Effects of bisphosphonates on mandibular condyle of ovariectomized osteoporotic rats using micro-ct and histomorphometric analysis.

OBJECTIVE: To evaluate microarchitectural changes in condylar cartilage and associated subchondral bone after bisphosphonates treatment using an ovariectomized (OVX) osteoporosis rat model. METHODS: Thirty six-month-old female Sprague-Dawley rats were randomly divided into sham, OVX, and risedronate (RIS)-treated groups. Both OVX and RIS groups received bilateral ovariectomy. OVX group was treated subcutaneously with saline, whereas RIS group received risedronate treatment (2.4 µg/kg) subcutaneously for 3 months. At the end of 3 months, animals were sacrificed and the entire condyles were harvested for micro-CT and histological analyses. Immunohistochemistry (IHC) was performed to assess the expression of type I/II collagen protein by semiquantitative imaging analysis. RESULTS: Micro-CT analysis showed OVX group had significant condylar subchondral bone loss compared to sham as shown by significant decrease in bone volume fraction (P = 0.028), trabecular thickness (P = 0.041), and significant increase in trabecular spacing (P = 0.003). In RIS group, partial inhibition of OVX-induced bone loss was detected. HE staining showed proliferative layer of condylar cartilage reduced, while hypertrophic chondrocyte layer increased significantly in RIS group compared to sham and OVX groups. IHC showed reduced expression of Col I in both the OVX and RIS groups, whereas expression of Col II was reduced in the OVX group but increased in the RIS group. CONCLUSION: Our findings suggest that systemic bisphosphonate treatment influences the structure and ossification of condylar cartilage and it has a dual action on condyle in a postmenopausal osteoporosis rat model which raises the concerns for the potential side effects of BPs on condyle to elder patients.

Combined toxic effects of nanoplastics and norfloxacin on antioxidant and immune genes in mussels.

Nanoplastics (NPs) and antibiotics (ABs) are two of the emerging marine contaminants that have drawn the most attention in recent years. Given the necessity of figuring out the effects of plastic and antibiotic contamination on marine organism life and population in the natural environment, it is essential to apply rapid and effective biological indicators to evaluate their comprehensive toxic effects. In this study, using mussel (Mytilus coruscus) as a model, we investigated the combined toxic effects of NP (80 nm polystyrene beads) and AB (Norfloxacin, NOR) at environmental-relevant concentrations on antioxidant and immune genes. In terms of the antioxidant genes, NPs significantly increased the relative expression of Cytochrome P450 3A-1 (CYP3A-1) under various concentrations of NOR conditions, but they only significantly increased the relative expression of CYP3A-2 in the high concentration (500 µg L-1 NOR) co-exposure group. In the NP-exposure group which exposed to no or low concentrations of NOR, nuclear factor erythroid 2-related factor 2 (Nrf2) was upregulated. In terms of the immune genes, interleukin-1 receptor-associated kinase (IRAK) -1 showed a significant increase in the low-concentration NOR group while a significant inhibition in the high-concentration NOR group. Due to the presence of NPs, exposure to NOR resulted in a significant increase in both IRAK-4 and heat shock protein (HSP) 70. Our findings indicate that polystyrene NPs can exacerbate the effects of NOR on the anti-oxidant and immune defense performance of mussels. This study delves into the toxic effects of NPs and ABs from a molecular perspective. Given the expected increase in environmental pollution due to NPs and ABs, future research is needed to investigate the potential synergistic effect of NPs and ABs on other organisms.

[Effect of different kinds of gingival retraction agents on the polymerization inhibition of polyvinyl siloxane impression materials].

PURPOSE: To evaluate the effect of different kinds of gingival retraction agents after directly contacted with polyvinyl siloxane impression materials on polymerization inhibition and the inhibition degree. METHODS: Five kinds of gingival retraction agents (0.1% epinephrine hydrochloride, 0.05% oxymetazoline, 15.5% ferric sulfate, 25% aluminum chloride and 5% aluminum chloride) were chosen, normal saline was as control group, and two kinds of polyvinyl siloxane impression materials (ExpressTM, ImprintTM Ⅱ) were combined into 12 groups. There were 12 specimens in each group and 144 specimens in total. Silicone rubber impression materials were mixed by the same operator using a dispensing gun into the acrylic mold, so that they could directly contact the gingival retraction agents on the densely woven cotton fabrics. The samples were removed when the polymerization time arrived according to the manufactures' recommendations and then placed under a stereomicroscope with a magnification of 10 times to observe whether polymerization inhibition occurred, the degree of inhibition was compared afterwards. SPSS 22.0 software package was used for statistical analysis. RESULTS: The polymerization inhibition of two kinds of silicone rubber impression materials occurred in 15.5% ferric sulfate group and 25% aluminum chloride group, and the inhibition occurrence rate was 100%, the difference was statistically significant (P＜0.05) compared with normal saline group. Inhibition was not found in 0.1% epinephrine hydrochloride group, 0.05% oxymetazoline group and 5% aluminum chloride. The effect of 15.5% ferric sulfate and 25% aluminum chloride on polymerization inhibition degree of ImprintTM Ⅱ was greater than ExpressTM, and the difference was statistically significant(P＜0.05). CONCLUSIONS: When silicone rubber impression material is used during impression procedure, attention should be paid to the effect of the gingival retraction agent containing 15.5% ferric sulfate and 25% aluminum chloride on its polymerization. The gingival retraction agent should be washed before impression to avoid the residue directly contacting the silicone rubber to prevent polymerization.

