Deep learning for virtual orthodontic bracket removal: tool establishment and application.

OBJECTIVE: We aimed to develop a tool for virtual orthodontic bracket removal based on deep learning algorithms for feature extraction from bonded teeth and to demonstrate its application in a bracket position assessment scenario. MATERIALS AND METHODS: Our segmentation network for virtual bracket removal was trained using dataset A, containing 978 bonded teeth, 20 original teeth, and 20 brackets generated by scanners. The accuracy and segmentation time of the network were tested by dataset B, which included an additional 118 bonded teeth without knowing the original tooth morphology. This tool was then applied for bracket position assessment. The clinical crown center, bracket center, and orientations of separated teeth and brackets were extracted for analyzing the linear distribution and angular deviation of bonded brackets. RESULTS: This tool performed virtual bracket removal in 2.9 ms per tooth with accuracies of 98.93% and 97.42% (P < 0.01) in datasets A and B, respectively. The tooth surface and bracket characteristics were extracted and used to evaluate the results of manually bonded brackets by 49 orthodontists. Personal preferences for bracket angulation and bracket distribution were displayed graphically and tabularly. CONCLUSIONS: The tool's efficiency and precision are satisfactory, and it can be operated without original tooth data. It can be used to display the bonding deviation in the bracket position assessment scenario. CLINICAL SIGNIFICANCE: With the aid of this tool, unnecessary bracket removal can be avoided when evaluating bracket positions and modifying treatment plans. It has the potential to produce retainers and orthodontic devices prior to tooth debonding.

Manganese Enhances the Osteogenic Effect of Silicon-Hydroxyapatite Nanowires by Targeting T Lymphocyte Polarization.

Biomaterials encounter considerable challenges in extensive bone defect regeneration. The amelioration of outcomes may be attainable through the orchestrated modulation of both innate and adaptive immunity. Silicon-hydroxyapatite, for instance, which solely focuses on regulating innate immunity, is inadequate for long-term bone regeneration. Herein, extra manganese (Mn)-doping is utilized for enhancing the osteogenic ability by mediating adaptive immunity. Intriguingly, Mn-doping engenders heightened recruitment of CD4+ T cells to the bone defect site, concurrently manifesting escalated T helper (Th) 2 polarization and an abatement in Th1 cell polarization. This consequential immune milieu yields a collaborative elevation of interleukin 4, secreted by Th2 cells, coupled with attenuated interferon gamma, secreted by Th1 cells. This orchestrated interplay distinctly fosters the osteogenesis of bone marrow stromal cells and effectuates consequential regeneration of the mandibular bone defect. The modulatory mechanism of Th1/Th2 balance lies primarily in the indispensable role of manganese superoxide dismutase (MnSOD) and the phosphorylation of adenosine 5'-monophosphate-activated protein kinase (AMPK). In conclusion, this study highlights the transformative potential of Mn-doping in amplifying the osteogenic efficacy of silicon-hydroxyapatite nanowires by regulating T cell-mediated adaptive immunity via the MnSOD/AMPK pathway, thereby creating an anti-inflammatory milieu favorable for bone regeneration.

Silicon-Based Biomaterials Modulate the Adaptive Immune Response of T Lymphocytes to Promote Osteogenesis/Angiogenesis via Epigenetic Regulation.

Silicon (Si)-based biomaterials are widely applied for bone regeneration. However, the underlying mechanisms of the materials function remain largely unknown. T lymphocyte-mediated adaptive immune response plays a vital role in the process of bone regeneration. In the current study, mesoporous silica (MS) is used as a model material of Si-based biomaterials. It shows that the supernatant of CD4+ T lymphocytes pretreated with MS extract significantly promotes the vascularized bone regeneration. The potential mechanism is closely related to the fact that MS extract can reduce the expression of regulatory factor X-1 (RFX-1) in CD4+ T lymphocytes. This may result in the overexpression of interleukin-17A (IL-17A) by boosting histone H3 acetylation and lowering DNA methylation and H3K9 trimethylation. Importantly, the in vivo experiments further reveal that MS particles significantly enhance bone regeneration with improved angiogenesis in the critical-sized calvarial defect mouse model accompanied by upregulation of IL-17A in peripheral blood and the proportion of Th17 cells. This study suggests that modulation of the adaptive immune response of T lymphocytes by silicate-based biomaterials plays an important role for bone regeneration.

