Molecular signaling and mechanisms of low-level laser-induced gene expression in cells involved in orthodontic tooth movement.

BACKGROUND: The study aimed to observe molecular signaling, including reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm), to evaluate the alteration of gene expression by low-level laser therapy (LLLT) and the correlation between its mechanisms and the NF-kB pathway in cells involved in orthodontic tooth movement. METHODS: Osteoblast-like cells (MG63), immortalized periodontal ligament cells (iPDL), and M1 macrophage-like cells were irradiated by 980-nm LLLT with energy densities of 1 and 10 J/cm2 ΔΨm and intracellular ROS were monitored using fluorescent probes. The changes of mRNA expression were assessed using reverse transcription polymerase chain reaction (RT-PCR). NF-kB inhibitor, ROS scavenger, and ΔΨm suppressor were used to analyze signals associated with the regulation of gene expression. Finally, Western blot analysis was performed to confirm NF-kB signaling after LLLT. RESULTS: We found the increases of ΔΨm and ROS in all three cell types after LLLT, but no significant difference was observed between 1 and 10 J/cm2 LLLT. Regarding gene expression, some target genes were upregulated in MG63 6 h, 12 h, and 1 day after LLLT and in iPDL cells 12 h and 1 day after LLLT. However, no changes occurred in M1 cells. The inhibitor that significantly reduced most changes in gene expression was NF-kB inhibitor. Western blot analysis showed the increase in p-IkBα level after LLLT in iPDL and MG63, but not in M1. CONCLUSION: The 980-nm LLLT increased ΔΨm and ROS production in all three cell types. However, changes in gene regulation were found only in MG63 and iPDL cells, which related to the NF-kB pathway.

Differential outcomes of jaw bone position after surgical-orthodontic treatment in three types of skeletal Class III asymmetry patients.

BACKGROUND/PURPOSE: Facial asymmetry is common in Class III patients requiring orthognathic surgery. This study aimed to analyze jaw bone position after surgical-orthodontic treatment in three types of skeletal Class III asymmetry patients. METHODS: The retrospective study included 30 Class III patients who underwent surgical-orthodontic treatment comprising LeFort I osteotomy and bilateral sagittal split osteotomy (BSSO) without genioplasty. Cone-beam computed tomography (CBCT) images obtained before surgery (T1) and after post-surgical orthodontic treatment (T2) were superimposed with voxel-based registration. Patients were classified into three groups based on T1 CBCT scans. Groups 1 and 2 exhibited menton and ramus deviated to the same side. Menton deviation was larger than ramus width asymmetry in group 1, while the reverse was true for group 2. Group 3 had menton deviation contralateral to the side with greater ramus width. RESULTS: Menton deviation after treatment was improved in all groups. Ramus width asymmetry and coronal ramus angle difference decreased in groups 1 and 2. Neither improvement nor deterioration of ramus width asymmetry was noted for group 3. Comparing to groups 1 and 2, group 3 had greater roll and yaw rotations of distal segment, more upward pitch of proximal segment on chin deviation side, and largest inward yaw as well as backward translation of proximal segment on non-deviation side. CONCLUSION: The positional changes of osteotomy segments differed among three types of mandibular asymmetry. Special attention should be given to the atypical mandibular asymmetry with mandibular body and ramus deviating to opposite directions during surgical correction of jaw deflection.

Orthodontic correction of acquired open bite with TMJ degeneration: A retrospective study of outcomes and stability.

BACKGROUND/PURPOSE: Newly developed temporary anchorage devices (TADs) serve a strong orthodontic anchorage to intrude molars for correction of anterior open bite (AOB). We measured cephalometric changes in skeletal open bite patients which developed subsequently to temporomandibular joint disorders with bilateral point contacts at terminal molars. METHODS: We retrospectively recruited 32 patients who had been treated their TMD before orthodontic correction (overbite: -3.14 ± 1.86 mm). Partial orthodontic appliances were used to intrude posterior teeth using TADs until positive OB obtained (T1). Full fixed appliances were then used to achieve proper overjet and overbite (T2). We collected lateral cephalograms before (T0), during (T1) and after (T2) treatment, and at follow-ups (T3). Using ANOVA, we analyzed the differences among these time points to determine treatment changes and stability of orthodontic results. RESULTS: In this group predominantly comprising young adult women, orthodontic treatment with TADs significantly reduced upper posterior dental heights (T2-T0:-1.84 ± 0.66 mm) and facilitated the retraction and uprighting upper incisors (T2-T0: -9.92 ± 1.72°), to achieve appropriate OJ (T2-T0: -3.21 ± 0.49 mm) and OB (T2-T0: 4.10 ± 0.28 mm) with p < 0.05. Except upper posterior dental height, most of cephalometric changes including OJ, OB, and upper incisal axis remained significant at follow-ups with retention time of 3.7 ± 2.6 years. Only three out of 30 patients experienced small amount of open bite at T3. CONCLUSION: Orthodontic correction of OJ remained relatively stable among 90 % of patients with TMJ degeneration by intrusion via TADs. This modern but conservative orthodontic approach can improve occlusal functions in skeletal open bites.

