Anti-angiogenic drug aggravates the degree of anti-resorptive drug-based medication-related osteonecrosis of the jaw by impairing the proliferation and migration function of gingival fibroblasts.

BACKGROUND: Long-term use of anti-resorptive or anti-angiogenic drugs in cancer patients with odontogenic infections may lead to medication-related osteonecrosis of the jaw (MRONJ). This study investigated whether anti-angiogenic agents aggravate MRONJ occurrence in anti-resorptive-treated patients. METHODS: The clinical stage and jawbone exposure of MRONJ patients caused by different drug regimens were analyzed to ascertain the aggravation effect of anti-angiogenic drugs on anti-resorptive drug-based MRONJ. Next, a periodontitis mice model was established, and tooth extraction was performed after administering anti-resorptive and/or anti-angiogenic drugs; the imaging and histological change of the extraction socket were observed. Moreover, the cell function of gingival fibroblasts was analyzed after the treatment with anti-resorptive and/or anti-angiogenic drugs in order to evaluate their effect on the gingival tissue healing of the extraction socket. RESULTS: Patients treated with anti-angiogenic and anti-resorptive drugs had an advanced clinical stage and a bigger proportion of necrotic jawbone exposure compared to patients treated with anti-resorptive drugs alone. In vivo study further indicated a greater loss of mucosa tissue coverage above the tooth extraction in mice treated with sunitinib (Suti) + zoledronate (Zole) group (7/10) vs. Zole group (3/10) and Suti group (1/10). Micro-computed tomography (CT) and histological data showed that the new bone formation in the extraction socket was lower in Suti + Zole and Zole groups vs. Suti and control groups. In vitro data showed that the anti-angiogenic drugs had a stronger inhibitory ability on the proliferation and migration function of gingival fibroblasts than anti-resorptive drugs, and the inhibitory effect was obviously enhanced after combining zoledronate and sunitinib. CONCLUSION: Our findings provided support for a synergistic contribution of anti-angiogenic drugs to anti-resorptive drugs-based MRONJ. Importantly, the present study revealed that anti-angiogenic drugs alone do not induce severe MRONJ but aggravate the degree of MRONJ via the enhanced inhibitory function of gingival fibroblasts based on anti-resorptive drugs.

Bone morphogenetic protein receptor 1α promotes osteolytic lesion of oral squamous cell carcinoma by SHH-dependent osteoclastogenesis.

Oral squamous cell carcinoma (OSCC) is an aggressive tumor that usually invades the maxilla or mandible. The extent and pattern of mandibular bone invasion caused by OSCC are the most important factors determining the treatment plan and patients' prognosis. Yet, the process of mandibular invasion is not fully understood. The following study explores the molecular mechanism that regulates the mandibular invasion of OSCC by focusing on bone morphogenetic protein receptor 1α (BMPR1α) and Sonic hedgehog (SHH) signals. We found that BMPR1α was positively correlated to bone defect of OSCC patients. Mechanistically, BMPR1α signaling regulated the differentiation and resorption activity of osteoclasts through the interaction of OSCC cells and osteoclast progenitors, and this process was mediated by SHH secreted by tumor cells. The inhibition of SHH protected bone from tumor-induced osteolytic activity. These results provide a potential new treatment strategy for controlling OSCC from invading the jawbones.

Fibronectin Facilitates Enterovirus 71 Infection by Mediating Viral Entry.