Combined effects of microfibers and polychlorinated biphenyls on the immune function of hemocytes in the mussel Mytilus coruscus.

Numerous studies have shown that microplastics can interact with other pollutants in the environment to produce synergistic effects, leading to more serious impacts. To date, there is little consensus on the combined effects of microfibers (MFs) and polychlorinated biphenyls (PCBs, Aroclor 1254), two legacy and alarming environmental pollutants. There is an urgent need to assess the impact of combined exposures on bivalve immune defences. In this study, we assessed the immune response of the mussels (Mytilus coruscus) hemocyte to MFs and PCBs alone and in combination by using flow cytometry. M. coruscus were exposed to MFs (1000 pieces/L) and PCBs (PCBs) (100 ng/L and 1000 ng/L) alone or in combination for 14 consecutive days and recovered for 7 days. The hemocyte of M. coruscus was collected on day 7, 14 and 21. MF exposure alone had no effect on the hemocyte. The total hemocyte count (THC), esterase (EA), lysosomal contents (LC), mitochondrial number (MN) and mitochondrial membrane potential (MMP) of mussels showed a decreasing trend with increasing PCB concentrations, both individually and in combination; The decreases in EA, MN and MMP were associated with the induction of reactive oxygen species (ROS). Hemocyte mortality (HM) was associated with a decrease in THC. Combined exposure to MFs and PCBs would exacerbate the effects on hemocyte immunity. These new findings improve our understanding of the toxic effects of MFs and organic chemical pollutants, and demonstrate the potential mechanism of PCBs to bivalves through changes in hemolymph immunity-related indicators.

Biomechanical Assessment of Tilted Mandibular Second Molars with Full-Crown Adjacent to Implant-Supported Restoration: 3D Finite Element Analysis.

Purpose: The aim of this study was to investigate the effects of tooth root inclination and crown preparation angulation on the stress distribution of tilted second molars, supporting structures and adjacent implant by using the finite element analysis method. Materials and Methods: 3D finite element models of tilted second molar and tooth-supporting structures, including the two designs with three different angles of root inclination and crown preparation angulations, were constructed for full-crown restoration. For all models, the stress distribution was analyzed under vertical and oblique loading conditions. Results: The maximum equivalent stress (MES) increased as root inclination increased, and the highest stress value occurred in the tooth root furcation of the model with 30° root inclination under oblique loading. When root inclination was the same, the MES of each structure was approximate under the same direction load regardless of crown preparation angulation. Higher stress values were found on the tooth root, periodontal ligament, and cortical bone of all models under oblique load compared with vertical load. The highest stress value occurred in the distal adjacent area of implant. Conclusion: Tooth roots with less than 30° inclination, occlusal preparation parallel to the bite plane and small oblique force loading are recommended as significant considerations for full-crown restoration of a mesial inclined mandibular second molar.

Multifeature superposition analysis of the effects of microplastics on microbial communities in realistic environments.

Microplastic (MP) contamination has become an increasingly serious environmental problem. However, the risks of MP contamination in complex global climatic and geographic scenarios remain unclear. We established a multifeature superposition analysis boosting (MFAB) machine learning (ML) approach to address the above knowledge gap. MFAB-ML identified and predicted the importance, interaction networks and superposition effects of multiple features, including 34 characteristic variables (e.g., MP contamination and climatic and geographic variables), from 1354 samples distributed globally. MFAB-ML analysis achieved realistic and significant results, in some cases even opposite to those obtained using a single or a few features, revealing the importance of considering complicated scenarios. We found that the microbial diversity in East Asian seas will continually decrease due to the superposition effects of MPs with ocean warming; for example, the Chao1 index will decrease by 10.32% by 2065. The present work provides a powerful approach to identify and predict the multifeature superposition effects of pollutants on realistic environments in complicated climatic and geographic scenarios, overcoming the bias from general studies.

