Cytotoxicity, Oxidative Stress, and Autophagy Effects of Tantalum Nanoparticles on MC3T3-E1 Mouse Osteoblasts.

As a bone implant material, porous tantalum (Ta) has better corrosion resistance and more suitable elastic modulus than titanium. Surface nanomodification can accelerate the integration of Ta implants with bone tissue, which has broad application prospects in the field of dental implantology. Due to mechanical stress and load wear, nanoscale Ta fragments are inevitably exfoliated from the implant surface and brought into direct contact with osteoblasts surrounding the implant. These wear fragments may affect the biological characteristics of osteoblasts and thus the stability of implants. To date, the interaction of nanoscale Ta fragments with osteoblasts has not been clearly investigated. In the current study, we used the mouse osteoblast cell line MC3T3-E1 to explore the effects of Ta nanoparticles (Ta-NPs) on the cytotoxicity, oxidative stress and autophagy of osteoblasts. We found that a low concentration (12.5 µg/mL) of Ta-NPs can promote the proliferation of osteoblasts, while the Ta-NPs began to induce a decrease in cell viability at concentrations ≥25 µg/mL. Increased cell mortality, reactive oxygen species (ROS) production and decreased mitochondrial membrane potential (MMP) occurred in a dose-dependent manner after Ta-NP treatment. Moreover, with Ta-NP stimulation, the ratio of LC3-II/LC3-I increased, and the level of p62 protein was reduced. However, the degradation of p62 was not continuously increased when the concentration of Ta-NPs was ≥25 µg/mL. These results indicate that Ta-NPs induced osteoblast damage via oxidative stress. Autophagy activation may be a key factor in the cellular response to Ta-NP toxicity and could have an important impact on determining the survival or death of osteoblasts.

Psoriasis Patients Suffer From Worse Periodontal Status-A Meta-Analysis.

Background and Objective: Patients with psoriasis have a significantly elevated risk of periodontitis compared with the nonpsoriasis controls. However, the data regarding the difference in the periodontal health status of the psoriasis patients and the nonpsoriasis controls are limited and inconsistent; hence, a specialized meta-analysis that quantitatively compared the periodontal status between the psoriasis and nonpsoriasis subjects by evaluating the related clinical periodontal indexes was needed. The aim of this meta-analysis was to quantitatively evaluate whether the periodontal status of psoriasis patients is worse than that of nonpsoriasis subjects. Methods: We searched PubMed and EMBASE for all eligible studies that compared the periodontal status between psoriasis patients and nonpsoriasis subjects. The studies were screened based on pre-established inclusion criteria. After extracting the available periodontal indexes from the included studies, the weighted mean difference (WMD) with 95% confidence intervals (CIs) was calculated by pooling the mean and standard deviations (SD) of each index. Results: In total, 8 studies, including 812 psoriasis patients and 772 nonpsoriasis subjects, were included in our meta-analysis, and the publication dates ranged from 2013 to 2019; eight periodontal indexes were analyzed. The WMD (95% CIs) for each index were: bleeding on probing (%), 9.188 (4.046-14.330, P < 0.001); probing depth (mm), 0.524 (0.183-0.865, P = 0.003); clinical attachment loss (mm), 0.408 (0.051-0.765, P = 0.025); plaque index, 0.186 (-0.170 to 0.543, P = 0.306); gingival index, 0.458 (-0.413 to 1.328, P = 0.303), remaining teeth, -1.709 (-2.106 to -1.312, P < 0.001); missing teeth, 1.130 (0.275-1.985, P = 0.010); the level of alveolar bone loss (mm), 0.400 (0.102-0.698, P = 0.008). Conclusion: In summary, our meta-analysis revealed that psoriasis patients suffer from worse periodontal health than do nonpsoriasis subjects, mainly characterized by worse gingival inflammation, more alveolar bone loss, fewer remaining teeth and more missing teeth. Considering the limitations of this meta-analysis, more high-quality and well-designed studies are needed to validate our conclusions in the future.

Osseointegration of layer-by-layer polyelectrolyte multilayers loaded with IGF1 and coated on titanium implant under osteoporotic condition.

