics. Porous silicon can provide controlled silicic acid release, inducing osteogenesis to support bone regeneration. Prelithiation, a strategy developed for battery technology, can introduce large, controllable amounts of lithium within porous silicon, but yields a highly reactive material, unsuitable for biomedicine. This work debuts a strategy to lithiate porous silicon nanowires (LipSiNs) which generates a biocompatible and bioresorbable material. LipSiNs incorporate lithium to between 1% and 40% of silicon content, releasing lithium and silicic acid in a tailorable fashion from days to weeks. LipSiNs combine osteogenic, cementogenic and Wnt/ß-catenin stimuli to regenerate bone, cementum and periodontal ligament fibres in a murine periodontal defect.

Acellular Extrinsic Fiber Cementum Is Invariably Present in the Superficial Layer of Apical Cementum in Mouse Molar.

The cementum is a highly mineralized tissue that covers the tooth root. The regional differences among the types of cementum, especially in the extrinsic fibers that contribute to tooth support, remain controversial. Therefore, this study used second harmonic generation imaging in conjunction with automated collagen extraction and image analysis algorithms to facilitate the quantitative examination of the fiber characteristics and the changes occurring in these fibers over time. Acellular extrinsic fiber cementum (AEFC) was invariably observed in the superficial layer of the apical cementum in mouse molars, indicating that this region of the cementum plays a crucial role in supporting the tooth. The apical AEFC exhibited continuity and fiber characteristics comparable with the cervical AEFC, suggesting a common cellular origin for their formation. The cellular intrinsic fiber cementum present in the inner layer of the apical cementum showed consistent growth in the apical direction without layering. This study highlights the dynamic nature of the cementum in mouse molars and underscores the requirement for re-examining its structure and roles. The findings of the present study elucidate the morphophysiological features of cementum and have broader implications for the maintenance of periodontal tissue health.

Hyaluronic acid enhances cell migration, viability, and mineralized tissue-specific genes in cementoblasts.

BACKGROUND/OBJECTIVES: It has been repeatedly demonstrated that cementum formation is a crucial step in periodontal regeneration. Hyaluronic acid (HA) is an important component of the extracellular matrix which regulates cells functions and cell-cell communication. Hyaluronic acid/derivatives have been used in regenerative periodontal therapy, but the cellular effects of HA are still unknown. To investigate the effects of HA on cementoblast functions, cell viability, migration, mineralization, differentiation, and mineralized tissue-associated genes and cementoblast-specific markers of the cementoblasts were tested. MATERIALS AND METHODS: Cementoblasts (OCCM-30) were treated with various dilutions (0, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128) of HA and examined for cell viability, migration, mineralization, and gene expressions. The mRNA expressions of osteocalcin (OCN), runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP), collagen type I (COL-I), alkaline phosphatase (ALP), cementum protein-1 (CEMP-1), cementum attachment protein (CAP), and small mothers against decapentaplegic (Smad) -1, 2, 3, 6, 7, ß-catenin (Ctnnb1) were performed with real-time polymerase chain reaction (RT-PCR). Total RNA was isolated on days 3 and 8, and cell viability was determined using MTT assay on days 1 and 3. The cell mineralization was evaluated by von Kossa staining on day 8. Cell migration was assessed 2, 4, 6, and 24 hours following exposure to HA dilutions using an in vitro wound healing assay (0, 1:2, 1:4, 1:8). RESULTS: At dilution of 1:2 to 1:128, HA importantly increased cell viability (p < .01). HA at a dilution of 1/2 increased wound healing rates after 4 h compared to the other dilutions and the untreated control group. Increased numbers of mineralized nodules were determined at dilutions of 1:2, 1:4, and 1:8 compared with control group. mRNA expressions of mineralized tissue marker including COL-I, BSP, RunX2, ALP, and OCN significantly improved by HA treatments compared with control group both on 3 days and on 8 days (p < .01). Smad 2, Smad 3, Smad 7, and ß-catenin (Ctnnb1) mRNAs were up-regulated, while Smad1 and Smad 6 were not affected by HA administration. Additionally, HA at dilutions of 1:2, 1:4, and 1:8 remarkably enhanced CEMP-1 and CAP expressions in a dilution- and time-dependent manner (p < .01). CONCLUSIONS: The present results have demonstrated that HA affected the expression of both mineralized tissue markers and cementoblast-specific genes. Positive effects of HA on the cementoblast functions demonstrated that HA application may play a key role in cementum regeneration.

