Low translational and rotational movements with 2-point stainless-steel retainers over a period of 1 and 3 years.

OBJECTIVES: Long-term stabilization of orthodontic treatment outcomes is an everyday challenge in orthodontics. The use of permanently attached lingual retainers has become gold standard. However, in some cases, patients with fixed lingual retainers show retainer-associated side effects. Aiming to reduce these side effects, clinical knowledge about how tooth and arch form stability adaption takes place over time is important to improve long-term retention protocols. Therefore, the present study aimed to investigate occlusion stability and risks for a newly developing malocclusion in a time-dependent manner in patients being treated with permanent 2­point steel retainers. MATERIALS AND METHODS: In this retrospective cohort study, a total of 66 consecutive patients with round stainless-steel retainers were analyzed for postorthodontic occlusion changes after 1 year (group 1, n = 33) and 3 years (group 2, n = 33). Digital Standard Tessellation Language (STL) datasets of the lower jaw were obtained before retainer insertion (T0), and after a 1- (T1) or 3­year (T2) retention period. Using superimposition software, T1 and T2 situations were compared to T0 regarding rotational and translational changes in tooth positions in all three dimensions. RESULTS: Occlusion changes were low in both groups. The investigated lower canines were nearly stable in the 1­ and 3­year group, although a retention-time-dependent increase in tooth position change of the central and lateral incisors could be observed. CONCLUSION: The present data provide evidence for time-dependent development of posttherapeutic occlusal adaption limited to central and lateral incisors in patients treated with a 2-point retainer. The observed occlusal changes should be interpreted as an occlusal adaption process rather than severe posttreatment changes associated with the orthodontic retainer.

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.

The Contribution of the Nrf2/ARE System to Mechanotransduction in Musculoskeletal and Periodontal Tissues.

Mechanosensing plays an essential role in maintaining tissue functions. Across the human body, several tissues (i.e., striated muscles, bones, tendons, ligaments, as well as cartilage) require mechanical loading to exert their physiological functions. Contrary, mechanical unloading triggers pathological remodeling of these tissues and, consequently, human body dysfunctions. At the cellular level, both mechanical loading and unloading regulate a wide spectrum of cellular pathways. Among those, pathways regulated by oxidants such as reactive oxygen species (ROS) represent an essential node critically controlling tissue organization and function. Hence, a sensitive balance between the generation and elimination of oxidants keeps them within a physiological range. Here, the Nuclear Factor-E2-related factor 2/Antioxidant response element (Nrf2/ARE) system plays an essential role as it constitutes the major cellular regulation against exogenous and endogenous oxidative stresses. Dysregulations of this system advance, i.a., liver, neurodegenerative, and cancer diseases. Herein, we extend our comprehension of the Nrf2 system to the aforementioned mechanically sensitive tissues to explore its role in their physiology and pathology. We demonstrate the relevance of it for the tissues' functionality and highlight the imperative to further explore the Nrf2 system to understand the physiology and pathology of mechanically sensitive tissues in the context of redox biology.

Knockout of bone sialoprotein in cementoblasts cell lines affects specific gene expression in unstimulated and mechanically stimulated conditions.

One of the major components in cementum extracellular matrix is bone sialoprotein (BSP). BSP knockout (Ibsp) mice were reported to have a nonfunctional hypo-mineralized cementum, as well as detachment and disorganization of the periodontal ligament tissue. However, studies investigating the influence of Ibsp in cementoblasts are missing yet. This study investigates the influences of Bsp in three cementoblasts cell lines (OCCM.30-WT,IbspΔNterm, and IbspKAE). The mRNA expression of cementoblast and osteoclast markers (Col1a1, Alpl, Ocn, Runx2, Ctsk, Rankl and Opg) and the cell morphology were compared. Additionally, a functional monocyte adhesion assay was performed. To understand the influence of external stimuli, the effect of Ibsp was investigated under static compressive force, mimicking the compression side of orthodontic tooth movement. Cementoblasts with genotype IbspΔNterm and IbspKAE showed slight differences in cell morphology compared to OCCM.30-WT, as well as different gene expression. Under compressive force, the Ibsp cell lines presented expression pattern markers similar to the OCCM.30-WT cell line. However, Cathepsin K was strongly upregulated in IbspΔNterm cementoblasts under compressive force. This study provides insight into the role of BSP in cementoblasts and explores the influence of BSP on periodontal ligament tissues. BSP markers in cementoblasts seem to be involved in the regulation of cementum organization as an important factor for a functional periodontium. In summary, our findings provide a basis for investigations regarding molecular biology interactions of BSP in cementoblasts, and a supporting input for understanding the periodontal and cellular cementum remodeling.

