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.

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.

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.

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.

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.

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.

Assessing the Influence of Chin Asymmetry on Perceived Facial Esthetics With 3-Dimensional Images.

PURPOSE: We used 3-dimensinonal (3D) images to assess the influence of chin asymmetry on perceived facial esthetics, investigate the cognitive boundaries of chin asymmetry among orthodontists, general dentists, and laypersons, and provide quantitative reference for clinical treatment. MATERIALS AND METHODS: A 3D facial image was obtained using the 3dMD imaging system (3dMD, Atlanta, GA). The chin was altered in 2-mm increments from 0 to 12 mm and to the left and right using the software program. These images were rated by 66 orthodontists, 89 general dentists, and 66 laypersons as 1) chin symmetry; 2) slight chin asymmetry but acceptable; and 3) serious asymmetry and treatment needed. Multivariate mixed linear regression and multivariate mixed logistic regression analyses were used for statistical data analysis. Descriptive and bivariate statistics were calculated, and statistical significance was considered present at P = .05. RESULTS: The observers progressively increased the grade ratings and the desire for surgery for greater asymmetries (P < .001). Orthodontists and general dentists could perceive a chin deviation of 4 mm to the right and 2 mm to the left, with 8-mm deviation considered to require treatment. Laypersons had a clear perception of a 4-mm chin deviation, with 8 mm to the right and 10 mm to the left considered to require treatment. The right chin deflection was more easily perceived than was the left (P < .05). CONCLUSIONS: Chin asymmetry has a great influence on facial esthetics. We found a statistically significant difference between clinicians (orthodontists and general dentists) and laypersons in the cognition of chin asymmetry and the recommendations for treatment.

Accuracy of different tooth surfaces on 3D printed dental models: orthodontic perspective.

BACKGROUND: Few studies have been reported regarding the accuracy of 3D-printed models for orthodontic applications. The aim of this study was to assess the accuracy of 3D-printed dental models of different tooth surfaces. METHODS: Thirty volunteers were recruited from the hospital, and then their dental models were produced by means of oral scanning and a stereolithography-based 3D printer. Each printed model was digitally scanned and compared with the oral-scanned STL file via superimposition analysis. A color map was used to assess the accuracy of different surfaces (occlusal, buccal, lingual) of anterior and posterior teeth. The Tukey test was used to evaluate the differences between the superimposition. RESULTS: Statistically significant differences were found in the average deviations of different tooth surfaces (P < 0.05). The mean average absolute deviations of the occlusal surfaces of posterior teeth were greater than those of other surfaces. Percentages of points beyond the upper and lower limits of different tooth surfaces displayed the same results (P < 0.05). CONCLUSIONS: Occlusal surfaces, especially pits and fissures of posterior teeth on 3D printed maxillary dental models, showed greater distortions than those of other teeth and regions.

Hemifacial microsomia treated with a hybrid technique combining distraction osteogenesis and a mandible-guided functional appliance: Pilot study.

INTRODUCTION: The purpose of this study was to evaluate the therapeutic effect of a hybrid treatment for hemifacial microsomia that combines distraction osteogenesis and a mandible-guided functional appliance to correct mandibular asymmetry. METHODS: This was a retrospective analysis of 10 patients with unilateral hemifacial microsomia who underwent mandibular ramus distraction osteogenesis in our hospital from February 2013 to July 2015. The cases were classified into 2 comparison groups: 5 patients were in the MG-DO group (distraction osteogenesis combined with an mandible-guided functional appliance) and 5 in the control group (distraction osteogenesis only). Anteroposterior cephalometric analyses were conducted before and after treatment. Soft tissue symmetry and the occlusal relationship were observed from facial and intraoral photographs. Statistical analyses were performed to determine changes between before and after treatment as well as intergroup differences. RESULTS: The MG-DO group showed greater vertical elongation of the mandibular ramus and less overcorrection and mandibular deviation than the control group. Occlusal reconstruction was enabled by the mandible-guided functional appliance owing to a decrease in lateral shifting. The symmetry of both skeletal and soft tissues was significantly improved in the MG-DO group. CONCLUSIONS: The hybrid technique combining distraction osteogenesis and the mandible-guided functional appliance proved to be effective in correcting canting and deviation during mandibular elongation, which improved facial symmetry and occlusal balance in patients with hemifacial microsomia.