PURPOSE: Titanium implant is a widely used method for dental prosthesis restoration. Nevertheless, in patients with systemic diseases, including osteoporosis, diabetes, and cancer, the success rate of the implant is greatly reduced. This study investigates a new implant material loaded with insulin-like growth factor 1 (IGF1), which could potentially improve the implant success rate, accelerate the occurrence of osseointegration, and provide a new strategy for implant treatment in osteoporotic patients. MATERIALS AND METHODS: Biofunctionalized polyelectrolyte multilayers (PEMs) with polyethylenimine as the excitation layer and gelatin/chitosan loaded with IGF1 were prepared on the surface of titanium implant by layer-by-layer self-assembly technique. The physical and chemical properties of the biofunctionalized PEMs, the biological characteristics of bone marrow mesenchymal stem cells (BMMSCs), and bone implant contact correlation test indexes were detected and analyzed in vitro and in vivo using osteoporosis rat model. RESULTS: PEMs coatings loaded with IGF1 (TNS-PEM-IGF1-100) implant promoted the early stage of BMMSCs adhesion. Under the action of body fluids, the active coating showed sustained release of growth factors, which in turn promoted the proliferation and differentiation of BMMSCs and the extracellular matrix. At 8 weeks from implant surgery, the new bone around the implants was examined using micro-CT and acid fuchsin/methylene blue staining. The new bone formation increased with time in each group, while the TNS-PEM-IGF1-100 group showed the highest thickness and continuity. CONCLUSION: TNS-PEM-IGF1-100 new implants can promote osseointegration in osteoporotic conditions both in vivo and in vitro and provide a new strategy for implant repair in osteoporotic patients.

Chronic sleep deprivation alters the myosin heavy chain isoforms in the masseter muscle in rats.

To investigate the changes in myosin heavy chain (MyHC) isoforms of rat masseter muscle fibres caused by chronic sleep deprivation and a possible link with the pathogenesis of disorders of the temporomandibular joint (TMJ). A total of 180 male rats were randomly divided into three groups (n=60 in each): cage controls, large platform controls, and chronic sleep deprivation group. Each group was further divided into three subgroups with different observation periods (7, 14, and 21 days). We investigated he expression of MyHC isoforms in masseter muscle fibres by real-time quantitative polymerase chain reaction (PCR), Western blotting, and immunohistochemical staining. In rats with chronic sleep deprivation there was increased MyHC-I expression in layers of both shallow and deep muscles at 7 and 21 days compared with the control groups, whereas sleep deprivation was associated with significantly decreased MyHC-II expression. At 21 days, there were no differences in MyHC-I or MyHC-II expression between the groups and there were no differences between the two control groups at any time point. These findings suggest that chronic sleep deprivation alters the expression of MyHC isoforms, which may contribute to the pathogenesis of disorders of the TMJ.

Should aspirin be stopped before tooth extraction? A meta-analysis.

OBJECTIVE: To carry out a standard meta-analysis to determine if aspirin should be stopped before tooth extraction. STUDY DESIGN: The PubMed, ScienceDirect, EBSCOhost, and Science Citation Index databases were searched for studies published up to September 30, 2014. Eligible studies were restricted to randomized controlled trials (RCTs) and controlled, nonrandomized trials. RESULTS: Three RCTs and seven controlled trials met the inclusion criteria (covering 1752 patients: 529 on aspirin therapy and 1223 not on aspirin therapy). The results showed that the risk of postoperative hemorrhage was significantly higher in patients on aspirin therapy (relative risk [RR] = 2.46; 95% confidence interval [CI]: 1.45-4.81) but that bleeding time (BT) was not significantly different between the two groups (standardized mean difference [SMD] = 0.63; 95% CI: -0.04 to 1.31). Sensitivity analyses showed that the results were unstable. CONCLUSIONS: We could reach a conclusion that BT is prolonged or hemorrhage is exacerbated by long-term use of aspirin. We recommend not stopping long-term aspirin use before tooth extraction but enhancing hemostasis methods, if necessary.

Simultaneous detection of four common oral Candida species from blood samples by the fluorescence polarization assay.

The genus Candida is both the commensal microbe and the opportunistic pathogen, containing approximately 200 species inhabiting in oral cavity of 53 % of the general population. Candida species can cause the diseases from local mucosal infections to systemic mycoses, even life-threatening infections in immunocompromised individuals. The timely differentiation of Candida species is important for the guidance of clinical medication. Four common Candida species in Chinese population (Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei) were chosen as the targets to develop the rapid screening method in this work. Combined with amplification by asymmetric PCR, this parallel fluorescence polarization (FP) immunoassay is carried out in homogeneous solution phase. The limit of detection of the assay was shown to be 50 copies/mL in blood samples. The evaluation in multicenter manner showed excellent reproducibility and stability. The comparison between DNA sequencing and the FP immunoassay indicated that there was no significant difference between these methods. This molecular strategy-based method is simple, rapid, and feasible for identifying common Candida species and thereby holding great potential in the application of clinical laboratories.

