GDF15 Contributes to the Regulation of the Mechanosensitive Responses of PdL Fibroblasts through the Modulation of IL-37.

During orthodontic tooth movement (OTM), areas of compressive and tensile forces are generated in the periodontal ligament (PdL), a mechanoreactive connective tissue between the teeth and alveolar bone. Mechanically stimulated PdL fibroblasts (PdLFs), the main cell type of PdL, express significantly increased levels of growth differentiation factor 15 (GDF15). In compressed PdL areas, GDF15 plays a fundamental role in modulating relevant OTM processes, including inflammation and osteoclast activation. However, the specific function of this factor in tensile areas has not yet been investigated. Thus, the aim of this study was to investigate the role of GDF15 in the mechanoresponse of human PdLFs (hPdLFs) that were exposed to biaxial tensile forces in vitro. Using siRNA-mediated knockdown experiments, we demonstrated that GDF15 had no impact on the anti-inflammatory force response of elongated hPdLFs. Although the anti-inflammatory markers IL1RN and IL10, as well as the activation of immune cells remained unaffected, we demonstrated an inhibitory role of GDF15 for the IL-37 expression. By analyzing osteogenic markers, including ALPL and RUNX2, along with an assessment of alkaline phosphatase activation, we further showed that the regulation of IL-37 by GDF15 modulates the osteogenic differentiation potential of hPdLFs. Despite bone resorption in tensile areas being rather limited, GDF15 was also found to positively modulate osteoclast activation in those areas, potentially by adjusting the IL-37 levels. In light of our new findings, we hypothesize that GDF15 modulates force-induced processes in tissue and bone remodeling through its various intra- and extracellular signaling pathways as well as interaction partners. Potentially acting as a master regulator, the modulation of GDF15 levels may hold relevance for clinical implications.

GDF15 Modulates the Zoledronic-Acid-Induced Hyperinflammatory Mechanoresponse of Periodontal Ligament Fibroblasts.

Orthodontic tooth movement (OTM) is thought to be impeded by bisphosphonate (BP) therapy, mainly due to increased osteoclast apoptosis and changes in the periodontal ligament (PdL), a connecting tissue between the alveolar bone and teeth. PdL cells, mainly fibroblasts (PdLFs), are crucial regulators in OTM by modulating force-induced local inflammatory processes. Recently, we identified the TGF-ß/BMP superfamily member GDF15 as an important modulator in OTM, promoting the pro-inflammatory mechanoresponses of PdLFs. The precise impact of the highly potent BP zoledronate (ZOL) on the mechanofunctionality of PdLFs is still under-investigated. Therefore, the aim of this study was to further characterize the ZOL-induced changes in the initial inflammatory mechanoresponse of human PdLFs (hPdLFs) and to further clarify a potential interrelationship with GDF15 signaling. Thus, two-day in vitro treatment with 0.5 µM, 5 µM and 50 µM of ZOL altered the cellular properties of hPdLFs partially in a concentration-dependent manner. In particular, exposure to ZOL decreased their metabolic activity, the proliferation rate, detected using Ki-67 immunofluorescent staining, and survival, analyzed using trypan blue. An increasing occurrence of DNA strand breaks was observed using TUNEL and an activated DNA damage response was demonstrated using H2A.X (phosphoS139) staining. While the osteogenic differentiation of hPdLFs was unaffected by ZOL, increased cellular senescence was observed using enhanced p21Waf1/Cip1/Sdi1 and ß-galactosidase staining. In addition, cytokine-encoding genes such as IL6, IL8, COX2 and GDF15, which are associated with a senescence-associated secretory phenotype, were up-regulated by ZOL. Subsequently, this change in the hPdLF phenotype promoted a hyperinflammatory response to applied compressive forces with an increased expression of the pro-inflammatory markers IL1ß, IL6 and GDF15, as well as the activation of monocytic THP1 cells. GDF15 appeared to be particularly relevant to these changes, as siRNA-mediated down-regulation balanced these hyperinflammatory responses by reducing IL-1ß and IL-6 expression (IL1B p-value < 0.0001; IL6 p-value < 0.001) and secretion (IL-1ß p-value < 0.05; IL-6 p-value < 0.001), as well as immune cell activation (p-value < 0.0001). In addition, ZOL-related reduced RANKL/OPG values and inhibited osteoclast activation were enhanced in GDF15-deficient hPdLFs (both p-values < 0.0001; all statistical tests: one-way ANOVA, Tukey's post hoc test). Thus, GDF15 may become a promising new target in the personalized orthodontic treatment of bisphosphonatepatients.