Dexamethasone-loaded zeolitic imidazolate frameworks nanocomposite hydrogel with antibacterial and anti-inflammatory effects for periodontitis treatment.

Periodontitis is a bacterial-induced, chronic inflammatory disease characterized by progressive destruction of tooth-supporting structures. Pathogenic bacteria residing in deep periodontal pockets after traditional manual debridement can still lead to local inflammatory microenvironment, which remains a challenging problem and an urgent need for better therapeutic strategies. Here, we integrated the advantages of metal-organic frameworks (MOFs) and hydrogels to prepare an injectable nanocomposite hydrogel by incorporating dexamethasone-loaded zeolitic imidazolate frameworks-8 (DZIF) nanoparticles into the photocrosslinking matrix of methacrylic polyphosphoester (PPEMA) and methacrylic gelatin (GelMA). The injectable hydrogel could be easily injected into deep periodontal pockets, achieving high local concentrations without leading to antibiotic resistance. The nanocomposite hydrogel had high antibacterial activity and constructs with stable microenvironments maintain cell viability, proliferation, spreading, as well as osteogenesis, and down-regulated inflammatory genes expression in vitro. When evaluated on an experimental periodontitis rat model, micro-computed tomography and histological analyses showed that the nanocomposite hydrogel effectively reduced periodontal inflammation and attenuated inflammation-induced bone loss in a rat model of periodontitis. These findings suggest that the nanocomposite hydrogel might be a promising therapeutic candidate for treating periodontal disease.

Rapid mineralization of hierarchical poly(l-lactic acid)/poly(ε-caprolactone) nanofibrous scaffolds by electrodeposition for bone regeneration.

Introduction: Hierarchical nanofibrous scaffolds are emerging as a promising bone repair material due to their high cell adhesion activity and nutrient permeability. However, the existing method for hierarchical nanofibrous scaffolds fabrication is complicated and not perfectly suitable for further biomedical application in view of both structure and function. In this study, we constructed a hierarchical nanofibrous poly (l-lactic acid)/poly(ε-caprolactone) (PLLA/PCL) scaffold and further evaluated its bone healing ability. Methods: The hierarchical PLLA/PCL nanofibrous scaffold (PLLA/PCL) was prepared by one-pot TIPS and then rapidly mineralized at room temperature by an electrochemical deposition technique. After electrode-positioning at 2 V for 2 hrs, a scaffold coated with hydroxyapatite (M-PLLA/PCL) could be obtained. Results: The pore size of the M-PLLA/PCL scaffold was hierarchically distributed so as to match the biophysical structure for osteoblast growth. The M-PLLA/PCL scaffold showed better cell proliferation and osteogenesis activity compared to the PLLA/PCL scaffold. Further in vivo bone repair studies indicated that the M-PLLA/PCL scaffold could accelerate defect healing in 12 weeks. Conclusion: The results of this study implied that the as-prepared hydroxyapatite coated hierarchical PLLA/PCL nanofibrous scaffolds could be developed as a promising material for efficient bone tissue repair after carefully tuning the TIPS and electrodeposition parameters.

An enzyme-responsive membrane for antibiotic drug release and local periodontal treatment.

Periodontitis is a chronic, destructive inflammatory disease that injures tooth- supporting tissues, eventually leading to tooth loss. Complete eradication of periodontal pathogenic microorganisms is fundamental to allow periodontal healing and commonly precedes periodontal tissue regeneration. To address this challenge, we report a strategy for developing an enzyme-mediated periodontal membrane for targeted antibiotic delivery into infectious periodontal pockets; the unique components of the membrane will also benefit periodontal alveolar bone repair. In this approach, a chitosan membrane containing polyphosphoester and minocycline hydrochloride (PPEM) was prepared. Physical, morphological, and ultrastructural analyses were carried out in order to assess cellular compatibility, drug release and antibacterial activity in vitro. Additionally, the functionality of the PPEM membrane was evaluated in vivo with a periodontal defect model in rats. The results confirm that the PPEM membrane exhibits good physical properties with excellent antibacterial activity and successfully promotes periodontal tissue repair, making it promising for periodontal treatment.

Early effects of parathyroid hormone on vascularized bone regeneration and implant osseointegration in aged rats.