Fibronectin (FN) is a high-molecular-weight extracellular matrix protein that contains the RGDS motif, which is required to bind to integrins. Synthetic RGDS peptides have been reported to compete with FN to bind to the cell surface and inhibit the function of FN. Here, we identified that synthetic RGDS peptides significantly inhibit human enterovirus 71 (EV71) infection in cell cultures. In addition, mice treated with RGDS peptides and infected with EV71 had a significantly higher survival rate and a lower viral load than the control group. Because RGDS peptides affect the function of FN, we questioned whether FN may play a role in virus infection. Our study indicates that overexpression of FN enhanced EV71 infection. In contrast, knockout of FN significantly reduced viral yield and decreased the viral binding to host cells. Furthermore, EV71 entry, rather than intracellular viral replication, was blocked by FN inhibitor pretreatment. Next, we found that FN could interact with the EV71 capsid protein VP1, and further truncated-mutation assays indicated that the D2 domain of FN could interact with the N-terminal fragment of VP1. Taken together, our results demonstrate that the host factor FN binds to EV71 particles and facilitates EV71 entry, providing a potential therapy target for EV71 infection.IMPORTANCE Hand, foot, and mouth disease outbreaks have occurred frequently in recent years, sometimes causing severe neurological complications and even death in infants and young children worldwide. Unfortunately, no effective antiviral drugs are available for human enterovirus 71 (EV71), one of the viruses that cause hand, foot, and mouth disease. The infection process and the host factors involved remain unknown, although several receptors have been identified. In this study, we found that the host factor fibronectin (FN) facilitated EV71 replication by interacting with EV71 particles and further mediated their entry. The RGDS peptide, an FN inhibitor, significantly inhibited EV71 replication in both RD cells and mice. In conclusion, our research identified a new host factor involved in EV71 infection, providing a new potential antiviral target for EV71 treatment.

Polycarbonates with Potent and Selective Antimicrobial Activity toward Gram-Positive Bacteria.

The resistance developed by life-threatening bacteria toward conventional antibiotics has become a major concern in public health. To combat antibiotic resistance, there has been a significant interest in the development of antimicrobial cationic polymers due to the ease of synthesis and low manufacturing cost compared to host-defense peptides (HDPs). Herein, we report the design and synthesis of amphiphilic polycarbonates containing primary amino groups. These polymers exhibit potent antimicrobial activity and excellent selectivity to Gram-positive bacteria, including multidrug resistant pathogens. Fluorescence and TEM studies suggest that these polymers are likely to kill bacteria by disrupting bacterial membranes. These polymers also show low tendency to elicit resistance in bacteria. Their further development may lead to new antimicrobial agents combating drug-resistance.

Efficacy of submental island flap closing advanced mandibular MRONJ lesion in malignancy patients.

OBJECTIVE: This study evaluated the effectiveness of a submental island flap in closing advanced mandibular medication-related osteonecrosis of the jaw (MRONJ) wounds in patients with malignant tumors. SUBJECTS AND METHODS: A total of 85 patients with stage II and III MRONJ of mandible with malignant tumor as their primary disease were retrospectively analyzed. All patients underwent surgical treatment, and the soft tissue wound closure was performed either with a submental island flap (SIF) or mucoperiosteal flap (MF). Univariate and multifactorial models were applied to analyze the factors influencing patients' prognosis. RESULTS: Univariate analysis (p = 0.004, OR 0.075-0.575, 95% CI) and binary logistic regression (p = 0.017, OR 0.032-0.713, 95% CI) suggested that the surgical prognosis of SIF wound closure was significantly better than that of MF. CONCLUSION: Closure of wound after resection of mandibular MRONJ lesions in patients with malignant tumors using SIF had a better clinical prognosis compared with MF.

Pyrosequencing analysis of bacterial community changes in dental unit waterlines after chlorogenic acid treatment.

Introduction: The contamination of dental unit waterlines (DUWLs) poses a significant risk of cross-infection in dentistry. Although chemical disinfectants have been effective in reducing number of bacteria, they do have limitations. Methods: This study aimed to investigate the potential of chlorogenic acid, a natural substance with broadspectrum antibacterial properties, for treating DUWLs. Over a period of three months, we analyzed the microbial communities in 149 DUWLs samples collected from 5 dental units using high-throughput pyrophosphate sequencing. Results: The results revealed that chlorogenic acid treatment had a significant impact on the microbial community profile in the DUWLs, with the most significant changes occurring within the first 15 days and stabilization observed in the last 30 days. The predominant genera detected in the samples were Bacteroides, Lactobacillus, Streptococcus, Methylobacterium, and Phreatobacter. Additionally, the relative abundance of certain beneficial bacteria, such as Alloprevotella, Roseburia, and Blautia, increased, while the presence of opportunistic pathogens like Mycobacteria significantly decreased. The functional prediction analysis using the KEGG database indicated a decrease in the pathogenicity of the bacterial community in the DUWLs following chlorogenic acid treatment. Discussion: This study introduces a novel approach for the prevention and treatment of infections associated with dental care.

New insight into the effect of microplastics on antibiotic resistance and bacterial community of biofilm.