Influence of the orthodontic bonding procedure on biofilm formation.

INTRODUCTION: In orthodontics, white spot lesions are a persistent and widespread problem caused by the demineralization of buccal tooth surfaces around bonded brackets. The remaining adhesive around the brackets leads to surface roughness, which might contribute to demineralization. The present in vitro study aimed to compare a conventional and a modern adhesive system (APC Flash-Free technology) for orthodontic brackets with regard to the adhesion of Streptococcus sobrinus, a leading caries pathogen. METHODS: This in vitro study included 20 premolar teeth and compared 10 APC Flash-Free adhesive-coated ceramic brackets (FF)with 10 conventionally bonded (CB) ceramic clarity brackets. Specimens were incubated in an S. sobrinus suspension for 3 h. To evaluate the bacterial formation, samples were analysed with a scanning electron microscope (SEM). Imaging software was used to quantify and statistically compare percentage values of colonization (PVC) in both groups' adhesion and transition areas. RESULTS: We found a significant difference in biofilm formation between the groups for the adhesive and transition areas. PVC in the adhesive area was approximately 10.3-fold greater for the CB group compared with the FF group (median: 3.2 vs 0.31; P < 0.0001). For the transition area, median PVC was approximately 2.4-fold greater for the CB group compared with the FF group (median: 53.17 vs 22.11; P < 0.01). CONCLUSIONS: There was a significantly lower level of S. sobrinus formation around the FF bracket system than there was surrounding the conventionally bonded group. This study suggests that the FF adhesive bracket system can help reduce the occurrence of bacterial growth around orthodontic brackets.

Orthognathic Surgery with Interdisciplinary Digital Planning in Patients with Geroderma Osteodysplasticum: A Case Report.

Patients with geroderma osteodysplasticum (GO) often times have dentofacial deformities and benefit from orthognathic surgery. Because of generalized osteopenia, operations must be prepared even more meticulously than usual, and the higher risk of unfortunate fractures (bad splits) should be explained to the patients in detail. This case report is intended to portray a digital, interdisciplinary and patient-individualized planning of orthognathic surgery. It points out the individual steps that must be considered and how they can be advantageously used in patients with underlying diseases or syndromes such as GO. Through a careful digital representation of the surgical options, production of the digitally modeled splints, 3D printing and good manual surgical implementation, the quality of life of patients with GO can be increased through orthognathic surgery. Both the functions in the oral, maxillofacial region and the patient's appearance in the case presented here benefited from the interdisciplinary, individualized and digital treatment approach.

Immunometabolic capacities of nutritional fatty acids in regulation of inflammatory bone cell interaction and systemic impact of periodontal infection.