Cemental and alveolar bone defects after chronic exposure to amoxicillin in rats (histopathologic and radiographic study).

OBJECTIVES: This study aimed to shed new light on the potential detrimental effects on cementum and adjacent alveolar bone after chronic exposure to amoxicillin. METHODS: Six pregnant adult Albino rats were equally divided into two groups. Saline solution and amoxicillin (100 mg/Kg) were given to rats of control and amoxicillin group, respectively from the 13th to the 21st day of pregnancy. The same treatment was given to the pups till the 42nd day. The cementum of the first molar teeth and the surrounding alveolar bone were examined qualitatively by histopathological and scanning electron microscope, and quantitatively by energy dispersive X-ray spectroscopy and cone beam computed tomography. RESULTS: Amoxicillin group depicted cemental and alveolar bone defects along with resorption lacunae. Statistically significant decreases in calcium and calcium/phosphorus ratio in cementum and in calcium only in alveolar bone were evident (p ≤ 0.05). Overall cementum and alveolar bone densities also showed statistically significant decreases (p ≤ 0.05). CONCLUSION: Chronic amoxicillin administration displayed destructive effects on cementum and the surrounding alveolar bone which may disturb tooth attachment integrity. Therefore, it is recommended to minimize its haphazard usage during pregnancy and early childhood.

Unusual imaging appearance of cemental tear in the maxillary first molar on cone-beam computed tomography: A case report.

A cemental tear (CeT) is a definitive clinical entity and its radiographic appearance is well known in single-rooted teeth. However, the imaging features of CeT in multi-rooted teeth have not been clarified. We report a case of CeT which arose in the maxillary first molar and exhibited an unusual appearance in cone-beam computed tomography images. The torn structure was verified as cementum by micro-computed tomography and histological analysis. The hypercementosis, most likely induced by occlusal force, might have been torn from the root by a stronger occlusal force caused by the mandibular implant. An unusual bridging structure was created between the two buccal roots. These features may occur in multi-rooted teeth with long-standing deep pockets and abscesses that are resistant to treatment.

Functional defects in cementoblasts with disrupted bone sialoprotein functional domains, in vitro.

Bone sialoprotein (BSP) is a multifunctional extracellular matrix (ECM) protein present in bone and cementum. Global in vivo ablation of BSP leads to bone mineralization defects, lack of acellular cementum, and periodontal breakdown. BSP harbors three main functional domains: N-terminal collagen-binding domain, hydroxyapatite-nucleating domain, and C-terminal RGD integrin-binding signaling domain. How each of these domains contributes to BSP function(s) is not understood. We hypothesized that collagen-binding and RGD domains play distinct roles in cementoblast functions. Three CRISPR/Cas9 gene-edited cell lines were derived from control wild-type (WT) OCCM.30 murine immortalized cementoblasts: 1) deletion of the N-terminus of BSP after signal peptide, including entire collagen binding domain (Ibsp∆N-Term); 2) deletion of exon 4 (majority of collagen-binding domain; Ibsp∆Ex4); and 3) deletion of C-terminus of BSP including the integrin binding RGD domain (Ibsp∆C-Term). Compared to WT, Ibsp∆Ex4 and Ibsp∆C-Term cell lines showed reduced BSP secretion, in vitro. Abnormal cell morphology was observed in all mutant cell lines, with Ibsp∆C-Term showing highly disorganized cytoskeleton. All mutant cell lines showed significantly lower cell proliferation compared to WT at all timepoints. Ibsp∆N-Term cells showed reduced cell migration by 24 h. All mutants exhibited over 50 % significant reduced mineralization at days 6 and 10. While WT cells were largely unaffected by seeding density, mutant cells failed to mineralize at lower cell density. Mutant cell lines diverged from WT and from each other by dysregulated expression in 23 genes involved in mineralization, ECM, and cell signaling. In summary, disabling BSP functional domains led to profound and distinct changes in cementoblast cell functions, especially dysregulated gene expression and reduced mineralization, in a way did not align with a straightforward narrative where each functional domain caused specific, expected differences. Instead, the study uncovered a significant level of complexity in how different mutant forms of BSP affected cell functions, in vitro.

