Magnetic Graphene Oxide Nanocomposites Boosts Craniomaxillofacial Bone Regeneration by Modulating circAars/miR-128-3p/SMAD5 Signaling Axis.

Background: The ability of nanomaterials to induce osteogenic differentiation is limited, which seriously imped the repair of craniomaxillofacial bone defect. Magnetic graphene oxide (MGO) nanocomposites with the excellent physicochemical properties have great potential in bone tissue engineering. In this study, we aim to explore the craniomaxillofacial bone defect repairment effect of MGO nanocomposites and its underlying mechanism. Methods: The biocompatibility of MGO nanocomposites was verified by CCK8, live/dead staining and cytoskeleton staining. The function of MGO nanocomposites induced osteogenic differentiation of BMSCs was investigated by ALP activity detection, mineralized nodules staining, detection of osteogenic genes and proteins, and immune-histochemical staining. BMSCs with or without MGO osteogenic differentiation induction were collected and subjected to high-throughput circular ribonucleic acids (circRNAs) sequencing, and then crucial circRNA circAars was screened and identified. Bioinformatics analysis, Dual-luciferase reporter assay, RNA binding protein immunoprecipitation (RIP), fluorescence in situ hybridization (FISH) and osteogenic-related examinations were used to further explore the ability of circAars to participate in MGO nanocomposites regulation of osteogenic differentiation of BMSCs and its potential mechanism. Furthermore, critical-sized calvarial defects were constructed and were performed to verify the osteogenic differentiation induction effects and its potential mechanism induced by MGO nanocomposites. Results: We verify the good biocompatibility and osteogenic differentiation improvement effects of BMSCs mediated by MGO nanocomposites. Furthermore, a new circRNA-circAars, we find and identify, is obviously upregulated in BMSCs mediated by MGO nanocomposites. Silencing circAars could significantly decrease the osteogenic ability of MGO nanocomposites. The underlying mechanism involved circAars sponging miR-128-3p to regulate the expression of SMAD5, which played an important role in the repair craniomaxillofacial bone defects mediated by MGO nanocomposites. Conclusion: We found that MGO nanocomposites regulated osteogenic differentiation of BMSCs via the circAars/miR-128-3p/SMAD5 pathway, which provided a feasible and effective strategy for the treatment of craniomaxillofacial bone defects.

Estrogenic disruption effects and formation mechanisms of transformation products during photolysis of preservative parabens.

The ubiquitous presence of emerging contaminants (ECs) in the environment and their associated adverse effects has raised concerns about their potential risks. The increased toxicity observed during the environmental transformation of ECs is often linked to the formation of their transformation products (TPs). However, comprehension of their formation mechanisms and contribution to the increased toxicity remains an unresolved challenge. To address this gap, by combining quantum chemical and molecular simulations with photochemical experiments in water, this study investigated the formation of TPs and their molecular interactions related to estrogenic effect using the photochemical degradation of benzylparaben (BZP) preservative as a representative example. A non-targeted analysis was carried out and three previously unknown TPs were identified during the transformation of BZP. Noteworthy, two of these novel TPs, namely oligomers BZP-o-phenol and BZP-m-phenol, exhibited higher estrogenic activities compared to the parent BZP. Their IC50 values of 0.26 and 0.50 µM, respectively, were found to be lower than that of the parent BZP (6.42 µM). The binding free energies (ΔGbind) of BZP-o-phenol and BZP-m-phenol (-29.71 to -23.28 kcal·mol-1) were lower than that of the parent BZP (-20.86 kcal·mol-1), confirming their stronger binding affinities toward the estrogen receptor (ER) α-ligand binding domain. Subsequent analysis unveiled that these hydrophobic residues contributed most favorably to ER binding, with van der Waals interactions playing a significant role. In-depth examination of the formation mechanisms indicated that these toxic TPs primarily originated from the successive cleavage of ester bonds (OCH2C6H5 and COO group), followed by their combination with BZP*. This study provides valuable insight into the mechanisms underlying the formation of toxic TPs and their binding interactions causing the endocrine-disrupting effects. It offers a crucial framework for elucidating the toxicological patterns of ECs with similar structures.