Introduction: Novel preventive strategies in periodontal disease target the bacterial-induced inflammatory host response to reduce associated tissue destruction. Strategies focus on the modulation of tissue-destroying inflammatory host response, particularly the reduction of inflammation and promotion of resolution. Thereby, nutrition is a potent immunometabolic non-pharmacological intervention. Human studies have demonstrated the benefit of olive oil-containing Mediterranean-style diets (MDs), the main component of which being mono-unsaturated fatty acid (FA) oleic acid (OA (C18:1)). Hence, nutritional OA strengthened the microarchitecture of alveolar trabecular bone and increased circulating pro-resolving lipid mediators following bacterial inoculation with periodontal pathogen Porphyromonas gingivalis, contrary to saturated FA palmitic acid (PA (C16:0)), which is abundant in Western-style diets. Additionally, the generalized distribution of inflammatory pathway mediators can occur in response to bacterial infection and compromise systemic tissue metabolism and bone homeostasis distant from the side of infection. Whether specific FA-enriched nutrition and periodontal inoculation are factors in systemic pathology that can be immune-modulatory targeted through dietary substitution is unknown and of clinical relevance. Methods: Normal-weight C57BL/6-mice received OA-or PA-enriched diets (PA-ED, OA-ED, PA/OA-ED) or a normal-standard diet (n=12/group) for 16 weeks and were orally infected with P. gingivalis/placebo to induce periodontal disease. Using histomorphometry and LC-MS/MS, systemic bone morphology, incorporated immunometabolic FA-species, serological markers of bone metabolism, and stress response were determined in addition to bone cell inflammation and interaction in vitro. Results: In contrast to OA-ED, PA-ED reduced systemic bone microarchitecture paralleled by increased lipotoxic PA-containing metabolite accumulation in bone. Substitution with OA reversed the bone-destructive impact of PA, which was accompanied by reduced diacylglycerols (DAG) and saturated ceramide levels. Further, PA-associated reduction in mineralization activity and concomitant pro-inflammatory activation of primary osteoblasts were diminished in cultures where PA was replaced with OA, which impacted cellular interaction with osteoclasts. Additionally, PA-ED increased osteoclast numbers in femurs in response to oral P. gingivalis infection, whereas OA-ED reduced osteoclast occurrence, which was paralleled by serologically increased levels of the stress-reducing lipokine PI(18:1/18:1). Conclusion: OA substitution reverses the bone-destructive and pro-inflammatory effects of PA and eliminates incorporated lipotoxic PA metabolites. This supports Mediterranean-style OA-based diets as a preventive intervention to target the accumulation of PA-associated lipotoxic metabolites and thereby supports systemic bone tissue resilience after oral bacterial P. gingivalis infection.

A Newly Developed Orthodontic-Logopedic Screening Procedure for Myofunctional Dysfunctions-A Pilot Study.

Interdisciplinary, patient-specific cooperation between orthodontics and speech therapy plays an important role in the therapy of myofunctional dysfunctions. The following orthodontic-logopedic screening procedure is intended to objectify the diagnosis of such dysfunctions and the progress of therapy. A diagnostic questionnaire was prepared based on existing diagnostic questionnaires for myofunctional dysfunction. It contains 32 questions, with a clinical weighting of 0 to 50 points in total. This results in a point score. The lower the score is, the lower the need for therapy is. The study included 108 patients between the ages of 6 and 50 years. After screening, the patient population was divided into Group 0 (score < 15; no speech therapy need; n = 36) and Group 1 (score ≥ 15; a speech therapy need; n = 72). Group 1 was additionally randomized into Subgroups A (with speech therapy; n = 36) and B (without speech therapy; n = 36). After a treatment interval of 6 months, all patients in Group 1 were examined again with the help of the screening procedure. Statistical analysis (SPSS) and significance testing (Mann-Whitney U test) were performed. At baseline, there was no significant difference between patients in Subgroups A and B (p = 0.157). Subgroup A had a median score of 25, and Subgroup B had a median score of 30. However, after the treatment interval, a significant improvement (p = 0.001) for Subgroup A with a median score of 11 (mean score difference = 14.67) over Subgroup B with a median score 23 (mean score difference of 7.08) was observed. The developed screening procedure was shown to be equally applicable to all patients and treatment providers. With the help of the scores in point form, the need for speech therapy and the progress of such therapy can be objectified.

Influence of Different Bracket Adhesive Systems on Enamel Demineralization-An In Vitro Study.

BACKGROUND: enamel demineralization is a common side effect of orthodontic therapy with fixed braces. The aim of the present in vitro study was to compare a conventional adhesive system and a modern adhesive system (APC Flash-Free [FF] technology) with regard to the demineralization of enamel by Streptococcus sobrinus (S. sobrinus). METHODS: this in vitro study included premolar teeth and compared APC FF adhesive brackets (Group A, n = 15) with conventional adhesive brackets (Group B, n = 15) from the same company. Specimens were incubated with a positive control group (PCG, n = 5) and a negative control group (NCG, n = 5) in an S. sobrinus suspension for three weeks. To evaluate the grade of enamel demineralization, the samples were analyzed using a polarizing microscope. RESULTS: the test specimens of group B with conventionally bonded bracket adhesive showed significantly greater (+10.8 µm) demineralization with regard to the penetration depth of the demineralization than the PCG (p = 0.012). Thus, there was a difference from group A with the new bracket adhesive of the FF brackets (+7.29 µm). Significantly, demineralization was more pronounced cervically than coronally in both groups, and it occurred cervically more frequently than grade 3 demineralization (p = 0.001). CONCLUSIONS: it seems plausible that new orthodontic bracket adhesives and the modern FF adhesive system positively contribute to the reduction in enamel demineralization during orthodontic treatment.