Occlusal function and electromyographic activity of masticatory muscles in skeletal Class III patients with different patterns of mandibular asymmetry.

BACKGROUND: Although jaw asymmetry is commonly seen in skeletal Class III patients, its correlation with occlusal function and masticatory muscle activity has not been fully elucidated. OBJECTIVES: The purpose of this study was to investigate the occlusal function and masticatory muscle activity in skeletal Class III patients with various patterns of mandibular asymmetry. METHODS: Forty-two patients and 10 normal participants were examined. The patients were categorised into three groups. Groups 1 and 2 exhibited menton and ramus deviation to the same side. Menton deviation was larger than ramus deviation in Group 1, whereas Group 2 showed the inverse relation. Group 3 patients showed menton and ramus deviation in opposite directions. Occlusal contact area (OCA), relative bite force (RBF), and temporalis anterior (TA) and masseter muscle (MM) activity at maximum clenching were measured using T-Scan Novus system and Bio-EMG-III. Statistical analysis was performed using the t-test, one-way analysis of variance with Bonferroni correction and Spearman correlation (α = .05). RESULTS: Compared with normal participants, the patients had smaller OCA and greater asymmetry in the distribution of masticatory muscle activity. Greater ramus deviation was associated with smaller OCA in Group 1 but with larger OCA in Group 3. In Group 1, greater menton deviation was related to stronger TA activity on the non-deviation side. In Group 2, greater ramus deviation was related to stronger MM activity on the deviation side. CONCLUSION: Deviation of the menton and ramus was individually related to OCA and masticatory muscle activity, and this relationship varied according to the pattern of mandibular asymmetry.

Determining factors affecting job strain for nurse practitioners in acute care practice.

PURPOSE: To investigate nurse practitioners' (NPs) experience of job strain in acute care settings, and to examine the factors that may affect job strain in that context. DESIGN: Descriptive design using a national survey was employed. METHODS: A total of 1396 NPs completed online surveys that recorded demographic characteristics and included a Job Content Questionnaire (JCQ), a Condition for Work Effectiveness Questionnaire (CWEQ), and the Dempster Practice Behavior Scale (DPBS). Multiple logistic regressions were conducted to explore the factors associated with job strain types. FINDINGS: NPs were classified into job strain categories of passive (24.4%), active (31.4%), low (19.2%), or high (24.9%). The passive job strain type was associated with overtime hours, organizational empowerment, and autonomy. The active job type was associated with higher organizational empowerment and autonomy. The high-strain and low-strain job types were both associated with overtime hours and autonomy. CONCLUSION: Nearly 25% of acute care NPs are in a high job strain type. Organizational empowerment and autonomy were two major factors associated with the passive and active job strain types. Overtime hours and autonomy were both associated with the high-strain and low-strain job types. CLINICAL RELEVANCE: Supportive hospital/nursing leadership should acknowledge the impact of NP practice as they can contribute to the operational efficacy of their organization. Hospital administrators should provide a supportive practice environment by empowering NPs, enhancing autonomy, and addressing working conditions for NPs to decrease the odds of having a passive or high-strain job type in practice.

Biological properties of human periodontal ligament cell spheroids cultivated on chitosan and polyvinyl alcohol membranes.