Effect of artificial landmarks of the prefabricated auxiliary devices located at different arch positions on the accuracy of complete-arch edentulous digital implant scanning: An in-vitro study.

OBJECTIVES: To examine the effect of artificial landmarks of prefabricated auxiliary devices (PAD) located at different arch positions on the accuracy of complete-arch edentulous digital implant scanning. METHODS: A reference model containing four analogs and PAD were fabricated by a 3D printer (AccuFab-C1s, 3DShining). 10 digital scans were performed using an intraoral scanner (Aoralscan 3, 3DShining), sv 1.0.0.3115, with artificial landmarks located at different arch positions: group I, without any artificial landmarks; group II, with artificial landmarks at the anterior region; group III, with artificial landmarks at the posterior region. group IV: with artificial landmarks at both anterior and posterior regions. For group V: Conventional open-tray splinted impressions. The reference file and conventional stone casts were digitalized by using a dental laboratory scanner. The related files were imported into inspection software for trueness and precision assessment. Statistical analysis was performed with One-way ANOVA and Kruskal-Wallis test. The level of significance was set at α=0.05. RESULTS: For the global accuracy assessment, significantly higher global trueness was seen in group II (p < 0.01), III (p < 0.001), IV (p < 0.001) and V (p < 0.001) than group I. Significantly higher global precision was seen in group III (p < 0.001), IV (p < 0.001) and V (p < 0.001) than group I. For the local accuracy assessment, the PAD primarily improved accuracy on the linear deviations. CONCLUSIONS: Artificial landmarks of PAD at different arch positions significantly influenced the scanning accuracy. Applying the PAD in group IV could achieve comparable outcomes to conventional open-tray splinted impressions. Artificial landmarks on the posterior region may be more pivotal than those on the anterior region. CLINICAL SIGNIFICANCE: Group IV could achieve comparable accuracy to conventional open-tray splinted impressions.

Effect of prefabricated auxiliary devices and scanning patterns on the accuracy of complete-arch implant digital impressions.

OBJECTIVES: This study aimed to evaluate the impact of prefabricated auxiliary devices (PAD) and scanning patterns on the accuracy of complete-arch implant digital impressions. METHODS: An edentulous maxillary model was inserted with four parallel implant analogs and four PAD. The model was scanned with D2000 dental laboratory scanner as the reference scans. Test scans were obtained by 8 different scanning patterns (SP), which including SPA, SPB, SPC, SPD, SPE, SPF, SPG and SPH, with (test group) or without (control group) using the PAD by an intraoral scanner (Aoralscan 3, 3DShining). SPA was the scanning pattern recommended by the manufacturer. Each scanning time was recorded. The related files were imported into inspection software for assessment. Aligned Ranks Transformation ANOVA, Kruskal-Wallis and Mann-Whitney tests were used to evaluate the values. The level of significance was set at α = 0.05. RESULTS: The scanning patterns significantly influenced the linear accuracy in the test group and the scanning time for both groups. Lower linear trueness in the test group was found in SPF (p<0.05) and SPG (p<0.05). Longer scanning time was found in SPB and SPG for both groups. The test group demonstrated linear accuracy enhancement in all the scanning patterns; angular trueness enhancement was seen in SPA (p<0.05), SPC (p<0.01) and SPH (p<0.01). Significant longer scanning time was found in SPB (p<0.05), SPF (p<0.05), SPG (p<0.05) and SPH (p<0.05) when using PAD. CONCLUSION: The scanning patterns impact the accuracy differently depending on the PAD's existence. The scanning time can be significantly influenced by the scanning patterns and the PAD. CLINICAL SIGNIFICANCE: In daily clinical practice, selecting a suitable scanning pattern is significant in achieving accurate digital impressions. The PAD demonstrated effective linear accuracy enhancement in all the scanning patterns tested.

Erratum: EPHB4 Regulates the Proliferation and Metastasis of Oral Squamous Cell Carcinoma through the HMGB1/NF-κB Signalling Pathway: Erratum.

[This corrects the article DOI: 10.7150/jca.59331.].

Effect of novel prefabricated auxiliary devices attaching to scan bodies on the accuracy of intraoral scanning of complete-arch with multiple implants: An in-vitro study.

