Integration of metalloproteome and immunoproteome reveals a tight link of iron-related proteins with COVID-19 pathogenesis and immunity.

Increasing clinical data show that the imbalance of host metallome is closely associated with different kinds of disease, however, the intrinsic mechanisms of action of metals in immunity and pathogenesis of disease remain largely undefined. There is lack of multiplexed profiling system to integrate the metalloproteome-immunoproteome information at systemic level for exploring the roles of metals in immunity and disease pathogenesis. In this study, we build up a metal-coding assisted multiplexed proteome assay platform for serum metalloproteomic and immunoproteomic profiling. By taking COVID-19 as a showcase, we unbiasedly uncovered the most evident modulation of iron-related proteins, i.e., Ft and Tf, in serum of severe COVID-19 patients, and the value of Ft/Tf could work as a robust biomarker for COVID-19 severity stratification, which overtakes the well-established clinical risk factors (cytokines). We further uncovered a tight association of transferrin with inflammation mediator IL-10 in COVID-19 patients, which was proved to be mainly governed by the monocyte/macrophage of liver, shedding light on new pathophysiological and immune regulatory mechanisms of COVID-19 disease. We finally validated the beneficial effects of iron chelators as anti-viral agents in SARS-CoV-2-infected K18-hACE2 mice through modulation of iron dyshomeostasis and alleviating inflammation response. Our findings highlight the critical role of liver-mediated iron dysregulation in COVID-19 disease severity, providing solid evidence on the involvement of iron-related proteins in COVID-19 pathophysiology and immunity.

A prospective study on maternal periodontal diseases and neonatal adverse outcomes.

OBJECTIVE: It is evident that periodontitis is linked to various adverse pregnancy outcomes. This prospective study explored the potential link of maternal periodontal diseases to neonatal adverse outcomes. MATERIALS AND METHODS: A total of 193 generally healthy females in their third trimester (34-36 weeks) of pregnancy were enrolled. All subjects received full-mouth periodontal assessment, and the periodontal inflamed surface area (PISA) was calculated. Demographic data, lifestyles and anthropometric measurements of the neonates (e.g., body length and head circumference) were recorded. Herein, small-for-gestational age (SGA) referred to gender- and age-adjusted birth weight below the 10th percentile in line with the standard reference. Multivariable logistic regression analysis and restricted cubic spline were performed for examining the association of periodontal parameters with SGA.  Results: There were 8.3% (16/193) of neonates with SGA. Significantly positive correlation existed between the percentage of tooth sites with increased probing depth and an elevated risk of SGA (OR: 1.052; P < 0.05). Yet, the PISA was positively associated with the risk of SGA (OR: 1.002; P < 0.05) as well. No significant link occurred between maternal periodontal status and other neonatal outcome measures. CONCLUSION: Within the limitations of this study, the findings suggest that there could be a link between maternal periodontal diseases and neonatal adverse outcomes like SGA. Further investigation is required to clarify the current findings and potential implications for promoting maternal oral/periodontal health and newborn health.

Microbial persisters and host: recent advances and future perspectives.

Microbial persisters are defined as the tiny sub-population of microorganisms that develop intrinsic strategies for survival with high tolerance to various antimicrobials. Currently, persister research remains in its infancy, and it is indeed a great challenge to precisely distinguish persister cells from other drug tolerant ones. Notably, the existence of persisters crucially contributes to prolonged antibiotic exposure time and treatment failure, yet there is the formation of antibiotic-resistant mutants. Further understanding on persisters is of profound importance for effective prevention and control of chronic infections/inflammation. The past two decades have witnessed rapid advances on the science, technologies and methodologies for persister investigations, along with deep knowledge about persisters and numerous anti-persister approaches developed. Whereas, various critical issues remain unsolved, such as what are the potential interaction profiles of persisters and host cells, and how to apply what we know about persisters to translational studies and clinical practice. Importantly, it is highly essential to better understand the multifaceted and complex cross-talk of microbial persisters with the host to develop novel tackling strategies for precision healthcare in the near future.

Periodontal ligament cells from patients with treated stable periodontitis: Characterization and osteogenic differentiation potential.

BACKGROUND AND OBJECTIVE: Periodontal ligament progenitor cells (PDL cells) isolated from patients with inflammatory periodontitis have impaired regenerative capacity, but it is unknown whether this capacity can be recovered upon treatment and stabilization of the periodontal condition. The study aimed to investigate the expression of surface markers and the proliferation and osteogenic potential of PDL cells isolated from patients with treated stable periodontitis (S-PDL cells), periodontally healthy individuals (H-PDL cells), and patients with inflammatory periodontitis (I-PDL cells). METHODS: H-PDL, I-PDL, and S-PDL cells were isolated from the extracted teeth of individuals who (1) were periodontally healthy, (2) had inflammatory periodontitis, and (3) had treated stable periodontitis, respectively. The expression levels of surface markers and the proliferative and osteogenic capacities of the PDL cells were assessed. RESULTS: PDL cells derived from all three sources exhibited mesenchymal stem cell (MSC) characteristics. They were positive for MSC-related markers and negative for a hematopoiesis-related marker. However, S-PDL cells had higher proliferation rates, higher expression levels of osteogenic markers, higher alkaline phosphatase activity, and more calcium nodules than I-PDL cells. But all of these parameters remained lower in S-PDL cells than in H-PDL cells. CONCLUSIONS: S-PDL cells proliferated faster and had greater osteogenic potential than I-PDL cells, although these values remained lower than those in H-PDL cells.