The psc-CVM assessment system: A three-stage type system for CVM assessment based on deep learning.

BACKGROUND: Many scholars have proven cervical vertebral maturation (CVM) method can predict the growth and development and assist in choosing the best time for treatment. However, assessing CVM is a complex process. The experience and seniority of the clinicians have an enormous impact on judgment. This study aims to establish a fully automated, high-accuracy CVM assessment system called the psc-CVM assessment system, based on deep learning, to provide valuable reference information for the growth period determination. METHODS: This study used 10,200 lateral cephalograms as the data set (7111 in train set, 1544 in validation set and 1545 in test set) to train the system. The psc-CVM assessment system is designed as three parts with different roles, each operating in a specific order. 1) Position Network for locating the position of cervical vertebrae; 2) Shape Recognition Network for recognizing and extracting the shapes of cervical vertebrae; and 3) CVM Assessment Network for assessing CVM according to the shapes of cervical vertebrae. Statistical analysis was conducted to detect the performance of the system and the agreement of CVM assessment between the system and the expert panel. Heat maps were analyzed to understand better what the system had learned. The area of the third (C3), fourth (C4) cervical vertebrae and the lower edge of second (C2) cervical vertebrae were activated when the system was assessing the images. RESULTS: The system has achieved good performance for CVM assessment with an average AUC (the area under the curve) of 0.94 and total accuracy of 70.42%, as evaluated on the test set. The Cohen's Kappa between the system and the expert panel is 0.645. The weighted Kappa between the system and the expert panel is 0.844. The overall ICC between the psc-CVM assessment system and the expert panel was 0.946. The F1 score rank for the psc-CVM assessment system was: CVS (cervical vertebral maturation stage) 6 > CVS1 > CVS4 > CVS5 > CVS3 > CVS2. CONCLUSIONS: The results showed that the psc-CVM assessment system achieved high accuracy in CVM assessment. The system in this study was significantly consistent with expert panels in CVM assessment, indicating that the system can be used as an efficient, accurate, and stable diagnostic aid to provide a clinical aid for determining growth and developmental stages by CVM.

Treatment of skeletal class III malocclusion using a combined clear aligner and surgery-early approach : Assessment based on the American Board of Orthodontics Objective Grading System.

PURPOSE: To evaluate whether clear aligner therapy (CAT) combined with a surgery-early approach can achieve good therapeutic effects in patients with skeletal class III malocclusion. METHODS: Thirty consecutive skeletal class III malocclusion cases treated with clear aligners combined with early surgery were selected. Treatment time, lateral cephalograms and American Board of Orthodontics Objective Grading System (ABO-OGS) scores of the treatment models were measured to evaluate the treatment efficiency, facial profile, and occlusion. RESULTS: The results showed that early surgery was achieved after 7.71 months of presurgical orthodontics, on average. ANB decreased by 5.57° (P < 0.001), and STissue N Vert to Pog' decreased by 7.29 mm (P = 0.001), both reaching normal values. The posttreatment ABO-OGS scores were 26.600 on average, meeting its standards. CONCLUSIONS: With the assistance of CAT, early surgery can be accomplished in patients with skeletal class III malocclusion, improving their facial profile and achieving functional occlusion.

Stress and movement trend of lower incisors with different IMPA intruded by clear aligner: a three-dimensional finite element analysis.