Activation of periodontal ligament cell cytodifferentiation by juxtacrine signaling from cementoblasts.

BACKGROUND: New cementum forms from existing cementum during periodontal tissue regeneration, indicating that cementoblasts may interact with progenitor cells in the periodontal ligament to enhance cementogenesis. However, the molecular mechanisms of this process are currently unknown. This study aims to clarify the role of cell-cell interactions between cementoblasts and periodontal ligament cells in differentiation into cementoblasts. METHODS: To analyze the role of human cementoblast-like cells (HCEMs) on human periodontal ligament cells (HPDLs), we mixed cell suspensions of enhanced green fluorescent protein-tagged HPDLs and HCEMs, and then seeded and cultured them in single wells (direct co-cultures). We sorted co-cultured HPDLs and analyzed their characteristics, including the expression of cementum-related genes. In addition, we cultured HPDLs and HCEMs in a non-contact environment using a culture system composed of an upper insert and a lower well separated by a semi-permeable membrane (indirect co-cultures), and similar analysis was performed. Gene expression of integrin-binding sialoprotein (IBSP) in cementoblasts was confirmed in mouse periodontal tissues. We also investigated the effect of Wingless-type (Wnt) signaling on the differentiation of HPDLs into cementoblasts. RESULTS: Direct co-culture of HPDLs with HCEMs significantly upregulated the expression of cementoblast-related genes in HPDLs, whereas indirect co-culture exerted no effect. Wnt3A stimulation significantly upregulated IBSP expression in HPDLs, whereas inhibition of canonical Wnt signaling suppressed the effects of co-culture. CONCLUSION: Our results suggest that direct cell interactions with cementoblasts promote periodontal ligament cell differentiation into cementoblasts. Juxtacrine signaling via the canonical Wnt pathway plays a role in this interaction.

Preclinical evaluation of a new synthetic carbonate apatite bone substitute on periodontal regeneration in intrabony defects.

OBJECTIVE: To evaluate the potential of a novel synthetic carbonate apatite bone substitute (CO3 Ap-BS) on periodontal regeneration. BACKGROUND: The use of various synthetic bone substitutes as a monotherapy for periodontal regeneration mainly results in a reparative healing pattern. Since xenografts or allografts are not always accepted by patients for various reasons, a synthetic alternative would be desirable. METHODS: Acute-type 3-wall intrabony defects were surgically created in 4 female beagle dogs. Defects were randomly allocated and filled with CO3 Ap-BS (test) and deproteinized bovine bone mineral (DBBM) or left empty (control). After 8 weeks, the retrieved specimens were scanned by micro-CT, and the percentages of new bone, bone substitute, and soft tissues were evaluated. Thereafter, the tissues were histologically and histometrically analyzed. RESULTS: Healing was uneventful in all animals, and defects were present without any signs of adverse events. Formation of periodontal ligament and cementum occurred to varying extent in all groups without statistically significant differences between the groups. Residues of both bone substitutes were still present and showed integration into new bone. Histometry and micro-CT revealed that the total mineralized area or volume was higher with the use of CO3 Ap-BS compared to control (66.06 ± 9.34%, 36.11 ± 6.40%; p = .014, or 69.74 ± 2.95%, 42.68 ± 8.68%; p = .014). The percentage of bone substitute surface covered by new bone was higher for CO3 Ap-BS (47.22 ± 3.96%) than for DBBM (16.69 ± 5.66, p = .114). CONCLUSIONS: CO3 Ap-BS and DBBM demonstrated similar effects on periodontal regeneration. However, away from the root surface, more new bone, total mineralized area/volume, and higher osteoconductivity were observed for the CO3 Ap-BS group compared to the DBBM group. These findings point to the potential of CO3 Ap-BS for periodontal and bone regeneration.