BACKGROUND/PURPOSE: Multicellular spheroid cultures have attracted increasing attention in the field of periodontal regeneration. However, very few studies have reported the periodontal ligament (PDL) cell spheroid formation via biomaterials-induced processes. This study investigated the biological characteristics of human PDL cell spheroids formed on two hydrophilic polymer-based biomaterials, namely chitosan and polyvinyl alcohol. METHODS: The expressions of periostin, paxillin, hypoxia-inducible factor 1-α (HIF-1α), and vascular endothelial growth factor (VEGF) were analyzed. Cell migration ability was assessed using a scratch assay. Furthermore, PDL cell spheroids were cultured in 3D-printed polylactic acid scaffolds to evaluate mineralizing capability. RESULTS: Western blot analysis revealed increased expressions of periostin, HIF-1α, and VEGF in the 3D spheroids. After the spheroids were reseeded, the cells gradually migrated outward from the spheroids and time-dependent distribution of paxillin was observed. The cells migrating outward from the 3D spheroids demonstrated greater migration ability than that of 2D monolayer cells. Compared to the dissociated cells from a monolayer culture, the cell spheroids formed on the chitosan membrane exhibited elevated alkaline phosphatase activity and an increase in mineralized matrix deposition. CONCLUSION: The biomaterial-induced formation of PDL cell spheroids suggests a novel strategy for cell delivery in research and clinical applications of periodontal regeneration.

Differences of condylar changes after orthognathic surgery among Class II and Class III patients.

BACKGROUND/PURPOSE: The nature of susceptibility to condylar resorption after orthognathic surgery can be different between skeletal Class II and Class III populations, which was addressed by few in the past. The aim of the present study was to use cone-beam computed tomographic (CBCT) images to investigate the displacement and morphological changes of temporomandibular joints (TMJs) in patients received orthodontic treatment combined with orthognathic surgery. METHODS: Both Class III (n = 34) and Class II (n = 17) patients were compared through overall and regional superimpositions of the initial and posttreatment CBCTs. Two-sample t-test was used to identify significance between group differences. Pearson's correlation coefficient was used to address changes of TMJ and the amount of setback or advancement. RESULTS: The axial ramal angle increased significantly in Class III group and decreased in Class II groups after orthognathic surgery (p < FDR_p). For condylar dimensions, significant widths and lengths reductions were noted only in Class II group. However, no significant difference was found after comparing subgroup differences according to one-jaw and two-jaw options, nor any significant correlation found between the condylar changes and the amount of surgical movements. CONCLUSION: The nature of condylar susceptibility could result more from different skeletal patterns than the amount of surgical movements. However, the direction of mandibular surgery may contribute to different changes of condylar angle in axial section.

Experimentally-constrained biophysical models of tonic and burst firing modes in thalamocortical neurons.

Somatosensory thalamocortical (TC) neurons from the ventrobasal (VB) thalamus are central components in the flow of sensory information between the periphery and the cerebral cortex, and participate in the dynamic regulation of thalamocortical states including wakefulness and sleep. This property is reflected at the cellular level by the ability to generate action potentials in two distinct firing modes, called tonic firing and low-threshold bursting. Although the general properties of TC neurons are known, we still lack a detailed characterization of their morphological and electrical properties in the VB thalamus. The aim of this study was to build biophysically-detailed models of VB TC neurons explicitly constrained with experimental data from rats. We recorded the electrical activity of VB neurons (N = 49) and reconstructed morphologies in 3D (N = 50) by applying standardized protocols. After identifying distinct electrical types, we used a multi-objective optimization to fit single neuron electrical models (e-models), which yielded multiple solutions consistent with the experimental data. The models were tested for generalization using electrical stimuli and neuron morphologies not used during fitting. A local sensitivity analysis revealed that the e-models are robust to small parameter changes and that all the parameters were constrained by one or more features. The e-models, when tested in combination with different morphologies, showed that the electrical behavior is substantially preserved when changing dendritic structure and that the e-models were not overfit to a specific morphology. The models and their analysis show that automatic parameter search can be applied to capture complex firing behavior, such as co-existence of tonic firing and low-threshold bursting over a wide range of parameter sets and in combination with different neuron morphologies.

Clinical outcomes of adjunctive indocyanine green-diode lasers therapy for treating refractory periodontitis: A randomized controlled trial with in vitro assessment.

