NLRP3-mediated periodontal ligament cell pyroptosis promotes root resorption.

AIM: To investigate the mechanisms by which periodontal ligament cells (PDLCs) convert biomechanical stimulation into inflammatory microenvironment inducing root resorption (RR). MATERIALS AND METHODS: RNA sequencing was employed to explore mechanisms in force-inflammatory signal transduction. Then resorption volume, odontoclastic activity, PDLC pyroptotic ratio and NOD-like receptor protein 3 (NLRP3)-mediated pyroptosis pathway activation were analysed under force and pyroptosis inhibition. Further osteoclast formation, macrophage number and transwell polarization demonstrated the effects of PDLC pyroptosis on osteoclastogenesis and M1 polarization. RESULTS: RNA sequencing revealed that NLRP3-mediated PDLC pyroptosis induced by Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NFκB)/NLRP3 pathway may be involved in mechano-inflammatory signal transduction. PDLC pyroptosis under force and the expression of NLRP3-mediated pyroptosis pathway in force-enhanced PDLCs were significantly increased, both in vivo and in vitro. MCC950 administration was sufficient to reduce PDLC pyroptosis and alleviate RR, odontoclast formation and M1 polarization in vivo. Further in vitro exploration showed that MCC950 treatment reduced PDLC force-promoted pyroptosis and blocked NLRP3-mediated pyroptosis pathway. Moreover, by treating THP-1 with force-pretreated PDLCs or supernatants, NLRP3-mediated PDLC pyroptotic released products induced osteoclast formation and M1 polarization. CONCLUSIONS: NLRP3-mediated PDLC pyroptosis promotes RR. PDLCs transmit excessive force into inflammation signals through TLR4/NFκB/NLRP3 pathway, inducing PDLC pyroptosis, which directly promotes odontoclast formation and subsequent RR or promotes M1 polarization to indirectly trigger odontoclastogenesis and RR.

Pyroptosis in bone loss.

Pyroptosis could be responsible for the bone loss from bone metabolic diseases, leading to the negative impact on people's health and life. It has been shown that osteoclasts, osteoblasts, macrophages, chondrocytes, periodontal and gingival cells may be involved in bone loss linked with pyroptosis. So far, the involved mechanisms have not been fully elucidated. In this review, we introduced the related cells involved in the pyroptosis associated with bone loss and summarized the role of these cells in the bone metabolism during the process of pyroptosis. We also discuss the clinical potential of targeting mechanisms in the osteoclasts, osteoblasts, macrophages, chondrocytes, periodontal and gingival cells touched upon pyroptosis to treat bone loss from bone metabolic diseases as well as the challenges of avoiding potential side effects and producing efficient treatment methods.

Wnt5a-Ror2 signaling mediates root resorption.

INTRODUCTION: This study aimed to investigate the role of wingless-type MMTV integration site family member 5a (Wnt5a)-receptor tyrosine kinase-like orphan receptor 2 (Ror2) signaling in root resorption. METHODS: The messenger RNA (mRNA) expression of Wnt5a, Ror2, and RANKL in periodontal ligament cells (PDLCs) under compression force (CF) with or without Ror2 small interfering RNA (siRNA) were measured by quantitative reverse transcription-polymerase chain reaction, and these proteins released into culture supernatants were measured using enzyme-linked immunosorbent assay. Then these PDLC-conditioned media under CF with or without Ror2 siRNA were used to culture osteoclast precursors to detect osteoclastogenesis effects via tartrate-resistant acid phosphatase staining. In in vivo studies, the odontoclast number and the root resorption volume under excessive CF with or without Ror2 siRNA were investigated by tartrate-resistant acid phosphatase immunohistochemical staining and microcomputed tomography. The protein levels for Wnt5a, Ror2, and receptor activator of nuclear factor-kappa B ligand (RANKL) in the periodontal ligament tissues were also detected using immunohistochemical staining. Finally, the odontoclast number, root resorption volume, and the mRNA and protein expressions were compared between immature and mature teeth. RESULTS: The mRNA production and protein release level of Wnt5a, Ror2, and RANKL increased after CF, whereas they were significantly downregulated with Ror2 siRNA. The osteoclast number increased treating with culture medium from PDLC applying CF, but the increase was inhibited after adding Ror2 siRNA. In the animal model, the odontoclast number and root resorption volume significantly increased in the CF group but decreased in the CF with the Ror2 siRNA group. The protein levels of Wnt5a, Ror2, and RANKL in periodontal ligament were upregulated under excessive CF, and the pathway was inhibited with Ror2 siRNA. In the immature tooth group, the odontoclast number, root resorption volume, and the mRNA and protein expressions of Wnt5a-Ror2 signaling were reduced. CONCLUSIONS: Wnt5a-Ror2 signaling in PDLCs enhanced by excessive CF could promote RANKL release and induce precursor differentiation, partly leading to increased odontoclast activity and ultimate root resorption. The less resorption of the immature tooth may be due to odontoclastogenesis inhibition by decreased expression of Wnt5a-Ror2 signaling.