Runx2 heterozygosity alters homeostasis of the periodontal complex.

BACKGROUND AND OBJECTIVE: Haploinsufficiency of Runx2 (Runx2+/- ) causes dental anomalies. However, little is known about the involvement of Runx2 in the maintenance of dentin, cementum, and the periodontal ligament (PDL) during adulthood. This study aimed to observe the effects of Runx2+/- on homeostasis of the periodontal complex. MATERIALS AND METHODS: A total of 14 three-month-old Runx2+/- mice and their wild-type littermates were examined using micro-computed tomography, histology, and immunohistochemistry. Phenotypic alterations in the dentin, cementum, and PDL were characterized and quantified. RESULTS: Haploinsufficiency of Runx2 caused cellular changes in the PDL space including reduction of cell proliferation and apoptosis, and irregular attachment of the collagen fibers in the PDL space into the cementum. Absence of continuous thickness of cementum was also observed in Runx2+/- mice. CONCLUSION: Runx2 is critical for cementum integrity and attachment of periodontal fibers. Because of its importance to cementum homeostasis, Runx2 is essential for homeostasis of periodontal complex.

Variability in human tooth cementum thickness reflecting functional processes.

OBJECTIVE: The aim of this study was to investigate the thickness of acellular extrinsic fibre cementum (AEFC) at four root positions of anterior and posterior teeth with special focus on functional aspects. Furthermore, the correlations between cementum thickness and chronological age and sex are investigated. BACKGROUND: While numerous studies confirm continuous cementum apposition with age, masticatory forces as well as physiological and orthodontically induced tooth movements also have the potential to affect tooth cementum thickness. MATERIALS AND METHODS: Undecalcified teeth were embedded in resin and transverse-sectioned in the cervical third of the root. Two sections per root were selected, and digital images at four positions were obtained (mesial, distal, oral, and vestibular) using light microscopy. The AEFC thickness of 99 teeth (anterior = 66, posterior = 33, male = 54, female = 45) were measured in both sections. The differences in mean values between root positions and the association of root position variation with tooth type, age, sex, and subject as well as the overall effects of age and sex were analysed using a mixed model. RESULTS: First incisors and canines showed the greatest mean AFEC thickness, in contrast to premolars which had the lowest values. Differences were found across the four root positions, with a pattern varying considerably between anterior and posterior teeth and between maxilla and mandible in the anterior teeth. An interaction between root position and subject pointed to the existence of an individual component in the variation of AEFC thickness across the four root positions. There was an age trend with an almost linear increase in cementum thickness of 1 µm per year. Overall, females tended to exhibit a significantly lesser AEFC thickness compared to males. CONCLUSIONS: Distinct differences in the pattern of thickness values across the four root positions in anterior and posterior teeth support the assumption that the AEFC is strongly affected by functional processes. In addition to sex-specific differences and age-related trends, the root position variation of AEFC thickness varies from individual to individual.

Anatomical and Histological Characteristics of Prematurely Erupted Teeth in Newborns.

Purpose: To assess the characteristics of dental hard and soft tissue structures of prematurely erupted teeth in newborns. Methods: Extracted natal and neonatal teeth were assessed in ground sections for evaluation of enamel, dentin, dentino-enamel junction and cementoenamel junction. Soft tissue harvested was histologically analyzed for cellularity, vascularity and other characteristics of the dental pulp. Results: This study included 15 teeth from nine neonates, of which seven erupted at birth, eight erupted within the first month of birth. All erupted in the mandibular anterior region. The demineralized ground section revealed enamel cracks, a straight dentinoenamel junction, and S-shaped dentinal tubules along with the enamel lamellae, and enamel spindle. None of the samples showed enamel tuft or cementum. Histology of soft tissue revealed enhanced cellularity, vascularity without any fibrosis, calcification and inflammation when compared with the young healthy pulp in permanent teeth. Conclusion: The prematurely erupted teeth in newborns were almost exclusively rootless and exhibited the characteristic anatomical findings pertaining to enamel, dentin, dentinoenamel junction and cementoenamel junction. The histology of their dental pulp was characterized by increased cellularity and vascularity compared.