Influence of Root Post Materials and Aging on Fracture Strength and Marginal Gap Quality of Ceramic Crowns-An In Vitro Study.

The design of and materials for prosthodontic abutments and posts have significant influences on the fracture resistance of restored teeth. This in vitro study compared the fracture strength and marginal quality of full-ceramic crowns as a function of the inserted root posts via simulation of a five-year period of use. Test specimens were prepared from 60 extracted maxillary incisors using titanium L9 (A), glass-fiber L9 (B), and glass-fiber L6 (C) root posts. The circular marginal gap behavior, linear loading capacity, and material fatigue after artificial aging were investigated. The marginal gap behavior and material fatigue were analyzed using electron microscopy. The linear loading capacity of the specimens was investigated using the Zwick Z005 universal testing machine. None of the tested root post materials showed statistically significant differences in marginal width values (p = 0.921), except in the case of marginal gap location. For Group A, there was a statistically significant difference from the labial to the distal (p = 0.012), mesial (p = 0.000), and palatinal (p = 0.005). Similarly, Group B showed a statistically significant difference from the labial to the distal (p = 0.003), mesial (p = 0.000), and palatinal (p = 0.003). Group C showed a statistically significant difference from the labial to the distal (p = 0.001) and mesial (p = 0.009). Linear load capacity reached mean values of 455.8-537.7 N, and micro-cracks occurred after artificial aging, predominantly in Groups B and C. Through the chosen experimental design, it was shown that the root post material and root post length had no influence on the fracture strength of the test teeth before or after artificial aging. However, the marginal gap location depends on the root post material and its length, which is wider mesially and distally and also tends to be greater palatinally than labially.

GDF15 Promotes the Osteogenic Cell Fate of Periodontal Ligament Fibroblasts, thus Affecting Their Mechanobiological Response.

Periodontal ligament fibroblasts (PdLFs) exert important functions in oral tissue and bone remodeling following mechanical forces, which are specifically applied during orthodontic tooth movement (OTM). Located between the teeth and the alveolar bone, mechanical stress activates the mechanomodulatory functions of PdLFs including regulating local inflammation and activating further bone-remodeling cells. Previous studies suggested growth differentiation factor 15 (GDF15) as an important pro-inflammatory regulator during the PdLF mechanoresponse. GDF15 exerts its effects through both intracrine signaling and receptor binding, possibly even in an autocrine manner. The extent to which PdLFs are susceptible to extracellular GDF15 has not yet been investigated. Thus, our study aims to examine the influence of GDF15 exposure on the cellular properties of PdLFs and their mechanoresponse, which seems particularly relevant regarding disease- and aging-associated elevated GDF15 serum levels. Therefore, in addition to investigating potential GDF15 receptors, we analyzed its impact on the proliferation, survival, senescence, and differentiation of human PdLFs, demonstrating a pro-osteogenic effect upon long-term stimulation. Furthermore, we observed altered force-related inflammation and impaired osteoclast differentiation. Overall, our data suggest a major impact of extracellular GDF15 on PdLF differentiation and their mechanoresponse.

Radiation Exposure and Frequency of Dental, Bitewing and Occlusal Radiographs in Children and Adolescents.