OBJECTIVES: To examine the effect of novel prefabricated auxiliary devices with different geometric features called Scan Body Clasp (SBC) at different levels on the accuracy of intraoral scanning of complete-arch with multiple implants. METHODS: An edentulous maxilla 4-implant model and SBCs with different geometric features (flat or curved) were fabricated by a 3D printer (AccuFab-C1s, 3DShining, Hangzhou, China). Test scans were performed using an intraoral scanner (Aoralscan 3, 3DShining, Hangzhou, China) software version 1.0.0.3104 under different scenarios: group A (CO), without any SBCs; group B&C (LC&HC), with curved SBCs adjacent to and away from the mucosa; group D&E (LF&HF), with flat SBCs adjacent to and away from the mucosa. 20 scans were done for each group (CO, LC, HC, LF and HF). Reference Scans were obtained by digitizing the model in group A using a dental laboratory scanner (D2000, 3Shape, Copenhagen, Denmark). The related files were imported into inspection software for trueness and precision assessment. Statistical analysis was performed with One-way ANOVA, Independent-Sample T test for trueness values. Kruskal-Wallis test and Mann-Whitney test were used to assess the precision values. The level of significance was set at α=0.05. RESULTS: Groups with SBCs demonstrated trueness enhancement, among which LF revealed the best trueness. Significant differences were also found between LF and HC (p < .01), LF and HF (p < .001), LC and HF (p < .01). LF and HF showed precision enhancement. The best precision was LF, which was found to be more precise than LC (p < .001) and HC (p < .001). HF was more precise than LC (p < .001) and HC (p < .001). CONCLUSIONS: Attaching the scan bodies with SBCs at different levels significantly influenced the scanning accuracy. The SBCs near the mucosa result in superior trueness, while the flat morphology benefits the precision. CLINICAL SIGNIFICANCE: The results demonstrated the feasibility of the SBCs in enhancing intraoral complete-arch implant scanning accuracy. Among the configurations tested in the present study, low-level and flat surfaces of the artificial landmarks may be the potential pivotal elements to optimizing long-span scanning accuracy.

Accuracy of single-implant digital impression with various scanbody exposure levels at anterior and posterior regions.

OBJECTIVES: This in vitro study aimed to evaluate the effect of the exposure heights of the scanbody on the accuracy of digital implant impressions at different positions. METHODS: Four maxillary master models with one analog at the anterior and posterior region were fabricated by a 3-dimensional (3D) printer. The analogs were submerged from the gingival margin to ensure four exposure heights of the scanbody: 10, 8, 6, and 4 mm. . The master models were then scanned with D2000 dental laboratory scanner as the reference models. An intraoral scanner obtained ten test models for each group. After aligning the scanbody library file, the related files were imported into inspection software for superimposition by a local fit algorithm based on the adjacent teeth. RESULTS: 3D trueness was significantly decreased at 6 and 4 mm scanbody exposure at the anterior region. In comparison, a significant decrease was only seen at 4 mm scanbody exposure at the posterior region. 3D precision was significantly decreased at 4 mm scanbody exposure at both anterior and posterior regions. CONCLUSION: The exposure height of the scanbody influenced the accuracy of the digital implant impression, according to the implant positions. Scanbody exposure of less than 6 mm at the anterior region and 4 mm scanbody exposure at the posterior region could lead to increased deviations, but still in the tolerance range. CLINICAL SIGNIFICANCE: The scanbody exposure height less than 6 mm at the anterior region and 4 mm scanbody exposure height at the posterior region could lead to significantly increased deviations. Though these deviations may be still in the clinically acceptable range, caution should be taken.

Ebselen restores peri-implantitis-induced osteogenic inhibition via suppressing BMSCs ferroptosis.