Microtensile bond strength of resin cements to 3-D printed and milled temporary restorative resins / Resistencia de unión en microtracción de cementos resinosos a resinas para provisionales impresas y fresadas

Abstract To evaluate the microtensile bond strength (µTBS) of two resin cements to 3D printed and milled CAD/CAM resins used for provisional fixed partial dentures. Blocks (5 x 5 x 5 mm) of three 3D-printed resins (Cosmos3DTemp / Yller; Resilab3D Temp / Wilcos and SmartPrint BioTemp, / MMTech) were printed (Photon, Anycubic Technology Co.). A milled material (VitaCAD-Temp, VITA) was used as control. Half the specimens were sandblasted and the rest were untreated. Two blocks were bonded with the corresponding resin cement: PanaviaV5 (Kuraray Noritake) and RelyX Ultimate (3M Oral Care). After 24 hours, the bonded blocks were sectioned into 1 x 1 mm side sticks. Half the beams were tested for µTBS and the other half was thermocycled (5000 cycles, 30s dwell-time, 5s transfer time) before µTBS testing. A four way Generalized Linear Model (material*sandblasting*cement*aging) analysis was applied. VITA exhibited the lowest µTBS, regardless of the cement, sandblasting and thermocycling. Sandblasting significantly improved the µTBS of VIT, especially after aging, but did not improve the µTBS of 3D printed resins. Sandblasting was not beneficial for 3D printed resins, although is crucial for adhesive cementation of milled temporary resins. Airborne particle abrasion affects the integrity of 3D-printed resins, without producing a benefit on the microtensile bond strength of these materials. However, sandblasting is crucial to achieve a high bond strength on milled temporary resins.

Resumen Evaluar la resistencia adhesiva en microtracción (µTBS) de dos cementos resinosos a resinas CAD/CAM impresas y fresadas indicadas para restauraciones provisionales. Bloques (5 x 5 x 5mm) de tres resinas impresas (Cosmos3DTemp / Yller; Resilab3D Temp / Wilcos and SmartPrint BioTemp, / MMTech) y una resina fresada (VitaCAD-Temp, VITA) fueron fabricados. La mitad de los especímenes fueron arenados y el resto no recibió tratamiento mecánico. Dos bloques con condiciones de tratamiento iguales fueron cementados con cemento resinoso (PanaviaV5 / Kuraray Noritake y RelyX Ultimate / 3M Oral Care). Después de 24 horas los bloques fueron seccionados en palitos de 1 mm² de área. En la mitad de los especímenes se midió la TBS inmediatamente y el resto fue termociclado (5000 ciclos, 30s remojo, 5s transferencia) antes de la prueba de TBS. Se aplica un análisis estadístico por Modelo Linear General con 4 factores (material*arenado*cemento*termociclado). La resina VITA presentó la menor µTBS, independientemente del cemento usado, el arenado y el termociclado. Sin embargo, el arenado aumentó la µTBS de VIT, especialmente después del termociclado. Por otro lado, el arenado no resultó en un aumento significativo de la µTBS de las resinas impresas. El arenado no fue beneficiosos para las resinas impresas, aunque es un paso crucial para la cementación adhesive de las resinas fresadas. El arenado afecta la integridad de las capas de las resinas impresas, sin generar un beneficio en la TBS.

3D finite element analysis of stress distribution as a result of oblique and horizontal forces after regenerative endodontic treatment part II: comparison of material thickness.