Intermittent parathyroid hormone improves orthodontic retention via insulin-like growth factor-1.

OBJECTIVES: This study aimed to investigate the effects of intermittent parathyroid hormone (iPTH) on the stability of orthodontic retention and to explore the possible regulatory role of insulin-like growth factor-1 (IGF-1) in this process. METHODS: Forty-eight 6-week-old male Wistar rats were adopted in this study. An orthodontic relapsing model was established to investigate the effects of iPTH on orthodontic retention. In vitro, an immortalized mouse cementoblast cell line OCCM-30 was detected by flow cytometry to study the effects of iPTH on cell proliferation and apoptosis. By application of a specific IGF-1 receptor inhibitor, the role of IGF-1 was also explored. RESULTS: In vivo study found that daily injection of PTH significantly reduced the relapsing distance. Histological staining and ELISA assay showed faster periodontal regeneration during retention period in PTH group with increased RANKL/OPG ratio and greater amount of OCN, ALP, and IGF-1 in gingival cervical fluid (GCF). Cell experiment revealed that iPTH promoted proliferation and suppressed apoptosis of cementoblast. IGF-1 receptor inhibitor significantly restrained the anabolic effect of iPTH on OCCM-30 cells. CONCLUSIONS: These findings suggest that iPTH could improve the stability of tooth movement by promoting periodontal regeneration. IGF-1 is essential in mediating the anabolic effects of iPTH.

Notch signaling inhibition protects against root resorption in experimental immature tooth movement in rats.

INTRODUCTION: This study aimed to build an experimental immature tooth movement model and verify less resorption of incompletely developed roots than those fully developed during the same orthodontic treatment, followed by investigating the cellular and molecular mechanism. METHODS: The development of Wistar rat tooth was investigated using in vivo microcomputed tomography and hematoxylin and eosin staining to decide the optimal ages of rats for immature tooth and mature tooth groups. The rats in the immature tooth and mature tooth groups were divided into experimental, sham control, and blank control groups. After orthodontic treatment for 3 weeks, the mesial root volume, crown movement distance, neck movement distance, root inclination, and apical distance were measured by microcomputed tomography. The expressions of TRAP, Jagged1, Notch2, IL-6, and RANKL were analyzed by immunohistochemical staining and real-time polymerase chain reaction. The repair of root resorption was also investigated after removing orthodontic force for 3 and 6 weeks. RESULTS: The root achieved the development stage around 10 weeks, so 4-week-old rats and 10-week-old rats were used in the immature tooth group and mature tooth group, respectively. The volume of root resorption in the experimental immature tooth group was 0.0869 ± 0.0244 mm3, which was less than that in the mature tooth group (0.1218 ± 0.0123 mm3) (P <0.001). Immature tooth movement decreased TRAP-positive odontoclasts on the compression side while having no statistically significant effect on osteoclasts. The protein expression of Jagged1, Notch2, IL-6, and RANKL in the mature tooth group increased significantly compared with the immature tooth group, not only on the compression side but also on the tension sides. The mRNA expression of Jagged1, Notch2, and RANKL was significantly lower in the immature tooth group, whereas the expression of IL-6 had no significance but a strong tendency. The root volume after repairing for 3 weeks was still less than that of blank control, whereas after repairing for 6 weeks, the difference was not statistically significant. CONCLUSIONS: The experimental immature tooth movement model for the Wistar rat was achieved for the first time. The immature tooth will suffer less root resorption than the mature tooth, which may be due to odontoclastogenesis inhibition by decreased expression of Jagged1/Notch2/IL-6/RANKL signaling.