BACKGROUND/PURPOSE: It is still challengeable to treat periodontal pockets refractory to mechanical debridement. This study is to evaluate the potential of indocyanine green (ICG)-diode laser-based photothermal therapy (PTT) for solving this dilemma. METHODS: Bone marrow-derived mesenchymal stem cells (BMSCs) and periodontal ligament cells (PDLCs) were incubated with phosphate-buffered saline, chlorhexidine, or ICG, non-irradiated or irradiated with 810-nm diode lasers, and the cell viability was evaluated. Patients with teeth refractory to mechanical periodontal debridement on different quadrants were recruited. At baseline (T0), all examined teeth received scaling and root planing, and those on the test quadrant (PTT group) received ICG-diode laser treatment. The outcome was evaluated using clinical parameters and cytokines in the gingival crevicular fluids at 4-6 weeks (T1) and 6 months (T2). RESULTS: In ICG-treated cultures, the viability of BMSCs and PDLCs was recovered on day 4, and laser irradiation inhibited the metabolic activities of BMSCs. 22 patients with 30 control teeth and 35 PTT-treated teeth were examined. All examined teeth showed modest reductions in probing pocket depth (PPD), clinical attachment loss (CAL), bleeding upon probing (BOP), and plaque score at T1 and T2 and significant reductions in IL-1ß and MMP-8 at T2. Compared with controls, BOP was reduced more prominently, IL-1ß and MMP-8 were significantly lower, and reductions in PPD and CAL were slightly greater in the PTT group at T1 (0.05-0.19 mm). CONCLUSION: ICG-diode laser-based PTT is compatible to periodontium and assists in faster resolution of gingival inflammation in periodontal pockets refractory to mechanical debridement.

Analysis of root canal system of maxillary first and second molars and their correlations by cone beam computed tomography.

BACKGROUND/PURPOSE: Understanding the root canal systems of molars and the association of root canal system in adjacent or contralateral molars is important for dental practice. This study aimed to use cone-beam computed tomography (CBCT) to analyze the morphology similarity of root canal systems in the maxillary first and second molars. METHODS: CBCT images of 1741 maxillary molars in a total of 519 patients were blindly examined to analyze the correlation of root canal systems between maxillary first and second molars as well as the bilateral first and second molars. RESULTS: The most common type in maxillary first molars is 3R4C (3 roots/4 canals), whereas in maxillary second molars is 3R3C.The symmetry in type of root canals in bilateral maxillary first and second molars were 87.36% and 79.85%, respectively. The similarities of root canal system in adjacent maxillary first and second molars were 53.07% (right side) and 52.58% (left side). The concurrence of MB2 canal in bilateral maxillary first molars is 77.8%, and 35.97% in maxillary second molars. In the 110 patients with MB2 canal in bilateral maxillary second molars, the chance of bilateral MB2 canals in their maxillary first molar is almost 100%. CONCLUSION: Maxillary first molars have higher prevalence of 3R4C than second molars. The symmetry in bilateral maxillary molars is higher than the similarity in adjacent maxillary first and second molars. Application of CBCT analysis of root canal system can improve endodontic treatment outcomes. The correlation of root canal system between teeth is useful for genetic linkage.

Aggregation of human dental pulp cells into 3D spheroids enhances their migration ability after reseeding.

Multicellular three-dimensional (3D) spheroids allow intimate cell-cell communication and cell-extracellular matrix interaction. Thus, 3D cell spheroids better mimic microenvironment in vivo than two-dimensional (2D) monolayer cultures. The purpose of this study was to evaluate the behaviors of human dental pulp cells (DPCs) cultured on chitosan and polyvinyl alcohol (PVA) membranes. The protein expression of hypoxia-inducible factor 1-α (HIF-1α) and vascular endothelial growth factor (VEGF), and the migration ability of the DPCs from 2D versus 3D environments were investigated. The results showed that both chitosan and PVA membranes support DPCs aggregation to form multicellular spheroids. In comparison to 2D cultures on tissue culture polystyrene, DPC spheroids exhibited higher protein expression of HIF-1α and VEGF. The treatment with YC-1 (inhibitor to HIF-1α) blocked the upregulation of VEGF, indicating a downstream event to HIF-1α expression. When DPC spheroids were collected and subjected to the transwell assay, the cells growing outward from 3D spheroids showed greater migration ability than those from 2D cultures. Moreover, DPCs aggregation and spheroid formation on chitosan membrane were abolished by Y-27632 (inhibitor to Rho-associated kinases), whereas the inhibitory effect did not exist on PVA membrane. This suggests that the mechanism regulating DPCs aggregation and spheroid formation on chitosan membrane is involved with the Rho-associated kinase signaling pathway. In summary, the multicellular spheroid structure was beneficial to the protein expression of HIF-1α and VEGF in DPCs and enhanced the migration ability of the cells climbing from spheroids. This study showed a new perspective in exploring novel strategies for DPC-based research and application.

