Effects of continuous and released compressive force on osteoclastogenesis invitro.

Objective: Compressive force has been found to be catabolic to alveolar bone during orthodontic tooth movement. This study quantified the fusion of mononuclear RAW 264.7 cells (a murine osteoclastic-like cell line) into multinucleated osteoclasts under a hydrostatic pressure-generated mechanical compression-the new model of various magnitudes and durations. Methods: RAW 264.7 cells were subjected to 0.3, 0.6 or 0.9 g/cm2 of compressive force by an acrylic cylinder custom-made by laser cutting or no compressive force for 4 days during osteoclastogenic induction. TRAP-positive multinucleated cells were quantified. For the release from force experiment, osteoclastogenesis was induced by 0.6 g/cm2 mechanical stimuli for 0, 1, 2, 3 or 4 days. Cell viability, TRAP-positive multinucleated cells, DCSTAMP and Cathepsin K (CTSK) gene expression were evaluated 4 days after release from force. Results: Compressive force at 0.6 and 0.9 g/cm2 significantly increase the number of TRAP-positive multinucleated cells (P < 0.05). Release from continuous mechanical compression after 4 days significantly elevated the number of TRAP-positive multinucleated cells and DCSTAMP and CTSK mRNA expression, with no adverse effects on cell viability (P < 0.05). Conclusions: Continuous stimulation with compressive force induced osteoclastogenesis in RAW 264.7 cells by enhancing DCSTAMP and CTSK expression, which provides new understanding of bone remodeling during orthodontic treatment.

Fabrication of vascularized tissue-engineered bone models using triaxial bioprinting.

Bone tissue is a highly vascularized tissue. When constructing tissue-engineered bone models, both the osteogenic and angiogenic capabilities of the construct should be carefully considered. However, fabricating a vascularized tissue-engineered bone to promote vascular formation and bone generation, while simultaneously establishing nutrition channels to facilitate nutrient exchange within the constructs, remains a significant challenge. Triaxial bioprinting, which not only allows the independent encapsulation of different cell types while simultaneously forming nutrient channels, could potentially emerge as a strategy for fabricating vascularized tissue-engineered bone. Moreover, bioinks should also be applied in combination to promote both osteogenesis and angiogenesis. In this study, employing triaxial bioprinting, we used a blend bioink of gelatin methacryloyl (GelMA), sodium alginate (Alg), and different concentrations of nano beta-tricalcium phosphate (nano ß-TCP) encapsulated MC3T3-E1 preosteoblasts as the outer layer, a mixed bioink of GelMA and Alg loaded with human umbilical vein endothelial cells (HUVEC) as the middle layer, and gelatin as a sacrificial material to form nutrient channels in the inner layer to fabricate vascularized bone constructs simulating the microenvironment for bone and vascular tissues. The results showed that the addition of nano ß-TCP could adjust the mechanical, swelling, and degradation properties of the constructs. Biological assessments revealed the cell viability of constructs containing different concentrations of nano ß-TCP was higher than 90% on day 7, The cell-laden constructs containing 3% (w/v) nano ß-TCP exhibited better osteogenic (higher Alkaline phosphatase activity and larger Osteocalcin positive area) and angiogenic (the gradual increased CD31 positive area) potential. Therefore, using triaxial bioprinting technology and employing GelMA, Alg, and nano ß-TCP as bioink components could fabricate vascularized bone tissue constructs, offering a novel strategy for vascularized bone tissue engineering.

The role of autophagy in SIM mediated anti-inflammatory osteoclastogenesis through NLRP3 signaling pathway.