Expression of proteins of elastic fibers and collagen type I in orthodontically rotated teeth in rats.

INTRODUCTION: The aims of this study were to investigate the expression of proteins of elastic fibers and collagen type I in the supra-alveolar structure of orthodontically rotated teeth in rats and to elucidate whether circumferential supracrestal fiberotomy diminishes relapse. METHODS: The rats' maxillary left first molars were rotated by couple of force. Specimens were divided into groups according to different orthodontic procedures. A1-3 and B1-3 were blank control groups and operation control groups. Group C underwent rotation only, and group D was treated with rotation and retention. Groups E and F were treated with rotation, retention, and release of retention; additionally, circumferential supracrestal fiberotomy was performed in group F before the release of retention. The animals were killed, and the jaws were processed for histologic evaluation using the immunohistochemical method to evaluate the protein expressions of elastin, fibrillin-1, fibrillin-2, and collagen type I in supra-alveolar structures (around and below the gingival sulcus) between the maxillary left first and second molars. The degree and percentage of relapse were measured by a series of impressions. RESULTS: The degree and percentage of relapse in group F were much lower than those in group E (P <0.05). Collagen type I was increased in group C (P <0.05) and at normal levels in groups D, E, and F. Elastin below the gingival sulcus and fibrillin-1 showed the same patterns of expression and were consistently elevated in groups C, D, E, and F (P <0.05). No positive staining for elastin was found around the gingival sulcus in any specimen. The difference in the expression of fibrillin-2 between the experimental groups (C, D, E, and F) and their matching control groups was not statistically significant (P >0.05). CONCLUSIONS: Circumferential supracrestal fiberotomy can alleviate the relapse of rotated teeth. Collagen fibers of supra-alveolar structures might contribute to relapse in a short time, whereas elastic fibers may be the reason that rotated teeth relapse to their original positions after retention.

Protein and mRNA expressions of IL-6 and its key signaling factors under orthodontic forces in mice: An in-vivo study.

INTRODUCTION: The purpose of this study was to investigate the mechanical loading-induced changes in protein and mRNA expressions of interleukin-6 (IL-6) and its key signaling factors glycoprotein 130 (gp130), signal transducer and activator of transcription 3 (STAT3), and the Src homology phosphotyrosine phosphatase (SHP2) at the tension and compression sides of the teeth in mouse models. METHODS: A total of 55 C57B/6 mice (10 weeks old) were divided into 3 groups. Orthodontic force was applied in group A (experimental group, n = 30); the tooth movement device was placed without activation in group B (sham control group, n = 15), and group C (blank control group, n = 10). Tooth movement was induced by a nickel-titanium coil spring inserted between the maxillary left incisor and the first molar with a force of approximately 4 g. The animals were killed 12 days after the interventions; protein and mRNA expressions of IL-6, gp130, STAT3, and SHP2 in the periodontal tissues were observed with immunohistochemistry and in-situ hybridization, respectively. RESULTS: In contrast with the control groups, we observed enhanced expressions of IL-6, gp130, STAT3, and SHP2 protein and mRNA at the mesial and distal sides of the teeth with application of orthodontic forces in the experimental group. In contrast with the distal side, we observed enhanced expression of gp130 protein and mRNA at the mesial side in the experimental group. CONCLUSIONS: We observed enhanced expression of IL-6 and its key signaling factors gp130, STAT3, and SHP2 protein and mRNA at the tension and compression sides of the teeth with application of orthodontic forces. The mechanical loading applied for orthodontic tooth movement might induce changes in protein localization and mRNA expression patterns of IL-6 and its key signaling factors gp130, STAT3, and SHP2 at the tension and compression sides of the periodontal ligaments of the teeth in mouse models. The result might demonstrate the special role of IL-6 and its key signaling factors in the alveolar bone-modeling process.