The decreased bone mass and impaired osteogenesis capacities that occur with aging may influence the outcome of dental implants. Parathyroid hormone (PTH) (1-34) is an anabolic agent for the treatment of osteoporosis. However, little is known about its effects and mechanisms on vascularized bone regeneration and implant osseointegration in aging. In current study, we adopted both in vivo and in vitro approaches to explore the mechanisms of early actions of PTH (1-34) on the angiogenic and osteogenic microenvironment to enhance implant osseointegration in aged rats. Daily subcutaneous injections of 30 µg/kg PTH (1-34) were given to female rats aged 20 months beginning on next day of implantation and lasting for 5 weeks. Radiological and histological analysis confirmed that PTH (1-34) improved new bone formation, angiogenesis and implant osseointegration in aged rats in the early stage. The osteogenic potential of aged bone mesenchymal stem cells (BMSCs) was enhanced, while their adipogenesis capacity was attenuated. Furthermore, PTH (1-34) was shown to promote angiogenesis directly via endothelial cell migration and blood vessel formation in vitro. Meanwhile, PTH (1-34) stimulated more osteoclasts participation in bone remodeling by secreting angiogenic and osteogenic growth factors to induce early vascularization and stimulate the migration or differentiation of BMSCs indirectly. Together, these results demonstrate mechanistic insight into how PTH (1-34) regulates the angiogenic and osteogenic microenvironment to result in more active bone remodeling and new bone formation, making it an excellent potential therapeutic agent for rapid vascularized bone regeneration and implant osseointegration in the aged population.

iTRAQ-based quantitative proteomic analysis on differentially expressed proteins of rat mandibular condylar cartilage induced by reducing dietary loading.

As muscle activity during growth is considerably important for mandible quality and morphology, reducing dietary loading directly influences the development and metabolic activity of mandibular condylar cartilage (MCC). However, an overall investigation of changes in the protein composition of MCC has not been fully described in literature. To study the protein expression and putative signaling in vivo, we evaluated the structural changes of MCC and differentially expressed proteins induced by reducing functional loading in rat MCC at developmental stages. Isobaric tag for relative and absolute quantitation-based 2D nano-high performance liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) technologies were used. Global protein profiling, KEGG and PANTHER pathways, and functional categories were analyzed. Consequently, histological and tartrate-resistant acid phosphatase staining indicated the altered histological structure of condylar cartilage and increased bone remodeling activity in hard-diet group. A total of 805 differentially expressed proteins were then identified. GO analysis revealed a significant number of proteins involved in the metabolic process, cellular process, biological regulation, localization, developmental process, and response to stimulus. KEGG pathway analysis also suggested that these proteins participated in various signaling pathways, including calcium signaling pathway, gap junction, ErbB signaling pathway, and mitogen-activated protein kinase signaling pathway. Collagen types I and II were further validated by immunohistochemical staining and Western blot analysis. Taken together, the present study provides an insight into the molecular mechanism of regulating condylar growth and remodeling induced by reducing dietary loading at the protein level.

[Correlation between platelet aggregation rates of aspirin users and intra-socket clotting after extraction of a maxillary tooth with periodontitis].

PURPOSE: To explore the optimal platelet aggregation rates (PAgTs) of aspirin users, which can ensure normal intra-socket clotting after extraction of a maxillary tooth with serious periodontitis, and to determine the reasonable time of pre-extraction aspirin suspension. METHODS: Ninety aspirin users (100mg/d) requiring extraction of a maxillary tooth with serious periodontitis were enrolled and divided into anterior tooth group (group A), premolar group (group B) and molar group (group C). Each group contained 10 cases with aspirin continuation, 10 cases with aspirin withdrawal 3 days pre-extraction and 10 cases with 5-day of aspirin withdrawal. After preoperative light transmission aggregometry (LTA) using arachidonic acid (AA) as the inducer to each case, the extractions were preformed. According to the intra-socket clotting classification table (class I indicated normal clotting, class II-III indicated successively increasing bleeding amount), the evaluation for each case was implemented and the analysis of the impacts caused by pre-extraction aspirin continuation and suspension on intra-socket clotting was conducted. The research on the best cut-off value of AA-induced PAgT to predict normal post-extraction clotting was also carried out by using receiver operating characteristic curve (ROC curve, SPSS11.6). RESULTS: (1)There were 4 cases of Class II in group A (3 without aspirin cessation and 1 with 3-day suspension), 8 in group B (5 without aspirin cessation, 2 with 3-day suspension and 1 with 5-day suspension) and 11 in group C (6 without aspirin cessation, 4 with 3-day suspension and 1 with 5-day suspension). The other cases in 3 groups were Class I. (2) The best cut-off value of AA induced PAgT was 10.15%, 13.25%, 16.5% for group A, B, C, respectively. CONCLUSIONS: For aspirin users (100mg/d) who will accept extraction of a maxillary tooth with serious periodontitis, a detection of pre-extraction LTA-AA is profitable, AA induced PAgT≥10.15% for an anterior tooth extraction, PAgT≥13.25% for a premolar extraction and PAgT≥16.5% for a molar extraction indicate normal postoperative intra-socket clotting, so aspirin continuation is feasible under such conditions. Otherwise, post-extraction bleeding amount can increase and preoperative aspirin cessation should be considered. The reasonable time of aspirin suspension is 3 days for an anterior tooth extraction and 5 days for a premolar or molar extraction. Additionally, hemostatic measures on the surgical site should be strengthened.