BACKGROUND: Inflammatory bone resorption is a prominent risk factor for implantation failure. Simvastatin (SIM) has anti-inflammatory effects independent of cholesterol lowering and reduces osteoclastogenesis by decreasing both the number and activity of osteoclasts. However, the specific mechanism of inflammatory bone loss alleviation by SIM remains to be elucidated. We hypothesized that SIM relieves inflammatory bone loss by modulating autophagy and suppressing the NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) signaling pathway. METHODS AND RESULTS: RAW264.7 cells were stimulated by lipopolysaccharide (LPS) after being pretreated with various concentrations of SIM. Osteoclast (OC) differentiation, formation and activity were evaluated by tartrate-resistant acid phosphatase staining, F-actin ring staining and bone resorption pit assays, respectively. We observed autophagosomes by transmission electron microscopy. Then NLRP3 inhibitor MCC950 was used to further explore the corresponding molecular mechanism underlying anti-inflammatory bone resorption, the expression of autophagy-related proteins and NLRP3 signaling pathway factors in pre-OCs were evaluated by western blot analysis, and the expression of OC-specific molecules was analyzed using reverse transcription-quantitative polymerase chain reaction. The results showed that SIM decreased the expression of tumor necrosis factor-α, whereas increased Interleukin-10. In addition, SIM inhibited LPS-induced OC differentiation, formation, bone resorption activity, the level of autophagosomes, and OC-specific markers. Furthermore, SIM significantly suppressed autophagy by downregulating LC3II, Beclin1, ATG7, and NLRP3-related proteins expression while upregulating P62 under inflammatory conditions. CONCLUSIONS: SIM may reduce autophagy secretion to attenuate LPS-induced osteoclastogenesis and the NLRP3 signaling pathway participates in this process, thus providing theoretical basis for the application of this drug in peri-implantitis.

LRP4 mutations, dental anomalies, and oral exostoses.

BACKGROUND: In order to generate a normal set of teeth, fine-tuning of Wnt/ß-catenin signaling is required, in which WNT ligands bind to their inhibitors or WNT inhibitors bind to their co-receptors. Lrp4 regulates the number of teeth and their morphology by modulating Wnt/ß-catenin signaling as a Wnt/ß-catenin activator or inhibitor, depending on its interactions with the partner proteins, such as Sostdc1 and Dkk1. AIM: To investigate genetic etiologies of dental anomalies involving LRP4 in a Thai cohort of 250 children and adults with dental anomalies. DESIGN: Oral and radiographic examinations and whole exome sequencing were performed for every patient. RESULTS: Two novel (p.Leu1356Arg and p.Ala1702Gly) and three recurrent (p.Arg263His, p.Gly1314Ser, and p.Asn1385Ser) rare variants in low-density lipoprotein receptor-related protein 4 (LRP4: MIM 604270) were identified in 11 patients. Oral exostoses were observed in five patients. CONCLUSION: Antagonism of Bmp signaling by Sostdc1 requires the presence of Lrp4. Mice lacking Lrp4 have been demonstrated to have alteration of Wnt-Bmp-Shh signaling and an abnormal number of incisors. Therefore, the LRP4 mutations found in our patients may disrupt Wnt-Bmp-Shh signaling, thereby resulting in dental anomalies and oral exostoses. Root maldevelopment in the patients suggests an important role of LRP4 in root morphogenesis.

Root resorption during maxillary molar intrusion with clear aligners: a randomized controlled trial.

OBJECTIVES: To compare changes in maxillary molar root resorption, intrusion amount, dentoskeletal measures, and maximum bite force (MBF) between clear aligners (CA) and fixed appliances with miniscrew (FM) during molar intrusion. MATERIALS AND METHODS: Forty adults with anterior open bite were randomized into either CA or FM groups. Lateral cephalograms, cone-beam computed tomography (CBCT), and MBF were collected at pretreatment (T0) and 6 months of treatment (T1). Maxillary molar intrusion in FM were intruded by nickel-titanium (NiTi) closed-coil spring delivered force (150 grams/side) while clear aligners combined with squeezing exercise were performed in CA. Parametric tests were used for statistical analysis. RESULTS: After 6 months of treatment, significant root resorption of 0.21-0.24 mm in CA and 0.38-0.47 mm in FM were found while maxillary molars were intruded 0.68 and 1.49 mm in CA and FM, respectively. CA showed significant less root resorption and intrusion than FM. Overbite, bite closing, and MBF increased significantly. CA showed significantly less overbite and SN-MP changes but more MBF increase than FM. MBF in CA was correlated with the amount of maxillary molar intrusion (r = 0.736, P < .05). CONCLUSIONS: Maxillary molar intrusion and root resorption in CA were half the amount in FM in 6 months. The amount of maxillary molar root resorption was one-third of the intrusion distance. CA displayed less overbite increase and bite closing but more MBF increase than FM. MBF in CA was positively correlated with the molar intrusion amount.

Triaxial bioprinting large-size vascularized constructs with nutrient channels.

