The dynamics of bacterial proliferation, viability, and extracellular polymeric substances in oral biofilm development.

OBJECTIVES: This study investigated the relationship between bacterial growth, viability, and extracellular polymeric substances (EPS) formation in biofilms, particularly regarding resistance development. It also examined the impact of chemical factors on the EPS matrix and bacterial proliferation in oral biofilms. METHODS: Three multi-species oral biofilms were incubated in anaerobic conditions. Three strains of Enterococcus faecalis were incubated in aerobic conditions. The incubation periods ranged from 0 h to 7 days for short-term biofilms, and from 3 to 90 days for long-term biofilms. Fluorescent labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) and flow cytometry were used to track EPS and bacterial growth. Confocal laser scanning microscopy (CLSM) assessed bacterial viability and EPS structure. Biofilms aged 7, 14, and 21 days were treated with 2 % chlorhexidine (CHX) and 1 % sodium hypochlorite (NaOCl) to evaluate their effects on EPS and bacterial proliferation. RESULTS: Short-term biofilms showed rapid bacterial proliferation and a gradual increase in EPS, maintaining stable viability. In the first two weeks, a significant rise in CFSE indicated growing maturity. From 14 to 90 days, EPS and CFSE levels stabilized. Following treatment, CHX significantly reduced bacterial proliferation, while NaOCl decreased EPS volume. CONCLUSIONS: Biofilm development involves a balance between bacterial proliferation and EPS production. The complexity of this process poses challenges in treating biofilm-associated infections, requiring strategies tailored to the biofilm's developmental stage. CLINICAL SIGNIFICANCE: For effective root canal treatment, it is imperative to focus on reducing bacterial proliferation during the early stages of oral infections. In contrast, strategies aimed at minimizing EPS production could be more beneficial for long-term management of these conditions.

Effect of miniscrew-assisted rapid palatal expansion on mandibular position / 口腔医学

Objective@#To explore the effect of miniscrew-assisted rapid palatal expansion (MARPE) on mandible position in the treatment of adult skeletal Class Ⅰ malocclusion with maxillary transverse deficiency. @*Methods@#In this retrospective study, 20 cases of adult skeletal Class Ⅰ malocclusion with maxillary transverse deficiency treated with MARPE in our hospital from July 2019 to March 2022 were selected as research objects. CBCT data of three time points before treatment (T0), immediately after expansion (T1) and six months after retention (T2) were collected. The head position was standardized and calibrated by Dolphin software, and then mandible landmarks (left and right Condylion, left and right Gonion, Menton) were positioned. The linear distance changes of each landmark relative to the reference plane of coronal plane, axial plane and sagittal plane were measured, which represented the sagittal, vertical and horizontal displacement of mandible respectively. Repeated measurement ANOVA and LSD multiple comparison were used to evaluate the position change of each landmark.@*Results @#The Menton and right Gonion rotated clockwise at T1, and relapsed to the initial position at T2. No lateral displacement of Menton was found.@*Conclusion@#When MARPE is used to treat skeletal Class Ⅰ malocclusion with maxillary transverse deficiency, it causes a transient clockwise rotation of the mandiblar. The mandible does not show sagittal, vertical and horizontal position changes in long-term evaluation.

Adenosine A2A Receptor Antagonist Improves Cognitive Impairment by Inhibiting Neuroinflammation and Excitatory Neurotoxicity in Chronic Periodontitis Mice.

The adenosine A2A receptor antagonist SCH58261 has been reported to have anti-inflammatory effects. However, its role in chronic periodontitis (CP)-induced cognitive impairment, which is associated with Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS), remains unclear. This study investigated the role of SCH58261 in mice with CP-induced cognitive impairment. C57BL/6J mice were used to develop CP model by injecting 0.5 mg/kg P. gingivalis LPS into the palatal gingival sulcus of maxillary first molars twice a week for four weeks. The mice were divided into control, P. gingivalis LPS (P-LPS), P-LPS + SCH58261, and SCH58261 groups. The passive avoidance test (PAT) and Morris water maze (MWM) were used to assess cognition in mice. Furthermore, CD73/adenosine, neuroinflammation, glutamate transporters, and glutamate were assessed. Compared with the P-LPS group, 0.1 and 0.5 mg/kg SCH58261 increased latency and decreased error times in PAT, but increased platform crossing number in MWM. SCH58261 inhibited microglial activation, and decreased pro-inflammatory cytokines and glutamate levels, but increased GLT-1 and PSD95 expression in the hippocampus. This was the first report of SCH58261 treatment for CP-induced cognitive impairment, which may be related to its anti-inflammatory activities and anti-glutamate excitatory neurotoxicity. This suggests that SCH58261 can be used as a novel agent to treat cognitive impairment.