Genetic and clinical risk factors of root resorption associated with orthodontic treatment.

INTRODUCTION: External apical root resorption (EARR) is a common complication in orthodontic treatment. Despite many studies on EARR, great controversies remain with regard to its risk factors. The objective of this study was to explore the relationship among sex, root movement, IL-1RN single nucleotide polymorphism (SNP) rs419598, IL-6 SNP rs1800796, and EARR associated with orthodontic treatment. METHODS: Altogether 174 patients (with 174 maxillary left central incisors) were selected for this study. Cone-beam computed tomography was performed before the start of the treatment and at the end of the treatment. Cone-beam computed tomography data were used to reconstruct a 3-dimensional image of each tooth; the volume and the root resorption volume of each tooth were calculated. Three-dimensional matching was used to measure the amount of movement of each root. Genomic DNA was extracted from buccal swabs, and genotypes of SNP rs419598 and SNP rs1800796 of each subject were determined using TaqMan polymerase chain reaction genotyping (Applied Biosystems, Foster City, Calif). The data were analyzed with multiple linear regression analysis. RESULTS: The statistical analysis indicated no relationship between sex, tooth movement amount, and IL-1RN SNP rs419598 with EARR. The IL-6 SNP rs1800796 GC was associated with EARR, and root resorption differed significantly between SNP rs1800796 GC and CC. CONCLUSIONS: IL-6 SNP rs1800796 GC is a risk factor for EARR. The amount of root movement, IL-1RN SNP rs419598, and sex as risk factors for EARR need further study.

Association between CYP19A1 genotype and pubertal sagittal jaw growth.

INTRODUCTION: Sagittal jaw growth is influenced during puberty by a ratio of androgens and estrogens. The CYP19A1 (formerly CYP19) gene encodes the cytochrome P450 enzyme aromatase (estrogen synthetase), which converts testosterone to estrogen. Genetic variations including single nucleotide polymorphisms might regulate CYP19A1 gene expression or the function of the aromatase protein and thus influence sagittal jaw growth. METHODS: The annual sagittal jaw growth in 92 pubertal orthodontic patients was determined by using pretreatment and posttreatment cephalometric radiographs. Single nucleotide polymorphisms rs2470144 and rs2445761 were genotyped and haplotypes constructed. Associations between genotypes or haplotypes and the annual sagittal growth were estimated by using JMP (version 9.0; SAS Institute, Cary, NC). RESULTS: Two single nucleotide polymorphisms were significantly associated with average differences in annual sagittal jaw growth in boys. Haplotype analysis demonstrated that haplotypes T(rs2470144)T(rs2445761) and C(rs2470144)T(rs2445761) had significant effects on annual sagittal maxillary growth and on mandibular growth in boys. No association was found in girls. CONCLUSIONS: A quantitative trait locus that influences male pubertal sagittal jaw growth might exist in the CYP19A1 gene, and single nucleotide polymorphisms rs2470144 and rs2445761 might be inside this quantitative trait locus or be linked to it.

Distalization of the mandibular dentition of an adult with a skeletal Class III malocclusion.

This case report describes the orthodontic treatment of an 18-year-old woman with a skeletal Class III malocclusion and a midline deviation. The treatment plan consisted of distalizing the mandibular dentition asymmetrically and producing space for retraction of the mandibular anterior teeth. Short Class III elastics, an open-coil spring, and the mulitloop edgewise archwire technique were used, combining the entire maxillary dentition as integrated anchorage. The active treatment period was 26 months. Normal overbite and overjet were obtained, and facial balance was improved.