Bioprinting has demonstrated great advantages in tissue and organ regeneration. However, constructing large-scale tissue and organsin vitrois still a huge challenge due to the lack of some strategies for loading multiple types of cells precisely while maintaining nutrient channels. Here, a new 3D bioprinting strategy was proposed to construct large-scale vascularized tissue. A mixture of gelatin methacrylate (GelMA) and sodium alginate (Alg) was used as a bioink, serving as the outer and middle layers of a single filament in the triaxial printing process, and loaded with human bone marrow mesenchymal stem cells and human umbilical vein endothelial cells, respectively, while a calcium chloride (CaCl2) solution was used as the inner layer. The CaCl2solution crosslinked with the middle layer bioink during the printing process to form and maintain hollow nutrient channels, then a stable large-scale construct was obtained through photopolymerization and ion crosslinking after printing. The feasibility of this strategy was verified by investigating the properties of the bioink and construct, and the biological performance of the vascularized construct. The results showed that a mixture of 5% (w/v) GelMA and 1% (w/v) Alg bioink could be printed at room temperature with good printability and perfusion capacity. Then, the construct with and without channels was fabricated and characterized, and the results revealed that the construct with channels had a similar degradation profile to that without channels, but lower compressive modulus and higher swelling rate. Biological investigation showed that the construct with channels was more favorable for cell survival, proliferation, diffusion, migration, and vascular network formation. In summary, it was demonstrated that constructing large-scale vascularized tissue by triaxial printing that can precisely encapsulate multiple types of cells and form nutrient channels simultaneously was feasible, and this technology could be used to prepare large-scale vascularized constructs.

Light orthodontic force with high-frequency vibration accelerates tooth movement with minimal root resorption in rats.

OBJECTIVES: To determine and compare the effects of high-frequency mechanical vibration (HFV) with light force and optimal force on the tooth movement and root resorption in rat model. MATERIALS AND METHODS: Seventy-two sites in 36 male Wistar rats were randomly assigned using a split-mouth design to control (no force/no vibration) or experimental groups: HFV (125 Hz), light force (5 g), optimal force (10 g), light force with HFV, and optimal force with HFV for 14 and 21 days. The amount of tooth movement, 3D root volume, and root resorption area were assessed by micro-computed tomography and histomorphometric analysis. RESULTS: Adjunction of HFV with light force significantly increased the amount of tooth movement by 1.8-fold (p = 0.01) and 2.0-fold (p = 0.01) at days 14 and 21 respectively. The HFV combined with optimal force significantly increased the amount of tooth movement by 2.1-fold (p = 0.01) and 2.2-fold (p = 0.01) at days 14 and 21 respectively. The root volume in control (distobuccal root (DB): 0.60 ± 0.19 mm3, distopalatal root (DPa): 0.60 ± 0.07 mm3) and HFV (DB: 0.60 ± 0.08 mm3, DPa: 0.59 ± 0.11 mm3) were not different from the other experimental group (range from 0.44 ± 0.05 to 0.60 ± 0.1 mm3) with the lowest volume in optimal force group. CONCLUSIONS: Adjunction of HFV with orthodontic force significantly increased tooth movement without causing root resorption. CLINICAL RELEVANCE: Using light force with HFV could help to identify alternative treatment option to reduce the risk of root resorption.

Overbite recognition and factors affecting esthetic tolerance among laypeople.

OBJECTIVES: To determine recognition ability and the levels of esthetic tolerance of deep bite and anterior open bite (AOB) among laypeople and investigate the factors affecting levels of tolerance. MATERIALS AND METHODS: Using a questionnaire, laypeople (N = 100) were examined, and overbite was measured. They were tested for whether they recognized deep bite and AOB. Esthetic tolerance thresholds for deep bite and AOB were selected by incremental depiction in grayscale images. Stepwise logistic regression analyses were used to quantify the effect of recognition and other factors (age, sex, education level, occupation, history of orthodontic treatment, interest in orthodontic treatment or retreatment, and overbite presence) affecting the tolerance of overbite problems (α = 0.05). RESULTS: Of the participants, 55% and 94% recognized deep bite and AOB, respectively. Participants with a deep bite were significantly more likely to esthetically tolerate deep bite compared with those without a deep bite (odds ratio [OR], 3.57; 95% confidence interval [CI], 1.29-9.89). Participants who recognized a deep bite problem had significantly lower esthetic tolerance to deep bite compared with participants who did not recognize a deep bite (OR, 0.17; 95% CI, 0.06-0.45). None of the other eight chosen factors significantly affected the tolerance level of AOB (P > .05). CONCLUSIONS: Participants with a deep bite or those who did not recognize a deep bite had significantly higher esthetic tolerance of deep bite than those without or those who recognized the problem (P < .05).