It is hard to reconstruct bone defects in peri-implantitis due to osteogenesis inhibited by excessive reactive oxygen species (ROS). Ferroptosis, a recently identified regulated cell death characterized by iron- and ROS- dependent lipid peroxidation, provides us with a new explanation. Our study aims to explore whether ferroptosis is involved in peri-implantitis-inhibited osteogenesis and confirm ebselen, an antioxidant with glutathione peroxidase (GPx)-like activity, could inhibit ferroptosis and promote osteogenesis in peri-implantitis. In this study, we used LPS to mimic the microenvironment of peri-implantitis. The osteogenic differentiation of bone-marrow-derived mesenchymal stem cells (BMSCs) was assessed by alkaline phosphatase (ALP), Alizarin Red S, and mRNA and protein expression of osteogenic-related markers. Ferroptosis index analysis included iron metabolism, ROS production, lipid peroxidation and mitochondrial morphological changes. Iron overload, reduced antioxidant capability, excessive ROS, lipid peroxidation and the characteristic mitochondrial morphological changes of ferroptosis were observed in LPS-treated BMSCs, and adding Ferrostatin-1 (Fer-1) restored the inhibitory effect of ferroptosis on osteogenic differentiation of BMSCs. Furthermore, ebselen ameliorated LPS-induced ferroptosis and osteogenic inhibition, which were reversed by erastin. Our results demonstrated that ferroptosis is involved in osteogenic inhibition in peri-implantitis and ebselen could attenuate osteogenic dysfunction of BMSCs via inhibiting ferroptosis.

Estrogenic Effect Mechanism and Influencing Factors for Transformation Product Dimer Formed in Preservative Parabens Photolysis.

The environmental transformation and health effects of endocrine disruptors (EDCs) need urgent attention, particularly the formation of transformation products with higher toxicity than parent EDCs. In this paper, an important transformation product dimer (short for ethyl 4-hydroxy-3-(2-((4-hydroxybenzoyl) oxy) ethyl) benzoate) with estrogenic activity was investigated and detected in the photolysis of preservative ethyl-paraben (EPB) dissolved in actual water. The environmental factors, such as the higher initial concentration of EPB, the stronger optical power and the lower pH could stimulate the formation of the dimer. Simultaneously, the interaction of multiple environmental factors was significant, especially the initial concentration and pH using the response surface methodology. Furthermore, the relationship between the environmental factors and the formation of the product dimer was further explained and the empirical model equation was built for predicting the amount of dimer in actual water. Quantum chemical and toxicological calculations showed the estrogenic effect mechanism of the product dimer and it was revealed further that the hydrogen bonds of the dimer and ERα proteins (ARG-394, Glu-353, His-524, GYY-521) were formed, with a lowest binding energy of -8.38 Kcal/mol during molecular docking. In addition, the health effect risk of the product dimer was higher than the parent compound in the blood, cardiovascular system, gastrointestinal system, kidney and liver. In short, the present study was of great significance for the transformation product in pollution control and health effects in the photolysis of EDCs.

The influences of PEG-functionalized graphdiyne on cell growth and osteogenic differentiation of bone marrow mesenchymal stem cells.

Guided bone regeneration (GBR) is a frequently used technique for patients with insufficient alveolar bone. The discovery of bone substitutes that can enhance osteogenesis is critical for GBR. Graphdiyne (GDY), a newly discovered carbon-based nanomaterial, has been recognized as the most stable allotrope of acetylene carbon and is anticipated to be able to promote osteogenesis. Whereas it still remains unknown whether it could enhance osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). In this study, GDY was modified with polyethylene glycol (PEG) and the influences of GDY-PEG at different concentrations on BMSCs cell growth and osteogenic differentiation were researched for the first time. In this study, we found that GDY-PEG at low concentration possessed premium bio-compatibility and revealed evident facilitation of BMSCs osteogenic differentiation. The cell growth and osteogenic differentiation of BMSCs treated with GDY-PEG were dose-dependent. GDY-PEG at 1 µg/mL demonstrated the optimal promoting effects of BMSCs osteogenic differentiation. Moreover, the regulating effect of BMSCs osteogenic differentiation by GDY-PEG might be associated with the Wnt/ß-catenin signaling pathway. In all, the present study indicated a novel application of GDY in promoting bone tissue regeneration, providing a novel biomaterial for bone augmentation in clinics.

Mixed bromine/chlorine transformation products of tetrabromobisphenol A formed in the combustion of printed circuit boards: Emission characteristics and transformation pathways.