Microplastics (MPs) could serve as substrates for microbial colonization and biofilm formation. However, research on the effects of different types of microplastics and natural substrates on biofilm formation and community structure in the presence of antibiotic-resistant bacteria (ARB) is limited. In this study, we employed by means of microcosm experiments to analyze the situation of biofilms conditions, bacterial resistance patterns, antibiotic resistance genes (ARGs) distribution, and bacterial community on different substrates using microbial cultivation, high throughtput sequencing and PCR. The result showed that biofilms on different substrates markedly increased with time, with MPs surfaces formed more biofilm than stone. Analyses of antibiotic resistant showed negligible differences in the resistance rate to the same antibiotic at 30 d, but tetB would be selectively enriched on PP and PET. The microbial communities associated with biofilms on MPs and stones exhibited variations during different stages of formation. Notably, phylum WPS-2 and Epsilonbacteraeota were identified as the dominant microbiomes of biofilms on MPs and stones at 30 d, respectively. Correlation analysis suggested that WPS-2 could potentially be a tetracycline-resistant bacterium, while Epsilonbacteraeota did not correlate with any detected ARB. Our results emphasized the potential threat posed by MPs as attachment carriers for bacteria, particularly ARB, in aquatic environments.

The Therapeutic Effectiveness Using Fluorescence-Guided Surgery for MRONJ.

Background: Long-term application of antiresorptive and/or antiangiogenic agents may cause oral disorders, including medication-related osteonecrosis of the jaw (MRONJ), which remains an incurable disease. Surgical treatment can help alleviate infection of the jaw and block the progress of the disease, but postoperative recurrence is often caused by incomplete resection of necrotic bone during surgery. The traditional method for determining the boundary of necrotic bone resection is primarily based on the color, geology, and microcirculation-based bleeding state according to the bone tissue, which is easily affected by the surgeon's clinical experience and can cause insufficient resection of osteonecrosis bone. Recent studies have proposed using fluorescence technology-assisted necrotic bone resection. Objective: Systematic literature review was conducted to evaluate the therapeutic effectiveness of fluorescence-guided MRONJ surgery. Design: PubMed/MedLine, Scopus, and Web of Science databases were searched from inception to February 7, 2022. Randomized controlled trial (RCT) studies were evaluated according to the Cochrane risk of bias tool ROB 2, and non-RCT (N-RCT) studies were evaluated according to the ROBINS-I tool. Results: A total of 6 articles were included in the systematic review, including 4 N-RCT studies (1 retrospective study and 3 prospective studies) and 2 RCT studies, with 240 patients and 280 MRONJ lesions. The vast majority of studies were with moderate risk of bias, and the quality of the evidence was moderate. Conclusion: Evidence of moderate strength suggests that fluorescence-assisted techniques effectively determine the bone resection boundaries in MRONJ surgery. However, whether the prognosis of patients treated with fluorescence-guided surgery is significantly better than that of traditional surgery must be proved by randomized controlled studies with larger sample sizes and higher quality.

[Correlation analysis of supragingival microbiome and host blood lipid levels].

PURPOSE: To analyze the correlation between microbiome of supragingival plaque and host blood lipid levels. METHODS: Samples of supragingival plaques from 68 volunteers aged 45-60 years were collected. The total DNA was extracted, and 16S rDNA V3-V4 regions were amplified via PCR. The amplified products were sequenced by Illumina MiSeq PE300. After that, OTU clustering and species annotation were carried out. Then, the correlation of annotated species (genus level) and host blood lipid level were calculated with Spearman correlation analysis. SPSS 16.0 software package was used for statistical analysis. RESULTS: The average number of sequences obtained from the supragingival plaque samples was 41 929, and the number of OTUs obtained by clustering was 1 037. A total of 25 phyla, 45 classes, 92 orders, 155 families and 330 genera were annotated. Species heat maps showed a high degree of consistency in the species composition abundance of 68 samples. Spearman correlation analysis showed that among the supragingival microorganisms, Alloprevotella spp., Dialister spp., Peptostreptococcus spp. and Prevotella 7 spp., were negatively correlated with the host serum total cholesterol/low density lipoprotein cholesterol level. Neisseria spp. was positively correlated with host serum high-density lipoprotein cholesterol, but negatively correlated with Prevotella 2 spp.. CONCLUSIONS: The compositional structure of the supragingival microbiome is relatively stable and has a correlation with the host blood lipid levels. Profiles of supragingival microbiome can be promising biomarkers of the lipid metabolism of the host.