MicroRNA expression profiles in peri-miniscrew implant crevicular fluid in orthodontics: a pilot study.

BACKGROUND: This study systematically evaluated microRNA (miRNA) expression patterns in peri-miniscrew implant crevicular fluid (PMICF) in orthodontic patients. METHODS: Next-generation sequencing (NGS) was performed to obtain miRNA profiles in PMICF or gingival crevicular fluid (GCF) collected from 3 healthy volunteers (H), 3 peri-implantitis patients (PMSII) and 5 periodontitis patients (P). MiRNA expression patterns were compared between normal and orthodontic PMICF and GCF. Differentially expressed miRNAs were estimated by quantitative real-time PCR (qRT-PCR). Enrichment analyses of the gene targets controlled by these miRNAs were conducted by Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. RESULTS: Compared with healthy donors, in PMSII patients, a total of 206 upregulated miRNAs and 152 downregulated miRNAs were detected in PMICF, while periodontitis patients had 333 upregulated miRNAs and 318 downregulated miRNAs. MiR-544a, miR-1245b-3p, miR-1825, miR-4291, miR-3689e, and miR-4477a were chosen randomly for further examination. qRT-PCR examination confirmed that the expression levels of miR-1245b-3p and miR-4291 were higher in PMSII than in H samples and that the expression levels of miR-1825 were higher in PMSII than in P samples. However, contrary to the NGS results, qRT-PCR analysis showed decreased expression of miR544a in PMSII. MiR3689e and miR4477a expression did not differ significantly among all samples. According to GO and KEGG pathway analyses of miR-1825, miR-4291, and miR-1245b-3p high enrichment of target genes involved in the PI3K-AKT signalling pathway was observed. CONCLUSIONS: The NGS analysis of normal and orthodontic PMICF/CGF showed different miRNA profiles, which may lay the foundation for future research on the molecular mechanism of PMSII. miR-4291, miR-1245b-3p and miR-1825 may be used as diagnostic markers and potential therapeutic targets for PMSII.

Profiles of inflammation factors and inflammatory pathways around the peri-miniscrew implant.

BACKGROUND: Peri-miniscrew implant is a temporary assistant armamentarium for the treatment of severe malocclusion and complex tooth movement, the inflammation around it is the main reason for the failure of orthodontic treatment due to the implant loosening and falling out. Inflammation around the peri-miniscrew implant is associated with the release of pro-inflammatory cytokines. These pro-inflammatory cytokines, in turn, recruit immune cells (such as macrophages, dendritic cells, T cells, and B cells), which can produce and release inflammatory biomarkers, regulate the interaction between immune cells, periodontal ligament cells, osteoblasts, and so on. However, there is currently no effective clinical treatment plan to prevent inflammation around implants. PURPOSE: To investigate the potentially essential factors in the inflammatory response around the peri-miniscrew implant and explore the signaling pathways involved. METHODS: Here, we review the studies focused on inflammatory biomarkers (Interleukins, tumor necrosis factor-α (TNF-α), receptor activator of NF-κB ligand (RANKL), matrix metalloproteinases (MMPs), and cellular adhesion molecules (CAMs)) in peri-miniscrew implant crevicular fluid (PMICF), as well as inflammatory signaling pathways (Wnt5a, JNK, Erk1/2, NF-κBp65 and TAB/TAK) in periodontal cells from 1998 to 2020. RESULTS: A literature search revealed TLR-2, TLR-4, LOX-1, and BMPs are involved in regulating ILs (IL-1ß, IL-6, IL-8, and IL-17), TNF-α, RANKL, MMP-2, MMP-9 expression via JNK, Erk1/2, Wnt5a, NF-κBp65, OPN, and TAB/TAK signaling pathways. Among them, IL-1ß and IL-6 are the critical inflammation factors in the signaling pathways inducing the inflammatory reaction surrounding implants. Besides, CAM-1 was also regulated by MMP-9 and IL-17. CONCLUSION: There are considerable potential factors involving regulating inflammatory biomarkers on downstream signaling pathways in peri-minisrew implant crevicular fluid. CLINICAL SIGNIFICANCE: This review provides the substantiation of these cell factors and signaling pathways around peri-miniscrew implants, proposes more practical clinical therapeutic ideas and schemes for improving the stability and clinical efficacy of peri-miniscrew implants.