Recently, mixed bromine/chlorine transformation products of tetrabromobisphenol A (ClyBrxBPAs) were found to be possibly related to the thermal treatment processes of electronic wastes. To explore their emission characteristics and formation mechanism, printed circuit board scraps were combusted in a tube furnace, under the temperature from 25 °C to 600 °C. The emission factor of the debromination products of tetrabromobisphenol A (BrxBPAs) was the highest, whereas that of ClyBrxBPAs was the lowest. Among three phases, most of the target compounds were partitioned into the oil and particle phases, and only negligible gaseous 2-BrBPA and bisphenol A were detected. The emission rates of most compounds were fastest at 300 °C, although 2-BrBPA, 2,6-Br2BPA, and 2-Cl-6-BrBPA peaked at 350 °C. Among the chemicals in total emission, 2-BrBPA was the dominant congener in BrxBPAs, whereas 2-Cl-2',6,6'-Br3BPA, 2-Cl-2',6#-Br2BPA, and Σ2Cl1Br1BPAs shared similar proportions in ClyBrxBPAs. Meanwhile, the composition profiles at 300 °C showed that 2,2',6-Br3BPA and 2-Cl-2',6,6'-Br3BPA occupied the largest proportions in BrxBPAs and ClyBrxBPAs, respectively. To reveal the possible transformation pathways, the Gibbs free energy was calculated based on a radical substitution reaction. After "•Br" removal from tetrabromobisphenol A or other BrxBPAs, the intermediate was more easily combined with "•H" than with "•Cl." In addition, the ClyBrxBPA formation via "-•H + •Cl" by BrxBPAs is nonspontaneous, thus limiting the further generation of ClyBrxBPAs. This study not only provides ideas for the study of other mixed halogenated products, but also provides constructive suggestions for environmental source analysis by combining previous research on the occurrence of ClyBrxBPAs in various environmental matrices.

Nanostructured Graphdiyne: Synthesis and Biomedical Applications.

Graphdiyne (GDY) is a 2D carbon allotrope that features a one-atom-thick network of sp- and sp2-hybridized carbon atoms with high degrees of π conjugation. Due to its distinct electronic, chemical, mechanical, and magnetic properties, GDY has attracted great attention and shown great potential in various fields, such as catalysis, energy storage, and the environment. Preparation of GDY with various nanostructures, including 0D quantum dots, 1D nanotubes/nanowires/nanoribbons, 2D nanosheets/nanowalls/ordered stripe arrays, and 3D nanospheres, greatly improves its function and has propelled its applications forward. High biocompatibility and stability make GDY a promising candidate for biomedical applications. This review introduces the latest developments in fabrication of GDY-based nanomaterials with various morphologies and summarizes their propective use in the biomedical domain, specifically focusing on their potential advantages and applications for biosensing, cancer diagnosis and therapy, radiation protection, and tissue engineering.

The Differential Metabolic Response of Oral Squamous Cell Carcinoma Cells and Normal Oral Epithelial Cells to Cisplatin Exposure.

Metabolic reprogramming is one of the hallmarks of a tumor. It not only promotes the development and progression of tumor but also contributes to the resistance of tumor cells to chemotherapeutics. The difference in the metabolism between drug-resistant and sensitive tumor cells indicates that drug-resistant tumor cells have experienced metabolic adaptation. The metabolic response induced by chemotherapy is dynamic, but the early metabolic response of tumor cells to anticancer drugs and the effect of an initial response on the development of drug resistance have not been well studied. Early metabolic intervention may prevent or slow down the development of drug resistance. The differential metabolic responses of normal cells and tumor cells to drugs are unclear. The specific metabolites or metabolic pathways of tumor cells to chemotherapeutic drugs can be used as the target of metabolic intervention in tumor therapy. In this study, we used comparative metabolomics to analyze the differential metabolic responses of oral cancer cells and normal oral epithelial cells to short-term cisplatin exposure, and to identify the marker metabolites of early response in oral cancer cells. Oral cancer cells showed a dynamic metabolic response to cisplatin. Seven and five metabolites were identified as specific response markers to cisplatin exposure in oral cancer cell SCC-9 and normal oral epithelial cell HOEC, respectively. Glyoxylate and dicarboxylate metabolism and fructose, malate, serine, alanine, sorbose and glutamate were considered as specific enriched metabolic pathways and biomarkers of SCC-9 cells in response to cisplatin, respectively. The existence of differential metabolic responses lays a foundation for tumor chemotherapy combined with metabolic intervention.