AIM: This study aimed to evaluate the stress distribution caused by secondary trauma forces after regenerative endodontic treatment (RET) using different thicknesses of coronary barrier material with three-dimensional finite element analysis(FEA). METHOD: A control model was created using the tomography image of the immature maxillary central tooth with computer software.Study models were created with the modulus of elasticity and Poisson's ratio of the materials used in RET.Enamel, dentin, cementum, periodontal ligament, cortical, and cancellous bone were modeled. Coronary barrier materials were applied in 3 mm and 5 mm thicknesses (Model 1: control model, model 2:3 mm/Calcium Enriched Mixture(CEM), model 3:3 mm/Mineral Trioxide Aggregate(MTA), model 4:3 mm/Biodentin, model 5:5 mm/CEM, model 6:5 mm/MTA, model 7:5 mm/Biodentin). For the trauma force simulation, 300 N force in the horizontal direction was applied to the buccal surface of the tooth in the first scenario. For the second scenario, maximum bite force simulation, a force of 240 N in the oblique direction was applied to the palatal surface of the tooth. FEA was performed with Algor Fempro. The resulting stresses were recorded as Von Mises, maximum, and minimum principal stresses. RESULTS: Lower stress values were obtained in 5 mm models compared to 3 mm models. However, the difference between them was insignificant. Lower stress values were obtained in all RET models compared to the control model. The lowest stress values in dental tissues and bone tissue were obtained in the CEM models. CONCLUSION: This is the first study in which the stress caused by different thicknesses of CEM on dental tissues was evaluated with FEA. RET strengthens immature teeth biomechanically. CEM and Biodentin are more successful materials in stress distribution than MTA. Considering the cost of treatment, 3 mm material thickness is ideal for RET since there is no significant difference between the stress values resulting from the use of 5 mm and 3 mm coronary barrier material.

Glycogen synthase kinase 3 inhibition enhances mineral nodule formation by cementoblasts in vitro.

This study aimed to investigate whether GSK-3 inhibition (CHIR99021) effectively promoted mineralization by cementoblasts (OCCM-30). OCCM-30 cells were used and treated with different concentrations of CHIR99021 (2.5, 5, and 10 mM). Experiments included proliferation and viability, cellular metabolic activity, gene expression, and mineral nodule formation by Xylene Orange at the experimental time points. In general, CHIR99021 did not significantly affect OCCM-30 viability and cell metabolism (MTT assay) (p > 0.05), but increased OCCM-30 proliferation at 2.5 mM on days 2 and 4 (p < 0.05). Data analysis further showed that inhibition of GSK-3 resulted in increased transcript levels of Axin2 in OCCM-30 cells starting as early as 4 h, and regulated the expression of key bone markers including alkaline phosphatase (Alp), runt-related transcription factor 2 (Runx-2), osteocalcin (Ocn), and osterix (Osx). In addition, CHIR99021 led to an enhanced mineral nodule formation in vitro under both osteogenic and non-osteogenic conditions as early as 5 days after treatment. Altogether, the results of the current study suggest that inhibition of GSK-3 has the potential to promote cementoblast differentiation leading to increased mineral deposition in vitro.

Glycometabolic reprogramming in cementoblasts: A vital target for enhancing cell mineralization.

Cementum, a constituent part of periodontal tissues, has important adaptive and reparative functions. It serves to attach the tooth to alveolar bone and acts as a barrier delimit epithelial growth and bacteria evasion. A dynamic and highly responsive cementum is essential for maintaining occlusal relationships and the integrity of the root surface. It is a thin layer of mineralized tissue mainly produced by cementoblasts. Cementoblasts are osteoblast-like cells essential for the restoration of periodontal tissues. In recent years, glucose metabolism has been found to be critical in bone remodeling and osteoblast differentiation. However, the glucose metabolism of cementoblasts remains incompletely understood. First, immunohistochemistry staining and in vivo tracing with 18 F-fluorodeoxyglucose (18 F-FDG) revealed significantly higher glucose metabolism in cementum formation. To test the bioenergetic pathways of cementoblast differentiation, we compared the bioenergetic profiles of mineralized and unmineralized cementoblasts. As a result, we observed a significant increase in the consumption of glucose and production of lactate, coupled with the higher expression of glycolysis-related genes. However, the expression of oxidative phosphorylation-related genes was downregulated. The verified results were consistent with the RNA sequencing results. Likewise, targeted energy metabolomics shows that the levels of glycolytic metabolites were significantly higher in the mineralized cementoblasts. Seahorse assays identified an increase in glycolytic flux and reduced oxygen consumption during cementoblast mineralization. Apart from that, we also found that lactate dehydrogenase A (LDHA), a key glycolysis enzyme, positively regulates the mineralization of cementoblasts. In summary, cementoblasts mainly utilized glycolysis rather than oxidative phosphorylation during the mineralization process.