Dental radiographs are valuable diagnostic aids for oral healthcare, but exposure to ionizing radiation carries health risks, especially in children due to their high radio-sensitivity. Valid reference values for intraoral radiographs in children and adolescents are still missing. This study aimed to investigate the radiation dose values and underlying justifications of dental, bitewing and occlusal X-rays in children and adolescents. Data from routinely executed intraoral radiographs between 2002 and 2020 with conventional and digital tube-heads were extracted from the Radiology Information System. The effective exposure was calculated from technical parameters and statistical tests performed. A total number of 4455 intraoral (3128 dental, 903 bitewing and 424 occlusal) radiographs were investigated. For dental and bitewing radiographs, the dose area product (DAP) was 2.57 cGy × cm2 and the effective dose (ED) 0.77 µSv. For occlusal radiographs, the DAP was 7.43 cGy × cm2 and the ED 2.22 µSv. Overall, 70.2% of all intraoral radiographs were dental, 20.3% bitewing and 9.5% occlusal radiographs. The most frequent indication for intraoral radiographs was trauma (28.7%), followed by caries (22.7%) and apical diagnostics (22.7%). Moreover, 59.7% of all intraoral radiographs were taken in boys, especially for trauma (66.5%) and endodontics (67.2%) (p ≤ 0.00). Girls were significantly more frequently X-rayed for caries diagnostics than boys (28.1% vs. 19.1%, p ≤ 0.00). The average ED of 0.77 µSv for intraoral dental and bitewing radiographs in this study was within the range of other reported values. The technical parameters of the X-ray devices were found at the lowest recommended levels to best limit the radiation exposure and to assure acceptable diagnostic efficacy. Intraoral radiographs were performed predominantly for trauma, caries and apical diagnostics-reflecting general recommendations for the use of X-rays in children. For improved quality assurance and radiation protection, further studies are necessary to determine the meaningful dose reference level (DRL) for children.

Effects of reducing excess dental adhesive on bacterial adhesion in the bracket periphery.

OBJECTIVES: White spot lesions are one of the most common side effects of orthodontic therapy with a multibracket appliance and may indicate a preliminary stage of caries, also known as initial caries. Several approaches may be utilized to prevent these lesions, such as reducing bacterial adhesion in the area surrounding the bracket. This bacterial colonization can be adversely affected by a number of local characteristics. In this context, the effects of excess dental adhesive in the bracket periphery were investigated by comparing a conventional bracket system with the APC flash-free bracket system. MATERIALS AND METHODS: Both bracket systems were applied to 24 extracted human premolars, and bacterial adhesion with Streptoccocus sobrinus (S. sobrinus) was performed for 24 h, 48 h, 7 d, and 14 d. After incubation, bacterial colonization was examined in specific areas by electron microscopy. RESULTS: Overall, significantly fewer bacterial colonies were found in the adhesive area around the APC flash-free brackets (n = 507 ± 13 bacteria) than the conventionally bonded bracket systems (n = 850 ± 56 bacteria). This is a significant difference (**p = 0.004). However, APC flash-free brackets tend to create marginal gaps with more bacterial adhesion in this area than conventional bracket systems (n = 265 ± 31 bacteria). This bacterial accumulation in the marginal-gap area is also significant (*p = 0.029). CONCLUSION: A smooth adhesive surface with minimal adhesive excess is beneficial for reducing bacterial adhesion but also poses a risk of marginal gap formation with subsequent bacterial colonization, which can potentially trigger carious lesions. CLINICAL RELEVANCE: To reduce bacterial adhesion, the APC flash-free bracket adhesive system with low adhesive excess might be beneficial. APC flash-free brackets reduce the bacterial colonization in the bracket environment. A lower number of bacteria can minimize white spot lesions in the bracket environment. APC flash-free brackets tend to form marginal gaps between the bracket adhesive and the tooth.

Frequency of Dental X-ray Diagnostics in Children and Adolescents: What Is the Radiation Exposure?

Children are exposed to ionizing radiation through radiographs during their development for various reasons. At present, there are no officially valid reference values for dental X-rays in children and adolescents for dental X-ray diagnostics. This study retrospectively examined 9680 extraoral dental radiographs in pediatric patients between 2002 and 2020. The aim was to analyze the radiation doses in pediatric patients, which indications were used, and whether there were specific age and gender differences. The evaluation showed that radiation doses were considered low, with dose area products of 2.2 cGy × cm2 for a lateral cephalogram, 14 cGy × cm2 for an orthopantomogram (OPG), and 45 cGy × cm2 for cone beam computer tomography (CBCT). This corresponds to an effective dose of 1.5 µSv for a lateral cephalogram, 7 µSv for an OPG, and 33.8 µSv for CBCT. Of the 9680 images, 78% were orthopantomograms, and only 0.4% were CBCT images. OPG has become more important over the years, as reflected in the indication. Approximately one-third of all extraoral exposures are orthodontic indications. Overall, the indications were similar for both genders. According to the dental indications, boys were X-rayed slightly more frequently than girls (54.5-45.5%). A future publication of dose guide values and corresponding guidelines is of high priority.