Effect of the use of stabilization splint on masticatory muscle activities in TMD patients with centric relation-maximum intercuspation discrepancy and absence of anterior/lateral guidance.

Objective: To assess the masticatory muscle activities in patients with temporomandibular disorders (TMD) with centric relation (CR)-maximum intercuspation (MI) discrepancy and absence of anterior/lateral guidance by electromyography (EMG).Methods: Forty-six patients with TMD and 34 controls underwent clinical evaluations and EMG recordings at baseline. Twenty-three patients and 17 controls wore a stabilization splint. The remaining subjects were followed up without any intervention. After 3 months, the EMG recordings and clinical evaluations were repeated.Results: At baseline, the EMG values at rest and EMG value variations after movements in patients were higher than those in controls. After treatment, no difference was observed between EMG values of patients and controls.Conclusion: In the present study, patients with TMD had abnormal muscular activities. The stabilization splint is effective for functional recovery of the masticatory muscles, possibly by eliminating the CR-MI discrepancy and establishing anterior/lateral guidance.

The use of a dynamic real-time jaw tracking device and cone beam computed tomography simulation.

BACKGROUND: The aim was to introduce and preliminarily evaluate a new software application, SICAT Function, which can directly combine and merge three-dimensional cone beam computed tomography (CBCT) and electronic SICAT jaw motion tracking (JMT) data. METHODS: A detailed description of the methods and dynamic clinical simulation of mandibular movements of a patient are demonstrated. Functional jaw movements on 3 days were recorded by JMT tracking system. The simulation was performed by merging CBCT and JMT data in the software SICAT Function suite. The condylar position simulated by SICAT Function suite was compared with real condyle position showed by a CBCT of the patient. RESULTS: The incisor ranges of functional movements were displayed by JMT tracking system. The visualization of patient-specific mandibular movement including the translation of the condyles was displayed after data merge. The recordings of mandibular movements of the patient were similar on 3 different days. The condylar position simulated by SICAT was coincident with real condyle position by CBCT data with the same amount of mouth opening. CONCLUSIONS: The SICAT Function software is a system capable of measuring and visualizing patient-specific jaw movement relative to the patient-specific anatomy of the jaw. Further studies are needed to validate its accuracy and its potential for future use.

Temporomandibular disorders with skeletal open bite treated with stabilization splint and zygomatic miniplate anchorage: a case report.

This case report describes the treatment of a patient with temporomandibular disorder (TMD) and skeletal open bite. First, the patient was treated with a stabilization splint to stabilize the condyles in centric relation and to alleviate TMD signs and symptoms. After making a definitive diagnosis from postsplint records, orthodontic treatment was initiated. Titanium miniplates were placed at bilateral zygomatic buttresses and used as orthodontic anchorage for molar intrusion and distalization. The treatment was completed after 30 months. Satisfactory appearance and function were achieved for this patient.

Camouflage treatment of skeletal Class III malocclusion with multiloop edgewise arch wire and modified Class III elastics by maxillary mini-implant anchorage.

OBJECTIVE: To evaluate the effect of the multiloop edgewise arch wire (MEAW) technique with maxillary mini-implants in the camouflage treatment of skeletal Class III malocclusion. MATERIALS AND METHODS: Twenty patients were treated with the MEAW technique and modified Class III elastics from the maxillary mini-implants. Twenty-four patients were treated with MEAW and long Class III elastics from the upper second molars as control. Lateral cephalometric radiographs were obtained and analyzed before and after treatment, and 1 year after retention. RESULTS: Satisfactory occlusion was established in both groups. Through principal component analysis, it could be concluded the anterior-posterior dental position, skeletal sagittal and vertical position, and upper molar vertical position changed within groups and between groups; vertical lower teeth position and Wits distance changed in the experimental group and between groups. In the experimental group, the lower incisors tipped lingually 2.7 mm and extruded 2.4 mm. The lingual inclination of the lower incisors increased 3.5°. The mandibular first molars tipped distally 9.1° and intruded 0.4 mm. Their cusps moved 3.4 mm distally. In the control group, the upper incisors proclined 3°, and the upper first molar extruded 2 mm. SN-MP increased 1.6° and S-Go/N-ME decreased 1. CONCLUSIONS: The MEAW technique combined with modified Class III elastics by maxillary mini-implants can effectively tip the mandibular molars distally without any extrusion and tip the lower incisors lingually with extrusion to camouflage skeletal Class III malocclusions. Clockwise rotation of the mandible and further proclination of upper incisors can be avoided. The MEAW technique and modified Class III elastics provided an appropriate treatment strategy especially for patients with high angle and open bite tendency.