Selenomethionine protects oxidative-stress-damaged bone-marrow-derived mesenchymal stem cells via an antioxidant effect and the PTEN/PI3K/AKT pathway.

Dental implant surgery is currently a routine therapy for the repair of missing dentition or dentition defects. Both clinical and basic research have elucidated that oxidative stress caused by the accumulation of reactive oxygen species (ROS) for various reasons impairs the process of osteointegration after dental implantation. Therefore, the osteogenic micro-environment must be ameliorated to decrease the damage caused by oxidative stress. Selenomethionine (SEMET) has been reported to play an important role in alleviating oxidative stress and accelerating cell viability and growth. However, it remains unclear whether it exerts protective effects on bone-marrow-derived mesenchymal stem cells (BMSCs) under oxidative stress. In this study, we explored the influence of selenomethionine on the viability and osteogenic differentiation of BMSCs under oxidative stress and the underlying mechanisms. Results showed that 1 µM selenomethionine was the optimum concentration for BMSCs under H2O2 stimulation. H2O2-induced oxidative stress suppressed the viability and osteogenic differentiation of BMSCs, manifested by the increases in ROS production and cell apoptosis rates, and by the decrease of osteogenic differentiation-related markers. Notably, the aforementioned oxidative damage and osteogenic dysfunction induced by H2O2 were rescued by selenomethionine. Furthermore, we found that the PTEN expression level was suppressed and its downstream PI3K/AKT pathway was activated by selenomethionine. However, when PTEN was stimulated, the PI3K/AKT pathway was down-regulated, and the protective effects of selenomethionine on BMSC osteogenic differentiation diminished, while the inhibition of PTEN up-regulated the protective effects of selenomethionine. Together, these results revealed that selenomethionine could attenuate H2O2-induced BMSC dysfunction through an antioxidant effect, modulated via the PTEN/PI3K/AKT pathway, suggesting that selenomethionine is a promising antioxidant candidate for reducing oxidative stress during the process of dental implant osteointegration.

EPHB4 Regulates the Proliferation and Metastasis of Oral Squamous Cell Carcinoma through the HMGB1/NF-κB Signalling Pathway.

Background: Malignant proliferation and cervical lymphatic metastasis restrict the prognosis of oral squamous cell carcinoma (OSCC). Erythropoietin-producing human hepatocellular B4 (EPHB4) regulates a series of tumour functions involving tumourigenesis, cancer cell attachment and metastasis. However, the mechanism of EphB4 regulating the malignant progression of OSCC has not been fully elucidated. Methods: EPHB4 expression was analysed in 65 OSCC samples and adjacent noncancerous tissues through immunohistochemistry (IHC). siRNA and overexpression plasmids were transfected into OSCC cells to modify EPHB4 expression, and then, regulatory functions were explored in vitro and in vivo. Co-immunoprecipitation (Co-IP) and mass spectrometry were applied to detect proteins interacting with EPHB4. Subsequently, protein stability assays and NF-κB pathway inhibition assays were used to verify the regulation of EPHB4, high-mobility group box 1 (HMGB1) and nuclear factor-κB (NF-κB) activation. Results: EPHB4 was found to be highly expressed in OSCC tissues, which was related to tumour stage and lymphatic metastasis and resulted in a poor prognosis. Cellular experiments and mouse tongue xenograft models further confirmed that high EPHB4 expression promoted the proliferation and metastasis of OSCC tumours. Mechanistically, co-IP and mass spectrometry studies indicated that EPHB4 could bind to HMGB1 and maintain HMGB1 stability. Downregulation of HMGB1 inhibited the proliferation and metastasis of OSCC cells and inhibited NF-κB phosphorylation activation but did not affect EPHB4 expression. Conclusion: This study revealed the mechanism by which EPHB4 promotes the proliferation and metastasis of OSCC by activating the HMGB1-mediated NF-κB signalling pathway, which can be exploited as a novel marker or therapeutic target to control metastasis and improve the survival rate of OSCC.