Effects of compressive stress combined with mechanical vibration on osteoclastogenesis in RAW 264.7 cells.

OBJECTIVES: To investigate the effects of compressive force and/or mechanical vibration on NFATc1, DCSTAMP, and CTSK (cathepsin K) gene expression and the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells in RAW 264.7 cells, a murine osteoclastic-like cell line. MATERIALS AND METHODS: RAW 264.7 cells were subjected to mechanical vibration, compressive force, or compressive force combined with vibration. Cell viability and the numbers of TRAP-positive multinucleated cells were evaluated. NFATc1, DCSTAMP, and CTSK gene expressions were analyzed using real-time quantitative reverse transcription polymerase chain reaction. RESULTS: Compressive force combined with mechanical vibration significantly increased the numbers of TRAP-positive multinucleated cells but did not significantly affect cell viability. In addition, compressive force combined with mechanical vibration significantly increased NFATc1, DCSTAMP, and CTSK mRNA expression compared with compressive force or vibration alone. CONCLUSIONS: Compressive force combined with mechanical vibration induces osteoclastogenesis and upregulates NFATc1, DCSTAMP, and CTSK gene expression in RAW 264.7 cells. These results provide more insight into the mechanisms by which vibratory force accelerates orthodontic tooth movement.

Varied temporal expression patterns of trigeminal TRPA1 and TRPV1 and the neuropeptide CGRP during orthodontic force-induced pain.

OBJECTIVE: The aim of this study was to quantify the temporal changes in inflammation and TRPA1, TRPV1 and CGRP expression in the trigeminal ganglion during force-induced orthodontic pain. DESIGN: Orthodontic force was applied to both maxillary first molars in 8-week-old Wistar rats for 12 h, 24 h, 3 d or 7 d. The rat grimace scale (RGS) score and duration of face grooming were used to measure orthodontic pain. Western blotting was performed to assess TRPA1, TRPV1 and CGRP expression in trigeminal ganglia. NF-кB levels and colocalization of TRPA1, TRPV1 and CGRP were evaluated by immunofluorescent staining. RESULTS: Application of continuous force significantly increased pain behaviours at 1 and 3 d. NF-кB significantly increased in periodontal ligament at 12 h until 3 d. TRPV1 was significantly elevated within 1 d; TRPA1 significantly increased from 1-3 d; CGRP expression significantly increased from 12 h to 3 d. The TRPV1/TRPA1 expression ratio was highest at 12 h; the TRPA1/TRPV1 ratio peaked at 3 d. The percentages of trigeminal neurons co-expressing TRPA1/TRPV1, TRPA1/CGRP, and TRPV1/CGRP significantly increased by 12 h and peaked at 24 h. CGRP expression had a stronger positive correlation with TRPV1 than TRPA1. CONCLUSIONS: Inflammation induced by application of orthodontic force sensitizes trigeminal TRPV1 and TRPA1; TRPV1 is primarily activated as an early response, whereas TRPA1 is activated as a late response. Activation of both nociceptors results in CGRP release. Thus, blocking both TRPV1 and TRPA1 may represent a primary therapeutic target for relief of orthodontic pain.

Interval Vibration Reduces Orthodontic Pain Via a Mechanism Involving Down-regulation of TRPV1 and CGRP.

BACKGROUND/AIM: The transient receptor potential vanilloid 1 (TRPV1) ion receptor is involved in the release of calcitonin gene-related peptide (CGRP), a major contributor to orthodontic pain. Approaches that attenuate expression of TRPV1 and CGRP may reduce orthodontic pain. We explored the ability of high-frequency interval vibration to reduce orthodontic pain. MATERIALS AND METHODS: Orthodontic force (50 g) was applied to both maxillary first molars in 8-week-old Wistar rats (n=72). Vibration was applied at 125 Hz for 15 min/day. Duration of face grooming was assessed as a measure of orthodontic pain. Immunofluorescence and western blotting were used to assess TRPV1 and CGRP in the trigeminal ganglia. RESULTS: Compared to orthodontic force alone, application of vibration significantly decreased the duration of face grooming at 24 h and day 3 and reduced expression of TRPV1 and CGRP at 24 h. CONCLUSION: Vibration represents a promising mechanical approach to reduce orthodontic pain.