Post-treatment Stability in Orthodontic Retention with Twistflex Retainers-Do Patients Benefit from Additional Removable Retainers?

OBJECTIVES: To evaluate post-treatment movements of lower anterior teeth during orthodontic retention in patients with fixed twistflex retainers versus those with combined fixed and removable retainers. MATERIALS AND METHODS: This study was based on a retrospective data analysis of 57 adult patients during orthodontic retention. They were assigned to two groups: In group 1 (n = 30) the lower jaw was provided with twistflex retainers only and in group 2 (n = 27) with a twistflex combined with a removable retainer for night-time use. Orthodontic study models of the lower jaw were digitalized and superimposed. Tooth movements were analyzed at the retainer bonding (t0) and follow-up appointment ≥ six months later (t1). Rotational tooth movements (°) were measured around the x-axis (mesial/distal direction), the y-axis (buccal/lingual direction) and the z-axis (longitudinal direction, tooth axis). Translational tooth movements (mm) were registered along the x-axis (buccal/lingual direction), the y-axis (mesial/distal direction) and the z-axis (apical/coronal direction). RESULTS: Canine and incisor position changes during orthodontic retention were more pronounced in group 1 compared to group 2 except for canine rotations around the z-axis. In both groups in most of the cases stable lower incisor alignment could be found, but the proportion was significant higher in group 2 (group 1: 56.7% vs. group 2: 81.5%). Severe misalignment was present in 13.3% of the participants of group 1 and only in 7.4% of group 2. The extent of canine tipping and movements along the x- and y-axis in severe misalignment cases was significantly lower in group 2 compared to 1. CONCLUSIONS: Lower incisor alignment was more stable in patients with combined fixed and removable retainers compared to fixed retainers only. CLINICAL RELEVANCE: Based on the present findings, the routinely application of supplementary removable retainers can be recommended to enhance anterior tooth alignment in patients with fixed twistflex retainers.

Side effects of twistflex retainers-3D evaluation of tooth movement after retainer debonding.

PURPOSE: Evaluation of tooth movement after retainer debonding in retainer-associated misalignment cases. METHODS: This pilot study is based on a retrospective data analysis. Adult patients (age 25.5 ± 4.9 years) wearing fixed twistflex retainers and having visible retainer-associated misalignment were included and examined for tooth movement after retainer debonding. Orthodontic study models were taken at retainer debonding (t0) and 14 (±1) weeks later (t1). They were digitally superimposed using 2D/3D dental imaging software and tooth movement was analyzed in all three dimensions. RESULTS: A total of 23 teeth (12 upper teeth: 10 incisors, 2 canines; 11 lower teeth: 7 incisors, 4 canines) were analyzed. Mean overall tipping was 1.11 ± 0.82° in the mesial/distal direction (angulation, x­axis), 2.02 ± 1.9° in the buccal/lingual direction (inclination, y­axis) and 1.28 ± 0.99° around the tooth axis (z-axis). Mean overall bodily movement was 0.30 ± 0.31 mm in the mesial/distal direction (angulation, x­axis), 0.10 ± 0.13 mm in the buccal/lingual direction (inclination, y­axis), and mean in- or extrusion 0.22 ± 0.24 mm (z-axis). Mean tipping and bodily movement were more pronounced in the upper jaw. CONCLUSION: The present data shows that tooth movement after debonding of twistflex retainers can be expected in misalignment cases.