Effects of chronic sleep deprivation on the extracellular signal-regulated kinase pathway in the temporomandibular joint of rats.

OBJECTIVES: To examine the possible involvement and regulatory mechanisms of extracellular signal-regulated kinase (ERK) pathway in the temporomandibular joint (TMJ) of rats subjected to chronic sleep deprivation (CSD). METHODS: Rats were subjected to CSD using the modified multiple platform method (MMPM). The serum levels of corticosterone (CORT) and adrenocorticotropic hormone (ACTH) were tested and histomorphology and ultrastructure of the TMJ were observed. The ERK and phospho-ERK (p-ERK) expression levels were detected by Western blot analysis, and the MMP-1, MMP-3, and MMP-13 expression levels were detected by real-time quantitative polymerase chain reaction (PCR) and Western blotting. RESULTS: The elevated serum CORT and ACTH levels confirmed that the rats were under CSD stress. Hematoxylin and eosin (HE) staining and scanning electron microscopy (SEM) showed pathological alterations in the TMJ following CSD; furthermore, the p-ERK was activated and the mRNA and protein expression levels of MMP-1, MMP-3, and MMP-13 were upregulated after CSD. In the rats administered with the selective ERK inhibitor U0126, decreased tissue destruction was observed. Phospho-ERK activation was visibly blocked and the MMP-1, MMP-3, and MMP-13 mRNA and protein levels were lower than the corresponding levels in the CSD without U0126 group. CONCLUSION: These findings indicate that CSD activates the ERK pathway and upregulates the MMP-1, MMP-3, and MMP-13 mRNA and protein levels in the TMJ of rats. Thus, CSD induces ERK pathway activation and causes pathological alterations in the TMJ. ERK may be associated with TMJ destruction by promoting the expression of MMPs.

[Effect of dentin proteoglycans on the bonding properties of dentin].

OBJECTIVE: To investigate the potential effect of proteoglycan (PG) and glycosaminoglycan (GAG) on the bonding of etch and rinse adhesive to dentin, in order to improve the bonding effect of dentin. METHODS: Forty-two extracted molars were used to obtain standard dentin bonding surface, and the specimens were etched for 15 s with 37% phosphoric acid and divided into three groups using a table of random numbers. Then the three groups undergone different incubating procedures as follow: specimens in chondroitinase ABC (C-ABC) group were incubated with C-ABC at 37 °C for 48 h in vibrator. Specimens in trypsin (TRY) group were incubated with trypsin, and specimens in the control group were incubated with deionized water for 48 h in the oscillators. Then specimens in each group were randomly assigned into two subgroups, A (Adper(TM) Single Bond 2) and B (Prime & Bond NT) (n = 7). The microtensile bond strength (µTBS), fracture mode and bonding interface morphology of the specimens were evaluated via microtensile testing, stereo microscope and field emission scanning electron microscopy (FE-SEM) respectively after specimens being incubated in 37 °C water for 24 h. RESULTS: The immediate µTBS of C-ABC group bonding with adhesive A and B [(32.9±2.5) and (26.8±2.2) MPa] were significantly lower than that of the control group [(40.7±3.3) and (34.6±3.7) MPa] (P < 0.05). While the immediate µTBS of TRY group [(49.0 ± 3.6) and (44.5 ± 3.0) MPa] were significantly higher than that of the control group(P < 0.05). CONCLUSIONS: Dentin PG participates in the dentin bonding process. Removal of PG increased the immediate µTBS of dentin and total etching adhesives, while removal of GAG decreased the immediate µTBS.

[DNA extraction and sex determination for human teeth dated 3000 years ago unearthed in Xi'an].

OBJECTIVE: To extracted DNA from ancient human teeth dated 3000 years ago unearthed in Xi'an and determine the genders for the individuals. METHODS: Thirty five ancient human teeth were studied. A 'Reverse-root-canal' technique and a Chelex-100 solution were used to extract the DNA. Specific primers for Amelogenin gene were designed for PCR amplification. RESULTS: Genomic DNA was successfully extracted from 30 samples, for which 8 were determined to be males and 22 were females. CONCLUSION: The 'Reverse-root-canal' technique may be used for extracting DNA from ancient human teeth. Genetics method can supplement physical anthropology for determination of sex for ancient samples.