Shear Wave Elastography in Bruxism-Not Yet Ready for Clinical Routine.

Ultrasound shear wave elastography (SWE) is an emerging modality for the estimation of stiffness, but it has not been studied in relation to common disorders with altered stiffness, such as bruxism, which affects almost one-third of adults. Because this condition could lead to an increased stiffness of masticatory muscles, we investigated SWE in bruxism according to a proof-of-principle and feasibility study with 10 patients with known bruxism and an age- and gender-matched control group. SWE of the left and right masseter muscles was estimated under three conditions: relaxed jaw, 50% of the subjective maximal bite force, and maximal jaw opening. Rejecting the null hypothesis, SWE was significantly increased during relaxed jaw (bruxism 1.92 m/s ± 0.44; controls 1.66 m/s ± 0.24), whereas for maximal mouth opening, the result was vice versa increased with 2.89 m/s ± 0.93 for bruxism patients compared with 3.53 m/s ± 0.95 in the healthy control, which could be due to limited jaw movement in chronic bruxism patients (bruxism 4.46 m/s ± 1.17; controls 5.23 m/s ± 0.43). We show that SWE in bruxism is feasible and could be of potential use for diagnostics and monitoring, though we also highlight important limitations and necessary methodological considerations for future studies.

Mediterranean diet component oleic acid increases protective lipid mediators and improves trabecular bone in a Porphyromonas gingivalis inoculation model.

AIM: Therapeutic modulation of bacterial-induced inflammatory host response is being investigated in gingival inflammation and periodontal disease pathology. Therefore, dietary intake of the monounsaturated fatty acid (FA) oleic acid (OA (C18:1)), which is the main component of Mediterranean-style diets, and saturated FA palmitic acid (PA (C16:0)), which is a component of Western-style diets, was investigated for their modifying potential in an oral inoculation model of Porphyromonas gingivalis. MATERIALS AND METHODS: Normal-weight C57BL/6-mice received OA- or PA-enriched diets (PA-ED, OA-ED, PA/OA-ED) or normal standard diet for 16 weeks and were inoculated with P. gingivalis/placebo (n = 12/group). Gingival inflammation, alveolar bone structure, circulating lipid mediators, and in vitro cellular response were determined. RESULTS: FA treatment of P. gingivalis-lipopolysaccharide-incubated gingival fibroblasts (GFbs) modified inflammatory activation, which only PA exacerbated with concomitant TNF-α stimulation. Mice exhibited no signs of acute inflammation in gingiva or serum and no inoculation- or nutrition-associated changes of the crestal alveolar bone. However, following P. gingivalis inoculation, OA-ED improved oral trabecular bone micro-architecture and enhanced circulating pro-resolving mediators resolvin D4 (RvD4) and 4-hydroxydocosahexaenoic acid (4-HDHA), whereas PA-ED did not. In vitro experiments demonstrated significantly improved differentiation in RvD4- and 4-HDHA-treated primary osteoblast cultures and reduced the expression of osteoclastogenic factors in GF. Further, P. gingivalis infection of OA-ED animals led to a serum composition that suppressed osteoclastic differentiation in vitro. CONCLUSIONS: Our results underline the preventive impact of Mediterranean-style OA-EDs by indicating their pro-resolving nature beyond anti-inflammatory properties.

Oleic acid-related anti-inflammatory effects in force-stressed PdL fibroblasts are mediated by H3 lysine acetylation associated with altered IL10 expression.