Vibration synergistically enhances IL-1ß and TNF-α in compressed human periodontal ligament cells in the frequency-dependent manner.

OBJECTIVES: To investigate whether mechanical vibration at 30 or 60 Hz combined with compressive force alter IL-1ß and TNF-α expression in human periodontal ligament (hPDL) cells. METHODS: hPDL cells isolated from the roots of first premolar teeth extracted from four independent donors were cultured and exposed to vibration (0.3 g, 20 min per cycle, every 24 h for 3 cycles) at 30 or 60 Hz (V30 or V60), 2.0 g/cm2 compressive force for 2 days (CF), or a combination of compressive force and vibration at 30 Hz or 60 Hz (V30CF or V60CF). Quantitative real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assays (ELISAs) were used to determine IL-1ß and TNF-α mRNA and protein, respectively. RESULTS: The levels of IL-1ß and TNF-α did not alter in groups V30 and V60. While, they were upregulated in groups CF, V30CF and V60CF. In addition, IL-1ß mRNA and TNF-α mRNA and protein were expressed at significantly higher levels in group V30CF compared to CF group. However, IL-1ß protein levels between V30CF and CF groups did not reach statistical significance. CONCLUSIONS: 30 Hz vibration had the synergistic effects with compressive force on the upregulation of IL-1ß mRNA and TNF-α mRNA and protein in PDL cells, while 60 Hz vibration did not have this synergistic effect.

Vibration activates the actin/NF-κB axis and upregulates IL-6 and IL-8 expression in human periodontal ligament cells.

We previously reported that mechanical vibration-induced proinflammatory cytokines, interleukin-6 (IL-6) and IL-8, expression in human periodontal ligament (hPDL) cells, however, the underlying mechanism remained unclear. Mechanical stimuli are able to activate cellular responses by inducing the activation of several signaling pathways including cytoskeletal changes and inflammation. The actin cytoskeleton is a highly dynamic network and plays many important roles in intracellular events. Here, we aimed to investigate the involvement of a pivotal mediator of inflammatory responses, nuclear factor-κB (NF-κB), and actin polymerization in vibration-induced upregulation of IL-6 and IL-8 expression in hPDL cells. hPDL cells were pretreated with the NF-κB inhibitor BAY 11-7082 or cytochalasin D, respectively, before exposure to vibration. IL-6 and IL-8 messenger RNA (mRNA) and protein expression were quantified by quantitative polymerase chain reaction and enzyme-linked immunosorbent assays, respectively. Subcellular localization of the NF-κB p65 subunit was visualized by immunofluorescent staining. We found an increase in NF-κB nuclear translocation in vibrated cells compared with control cells. Pretreatment with BAY 11-7082 significantly inhibited vibration-induced IL-6 and IL-8 mRNA and protein expression in hPDL cells. Moreover, pretreatment with cytochalasin D inhibited NF-κB nuclear translocation and attenuated upregulation of IL-6 and IL-8 mRNA and protein in vibrated cells. Therefore, modulation of actin cytoskeletal polymerization in response to vibration may activate the NF-κB signaling pathway and subsequently upregulate IL-6 and IL-8 expression in hPDL cells.

The influence of leukocyte-platelet-rich plasma on accelerated orthodontic tooth movement in rabbits.

OBJECTIVE: To determine the effects of a local injection of leukocyte-platelet-rich plasma (L-PRP) on orthodontic tooth movement in rabbits. METHODS: Twenty-three male New Zealand white rabbits were included in a split-mouth design. Tooth movement with a 100-g nickel-titanium closed-coil spring was performed on the maxillary first premolars. L-PRP was injected submucosally at the buccal and lingual areas of the first premolar in one random side of the maxilla and the other side served as the control and received normal saline. The amount of tooth movement was assessed on three-dimensional digital models on days 0, 3, 7, 14, 21, and 28. Histological findings and osteoclast numbers were examined on day 0 as the baseline and on days 7, 14, and 28. RESULTS: The L-PRP group showed significantly greater cumulative tooth movement at all observed periods. However, a significantly higher rate of tooth movement was observed only on days 0-7 and 7-14. The osteoclast numbers were significantly increased in the L-PRP group on days 7 and 14. CONCLUSIONS: Local injection of L-PRP resulted in a transient increase in the rate of tooth movement and higher osteoclast numbers.