Influence of sleep deprivation on expression of MKK4 and c-fos in the mandibular condylar cartilage of rats.

The aim of this study was to investigate the changes in expression of mitogen-activated protein kinase kinase 4 (MKK4) and c-fos in the mandibular condylar cartilage of rats that had been subjected to sleep deprivation. One hundred and twenty female Wistar rats were randomly divided into 6 groups with 20 in each: sleep deprivation for 2 days, 4 days, 6 days, and 8 days, large-platform controls, and cage controls. After sleep deprivation by the modified multiple platform method the sleep-deprived rats were killed. The large-platform and cage control rats were killed at the same time as the rats deprived of sleep for 8 days. Haematoxylin and eosin were used to record the morphological changes in cartilage, and immunohistochemistry and real-time quantitative polymerase chain reaction (PCR) were used to detect the expression of MKK4 and c-fos. Pathological alterations were apparent after 6 and 8 days of sleep deprivation. Compared with control groups, the expression of MKK4 in the sleep-deprived groups was lower, while that of c-fos was higher. As the duration of sleep deprivation increased, the expression of MKK4 decreased. These results indicate that the variation in expression of MKK4 and c-fos may be correlated with pathological changes induced by sleep deprivation in mandibular condylar cartilage in rats.

The influence of psychological stress on the rat temporomandibular joint with the application of countermeasures.

OBJECTIVE: This study aimed to provide an experimental theoretical basis for the treatment of temporomandibular disorders by observing the effects of psychological stress and countermeasures on the rat temporomandibular joint (TMJ). METHODS: Rats were exposed to psychological stress via a communication box and the lateral pterygoid muscle and TMJ were observed with transmission electron microscopy and scanning electron microscopy. Furthermore, the expression of interleukin-1 and tumor necrosis factor-α was assessed in control animals and psychological stress (PS) and stress with diazepam (PS+DI) groups. RESULTS: Transmission electron microscopy of the lateral pterygoid muscle fibers in the PS showed vacuolar changes in the mitochondria, loss of cristae, and reduced matrix density to variable degrees after 1, 3, and 5 wk of stress. After 5 wk stress+recovery, the cristae and matrix were normal in the PS and PS+DI groups. Scanning electron microscopy of PS rats showed some synovial membranes were detached from the surface of the articular disc after 1 wk. After 3 wk, collagen fibers appeared to have wider waves and worn strips changing in size on the articular disc; after 5 wk, the distribution of collagen fibers was distorted. In PS+recovery and PS+DI rats, no obvious changes were observed on the surface of the articular disc after 1 to 5 wk stress. In PS rats, interleukin-1 and tumor necrosis factor-α expression increased significantly but was at control levels in the PS+DI and PS+recovery groups. CONCLUSION: Counteracting psychological stress can antagonize its effects on the TMJ and provide a reference for the treatment of stress-related temporomandibular disorders.

In vivo alveolar bone regeneration by bone marrow stem cells/fibrin glue composition.

The repair of alveolar bone defects caused by trauma, periodontal diseases and inflammation is still a challenge for both researchers and clinicians. Although there are many attempts to regenerate bone based on different seed cells and scaffolds, the results are still unsatisfactory. This study aims to clarify whether it could be efficient to reconstruct the alveolar bone by the combination of bone marrow stem cells (BMSCs) without pre-osteoinduction in vitro with fibrin glue (FG). The BMSCs were obtained from 2-week-old Sprague-Dawley (SD) rats and expanded in vitro with non-introduction. Afterwards, they were composited with FG for in vivo implantation. The animal models of traumatic alveolar bone defects were established bilaterally in the maxilla of 15 rats which were randomly divided into 3 groups. The BMSCs/FG composition was transplanted into 5 rats of the treated group. Another 5 rats in the negative control group were transplanted by pure FG without BMSCs. The rest 5 rats served as the blank control. Gross observation and histological analysis were made to evaluate the new bone formation 6 weeks after transplantation. Micro-CT was also used to estimate the bone healing through three-dimensional reconstruction and the bone density analysis. The amount of new bone formed in the treated group was significantly greater than the negative and blank control. Our results suggest that the strategy of combing BMSCs with FG is effective in the repair of alveolar bone defects. Its clinical application is promising.