The initiation of a spatially and temporally limited inflammation is essential for tissue and bone remodelling by the periodontal ligament (PdL) located between teeth and alveolar bone. Nutritional components may cause alterations in the inflammatory response of PdL fibroblasts to mechanical stress such as those occurring during orthodontic tooth movement (OTM). Recently, we reported an attenuated pro-inflammatory response of human PdL fibroblasts (HPdLFs) to compressive forces when stimulated with oleic acid (OA), a monounsaturated fatty acid particularly prominent in the Mediterranean diet. Fatty acids could serve as alternative source of acetyl-CoA, thereby affecting epigenetic histone marks, such as histone 3 lysine acetylation (H3Kac) in a lipid metabolism-dependent manner. In this study, we aimed to investigate the extent to which OA exerts its anti-inflammatory effect in compressed HPdLFs via changes in H3Kac. Six-hour compressed HPdLFs showed increased H3Kac when cultured with OA. Inhibition of histone deacetylases resulted in a comparable IL10-increase as observed in compressed OA-cultures. In contrast, inhibition of histone acetyltransferases, particularly p300/CBP, in compressed HPdLFs exposed to OA normalized the inflammatory response to control levels. OA-dependent increased association of H3Kac to IL10 promoter regions in compressed HPdLFs further strengthened the assumption that OA exhibits its anti-inflammatory properties via modulation of this epigenetic mark. In conclusion, our study strongly suggests that nutritional components can directly affect PdL cells via changes in their epigenetic code. Since epigenetic inhibitors are already widely used clinically, they may hold promise for novel approaches for personalized orthodontic treatment that incorporates nutritional and metabolism-related changes.

Palmitate-Triggered COX2/PGE2-Related Hyperinflammation in Dual-Stressed PdL Fibroblasts Is Mediated by Repressive H3K27 Trimethylation.

The interrelationships between periodontal disease, obesity-related hyperlipidemia and mechanical forces and their modulating effects on the epigenetic profile of periodontal ligament (PdL) cells are assumed to be remarkably complex. The PdL serves as a connective tissue between teeth and alveolar bone and is involved in pathogen defense and the inflammatory responses to mechanical stimuli occurring during tooth movement. Altered inflammatory signaling could promote root resorption and tooth loss. Hyperinflammatory COX2/PGE2 signaling was reported for human PdL fibroblasts (HPdLFs) concomitantly stressed with Porphyromonas gingivalis lipopolysaccharides and compressive force after exposure to palmitic acid (PA). The aim of this study was to investigate the extent to which this was modulated by global and gene-specific changes in histone modifications. The expression of key epigenetic players and global H3Kac and H3K27me3 levels were quantitatively evaluated in dual-stressed HPdLFs exposed to PA, revealing a minor force-related reduction in repressive H3K27me3. UNC1999-induced H3K27me3 inhibition reversed the hyperinflammatory responses of dual-stressed PA cultures characterized by increased COX2 expression, PGE2 secretion and THP1 adhesion. The reduced expression of the gene encoding the anti-inflammatory cytokine IL-10 and the increased presence of H3K27me3 at its promoter-associated sites were reversed by inhibitor treatment. Thus, the data highlight an important epigenetic interplay between the different stimuli to which the PdL is exposed.

Shear bond strength after using sealant before bonding: a systematic review and meta-analysis of in vitro studies.

OBJECTIVES: Decalcification during orthodontic treatment is significantly increased. To prevent this negative impact, new treatments with sealants before bonding brackets are commonly been used. This systematic review discusses current knowledge on shear bond strength when using sealant before bonding. MATERIALS AND METHODS: A systematic review and meta-analysis were performed to identify studies that address shear bond strength after using a sealant before bonding brackets. The search was carried out using common electronic databases in addition to individual searches. Both screening and study eligibility analysis were performed according to PRISMA and Cochrane Guidelines for systematic reviews. Several terms describing shear bond strength after using a sealant before bonding brackets were searched. Particular attention was paid to bond failure and bracket loss. For the statistical outcome, all results were shown in a forest plot based on standardized mean differences (SMD) with a random-effects model to respect heterogeneity of these studies. To assess the heterogeneity of the different trials, I2-value and the Q-Test were performed. RESULTS: The initial search identified 416 studies. After a thorough selection process, a total of 15 articles met the inclusion criteria. All 15 articles reported results of in vitro studies. Papers were divided into four subgroups according to their used product: ProSeal, Transbond bonding, the combination of Transbond bonding and ProSeal and Clearfil Protect Bond. The results of this review demonstrate a high heterogeneity of the studies. The SMD of the examined 15 articles show nearly no difference between the control and the intervention groups in shear bond strength (p < 0.0001; OR - 0.12; Cl - 0.47-0.23). Forest plots for comparison of the subgroups depict no difference in shear bond strength as well. CONCLUSIONS: This meta-analysis concludes that there is no additive benefit for shear bond strength when using sealant before bonding. However, additional randomized controlled studies should be performed to analyze impact of sealants on bonding strength and bracket loss in more detail. CLINICAL RELEVANCE: Using sealants before orthodontic bonding does not reduce shear bond strength.