DNA identification from dental pulp and cementum.

Teeth are one of the body tissues remaining after severe decomposition from which a DNA profile can be obtained to aid in human identification. Currently, the standard approach to isolate DNA from teeth requires pulverizing the entire tooth. This destructive approach compromises any further morphological or anthropological study. We report on two methods of DNA isolation that minimizes destruction of the tooth when accessing the DNA within pulp and cementum. Forty-nine teeth, removed as part of normal dental procedures, were buried for up to 92 days, with a further nine teeth acting as unburied controls. Additionally, four teeth samples collected during a forensic examination were included in this study. The two processes were: using a fine drill to access the pulp from the crown and then using endodontic files to collect the biological material; and using a sterile blade to scrape the cementum. It was found that the samples collected from the cementum had greater DNA quality compared to those samples obtained from the pulp. Microbial activity was found to play a role in the degradation of the nuclear material, reducing DNA yields from pulp. DNA profiling data from 24 loci, including 22 STR markers, indicated that multi-rooted teeth provided better DNA quantity and quality than those with a single root. The DNA quantity obtained from pulp samples of teeth which exhibited cavities was adversely affected, although this DNA loss was not from samples collected from the cementum of teeth in similar condition. Obtaining samples from DNA profiling from the cementum was found to be ideal if the morphological preservation of the tooth is required. Obtaining pathogen DNA is of interest when an occlusal approach to retrieve pulp may serve as a good alternative to prepare DNA without destruction of the tooth structure.

Development, Establishment, and Validation of a Model for the Mineralization of Periodontium Remodelling Cells: Cementoblasts.

Chronic kidney disease (CKD) patients undergoing dialysis are at high risk of bone fractures. CKD-induced mineral and bone disorder is extended to periodontal disease due to changes in the ionic composition of saliva in CKD patients, dysregulating mineralization, hindering regeneration and thereby promoting the progression of dental complications. Despite the importance of cementum for overall oral health, the mechanisms that regulate its development and regeneration are not well comprehended, and a lack of sufficient in vitro experimental models has hindered research progress. In this study, the impact of experimental conditions on the calcification of cementoblasts was systematically investigated, aimed at establishing a standardized and validated model for the calcification of cementoblasts. The effects of phosphate, calcium, ascorbic acid, ß-glycerolphosphate, dexamethasone, and fetal calf serum on the calcification process of cementoblasts were analyzed over a wide range of concentrations and time points by investigating calcium content, cell viability, gene expression and kinase activity. Cementoblasts calcified in a concentration- and time-dependent manner with higher concentrations of supplements cause a higher degree of calcification but decreased cell viability. Phosphate and calcium have a significantly stronger effect on cementoblast calcification processes compared to osteogenic supplements: ascorbic acid, ß-glycerolphosphate, and dexamethasone induce calcification over a wide range of osteogenic signalling pathways, with osteopontin being a central target of gene regulation. Conversely, treatment with ascorbic acid, ß-glycerolphosphate, and dexamethasone leads to activating only selected pathways, especially promoting bone sialoprotein expression. The developed and validated cementoblast calcification protocol, incubating up to 60% confluent cementoblasts with 1.9 mmol L-1 of phosphate supplementation for a reasonable, multi-pathway calcification induction and 10 mmol L-1 ß-glycerolphosphate, 75 µmol L-1 ascorbic acid and 10 nmol L-1 dexamethasone for a reasonable osteogenic differentiation-based calcification induction, provides standard in vitro experimental models for better understanding cementoblast function and regeneration.