BACKGROUND: During the intrusion of lower incisors with clear aligners (CAs), root disengagement from the alveolar bone often occurs, resulting in serious complications. This study aimed to determine the potential force mechanism of the mandibular anterior teeth under the pressure of CA, providing theoretical data for clinical practice. METHODS: In this study, a 3D finite element model was established, including the CA, periodontal ligament, and mandibular dentition. Incisor mandibular plane angles were set as 5 groups: 90°, 95°, 100°, 105°, and 110°. The 4 mandibular incisors were intruded by 0.2 mm, while the canines were the anchorage teeth. The stress, force systems, and potential movement trends of mandibular anterior teeth were obtained. RESULTS: The compressive stress of the incisors was concentrated in the lingual fossa, incisal ridge, and apex. With the increase in IMPA, the moment of central incisors changed from lingual crown moment to labial crown moment, with the turning point between 100° and 105°, but the center of resistance (CR) was always subjected to the force toward the lingual and intrusive direction. The force and moment toward the labial side of the lateral incisors were greater than those toward the central incisors. The canines always tipped distally and received extrusive force with no relationship with IMPA. CONCLUSIONS: With the increase in the initial IMPA, the direction of labiolingual force on the mandibular incisors was reversed. However, the root of the lower incisors always tipped labially, which indicated fenestration and dehiscence.

Balancing the antibacterial and osteogenic effects of double-layer TiO2 nanotubes loaded with silver nanoparticles for the osseointegration of implants.

The improvement of Ag nanoparticles (AgNPs), in particular, loaded titania nanotubes, includes not only the antibacterial effect but also balancing the side effects from the antibacterial effect and osteogenesis properties, which can lead to an increased success rate of implants. Herein, based on the various needs of the graft to inhibit bacteria at different stages in vivo, we used a special osteogenic honeycomb-like "large tube over small tube" double-layered nanotube structure and created ultra-small-sized silver nanoparticles uniformly loaded on the surface and the interior of double-layer nanotubes by an optimized sputter coating method to ensure the time-dependent controllable release of antibacterial Ag ions from grafts and achieve the balance of the antibacterial effect and osteogenesis properties. The release of Ag+ from DNT-Ag8 was determined by inductively coupled plasma spectrometry. The release rate of Ag was slow; it was 30% on the first day and plateaued by the 19th day. Porphyromonas gingivalis adhesion and live bacteria were less abundant on the surface of DNT-Ag8, reaching an antibacterial efficiency of 55.6% in vitro. DNT-Ag8 shows a significantly higher antibacterial effect in a rat model infected with Staphylococcus aureus. An in vitro study demonstrated that DNT-Ag8 had no adverse effects on the adhesion, viability, proliferation, ALP staining, or activity assays of rat BMSCs. In contrast, it increased the expression of osteogenic genes. In vivo, DNT-Ag8 promoted bone-implant osseointegration in a beagle mandibular tooth loss model. This study demonstrated that the uniform loading of small-diameter silver nanoparticles using a honeycomb bilayer nanotube template structure is a promising method for modifying titanium surfaces to improve both bacteriostasis and osseointegration.

Dynamic network biomarker identifies cdkn1a-mediated bone mineralization in the triggering phase of osteoporosis.

The identification of predictive markers to determine the triggering phase prior to the onset of osteoporosis is essential to mitigate further irrevocable deterioration. To determine the early warning signs before osteoporosis, we used the dynamic network biomarker (DNB) approach to analyze time-series gene expression data in a zebrafish osteoporosis model, which revealed that cyclin-dependent kinase inhibitor 1 A (cdkn1a) is a core DNB. We found that cdkn1a negatively regulates osteogenesis, as evidenced by loss-of-function and gain-of-function studies. Specifically, CRISPR/Cas9-mediated cdkn1a knockout in zebrafish significantly altered skeletal development and increased bone mineralization, whereas inducible cdkn1a expression significantly contributed to osteoclast differentiation. We also found several mechanistic clues that cdkn1a participates in osteoclast differentiation by regulating its upstream signaling cascades. To summarize, in this study, we provided new insights into the dynamic nature of osteoporosis and identified cdkn1a as an early-warning signal of osteoporosis onset.