Low magnitude high frequency vibration induces RANKL via cyclooxygenase pathway in human periodontal ligament cells in vitro.

OBJECTIVE: This study aimed to examine the effects of PGE2 on RANKL expression in response to vibration and vibration in combination with compressive stress and characterise this transduction pathway in periodontal ligament (PDL) cells. METHODS: Cultured human PDL cells obtained from extracted premolar teeth (from six individuals) were subjected to three cycles of vibration (0.3 g, 30 Hz for 20 min every 24 h; V), compressive stress (1.5 g/cm2, 48 h; C) or vibration in combination with compressive stress (VC). To investigate whether the expression of RANKL and PGE2 was COX-dependent, PDL cells were treated with indomethacin prior to the onset of mechanical stimulation. RANKL and OPG expressions were examined by quantitative real-time polymerase chain reaction (qPCR). Quantification of PGE2, soluble RANKL (sRANKL) and OPG productions were measured using enzyme-linked immunosorbent assay (ELISAs). RESULTS: All mechanical stresses (V, C and VC) significantly increased PGE2 and RANKL. OPG was not affected by vibration, but was downregulated in compressed cells (C and VC). Indomethacin abolished induction of RANKL and downregulated OPG in response to all mechanical stresses. CONCLUSION: These results suggest that vibration, compressive stress and vibration in combination with compressive stress induce RANKL expression in human PDL cells by activating the cyclooxygenase pathway.

The effect of compressive force combined with mechanical vibration on human alveolar bone osteoblasts.

OBJECTIVE: This study aimed to investigate the effects of compressive force combined with mechanical vibration on the expression of pro-inflammatory cytokines that promote osteoclastogenesis and related to orthodontic tooth movement acceleration in human alveolar bone osteoblasts in vitro. METHODS: Osteoblasts were subjected to compressive force (C), mechanical vibration (V), compressive force combined with mechanical vibration (CV), or no force as a control for 12, 24 and 48 h. Interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), receptor activator of nuclear factor kappa-Β ligand (RANKL) and osteoprotegerin (OPG) mRNA and protein expression were assessed using quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assays. RESULTS: In C and CV groups, IL-1ß and IL-6 mRNA and protein expression were significantly higher and OPG mRNA and protein expression were significantly lower than control and V groups. However, the expressions were not different between C and CV groups. RANKL mRNA and protein expression were not different between any groups. While, OPG mRNA and protein expression in V group were significantly higher than control group. CONCLUSIONS: Vibration neither enhanced nor inhibited the expression of IL-1ß, IL-6, RANKL and OPG in compressed human alveolar bone osteoblasts.

Periostin plays role in force-induced stem cell potential by periodontal ligament stem cells.

Mechanical stimuli have been shown to play an important role in directing stem cell fate and maintenance of tissue homeostasis. One of the functions of the mechanoresponsive tissue periodontal ligament (PDL) is to withstand the functional forces within the oral cavity. Periodontal ligament stem cells (PDLSCs) derived from periodontal tissue have been demonstrated to be able to respond directly to mechanical forces. However, the mechanisms of action of mechanical force on PDLSCs are not totally understood. The aim of this study was to investigate the mechanisms by which compressive force affects PDLSCs, especially their stemness properties. PDLSCs were established from extracted human third molars; their stem cell characteristics were validated by detecting the expression of stem cell markers and confirming their ability to differentiate into osteogenic and adipogenic lineages. PDLSCs were subjected to various magnitudes of static compressive force (0 [control], 0.5, 1.0, 1.5, or 2 g/cm2 ). Application of 1.0 g/cm2 compressive force significantly upregulated a panel of stem cell marker genes, including NANOG and OCT4. Conversely, higher force magnitudes downregulated these genes. Mechanical loading also upregulated periostin, a matrix protein that plays important roles in tissue morphogenesis. Interestingly, knockdown of periostin using siRNA abolished force-induced stem cell marker expression in PDLSCs. This study suggests a proper magnitude of compressive force could be one important factor involved in the modulation of the pluripotency of PDLSCs through the action of periostin. The precise mechanism by which periostin regulates stemness requires further detailed investigation.

Effects of two frequencies of vibration on the maxillary canine distalization rate and RANKL and OPG secretion: A randomized controlled trial.