Analysing the potential of hydrophilic adhesive systems to optimise orthodontic bracket rebonding.

INTRODUCTION: Bond failure during fixed orthodontic treatment is a frequently occurring problem. As bracket rebonding is associated with reduced shear bond strength, the aim of the present investigation is to analyse the effect of different innovative rebonding systems to identify optimised rebonding protocols for orthodontic patient care. METHODS: Metallic brackets were bonded to the frontal enamel surfaces of 240 bovine lower incisors embedded in resin bases. Teeth were randomly divided into two major experimental groups: in group 1 a hydrophilic primer (Assure™ PLUS) was compared to commonly used orthodontic adhesives (Transbond XT™, BrackFix®, Grengloo™) and a zero control. In group 2 different rebonding systems were analysed using a hydrophilic primer (Assure™ PLUS), a methyl methacrylate-consisting primer (Plastic Conditioner) and a conventional adhesive (Transbond XT™). All teeth were tested for shear bond strength according to the DIN-13990 standard, the Adhesive Remnant Index and enamel fracture rate. RESULTS: The hydrophilic primer enhanced shear bond strength at first bonding (Assure™ PLUS 20.29 ± 4.95 MPa vs. Transbond XT™ 18.45 ± 2.57 MPa; BrackFix® 17 ± 5.2 MPa; Grengloo™ 19.08 ± 3.19 MPa; Meron 8.7 ± 3.9 MPa) and second bonding (Assure™ PLUS 16.76 ± 3.71 MPa vs. Transbond XT™ 13.06 ± 3.19 MPa). Using Plastic Conditioner did not seem to improve shear bond strength at rebonding (13.57 ± 2.94). When enamel etching was left out, required shear bond strength could not be achieved (Plastic Conditioner + Assure™ PLUS 8.12 ± 3.34 MPa; Plastic Conditioner: 3.7 ± 1.95 MPa). Hydrophilic priming systems showed decreased ARI-scores (second bonding: 2.63) and increased enamel fracture rates (first bonding: 55%; second bonding 21,05%). CONCLUSIONS: Based on the present study we found that rebonding strength could be compensated by the use of hydrophilic priming systems. The additional use of a methyl methacrylate-consisting primer does not seem to enhance shear bond strength. No etching approaches resulted in non-sufficient bond strength.

Functional and molecular evidence for heteromeric association of P2Y1 receptor with P2Y2 and P2Y4 receptors in mouse granulocytes.

BACKGROUND: All hematopoietic cells express P2 receptors, however pharmacological characteristics such as expression and affinity in granulocytes are unknown. METHODS: Pharmacological characteristics of P2 receptors were evaluated by Ca(2+) measurements using Fura-2 fluorophore. P2 receptors expression were analyzed by flow cytometry and RT-PCR. P2 interaction were shown by coimmunoprecipitation, western blotting and FRET. RESULTS: Granulocytes were responsive to P2Y agonists, whereas P2X agonists were ineffective. Ca(2+) increase, elicited by ADP and UTP was dependent on intracellular stocks and sensitive to G-coupled receptor inhibition. Moreover, MRS2179, a specific antagonist of the P2Y1 receptor, abolished ADP response. Interestingly, ADP and UTP exhibited full heterologous desensitization, suggesting that these agonists interact with the same receptor. The heteromeric association between P2Y1 receptor and the P2Y2 and P2Y4 receptors was shown by immunoprecipitation and FRET analysis. CONCLUSION: Clear evidence of heteromeric association of P2Y receptors was found during the evaluation of P2 receptors present in mice granulocytes, which could impact in the classical pharmacology of P2Y receptors in granulocytes.

Anabolic properties of high mobility group box protein-1 in human periodontal ligament cells in vitro.