Lithium-Containing Biomaterials Stimulate Cartilage Repair through Bone Marrow Stromal Cells-Derived Exosomal miR-455-3p and Histone H3 Acetylation.

The repair of damaged cartilage still remains a great challenge in clinic. It is demonstrated that bone marrow stromal cells (BMSCs)-chondrocytes communication is of great significance for cartilage repair. Moreover, BMSCs have been confirmed to enhance biological function of chondrocytes via exosome-mediated paracrine pathway. Lithium-containing scaffolds have been reported to effectively promote cartilage regeneration; however, whether lithium-containing biomaterial could facilitate cartilage regeneration through regulating BMSCs-derived exosomes has not been illustrated. In the study, the model lithium-substituted bioglass ceramic (Li-BGC) is selected and regulatory effects of BMSCs-derived exosomes after Li-BGC treatment (Li-BGC-Exo) are systemically evaluated. The data reveal that Li-BGC-Exo notably promotes chondrogenesis, which attributes to the upregulated exosomal miR-455-3p transfer, consequently leads to suppression of histone deacetylase 2 (HDAC2) and enhanced histone H3 acetylation in chondrocytes. Notably, BMSCs-derived exosomes after LiCl treatment (LiCl-Exo) exhibits the similar regulatory effect with Li-BGC-Exo, indicating that the pro-chondrogenesis capability of them is mainly owing to the lithium ions. Furthermore, the in vivo study proves that LiCl-Exo remarkably facilitates cartilage regeneration. The research may provide novel possibility for the intrinsic mechanism of chondrogenesis trigged by lithium-containing biomaterials, and suggests that application of lithium-containing scaffolds may be a promising strategy for cartilage regeneration.

Carbon dots derived from folic acid attenuates osteoarthritis by protecting chondrocytes through NF-κB/MAPK pathway and reprogramming macrophages.

BACKGROUND: Osteoarthritis (OA) is a common joint disorder worldwide which causes great health and economic burden. However, there remains an unmet goal to develop an effective therapeutic method to prevent or delay OA. Chondrocytes, as the major cells involved in OA progression, may serve as a promising therapeutic target. RESULTS: A kind of carbon dots (CDs) with excellent biocompatibility was fabricated from folic acid via hydrothermal method and could effectively attenuate osteoarthritis. It was demonstrated that CDs treatment could rescue IL1ß-induced proinflammatory responses, oxidative stress, cartilage degeneration and extracellular matrix degradation. Moreover, CDs reprogrammed lipopolysaccharide (LPS)-induced macrophage inflammation and polarization. Conditioned medium (CM) from CDs-treated macrophages could attenuate IL1ß-induced chondrocyte injury. Also, CM from CDs-treated chondrocytes had immunoregulatory functions on macrophages. Mechanistically, CDs inhibited the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPK) signaling pathways in IL1ß-stimulated chondrocytes. In vivo, anterior cruciate ligament transection (ACLT) mice model was adopted and it was indicated that intra-articular injection of CDs effectively delays OA pathogenesis. CONCLUSIONS: Taken together, these findings indicated CDs could mediate OA via promoting cartilage repair and immunomodulating macrophages within local microenvironment, which may provide evidences for utilizing CDs as a novel nanomaterial for OA treatment.

Effects of the advanced mandibular spring on mandibular retrognathia treatment: a three-dimensional finite element study.