OBJECTIVES: To investigate the effects of 30 and 60 Hz vibratory stimulus on canine distalization and RANKL and OPG secretion. SETTING AND SAMPLE POPULATION: Sixty patients requiring canine distalization at the Orthodontic Clinic, Prince of Songkla University. MATERIALS AND METHODS: Patients were randomly assigned to 30 Hz vibration (n = 20), 60 Hz vibration (n = 20), or the control group (n = 20). Modified electric toothbrushes were used to apply vibration to the randomly selected canine for 20 min/day by the investigator combined with 60 cN continuous distalization force from day 1 to day 7. RANKL and OPG were analysed before (T1) and 24 hours (T2), 48 hours (T3) and 7 days (T4) after initiation of distalization. From day 8, vibratory devices were used by the subjects at home. Rate of canine distalization (T1 to 3 months after initiation [T5]) was calculated. Kruskal-Wallis tests were used for multiple comparisons (significance level, 0.05). RESULTS: Canine distalization rate was not different between groups (median; 0.82, 0.87, and 0.83 mm/month for 30, 60 Hz, and control group, respectively; P > 0.05). No within- or between-group differences in RANKL and OPG were observed (P > 0.05), except RANKL on the compression side of the control group was significantly higher at T2, T3 and T4 than T1 (P < 0.001). CONCLUSION: In the clinic, 30 and 60 Hz vibratory stimulus have no additive effect on rate of canine distalization rate, RANKL and OPG secretion or RANKL/OPG ratio.

Mandible and iliac osteoblasts exhibit different Wnt signaling responses to LMHF vibration.

OBJECTIVE: The jaw bones and long bones have distinct developmental origins and respond differently to mechanical stimuli. This study aimed to compare the Wnt signaling responses of human mandible osteoblasts and long bone osteoblasts to low-magnitude, high-frequency (LMHF) mechanical vibration in vitro. METHODS: Primary human osteoblast cultures were prepared from mandibular bone (n = 3) and iliac bone (n = 3) specimens (six individuals). Osteoblast cell lines were subjected to vibration (0, 30, 60, 90, or 120 Hz) for 30 min. After 24 h, cells were vibrated for 30 min again, then harvested immediately to quantify Wnt10b, Wnt5a and runt-related transcription factor 2 (RUNX2) mRNA expression, ß-catenin protein expression and alkaline phosphatase (ALP) activity. RESULTS: Mandible and iliac osteoblasts responded differently to LMHF vibration: Wnt10b mRNA was upregulated by the frequency range tested; Wnt5a, ß-catenin protein expression and RUNX2 mRNA expression were not altered. Furthermore, vibration upregulated ALP activity in mandible osteoblasts, but not in iliac osteoblasts. CONCLUSIONS: This study demonstrates mandible osteoblasts and long bone osteoblasts respond differently to LMHF mechanical vibration in terms of Wnt signaling expression and ALP activity. Therefore, the effects of whole-body vibration on the long bones cannot be generalized to the jaw bones. Furthermore, osteoblast-like cells mediate the cellular responses to vibration, at least in part, by secreting extracellular signaling molecules.

Comparison of clinical and histological characteristics of orthodontic tooth movement into recent and healed extraction sites combined with corticotomy in rats.

OBJECTIVE: This study was performed to investigate the rate of tooth movement and histological characteristics of extraction sockets those were subjected to corticotomy. METHODS: A split-mouth randomized controlled trial experiment was designed. Thirty-two adult, male Wistar rats were divided into 2 groups: healing extraction socket (H) and recent extraction socket (R); these groups were randomly classified into 4 subgroups (0/7/21/60 days). The first maxillary molar was extracted on 1 side and 2 months were allowed for complete bone healing; then, the corresponding molar was extracted on the other side and surgical intervention was performed at the mid-alveolar point of the first maxillary molar. Ten grams of continuous force was applied. The outcomes measured were rate of tooth movement, percentage of periodontal space and histological evaluation. The rate of tooth movement was calculated as the measured distance divided by the duration of molar movement. Histomorphometric evaluations were performed on the second and third maxillary molars. The Wilcoxon signed rank test was used to compare differences between the two groups. RESULTS: There were no significant differences in the rates of tooth movement between H and R groups at any of the 4 time points. The histological appearance and percentage of periodontal space between the R and H groups also demonstrated no significant differences. CONCLUSIONS: The rates of orthodontic tooth movement into recent and healed socket sites did not differ between the groups. Histological analysis of tooth movement revealed regional acceleration during every time period.