Impact of FGF1 on human periodontal ligament fibroblast growth, osteogenic differentiation and inflammatory reaction in vitro.

PURPOSE: To investigate in vitro the impact of fibroblast growth factor 1 (FGF1) in comparison to ascorbic acid (AscA) on human periodontal ligament fibroblast (HPdLF) growth, their osteogenic differentiation, and modulation of their inflammatory reaction to mechanical stress. METHODS: The influence of different concentrations of FGF1 (12.5-200 ng/mL) on growth and proliferation of HPdLF cells was analyzed over 20 days by counting cell numbers and the percentage of Ki67-positive cells. Quantitative expression analysis of genes encoding the osteogenic markers alkaline phosphatase (ALPL), Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OSP), as well as the fibroblast markers vimentin (VIM) and fibroblast-specific protein 1 (FSP1), was performed after 2 and 20 days of cultivation. Metabolic activity was determined by MTT assay. For comparison with AscA, 50 ng/mL FGF1 was used for stimulation for 2 and 20 days. Cell number, percentage of Ki67-positive cells, and expression of osteoblast- and fibroblast-specific genes were examined. Alkaline phosphatase activity was visualized by NBT/BCIP and calcium deposits were stained with alizarin red. Cytokine (IL­6, IL­8, COX2/PGE2) expression and secretion were analyzed by qPCR and ELISA in 6 h mechanically compressed HPdLF cultured for 2 days with FGF1 or ascorbic acid. RESULTS: Higher concentrations of FGF1 promoted cell proliferation upon short-term stimulation, whereas prolonged treatment induced the expression of osteogenic markers even with low concentrations. AscA promotes cell growth more markedly than FGF1 in short-term cultures, whereas FGF1 induced osteogenic cell fate more strongly in long-term culture. Both factors induced an increased inflammatory response of HPdLF to mechanical compression. CONCLUSION: Our data suggest that FGF1 promotes an osteogenic phenotype of HPdLF and limits inflammatory response to mechanical forces compared to AscA.

GDF15 Supports the Inflammatory Response of PdL Fibroblasts Stimulated by P. gingivalis LPS and Concurrent Compression.

Periodontitis is characterized by bacterially induced inflammatory destruction of periodontal tissue. This also affects fibroblasts of the human periodontal ligaments (HPdLF), which play a coordinating role in force-induced tissue and alveolar bone remodeling. Excessive inflammation in the oral tissues has been observed with simultaneous stimulation by pathogens and mechanical forces. Recently, elevated levels of growth differentiation factor 15 (GDF15), an immuno-modulatory member of the transforming growth factor (TGFB) superfamily, were detected under periodontitis-like conditions and in force-stressed PdL cells. In view of the pleiotropic effects of GDF15 in various tissues, this study aims to investigate the role of GDF15 in P. gingivalis-related inflammation of HPdLF and its effect on the excessive inflammatory response to concurrent compressive stress. To this end, the expression and secretion of cytokines (IL6, IL8, COX2/PGE2, TNFα) and the activation of THP1 monocytic cells were analyzed in GDF15 siRNA-treated HPdLF stimulated with P. gingivalis lipopolysaccharides alone and in combination with compressive force. GDF15 knockdown significantly reduced cytokine levels and THP1 activation in LPS-stimulated HPdLF, which was less pronounced with additional compressive stress. Overall, our data suggest a pro-inflammatory role for GDF15 in periodontal disease and demonstrate that GDF15 partially modulates the force-induced excessive inflammatory response of PdLF under these conditions.