Accuracy of in vitro mandibular volumetric measurements from CBCT of different voxel sizes with different segmentation threshold settings.

BACKGROUND: To determine the accuracy of volumetric measurements of the mandible in vitro by cone-beam computed tomography (CBCT) and to analyze the influence of voxel sizes and segmentation threshold settings on it. METHODS: The samples were obtained from pig mandibles and scanned with 4 voxel sizes: .125 mm, .20 mm, .30 mm, and .40 mm. The minimum segmentation thresholds in Hounsfield units (HU) were set as 0, 100, 200, 300, and 400, respectively, for each voxel size for 3D reconstruction. Laser scanning as the reference, the volumes of each CBCT scanning, the mean iterative distances of superimposition and total positive and negative deviations were recorded and compared. RESULTS: The volumes of CBCT-scan deviated from those of laser-scan by + 7.67% to - 3.05% with different HU and voxel sizes. The deviation increased with the voxel size. There was a more suitable minimum HU threshold of segmentation (HU100 for .125 mm, 200 for .20 mm, 300 for .30 mm, and 400 for .40 mm) for each voxel size. CONCLUSIONS: Voxel sizes and Hounsfield unit thresholds influence the accuracy of volumetric measurements in CBCT scanning. The volume increase with the voxel size, and different voxel sizes correspond to different optimal Hounsfield unit thresholds.

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.

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.

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.

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.

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.

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.

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.

Ectopic study of calcium phosphate cement seeded with pBMP-2 modified canine bMSCs mediated by a non-viral PEI derivative.

We have evaluated the ectopic new bone formation effects of CPC (calcium phosphate cement) seeded with pBMP-2 (plasmids containing bone morphogenetic protein-2 gene) transfected canine bMSCs (bone marrow stromal cells) mediated by a non-viral PEI (polyethylenimine) derivative (GenEscort™ II) in nude mice. Canine bMSCs were transfected with pBMP-2 or pEGFP (plasmids containing enhanced green fluorescent protein gene) mediated by GenEscort™ II in vitro, and the osteoblastic differentiation was explored by ALP (alkaline phosphatase) staining, ARS (alizarin red S) staining and RT-qPCR (real-time quantitative PCR) analysis. Ectopic bone formation effects of CPC/pBMP-2 transfected bMSCs were evaluated and compared with CPC/pEGFP transfected bMSCs or CPC/untransfected bMSCs through histological, histomorphological and immunohistochemical analysis 8 and 12 weeks post-operation in nude mice. Transfection efficiency was up ∼35% as demonstrated by EGFP (enhanced green fluorescent protein) expression. ALP and ARS staining were stronger with pBMP-2 gene transfection, and mRNA expression of BMP-2 (bone morphogenetic protein-2), Col 1 (collagen 1) and OCN (osteocalcin) in pBMP-2 group was significantly up-regulated at 6 and 9 days. Significantly higher NBV (new bone volume) was achieved in pBMP-2 group than in the control groups at 8 and 12 weeks (P<0.05). In addition, immunohistochemical analysis indicated higher OCN expression in pBMP-2 group (P<0.01). We conclude that CPC seeded with pBMP-2 transfected bMSCs mediated by GenEscort™ II could enhance ectopic new bone formation in nude mice, suggesting that GenEscort™ II mediated pBMP-2 gene transfer is an effective non-viral method and CPC is a suitable scaffold for gene enhanced bone tissue engineering.

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.