Particle-Attachment-Mediated and Matrix/Lattice-Guided Enamel Apatite Crystal Growth.

Tooth enamel is a hard yet resilient biomaterial that derives its unique mechanical properties from decussating bundles of apatite crystals. To understand enamel crystal nucleation and growth at a nanoscale level and to minimize preparation artifacts, the developing mouse enamel matrix was imaged in situ using graphene liquid cells and atomic resolution scanning transmission electron and cryo-fracture electron microscopy. We report that 1-2 nm diameter mineral precipitates aggregated to form larger 5 nm particle assemblies within ameloblast secretory vesicles or annular organic matrix subunits. Further evidence for the fusion of 1-2 nm mineral precipitates into 5 nm mineral aggregates via particle attachment was provided by matrix-mediated calcium phosphate crystal growth studies. As a next step, aggregated particles organized into rows of 3-10 subunits and developed lattice suprastructures with 0.34 nm gridline spacings corresponding to the (002) planes of apatite crystals. Mineral lattice suprastructures superseded closely matched organic matrix patterns, suggestive of a combination of organic/inorganic templates guiding apatite crystal growth. Upon assembly of 2-5 nm subunits into crystal ribbons, lattice fringes indicative of the presence of larger ordered crystallites were observed surrounding elongating crystal ribbons, presumably guiding the c-axis growth of composite apatite crystals. Cryo-fracture micrographs revealed reticular networks of an organic matrix on the surface of elongating enamel crystal ribbons, suggesting that protein coats facilitate c-axis apatite crystal growth. Together, these data demonstrate (i) the involvement of particle attachment in enamel crystal nucleation, (ii) a combination of matrix- and lattice-guided crystal growth, and (iii) fusion of individual crystals via a mechanism similar to Ostwald ripening.

Nonstructural protein 2A modulates replication and virulence of enterovirus 71.

Enterovirus 71 (EV71) can cause hand, foot, and mouth disease in children, and severe infections can induce neurological complications and even death. However, the pathogenesis of EV71 remains unknown. The 2A proteinase (2Apro) of EV71 plays an important role in segmenting the precursor polyprotein during viral replication, inhibiting host protein synthesis, and evading innate immunity. This study was to determine the function of EV71 2Apro in replication and virulence. A chimeric strain (SDLY 107-2A-1) was recombined by replacing 2Apro of a severe strain (SDLY107) with that of a mild strain (SDLY1) based on an infectious cDNA clone. The replication kinetics of the chimeric strain in vitro and in vivo were determined by qRT-PCR, which showed that the chimeric strain replicated slower and generated less viral RNA than the severe strain. The pathological change and viral load of chimeric strain infected mice were intermediate between severe strain infected mice and mild strain infected mice. Cellular cytotoxicity assays revealed that 2Apro was associated with the neurotoxicity of EV71. Histopathological and immunohistochemical assays detected tissue pathological damage in the lungs, muscles, brain, and intestinal tissues. Together, these results suggest that 2Apro modulates replication and virulence of EV71. This provides a theoretical basis for virulence determination of EV71.

pH-dependent surface electrostatic effects in retention on immobilized artificial membrane chromatography: Determination of the intrinsic phospholipid-water sorption coefficients of diverse analytes.

The passive membrane permeability of drugs may be potentially predicted by phospholipid-water sorption coefficient (KPLIPW) through immobilized artificial membrane (IAM) chromatography. However, for predominantly ionized compounds, unexpected confounding electrostatic interaction would influence the accurate determination of KPLIPW, accompanied by the mobile phase with varied pH and ionic strength. In order to measure the intrinsic phospholipid-water sorption coefficient (KIAM,intr) for diverse analytes, a mathematic model by characterizing the confounding electrostatic effects was developed in this study. Based on the developed electrostatic model, additional electrostatic repulsion/attraction interactions with the charged IAM surface were characterized for organic cations/anions. Considering that most of the IAM chromatography data was determined under physiological conditions, the KPLIPW values of all compounds were calculated with the ionic strength of 0.1 M and the pH value of 7. Generally, KPLIPW was hardly affected by the pH and ionic strength of mobile phase for those electroneutral compounds. KIAM,intr was 0.28 log units smaller than apparent phospholipid-water sorption coefficient (KIAM,app) for cations. In contrast, KIAM,intr was larger than KIAM,app by 0.28 log units for anions. Herein, KIAM,intr of diverse analytes can be easily calculated by the developed electrostatic model. For application, KIAM,intr was further compared with intrinsic Caco-2 permeability, and good sigmoidal correlations (R2 = 0.76) were established between them for organic cations and neutral compounds but not for organic anions.