BACKGROUND: The Advanced Mandibular Spring (AMS) was newly developed as a dentofacial orthopedic appliance in conjunctive use of clear aligners to treat Class II malocclusion with mandibular retrognathia in adolescents. This study aimed to launch a biomechanical assessment and evaluate whether the stress patterns generated by AMS promote mandibular growth. METHODS: A three-dimensional finite element model was constructed using images of CBCT and spiral CT. The model consisted of craniomaxillofacial bones, articular discs, retrodiscal elastic stratum, masticatory muscle, teeth, periodontal ligament, aligner and AMS. Mechanical effects were analyzed in three types of models: mandibular postural position, mandibular advancement with AMS, and mandibular advancement with only muscular force. RESULTS: The stress generated by AMS was distributed to all teeth and periodontal ligament, pushing mandibular teeth forward and maxillary teeth backward. In the temporomandibular joint area, the pressure in the superior and posterior aspects of the condyle was reduced, which conformed to the stress pattern promoting condylar and mandibular growth. Stress distribution became even in the anterior aspect of the condyle and the articular disc. Significant tensile stress was generated in the posterior aspect of the glenoid fossa, which conformed to the stress pattern stimulating the remodeling of the fossa. CONCLUSIONS: AMS created a favorable biomechanical environment for treating mandibular retrognathia in adolescents.

Fabrication and Effect of Strontium-Substituted Calcium Silicate/Silk Fibroin on Bone Regeneration In Vitro and In Vivo.

Achieving rapid osteogenesis and angiogenesis was the key factor for bone regeneration. In the present study, the strontium-substituted calcium silicate (SrCS)/silk fibroin (SF) composite materials have been constructed by combining the different functional component ratios of SrCS (12.5 wt%, 25 wt%) and SF. Then, the effects of SrCS/SF materials on proliferation, osteogenic differentiation, and angiogenic factor secretion of rat bone marrow-derived mesenchymal stromal cells (rBMSCs) were first evaluated in vitro. Moreover, the in vivo effect of osteogenesis was evaluated in a critical-sized rat calvarial defect model. In vitro studies showed that SrCS/SF significantly enhanced the cell proliferation, alkaline phosphatase (ALP) activity, and the expression of osteogenic and angiogenic factors of rBMSCs as compared with the SF and CS/SF, and the optimum proportion ratio was 25 wt%. Besides, the results also showed that CS/SF achieved enhanced effects on rBMSCs as compared with SF. The in vivo results showed that 25 wt% SrCS/SF could obviously promote new bone formation more than SF and CS/SF. The present study revealed that SrCS could significantly promote the osteogenic and angiogenic activities of SF, and SrCS/SF might be a good scaffold material for bone regeneration.

Titanium Nanobowl-Based Nest-Like Nanofiber Structure Prepared at Room Temperature and Pressure Promotes Osseointegration of Beagle Implants.

Nest-like nanofiber structures have potential applications in surface modifications of titanium implants. In this study, nest-like nanofiber structures were prepared on a titanium surface at room temperature and pressure by using the nanobowl template-assisted method combined with alkali etching. The characterization and biocompatibility of this material were analyzed by cellular adhesion, death, CCK-8, ALP, and RT-PCR assays in vitro, and osseointegration was evaluated by micro-CT and fluorescent labeling in vivo. The results showed that this nest-like nanofiber structure has a firmer and asperate surface than nanotubes, which leads to better cellular adhesion, proliferation, and differentiation capacity. In a beagle alveolar bone implant model, the nest-like nanofiber structure showed a better osseointegration capacity. In conclusion, this nest-like nanofiber structure has potential applications in dental implantology.

Accuracy of dental arch form in customized fixed labial orthodontic appliances.