[Diagnostic accuracy of orthopantomogram and periapical film in evaluating root resorption associated with orthodontic force].

OBJECTIVE: To test the diagnostic accuracy of qualitative and semi-quantitative evaluations on root resorption associated with orthodontic force (RRAOF) using orthopantomogram (OPG) and periapical film, through comparing with assessments with cone beam computed tomography (CBCT), which is considered as the gold standard. METHODS: Before and after treatment, OPG, periapical film, CBCT images of 121 upper anterior teeth in 32 orthodontic patients were obtained. Blind evaluation of RRAOF was performed according to Levander and Malmgren's method by three trained radiographers through examining three kinds of images of these teeth. The diagnostic accuracy of qualitative and semi-quantitative evaluations on RRAOF using OPG and periapical films were test through comparing with assessments with CBCT, which was considered as the gold standard. RESULTS: To the same sample, the RRAOF qualitative and semi-quantitative diagnostic classification results of OPG and periapical films compared with CBCT were significant differences (P<0.05). The sensitivity of periapical films and OPG was 71.6% and 78.4%. The specificity of periapical films and OPG was 25.5% and 51.1%. The accuracy of periapical films and OPG was 53.7% and 67.8%, respectively. CONCLUSION: Periapical films and OPG for evaluating RRAOF have a poor diagnostic accuracy.

[The centric relation-maximum intercuspation discrepancy in adult angle's class II pretreatment patients].

OBJECTIVE: This study investigates the differences in condylar position between centric relation (CR) and maximum intercuspation (MI) in Angle's Class II orthodontic patients before treatment. METHODS: 80 cases, who were Angle's Class II pretreatment patients, and 50 cases, who were normal(ideal) occlusion were accepted. Dental casts were mounted on Panadent articulator with CR bite record, taken by bilateral manipulation and load testing. The differences in condylar position between CR and MI in all three spatial planes were measured using the Panadent condylar position indication (CPI). RESULTS: (1) The positive rate of CR-MI discrepancy was 92.50%(74 cases) in the group of Angle's Class II malocclusion and 10.00% (5 cases) in the group of normal occlusion(P< 0.001). 2)74.32% (55 cases) CR-MI discrepancy in 74 cases in the group of Angle's Class II pretreatment patients were coincidence discrepancy. (3)91.25% patients in the group of Angle's Class 11 malocclusion and 66.00% in the group of normal occlusion present occlusion interferences which located at the posterior teeth. CONCLUSION: The results suggested that orthodontists should be aware of a high incidence of condylar displacement in Angle's Class ii pretreatment patients, and measure condylar displacement before the start of comprehensive orthodontic treatment to unmask real jaw relationships and avoid possible misdiagnoses.

[The measurement of utility value for skeletal malocclusion].

OBJECTIVE: To measure the utility value of different skeletal malocclusion for patients receiving orthodontic treatment. METHODS: Utility value of different skeletal malocclusion for patients was measured by rating scale and time trade-off. RESULTS: The youth group had higher utility values than adult group for skeletal malocclusion Class II (protruding facial type) with median mandibular angle. The utility value of skeletal malocclusion Class III (concave facial type) with low mandibular angle was the lowest, and the utility value of skeletal malocclusion Class II (protruding facial type) with median mandibular angle was the highest. There was no difference in the utility values by rating scale and by time trade-off. CONCLUSION: For some skeletal malocclusion, the youth had different utility values with the adult.