Periostin inhibits mechanical stretch-induced apoptosis in osteoblast-like MG-63 cells.

BACKGROUND/PURPOSE: Appropriate mechanical stress plays an important role in regulating the proliferation and differentiation of osteoblasts, whereas high-level mechanical stress may be harmful and compromise cell survival. Periostin, a matricellular protein, is essential in maintaining functional integrity of bone and collagen-rich connective tissue in response to mechanical stress. This study investigated whether or not high-level mechanical stretch induces cell apoptosis and the regulatory role of periostin in mechanical stretch-induced apoptosis in osteoblastic cells. METHODS: Osteoblast-like MG-63 cells were seeded onto Bio-Flex I culture plates and subjected to cyclic mechanical stretching (15% elongation, 0.1 Hz) in a Flexercell tension plus system-5000. The same process was applied to cells pre-treated with exogenous human recombinant periostin before mechanical stretching. We used a chromatin condensation and membrane permeability dead cell apoptosis kit to evaluate the stretch-induced cell responses. Expression of caspase-3 and cPARP was examined by immunofluorescent stain and flow cytometry. RESULTS: The expression of periostin in MG-63 cells is involved in the TGF-ß signaling pathway. High-level cyclic mechanical stretch induced apoptotic responses in MG-63 osteoblastic cells. The percentages of apoptotic cells and cells expressing cPARP protein increased in the groups of cells subjected to mechanical stretch, but these responses were absent in the presence of exogenous periostin. CONCLUSION: Our study revealed that high-level mechanical stretch induces apoptotic cell death, and that periostin plays a protective role against mechanical stretch-induced apoptosis in osteoblastic cells.

Airway increase after open bite closure with temporary anchorage devices for intrusion of the upper posteriors: Evidence from 2D cephalometric measurements and 3D magnetic resonance imaging.

OBJECTIVE: The aim of this study was to analyse morphological changes in the upper airways in patients with anterior open bite treated with temporary anchorage devices for intrusion of upper posterior teeth. MATERIALS AND METHODS: Twelve nonobese (body mass index: <25) anterior open bite patients between the ages of 19 and 44 years (mean age: 22.83 ± 8.19 years) were recruited for this study. Cephalometric radiographs and magnetic resonance imaging (MRI) scans before and after anterior bite closure without bracketing on anterior teeth were used to measure the upper airway, which was divided into retropalatal and retroglossal regions. RESULTS: The mandibular plane angle and lower facial height were significantly reduced by intrusion of the upper posteriors and autorotation of the mandible. The retroglossal airway width (AW2) and retroglossal area (RG area) measured on cephalometric radiographs both increased significantly after treatment. Retroglossal volume increased and the retroglossal width/length ratio decreased significantly in MRI analysis. All other measurements were not significantly changed. However, no statistically significant correlations were observed between all measurements in 2D and 3D images, with the exception of the AW2 linear measurement in 2D images correlating with the AP length in MRI axial view images (r = 0.56, P = 0.0430). CONCLUSION: Counterclockwise rotation of the mandible after anterior open bite closed using orthodontic treatment changed the airway morphology. Retroglossal volume significantly increased and the airway shape became less elliptical after bite closure.

Dual-effect liposomes encapsulated with doxorubicin and chlorin e6 augment the therapeutic effect of tumor treatment.

BACKGROUND AND OBJECTIVE: Long circulating doxorubicin (Dox)-loaded PEGylated liposomes are clinically safer than the free form due to the significant reduction of cardiac toxicity. However, the therapeutic efficacy of the PEGylated liposome could further be improved if poor diffusivity and slow drug release of the liposome in tumor interstitium can be overcome. In this study, a dual-effect liposome triggered by photodynamic effect was developed to improve the therapeutic efficacy of Dox-loaded PEGylated liposomes. MATERIALS AND METHODS: Dox and chlorin e6 (Ce6) were co-encapsulated in PEGylated liposomes (named as PL-Dox-Ce6). To induce the drug release, photodynamic effect was triggered by the light irradiation of a 662 nm diode laser. The cellular distribution of Dox and Ce6 was examined under confocal microscope. The in vitro and in vivo cytotoxicity of PL-Dox-Ce6 was determined via the colony formation assay and the synergistic C26 tumor model, respectively. RESULTS: The cellular distribution of PL-Dox-Ce6 was in the cytoplasmic area; while under light irradiation, Dox was co-localized with nuclear staining positive signals. The cellular cytotoxicity of PL-Dox-Ce6 was significantly higher than the controls including liposomes encapsulating either Dox (PL-Dox) or Ce6 (PL-Ce6). The in vivo treatment efficacy of PL-Dox-Ce6 determined in the C26 tumor model reveals a significant therapeutic effect compared to that of PL-Ce6 and PL-Dox alone or in combination. CONCLUSION: This study indicates that this dual-effect PEGylated liposome could provide clinical advantages in the combination regimen of photodynamic therapy and chemotherapy.