Selenomethionine-Modified Polyethylenimine-Based Nanoparticles Loaded with miR-132-3p Inhibitor-Biofunctionalized Titanium Implants for Improved Osteointegration.

Titanium and its alloys have been widely used as bone implants, but for reduced treatment span, improvements are urgently needed to achieve faster and better osteointegration. In this study, we found that miR-132-3p inhibited bone-marrow-derived stem cell (BMSC) osteogenic differentiation via targeting BMP2, and that inhibiting miR-132-3p could significantly improve the osteogenic capability of BMSCs. Moreover, we fabricated a biocompatible selenomethionine (SEMET)-modified polyethylene glycol (PEG)/polyethylenimine (PEI) nanoparticle (SeNP) cross-linked with 0.2% gelatin solutions and delivered miR-132-3p inhibitor to biofunctionalize alkali heat-treated titanium implants, resulting in the development of a novel coating for reverse transfection. The biological performances of PEG/PEI/miR-132-3p inhibitor and SeNP/miR-132-3p inhibitor-biofunctionalized titanium were compared. The biological effects, including cell viability, cytotoxicity, adhesion, cellular uptake, and osteogenic capacity of SeNP/miR-132-3p inhibitor-biofunctionalized titanium implants, were then assessed. Results showed that SeNPs presented appropriate morphology, diameter, and positive zeta potential for efficient gene delivery. The transfection efficiency of the SeNP/miR-132-3p inhibitor was comparable to that of the PEG/PEI/miR-132-3p inhibitor, but the former induced less reactive oxygen species (ROS) production and lower apoptosis rates. Confocal laser scanning microscopy (CLSM) demonstrated that SeNP/miR-132-3p inhibitor nanoparticles released from the titanium surfaces and were taken up by adherent BMSCs. In addition, the release profile showed that transfection could obtain a long-lasting silencing effect for more than 2 weeks. The cell viability, cytotoxicity, and cell spreading of SeNP/miRNA-132-3p inhibitor-biofunctionalized titanium were comparable with those of untreated titanium and the SeNP/miRNA-132-3p inhibitor negative control (NC)-biofunctionalized titanium but resulted in higher ALP activity and osteogenic gene expression levels. In vivo animal studies further certified that SeNP/miRNA-132-3p inhibitor nanoparticles from titanium surfaces promoted osteointegration, which was revealed by microcomputed tomography (micro-CT) and histological observations. Taken together, these findings suggested that selenomethionine-modified PEI-based nanoparticles could achieve better biocompatibility. Moreover, titanium implants biofunctionalized by SeNP/miRNA-132-3p inhibitor nanoparticles might have significant clinical potential for more effective osteointegration.

Effect of magnetic graphene oxide on cellular behaviors and osteogenesis under a moderate static magnetic field.

The biological behaviors of magnetic graphene oxide (MGO) in a static magnetic field (SMF) are unknown. The current study is to investigate the cellular behaviors, osteogenesis and the mechanism in BMSCs treated with MGO combined with an SMF. Results showed that the synthetic MGO particles were bio-compatible and could significantly improve the osteogenesis of BMSCs under SMFs, as verified by elevated alkaline phosphatase activity, mineralized nodule formation, and expressions of mRNA and protein levels. Under SMF at the same intensity, the addition of graphene oxide to Fe3O4 could increase the osteogenic ability of BMSCs. The Wnt/ß-catenin pathway was indicated to be related to the MGO-driven osteogenic behavior of the BMSCs under SMF. Taken together, our findings suggested that MGO under an SMF could promote osteogenesis in BMSCs through the Wnt/ß-catenin pathway and hence should attract more attention for practical applications in bone tissue regeneration.

Metabolomics study reveals the potential evidence of metabolic reprogramming towards the Warburg effect in precancerous lesions.