Heavy-metal contents in suspended solids of Meiliang Bay, Taihu Lake and its environmental significances.

Surface water was taken from river mouth to the central area of Meiliang Bay, Taihu Lake, a large shallow eutrophic lake in China. Suspended solids were condensed by centrifugation 25 L surface water samples from each selected site. Suspended solids and surface sediments were further freeze-dried and microwave digested before determining the metals by ICP-AES. Among the metals analyzed in suspended solids and sediments, contents of Cr, Cu, Mn, Ni, and Zn in suspended solids were significantly higher than those in sediments while contents of Al, Ba, Be, Ca, Co, Fe, K, Mg, Pb, and V in suspended solids were 10%-30% higher than those in sediments. Sr and Ti contents in suspended solids and sediments were very similar. Na content in suspended solids was lower than that in sediments. Heavy metals were significantly accumulated in suspended solids. From the river mouth to the center of Meiliang Bay, contents of Cr, Cu, Pb, and Zn in suspended solids showed a gradual decreasing trend indicating the river (Zhihugang River) still discharged large quantity of heavy metals to Meiliang Bay. The study suggests that the geochemical behaviors and ecological effects of heavy metals in suspended solids may serve as a good indicator for the pollution of lake.

High-resolution electron microscopy and spectroscopy of ferritin in biocompatible graphene liquid cells and graphene sandwiches.

Atomic and electronic structures of hydrated ferritin are characterized using electron microscopy and spectroscopy through encapsulation in single layer graphene in a biocompatible manner. Graphene's ability to reduce radiation damage levels to hydrogen bond breakage is demonstrated. A reduction of iron valence from 3+ to 2+ is measured at nanometer-resolution in ferritin, showing initial stages of iron release by ferritin.

[Turbidity removal of summer high algae content Taihu Lake raw water using composite coagulants of polyaluminum chloride and polydimethyldiallylammonium chloride].

A series of stable composite coagulants was prepared using polyaluminum chloride (PAC) and polydimethyldiallylammonium chloride (PDM) with different intrinsic viscosity as raw materials. The enhanced coagulation processes of summer high algae content Taihu lake raw water by using the composite coagulants were reported in this paper. Jar tests were used to study the influences of the composite ratios of PAC to PDM and the intrinsic viscosity of PDM on the turbidity removal and precipitating performance. The results showed that for the summer Taihu raw water with temperature of 28-30 degrees C and turbidity of 30-33 NTU, algae content of 2.6 x 10(7) cells/L. When 2NTU turbidity of supernatant from settlement tank was required according to the current process of potable water plant in Wuxi, the dosage of 7.00 mg/L PAC was necessary, the PAC/PDM composite coagulants with the ratios (m/m) of 20:1, 10:1, 5:1 only needed dosage ranges of 3.00-2.83 mg/L, 3.50-3.49 mg/L, 5.37-4.67 mg/L with the increasing of PDM intrinsic viscosity from 0.52 dL/g to 2.46 dL/g. And compared with PAC only used, composite coagulants could decrease the PAC dose by 57.14%-59.57%, 50%-50.14%, 23.29%-33.29% correspondingly. When residual turbidity of 1 NTU was required, the dosage of 10 mg/L were necessary for the treatment with PAC , composite coagulants with the ratios(m/m) of PAC to PDM of 20:1, 10:1, 5:1 and the different intrinsic viscocity of PDM needed dosage of 8.33-3.91 mg/L, and could decrease the PAC dose by 16.7%-60.9%. Therefore, the efficiency turbidity removal and precipitating performance of composite coagulants were remarkably, and the lower the ratios of PAC to PDM and the higher the intrinsic viscosity of PDM, the higher efficiency of turbidity removal.