INTRODUCTION: This study evaluated arch form accuracy with or without premolar extraction in customized fixed labial orthodontic appliance treatment. METHODS: Setup and posttreatment digital models of 27 samples (15 extractions and 12 nonextractions) were selected and superimposed by best-fit surface-based registration in both the maxilla and the mandible. The facial axis points were identified and converted into Cartesian coordinates. A sixth-order polynomial equation was used to fit dental arches. Arch discrepancies (the mean distance between 2 arch forms) and similarities were compared between extraction and nonextraction groups, maxilla and mandible, and anterior and posterior arches. RESULTS: The arch discrepancy between extraction and nonextraction groups showed no statistically significant difference, but a statistically significant difference in arch similarity was found in the mandible. There were statistically significant differences between anterior and posterior arch discrepancies in the extraction (mandible) and the nonextraction (maxilla and mandible) groups. However, no statistically significant correlation was shown between anterior and posterior arch discrepancies. The arch similarities were 96.18% and 97.38% in the maxilla and 96.01% and 97.49% in the mandible between extraction and nonextraction groups. Arch form discrepancies and similarities showed a moderate correlation but no statistically significant differences between the maxilla and the mandible. CONCLUSIONS: In customized fixed labial orthodontic appliance treatment, arch form setup can be accurately achieved with and without premolar extraction. Anterior arch form acquires fewer discrepancies than the posterior arch, and overcorrection should be added to the end of the customized archwire to reduce posterior arch discrepancies. The discrepancy of the maxillary and mandibular arches is interrelated, and adjustments should be made on both maxillary and mandibular archwires to correct single-jaw transverse malposition.

Synergistic Effect of Micro-Nano-Hybrid Surfaces and Sr Doping on the Osteogenic and Angiogenic Capacity of Hydroxyapatite Bioceramics Scaffolds.

BACKGROUND: The synergistic effect of chemical element doping and surface modification is considered a novel way to regulate cell biological responses and improve the osteoinductive ability of biomaterials. METHODS: Hydroxyapatite (HAp) bioceramics with micro-nano-hybrid (a mixture of microrods and nanorods) surfaces and different strontium (Sr) doping contents of 2.5, 5, 10, and 20% (Srx-mnHAp, x: 2.5, 5, 10 and 20%) were prepared via a hydrothermal transformation method. The effect of Srx-mnHAp on osteogenesis and angiogenesis of bone marrow stromal cells (BMSCs) was evaluated in vitro, and the bioceramics scaffolds were further implanted into rat calvarial defects for the observation of bone regeneration in vivo. RESULTS: HAp bioceramics with micro-nano-hybrid surfaces (mnHAp) could facilitate cell spreading, proliferation ability, ALP activity, and gene expression of osteogenic and angiogenic factors, including COL1, BSP, BMP-2, OPN, VEGF, and ANG-1. More importantly, Srx-mnHAp (x: 2.5, 5, 10 and 20%) further promoted cellular osteogenic activity, and Sr10-mnHAp possessed the best stimulatory effect. The results of calvarial defects revealed that Sr10-mnHAp could promote more bone and blood vessel regeneration, with mnHAp and HAp bioceramics (dense and flat surfaces) as compared. CONCLUSION: The present study suggests that HAp bioceramics with micro-nano-hybrid surface and Sr doping had synergistic promotion effects on bone regeneration, which can be a promising material for bone defect repair.

Biomimetic Hydroxyapatite Nanorods Promote Bone Regeneration via Accelerating Osteogenesis of BMSCs through T Cell-Derived IL-22.