High mobility group box protein-1 (HMGB1) is mainly recognized as a chemoattractant for macrophages in the initial phase of host response to pathogenic stimuli. However, recent findings provide evidence for anabolic properties in terms of enhanced proliferation, migration, and support of wound healing capacity of mesenchymal cells suggesting a dual role of the cytokine in the regulation of immune response and subsequent regenerative processes. Here, we examined potential anabolic effects of HMGB1 on human periodontal ligament (PDL) cells in the regulation of periodontal remodelling, for example, during orthodontic tooth movement. Preconfluent human PDL cells (hPDL) were exposed to HMGB1 protein and the influence on proliferation, migration, osteogenic differentiation, and biomineralization was determined by MTS assay, real time PCR, immunofluorescence cytochemistry, ELISA, and von Kossa staining. HMGB1 protein increased hPDL cell proliferation, migration, osteoblastic marker gene expression, and protein production as well as mineralized nodule formation significantly. The present findings support the dual character of HMGB1 with anabolic therapeutic potential that might support the reestablishment of the structural and functional integrity of the periodontium following periodontal trauma such as orthodontic tooth movement.

Downregulation of 14q32 microRNAs in Primary Human Desmoplastic Medulloblastoma.

Medulloblastoma (MB) is one of the most common pediatric cancers, likely originating from abnormal development of cerebellar progenitor neurons. MicroRNA (miRNA) has been shown to play an important role in the development of the central nervous system. Microarray analysis was used to investigate miRNA expression in desmoplastic MB from patients diagnosed at a young age (1 or 2 years old). Normal fetal or newborn cerebellum was used as control. A total of 84 differentially expressed miRNAs (64 downregulated and 20 upregulated) were found. Most downregulated miRNAs (32/64) were found to belong to the cluster of miRNAs at the 14q32 locus, suggesting that this miRNA locus is regulated as a module in MB. Possible mechanisms of 14q32 miRNAs downregulation were investigated by the analysis of publicly available gene expression data sets. First, expression of estrogen-related receptor-γ (ESRRG), a reported positive transcriptional regulator of some 14q32 miRNAs, was found downregulated in desmoplastic MB. Second, expression of the parentally imprinted gene MEG3 was lower in MB in comparison to normal cerebellum, suggesting a possible epigenetic silencing of the 14q32 locus. miR-129-5p (11p11.2/7q32.1), miR-206 (6p12.2), and miR-323-3p (14q32.2), were chosen for functional studies in DAOY cells. Overexpression of miR-129-5p using mimics decreased DAOY proliferation. No effect was found with miR-206 or miR-323 mimics.

Activation of P2Y1 receptor triggers two calcium signaling pathways in bone marrow erythroblasts.

In this study, we describe the presence of P2 receptor subtypes and Ca2+ signaling in erythroblasts. ATP and ADP produced a biphasic increase of intracellular Ca2+ concentration ([Ca2+]i), with an initial transient phase followed by a sustained phase. Reverse transcription polymerase chain reaction (RT-PCR) showed the expression of P2Y1, P2Y2 and P2Y12. The selective P2Y1 receptor antagonist 2'-deoxy-N6-methyl-adenosine-3',5'-diphosphate (MRS2179) and the G(i) protein inhibitor pertussis toxin blocked Ca2+ increase. The initial transient [Ca2+]i increase phase was sensitive to the 1,4,5-inositol trisphosphate (IP3) receptor blocker 2-aminoethoxy-diphenylborate (2-APB), while the sustained phase was sensitive to the protein kinase C (PKC) inhibitor 2-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimide (GF109203X) and calcium calmodulin kinase II (CaMKII) inhibitor 1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62). In addition, the PKC activator phorbol-12,13-dibutyrate (PDBu) produced increase of [Ca2+]i. Flow cytometry analysis showed the expression of Ca2+-dependent PKC alpha, betaI, gamma and phospho-CaMKII. These results suggest that the activation of the P2Y1 receptor triggers two different [Ca2+]i increase pathways, one IP3-dependent and the other kinase-dependent.