MicroRNA-125a-5p modulates macrophage polarization by targeting E26 transformation-specific variant 6 gene during orthodontic tooth movement.

OBJECTIVE: The aim of this study was to investigate the role of microRNA-125a-5p (miR-125a-5p) in macrophages during orthodontic tooth movement (OTM). DESIGN: Periodontal ligament tissues were collected from patients underwent OTM. Periodontal ligament cells were isolated from periodontal ligament tissues. Periodontal ligament stem cells were isolated from normal human impacted third molars. The miR-125-5p levels were measured by real-time quantitative polymerase chain reaction. The impact of miR-125-5p on macrophage polarization was tested by alizarin red staining assay. The effects of miR-125-5p and E26 transformation-specific variant 6 gene (ETV6) on M1/M2 macrophages phenotype markers were determined by real-time quantitative polymerase chain reaction, western blot, and flow cytometry analyses. The interaction between miR-125-5p and ETV6 was verified using luciferase reporter and RNA immunoprecipitation assays. RESULTS: Periodontal miR-125a-5p was upregulated under the force. Macrophage polarization facilitated osteogenesis by cocultured system. Moreover, miR-125a-5p was upregulated in macrophages polarized with M2 conditions. MiR-125a-5p downregulation promoted the expression of M1 phenotype markers, while suppressed the expression of M2 markers. Mechanistically, ETV6 was confirmed to be a target of miR-125a-5p. ETV6 overexpression increased the expression of M1 polarized markers, while decreased the expression of M2 polarized markers. Furthermore, ETV6 knockdown reversed the effects of miR-125a-5p inhibitor on both M1 macrophages and M2 macrophages. CONCLUSIONS: Overall, miR-125a-5p facilitates bone healing by targeting ETV6 to promote macrophage M2 polarization.

Study on the correlation parameters of Class II malocclusion in child tooth dentition early stage by using 3D technique.

OBJECTIVE: To record the dentition, jaw and facial growth and development of children with class II malocclusion at the age of 7-8 years old in the early dental transitional stage with 3D technology and to provide the study basis for the growth and development parameters of normal children and children with class II malocclusion. METHODS: Twenty-four children who were suffering class-II malocclusion in the early dental transitional stage and received treatment between July 2016 and July 2017 in our hospital were selected as the study group, and 20 healthy children were selected as the control group in the same period. SIRONA CEREC dentition scanning, 3D reconstruction of the lower mandible and 3d MD face scanning were performed on the children. Relevant data were recorded and compared. RESULTS: The dentition scanning results suggested that the study group had significantly larger anterior overbite and anterior overjet and smaller width of the upper arch than the control group; there was a significant difference between the two groups (P<0.05). The 3D reconstruction of the lower mandible suggested that the study group had smaller Go angle and SNB angle and shorter ANS-Me distance, Go-Me distance and N-Me distance compared to the control group; the differences had statistical significance (P<0.05). The face scanning results demonstrated that the nasolabial angle and facial convexity angle of the study group were significantly larger than those in the control group, and the difference was statistically significant (P<0.05). CONCLUSION: The dentition scanning results suggested that the study group had significantly larger anterior overbite and anterior overjet and smaller width of the upper arch than the control group; there was a significant difference between the two groups (P<0.05). The 3D reconstruction of the lower mandible suggested that the study group had smaller Go angle and SNB angle and shorter ANS-Me distance, Go-Me distance and N-Me distance compared to the control group; the differences had statistical significance (P<0.05). The face scanning results demonstrated that the nasolabial angle and facial convexity angle of the study group were significantly larger than those in the control group, and the difference was statistically significant (P<0.05).

Seeking for Correlative Genes and Signaling Pathways With Bone Metastasis From Breast Cancer by Integrated Analysis.