Manipulations of morphological properties of nanobiomaterials have been demonstrated to modulate the outcome of osteoimmunomodulation and eventually osteogenesis through innate immune response. However, the functions and mechanisms of adaptive immune cells in the process of nanobiomaterials-mediated bone regeneration have remained unknown. Herein, we developed bone-mimicking hydroxyapatite (HAp) nanorods with different aspect ratios as model materials to investigate the impacts of the nanoshape features on osteogenesis and to explore the underlying mechanisms focusing on the functions of T cells and T cell-derived cytokines. HAp nanorods with different aspect ratios (HAp-0, HAp-30, and HAp-100) were implanted into mouse mandibular defect models. Micro-CT and hematoxylin and eosin staining demonstrated that HAp-100 had the best osteogenic effects. Flow cytometry analysis revealed that HAp-100 increased the percentage of T cells in injured mandibles. The osteogenic effects of HAp-100 were significantly blunted in injured mandibles of TCRß-/- mice. The Luminex xMAP assay and ELISA showed that HAp-100 induced a marked increase of interleukin (IL)-22 in injured mandibles. In cultured T cells, HAp-100 manifested the best capacity to induce the production of IL-22. Conditioned media from HAp-100-primed T cells promoted osteogenesis and JAK1/STAT3 activation in bone marrow stromal cells, all of which were abolished by neutralizing antibodies against IL-22. In summary, bone-mimicking HAp nanorods with different aspect ratios could regulate osteogenesis through modulation of T cells and IL-22 in the bone regeneration process. These findings provided insights for mediation of the immune response of T cells by nanomaterials on osteogenesis and strategies for designing biomaterials with osteoimmunomodulative functions.

Low-intensity pulsed ultrasound promotes angiogenesis via the AKT pathway and DNA methylation in human umbilical vein endothelial cells.

Angiogenesis involves the activation of endothelial cells (ECs). Low-intensity pulsed ultrasound (LIPUS), which delivers ultrasound waves at a low intensity, can induce the angiogenic potential of ECs. However, the underlying cellular mechanisms remain to be elucidated. In this study, the LIPUS parameters were 1.5 MHz pulsed frequency, 200 us pulse duration, 1.0 kHz repetition rate, and 30 mW/cm2 energy intensity. First, we evaluated the effects of LIPUS on the proliferation and angiogenic differentiation of the EC line EA.hy926. The results showed that LIPUS could induce cell proliferation, promote migration, and increase mRNA level inKDR and CD144.Also, the mRNA level and secretion of VEGF were enhanced. We then investigated the role of the AKT signaling pathway in this process. We observed that the expression of p-AKT was upregulated which means that the AKT signaling pathway could be activated by LIPUS, while inhibitor LY294002 of the AKT signaling pathway effectively blocked LIPUS-induced angiogenesis. Finally,we applied confocal Raman microscopy to track biomolecular changes in cells after LIPUS treatment. Spectral analysis showed DNA methylation changes. An Infinium Methylation assay suggested that399 sites were significantly different. After KEGG enrichment analysis, we found seven genes (IRS1, GNG7, COL4A1, FOXO3, COL4A2, CDK4 and EGF) which were closely related to AKT signaling pathway. We verified that AKT signaling pathway inhibition partially blocked LIPUS-induced DNA methylation changes. Ourstudy demonstrated that LIPUS couldpromote the proliferation and angiogenic differentiation of ECs via the AKT signaling pathway. LIPUS could also alter DNA methylation of ECs via the activation of AKT signal.

Aberrant Fluid Shear Stress Contributes to Articular Cartilage Pathogenesis via Epigenetic Regulation of ZBTB20 by H3K4me3.