Influence of fiber posts on the fracture resistance of endodontically treated premolars with different dental defects.

This study aimed to evaluate the influence of quartz fiber post placement on the fracture resistance of endodontically treated premolars with different dental defects under dynamic loading. Fifty extracted single-rooted mandibular premolars were randomized into five groups. Each group was prepared according to numbers of residual walls ranged from 0 to 4. Then each group was divided into two subgroups with one restored with quartz fiber posts and the other without posts. In no-post groups, gutta percha point 2 mm below cemento-enamel junction was removed. Composite resin was adapted to the well and used to shape the core directly. Each tooth was restored with a complete metal crown. Dynamic loading was carried out in a masticatory simulator with a nominal load of 50 N at 2 Hz for 300 000 loading cycles. Then a quasi-statically load was applied in a universal testing machine 30° to the long axis with a crosshead speed of 1 mm⋅min(-1) until fracture. Data were analyzed with one-way analysis of variance and pairwise comparison (P<0.05). No specimens failed during dynamic loading. The fracture resistance enhanced with the increase of numbers of coronal walls and the differences were significant (P<0.05). Placement of fiber posts had a significant effect when fewer than two walls remained (P<0.05), but it had no significant influence in groups with two, three or four walls (P>0.05). Fiber post did not change failure mode, and the fracture pattern was mainly favorable. More dentin walls need to be retained in clinic. When no less than two walls remained, a fiber post is not always necessary.

[A clinical study on the stability of miniscrew anchorage in orthodontics].

PURPOSE: The aim of this retrospective study was to summarize the cases who used miniscrew anchorage and to evaluate the clinical factors influencing the stability of miniscrew. METHODS: The sample comprised 13 patients with 32 miniscrews, in whom the design, occlusogingival position, alveolar position, loading opportunity, gingival condition were investigated to assess the factors related to stability. The data was analyzed by univariate analysis and multivariate analysis using SAS 10.0 software package. RESULTS: The overall success rate was 90.6%. There was significant relationship between the patients' age and the stability of miniscrews (P=0.04). The success rate was 75% in adolescent patients, and 100% in adult patients. The osseointegration was affected by initial over-loading, but the long-term orthodontic loading (200g) was safe, if osseointegration was achieved. Perio-implant gingivitis, even the suppurative gingivitis (2 cases) didn't destroy the bone-implant interface. The success rate was independent on gender, occlusogingival position, jaw positioning, inserting times of the same miniscrew, and early loading. CONCLUSIONS: The primary stability is the crucial factor for long-term stability of the miniscrews.

[A study on the stability of miniscrew on different loading time as orthodontic anchorage].

PURPOSE: To evaluate the stability of the miniscrew on different loading time as orthodontic anchorage. METHODS: 2 healthy adult male Beagle dogs were used in this study. 48 mini-implants were implanted into the maxilla and mandible of the dogs. The miniscrews were divided into 8 different groups,1 group was loaded 0g as control group and the others were loaded 200g forces as experimental groups. 200g forces were loaded on the corresponding mini-implant anchorages immediately after implantation and at the time of 1w,2w,3w,4w,5w and 6w after implantation. The dogs were sacrificed at 12w after implantation. Histological progresses of implant-bone interfaces was examined with light microscope. Blue deposition and contact ratio were calculated and analyzed using SPSS13.0 software package. RESULTS: Fibrous and osseous-integration was noted in the interface, there was no significant difference in bone deposition ratio and bone contact ratio. CONCLUSION: Different force loading time does not affect the stability of the mini-implant anchorage.

[Progress of histological study on anchorage implants].

The study and the application of anchorage implants had attracted more and more attention from the orthodontists. This article reviewed the histological studies involving the implants used as orthodontic anchorage and found tendency to be micro-implants and self-drilling. The new concepts of osseointegration between micro-implant and bone, immediate loading, long-term loading and periodontal repair after injury from anchorage implant evoked challenges to the conventional understanding.