Background: Bone metastasis frequently occurs in advanced breast cancer patients, and it is one of major causes of breast cancer associated mortality. The aim of the current study is to identify potential genes and related signaling pathways in the pathophysiology of breast cancer bone metastasis. Methods: Three mRNA expression datasets for breast cancer bone metastasis were obtained from Gene Expression Omnibus (GEO) dataset. The differentially expressed genes (DEGs) were obtained. Functional analyses, protein-protein interaction (PPI) network, and transcription factors (TFs)-target genes network was constructed. Real-time PCR using clinical specimens was conducted to justify the results from integrated analysis. Results: A 749 DEGs were obtained. Osteoclast differentiation and rheumatoid arthritis were two significantly enriched signaling pathways for DEGs in the bone metastasis of breast cancer. SMAD7 (degree = 10), TGFBR2 (degree = 9), VIM (degree = 8), FOS (degree = 8), PDGFRB (degree = 7), COL5A1 (degree = 6), ARRB2 (degree = 6), and ITGAV (degree = 6) were high degree genes in the PPI network. ETS1 (degree = 12), SPI1 (degree = 12), FOS (degree = 10), FLI1 (degree = 5), KLF4 (degree = 4), JUNB (degree = 4), NR3C1 (degree = 4) were high degree genes in the TFs-target genes network. Validated by QRT-PCR, the expression levels of IBSP, MMP9, MMP13, TNFAIP6, CD200, DHRS3, ASS1, RIPK4, VIM, and PROM1 were roughly consistent with our integrated analysis. Except PROM1, the other genes had a diagnose value for breast cancer bone metastasis. Conclusions: The identified DEGs and signaling pathways may make contribution for understanding the pathological mechanism of bone metastasis from breast cancer.

Assessing the dyeing efficiency and irritation potentials of plant hair dyes: A multi-analytical in vitro approach.

BACKGROUND: The interest toward dyeing hairs with plant colorants has grown in popularity considering its low-toxic nature. However, researches reporting plant hair dyes are limited and the potential adverse effects of irritation are unclear. OBJECTIVES: This study is aimed to provide an avenue by which to more accurately assess the dyeing efficiency and irritation potentials of plant hair dyes. METHODS: Four extracted plant colorants were incorporated in hydrogel hair dyes that were directly applied on unbleached gray human hairs. Their dyeing performances and the effect of an iron (Ⅱ) mordant were photometrically measured in CIELab coordinates and color strength. The eye and skin irritancy was assessed by combining various in vitro methods, including bovine corneal opacity and permeability (BCOP) assay in combination with histopathological analysis, Hen's egg test on chick chorioallantoic membrane (HET-CAM) and a test on reconstructed human epidermis models. RESULTS: The investigated hair dyes exhibited desirable dyeing efficiency on human hairs. Post-treatment with the iron (Ⅱ) mordant caused a significant increase in color strength with subtle changes to the hue of dyed color. In the irritation testing, the four hair dyes were categorized as slight-to-mild eye irritants but possessed no skin irritation potential, while the mordant was determined as a non-irritant. CONCLUSIONS: The results demonstrate the efficacy of a multi-analytical approach for in vitro assessment of various plant colorants for hair dyeing. The investigated plant extracts are suitable for producing viable colors on human hairs and may serve as a low-irritating alternative to the synthetic hair dyes.

[Study on application of CPP-ACP on tooth mineralization during orthodontic treatment with fixed appliance].

PURPOSE: To determine the value of clinical application of CPP-ACP during orthodontic treatment with fixed appliance. METHODS: Seventy-five subjects were divided randomly into three groups, the control group, experimental group A and experimental group B. The control group were emphasized on daily oral hygiene. Experimental group A used fluor protector every three months under the dentist's guidance. Experimental group B used Casein phosphopeptide amorphous calcium phosphate(CCP-ACP) once a day. After finishing the orthodontic treatment, photos were taken under the same condition ,then the degree of the enamel's demineralization was examined and the enamel decalcification index(EDI) was calculated. SPSS11.0 software package was used for statistical analysis. RESULTS: (1)The incidence of three groups' enamel decalcification declined in sequence as the control group(60%),experimental group A (36%), experimental group B (32%).(2)The incidence of the teeth's enamel calcification of three groups declined in sequence as the control group(14.7%),experimental group A (8.46%), experimental group B (7.72%), the difference between the control group and two experimental groups was significant, while no significant difference was found between the two experimental groups.(3(EDI of the control group was 0.155+/-0.023, EDI of the experimental group A was 0.082+/-0.009,while EDI of the experimental group B was 0.078+/-0.006. The difference between the control group and two experimental groups was statistically significant, while was not statistically significant between the two experimental groups. CONCLUSIONS: The application of fluor protector and CPP-ACP can improve the mineralization of the teeth during orthodontic treatment with fixed appliance. CPP-ACP is more convenient for the patients to use. Supported by Research Fund of Bureau of Science and Technology of Futian District Shenzhen City(Grant No.FTWS056).