Revisiting the stability of mini-implants used for orthodontic anchorage.

BACKGROUND/PURPOSE: The aim of this study is to comprehensively analyze the potential factors affecting the failure rates of three types of mini-implants used for orthodontic anchorage. METHODS: Data were collected on 727 mini-implants (miniplates, predrilled titanium miniscrews, and self-drilling stainless steel miniscrews) in 220 patients. The factors related to mini-implant failure were investigated using a Chi-square test for univariate analysis and a generalized estimating equation model for multivariate analysis. RESULTS: The failure rate for miniplates was significantly lower than for miniscrews. All types of mini-implants, especially the self-drilling stainless steel miniscrews, showed decreased stability if the previous implantation had failed. The stability of predrilled titanium miniscrews and self-drilling stainless steel miniscrews were comparable at the first implantation. However, the failure rate of stainless steel miniscrews increased at the second implantation. The univariate analysis showed that the following variables had a significant influence on the failure rates of mini-implants: age of patient, type of mini-implant, site of implantation, and characteristics of the soft tissue around the mini-implants. The generalized estimating equation analysis revealed that mini-implants with miniscrews used in patients younger than 35 years, subjected to orthodontic loading after 30 days and implanted on the alveolar bone ridge, have a significantly higher risk of failure. CONCLUSION: This study revealed that once the dental surgeon becomes familiar with the procedure, the stability of orthodontic mini-implants depends on the type of mini-implant, age of the patient, implantation site, and the healing time of the mini-implant. Miniplates are a more feasible anchorage system when miniscrews fail repeatedly.

Increased Histone Deacetylase Activity Involved in the Suppressed Invasion of Cancer Cells Survived from ALA-Mediated Photodynamic Treatment.

Previously, we have found that cancer cells survived from 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) have abnormal mitochondrial function and suppressed cellular invasiveness. Here we report that both the mRNA expression level and enzymatic activity of histone deacetylase (HDAC) were elevated in the PDT-derived variants with dysfunctional mitochondria. The activated HDAC deacetylated histone H3 and further resulted in the reduced migration and invasion, which correlated with the reduced expression of the invasion-related genes, matrix metalloproteinase 9 (MMP9), paternally expressed gene 1 (PEG1), and miR-355, the intronic miRNA. Using chromatin immunoprecipitation, we further demonstrate the reduced amount of acetylated histone H3 on the promoter regions of MMP9 and PEG1, supporting the down-regulation of these two genes in PDT-derived variants. These results indicate that HDAC activation induced by mitochondrial dysfunction could modulate the cellular invasiveness and its related gene expression. This argument was further verified in the 51-10 cybrid cells with the 4977 bp mtDNA deletion and A375 ρ° cells with depleted mitochondria. These results indicate that mitochondrial dysfunction might suppress tumor invasion through modulating histone acetylation.

Mechanoregulation of osteoblast-like MG-63 cell activities by cyclic stretching.