Background: Most tumors have an enhanced glycolysis flux, even when oxygen is available, called the aerobic glycolysis or the Warburg effect. Metabolic reprogramming promotes cancer progression, and is even related to the tumorigenesis. However, it is not clear whether the observed metabolic changes act as a driver or a bystander in cancer development. Methods: In this study, the metabolic characteristics of oral precancerous cells and cervical precancerous lesions were analyzed by metabolomics, and the expression of glycolytic enzymes in cervical precancerous lesions was evaluated by RT-PCR and Western blot analysis. Results: In total, 115 and 23 metabolites with reliable signals were identified in oral cells and cervical tissues, respectively. Based on the metabolome, oral precancerous cell DOK could be clearly separated from normal human oral epithelial cells (HOEC) and oral cancer cells. Four critical differential metabolites (pyruvate, glutamine, methionine and lysine) were identified between DOK and HOEC. Metabolic profiles could clearly distinguish cervical precancerous lesions from normal cervical epithelium and cervical cancer. Compared with normal cervical epithelium, the glucose consumption and lactate production increased in cervical precancerous lesions. The expression of glycolytic enzymes LDHA, HK II and PKM2 showed an increased tendency in cervical precancerous lesions compared with normal cervical epithelium. Conclusions: Our findings suggest that cell metabolism may be reprogrammed at the early stage of tumorigenesis, implying the contribution of metabolic reprogramming to the development of tumor.

Host Immune Response Triggered by Graphene Quantum-Dot-Mediated Photodynamic Therapy for Oral Squamous Cell Carcinoma.

INTRODUCTION: With the innovation of photosensitizers, photodynamic therapy is now widely used in antitumor detection and treatment. Graphene quantum dots (GQDs) are proposed as a promising alternative photosensitizer due to their high biocompatibility, specific photoactivity, and strong tumor concentration. However, the changes in host immunity triggered by GQDs have only rarely been reported. METHODS: In this work, GQDs as photosensitizers were conjugated to polyethylene glycol (PEG) to enhance solubility and blood circulation. The phototoxicity of the resulting GQD-PEG nanomaterials was then detected in vitro and in vivo. The antitumor immunity triggered by GQD-PEG under irradiation was further evaluated in an oral squamous cell carcinoma animal model. RESULTS: The obtained GQD-PEG nanomaterials exhibited low cytotoxicity, good solution stability, and excellent endocytosis. Both in vitro and in vivo, all demonstrated strong ablation for oral squamous cell carcinoma under irradiation. Meanwhile, host-immunity-related CD8+ T cells (cytotoxic T lymphocytes) and proinflammatory cytokines, including IFN-γ and TNF-α, were significantly increased after photo-activated antitumor activity. CONCLUSION: These results highlight the dominant role of GQD-PEG in photodynamic therapy and could have significant implications for further combination therapy as a promising antitumor immune response strategy triggered by nanomaterials.

Upregulation of Circular RNA circATRNL1 to Sensitize Oral Squamous Cell Carcinoma to Irradiation.

Accumulating evidence has demonstrated that circular RNAs (circRNAs) play important roles in regulating gene expression involved in tumor development. However, the role of circRNAs in modulating the radiosensitivity of oral squamous cell carcinoma (OSCC) and its potential mechanisms have not been documented. We performed high-throughput RNA sequencing (RNA-seq) to investigate the circRNA expression profile in OSCC patients and discovered that the circATRNL1 expression was significantly downregulated and closely related to tumor progression. The circATRNL1 was structurally validated via Sanger sequencing, RNase R treatment, and specific convergent and divergent primer amplification. Importantly, the expression levels of circATRNL1 decreased after irradiation treatment, and upregulation of circATRNL1 enhanced the radiosensitivity of OSCC through suppressing proliferation and the colony survival fraction, inducing apoptosis and cell-cycle arrest. Moreover, we observed that circATRNL1 could directly bind to microRNA-23a-3p (miR-23a-3p) and relieve inhibition for the target gene PTEN. In addition, the tumor radiosensitivity-promoting effect of circATRNL1 overexpression was blocked by miR-23a-3p in OSCC. Further experiments also showed that PTEN can reverse the inhibitory effect of OSCC radiosensitivity triggered by miR-23a-3p. We concluded that circANTRL1 may function as the sponge of miR-23a-3p to promote PTEN expression and eventually contributes to OSCC radiosensitivity enhancement. This study indicates that circANTRL1 may be a novel therapeutic target to improve the efficiency of radiotherapy in OSCC.