PURPOSE: Osteoarthritis (OA) is a common disease for human beings, characterized by severe inflammation, cartilage degradation, and subchondral bone destruction. However, current therapies are limited to relieving pain or joint replacement and no effective treatment methods have been discovered to improve degenerative changes. Currently, a variety of evidences have indicated that aberrant mechanical stimuli is closely associated with articular joint pathogenesis, while the detailed underlying mechanism remains unelucidated. In the present study, we determined to investigate the impact of excessive high fluid shear stress (FSS) on primary chondrocytes and the underlying epigenetic mechanisms. MATERIALS AND METHODS: Phalloidin staining and EdU staining were used to evaluate cell morphology and viability. The mRNA level and protein level of genes were determined by qPCR, Western blot assay, and immunofluorescence staining. Mechanistic investigation was performed through RNA-sequencing and CUT&Tag sequencing. In vivo, we adopted unilateral anterior crossbites (UAC) mice model to investigate the expression of H3K4me3 and ZBTB20 in aberrant force-related cartilage pathogenesis. RESULTS: The results demonstrated that FSS greatly disrupts cell morphology and significantly decreased chondrocyte viability. Aberrant FSS induces remarkable inflammatory mediators production, leading to cartilage degeneration and degradation. In depth mechanistic study showed that FSS results in more than 10-fold upregulation of H3K4me3, and the modulatory effect of H3K4me3 on cartilage was obtained by directly targeting ZBTB20. Furthermore, Wnt signaling was strongly activated in high FSS-induced OA pathogenesis, and the negative impact of ZBTB20 on chondrocytes was also achieved through activating Wnt signaling pathway. Moreover, pharmacological inhibition of H3K4me3 activation by MM-102 or treatment with Wnt pathway inhibitor LF3 could effectively alleviate the destructive effect of FSS on chondrocytes. In vivo UAC mice model validated the dysregulation of H3K4me3 and ZBTB20 in aberrant force-induced cartilage pathogenesis. CONCLUSION: Through the combination of in vitro FSS model and in vivo UAC model, KMT2B-H3K4me3-ZBTB20 axis was first identified in aberrant FSS-induced cartilage pathogenesis, which may provide evidences for epigenetic-based therapy in the future.

In situ construction of a nano-structured akermanite coating for promoting bone formation and osseointegration of Ti-6Al-4V implants in a rabbit osteoporosis model.

With the aging population worldwide, osteoporosis, as an age-related bone metabolic disease, is becoming a hot issue in public health. However, it is still a great challenge to realize osteoporotic bone healing due to the alteration of the bone microenvironment in osteoporosis patients. In this study, a nano-structured akermanite (nAK) coating was in situ constructed on Ti-6Al-4V implants to improve osteoporotic bone repair. In vitro studies indicated that both the surface nano-topography and bioactive ions released from the nAK coatings promoted the proliferation, osteogenesis, angiogenesis and inhibited osteoclastogenesis of ovariectomy rabbit-derived bone marrow mesenchymal stem cells (OVX-rBMSCs). Furthermore, the nAK-coated Ti-6Al-4V implants improved new bone formation and osseointegration in an osteoporosis rabbit model in vivo. These results indicated that the AK coating with a nano-structured surface on the Ti-6Al-4V implant could synergistically promote bone formation and osseointegration for osteoporosis patients. This may be a promising strategy to improve the bone regeneration and osseointegration capability of orthopedic implants under osteoporosis conditions.

Establishment of a C57BL/6 Mandibular Critical-Size Bone Defect Model.

PURPOSE: The goal of this study was to identify bone defects of critical size in C57BL/6 mouse mandibles. MATERIALS AND METHODS: Twenty-four male mice were included in this study. All mice underwent surgeries on their left mandibles. Mandibular defects of 1.0 mm (n = 8), 1.6 mm (n = 8), and 2.3 mm (n = 8) were created. For the investigation of bone healing after an 8-week period, micro-computed tomography scans and histomorphology were performed. RESULTS: Mandibular bone nonunions were seen 0/8 in the 1.0-mm group, 6/8 in the 1.6-mm group, and 8/8 in the 2.3-mm group. The outcome of micro-computed tomography showed that, after 8 weeks, the bone mineral density and the bone volume to total volume ratio were significantly different among the 3 groups. The defect gaps in the nonunion 1.6- and 2.3-mm groups were filled with connective tissue, and no obvious bone formation was found. Additionally, in quantitative analysis, according to the new bone fill calculations, the percentages were 91.85% ±â€Š8.03% in the 1.0-mm group, 59.84% ±â€Š20.60% in the 1.6-mm group, and 15.36% ±â€Š8.28% in the 2.3-mm group, which indicated statistically significantly lower defect healing in the 2.3-mm group. CONCLUSIONS: The creation of 2.3-mm mandibular defects produces osseous nonunion in C57BL/6 mice.