BACKGROUND/PURPOSE: Mechanical loading plays an important role in regulating bone formation and remodeling. Relevant mechanical stretching can increase the proliferation and differentiation of osteoblastic cells in vitro. However, little is known about the effects of supraphysiological high-level mechanical stretching on the growth and cell cycle progression of osteoblastic cells. METHODS: Osteoblast-like MG-63 cells were seeded onto flexible-bottomed plates and subjected to cyclic mechanical stretching (15% elongation, 0.5 Hz) for 24 and 48 hours in a Flexercell FX-4000 strain unit. Cellular activities were measured by an assay based on the reduction of the tetrazolium salt, 3-[4,5-dimethyldiazol-2-yl]-2,5-diphenyl tetra-zolium bromide (MTT). The number of viable cells was also determined by the trypan blue dye exclusion technique. Cell cycle progression was checked by flow cytometry. mRNA expressions of apoptosis- and cell cycle-related genes (Bcl2, Bax, cdc2, cdc25C, and cyclin B1) were analyzed using an RT-PCR technique. RESULTS: The number of viable cells significantly decreased in osteoblast-like MG-63 cells subjected to cyclic mechanical stretching for 24 or 48 hours. The MTT activity of stretched cells did not change at 24 hours, whereas a significant decrease was noted at 48 hours in comparison to the unstretched controls. The flow cytometry showed that mechanical stretching induced S-phase cell cycle arrest. Furthermore, exposure to mechanical stretching led to apoptotic cell death, as shown by the increase in the hypodiploid sub-G0/G1 cell population. Furthermore, a decreased cdc25C mRNA level was consistently noted in stretched cells. However, the mRNA expressions of Bcl2, Bax, cdc2, and cyclin B1 genes were not significantly altered compared to the unstretched control cells. CONCLUSION: High-level mechanical stretching induced S-phase cell cycle arrest and apoptotic cell death in osteoblastic cells. The results suggest that heavy tensional force is a negative regulator of osteoblastic activities and should, therefore, be minimized if bone formation is attempted during orthodontic/orthopedic treatment.

Active vertical control with skeletal anchorage for optimizing facial profile in a severe Class II high-angle protrusion case.

This case report describes the nonsurgical management of a patient with a Class II skeletal pattern, retrognathic mandible, steep mandibular angle, maxillary vertical excess, and lip incompetence. The treatment approach involved orthodontic mechanics supported with skeletal anchorage to achieve maximal intrusion and retraction of the dentition. A novel elastic hanging rack appliance, supported by midpalatal miniscrews, was used. A maximal anchorage setup for active vertical control on both arches was illustrated. Significant improvement in the facial profile was achieved with optimal occlusion. Cephalometric analysis revealed successful incisor retraction and intrusion, as well as a forward rotation of the mandible. The treatment outcome illustrates the impact of active vertical control on orthodontic camouflage treatment for severe protrusion.

Functional architecture of intracellular oscillations in hippocampal dendrites.

Fast electrical signaling in dendrites is central to neural computations that support adaptive behaviors. Conventional techniques lack temporal and spatial resolution and the ability to track underlying membrane potential dynamics present across the complex three-dimensional dendritic arbor in vivo. Here, we perform fast two-photon imaging of dendritic and somatic membrane potential dynamics in single pyramidal cells in the CA1 region of the mouse hippocampus during awake behavior. We study the dynamics of subthreshold membrane potential and suprathreshold dendritic events throughout the dendritic arbor in vivo by combining voltage imaging with simultaneous local field potential recording, post hoc morphological reconstruction, and a spatial navigation task. We systematically quantify the modulation of local event rates by locomotion in distinct dendritic regions and report an advancing gradient of dendritic theta phase along the basal-tuft axis, then describe a predominant hyperpolarization of the dendritic arbor during sharp-wave ripples. Finally, we find spatial tuning of dendritic representations dynamically reorganizes following place field formation. Our data reveal how the organization of electrical signaling in dendrites maps onto the anatomy of the dendritic tree across behavior, oscillatory network, and functional cell states.

Craniofacial and olfactory sensory changes after long-term unilateral nasal obstruction-an animal study using MMP-3-LUC transgenic rats.

Nasal obstruction exerts considerable physiological effects on the respiratory system and craniofacial morphology during the developmental stage. This study used MMP-3-LUC transgenic rats for in vivo tracking of long-term expression in the rat nasal region after unilateral nasal obstruction. Skeletal changes of the craniofacial, nasal, and sinus regions were measured through micro-computed tomography examination and analysis with 3D image processing and calculation. Matrix metalloproteinase-3 and olfactory marker protein expression were also investigated through immunohistochemistry (IHC). Unilateral nasal obstruction significantly reduced the MMP-3 signal in the nasal region of MMP-3-LUC transgenic rats, which was mainly expressed in the respiratory epithelium. Long-term obstruction also caused morphological changes of the craniofacial hard tissue, such as nasal septal deviation, longer inter-jaw distance, and increased maxillary molar dental height. It also caused compensatory growth in olfactory nerve bundles and the olfactory epithelium, as confirmed by IHC. In our study, long-term unilateral nasal obstruction caused nasal septal deviation toward the unobstructed side, hyper divergent facial development including longer molar dental height, and reduced MMP-3 production. However, further investigation is necessary to explore the mechanism in depth.