Comparative analysis of serum and saliva samples using Raman spectroscopy: a high-throughput investigation in patients with polycystic ovary syndrome and periodontitis.

BACKGROUND: Polycystic ovary syndrome (PCOS) and periodontitis significantly affect women's oral and systemic health worldwide, and yet increase the risk of cardiovascular-metabolic diseases like diabetes and coronary heart disease. Regarding the PCOS-periodontitis connection, whether sex hormones, metabolic and inflammatory mediators could account for the underlying linking mechanism needs to be further investigated. This case-control study evaluated the hormonal, metabolic and inflammatory profiles in PCOS and non-PCOS subjects with various periodontal conditions, via assessing serum and saliva samples by Raman spectroscopy. METHODS: A total of 66 females with PCOS and 22 systemically healthy female volunteers were recruited in a single hospital. Full-mouth periodontal examination was undertaken for identifying the subjects with periodontal health, gingivitis or periodontitis. The datasets of sex hormones and metabolic indicators were retrieved from the hospital information system. Both serum and saliva samples were collected for detecting inflammatory mediators and Raman spectroscopic assessment. The subjects were categorized into four groups according to their conditions of PCOS and periodontitis for Raman spectroscopic analysis. Partial least squares discriminant analysis was performed to examine the inter-group differences in Raman spectra. RESULTS: PCOS patients exhibited greater mean probing depth (P < 0.05) and higher serum levels of triglycerides (P < 0.05) and matrix metalloproteinase-8 (P < 0.05) than those in non-PCOS participants. Both probing depth and triglyceride level were positively correlated with PCOS (P < 0.05). There was a significant difference in mean Raman spectra of saliva samples among the four groups with different conditions of PCOS and periodontitis (P < 0.05), while no significant inter-group difference existed in serum samples. CONCLUSIONS: The present study shows that periodontal condition may affect the biomolecular profiles of Raman spectra in serum and saliva of PCOS patients. It underscores the importance of the collaborative teamwork of dentists and gynecologists for enhancing women's oral health, general wellbeing and quality of life.

Maternal periodontal diseases affect the leukocyte profiles of umbilical cord blood: A cohort study.

AIM: This study evaluated the connection of periodontal status with the leukocyte profiles of maternal peripheral blood (MPB) and umbilical cord blood (UCB). MATERIALS AND METHODS: Ninety-nine pregnant females were recruited, and their data were collected via questionnaire and from medical records, including demographics, systemic conditions, complete blood count (CBC) and C-reaction protein (CRP) level in MPB. Full-mouth periodontal assessment was performed. CBC and CRP levels in UCB were measured after parturition. RESULTS: All subjects and their neonates were generally healthy. 30.3% of the participants presented with periodontal health condition, whereas 69.7% had different severities of periodontal diseases. The counts/percentages of eosinophils and monocytes in UCB from the subjects with periodontal diseases elevated, and the percentage of neutrophils decreased referencing to that from the counterparts (p < 0.05). There were positive correlations for total leukocyte count, neutrophils and lymphocytes counts/percentages in MPB and UCB among the periodontally healthy subjects (r > 0.4, p < 0.05), but such findings did not exist in those with periodontal diseases. Moreover, periodontal diseases independently accounted for the counts/percentages of neutrophils and eosinophils in UCB after controlling confounders in four testing models (ANCOVA, p < 0.05). CONCLUSION: Maternal periodontal diseases could to some extent disturb the leukocyte profiles of umbilical cord blood.

Restrictive Regulation of Ionic Liquid Quaternary Ammonium Salt in SBA-15 Pore Channel for Efficient Carbon Dioxide Conversion.

Herein, the synthesis of a new type of catalyst, SBA-M (Schiff complex of different metal types grafted on SBA-15) based on a quaternization reaction, is described. Various amounts of ionic liquid were grafted into the pore channels of SBA-15 using the post-grafting method, which allowed the ionic liquid to be grafted into the pore channels restrictively. Notably, over six cycles, SBA-Mn (0.2) has been shown to maintain its catalytic activity and stability. In addition, a reaction mechanism for the cycloaddition of CO2 with epoxides based on density-functional theory is proposed. The cycloaddition reaction of CO2 and epoxides is an efficient way of carbon fixation. It is demonstrated that the metal coordinated with the oxygen atom of the epoxides and that a halogen attacked the carbon of epoxides. Moreover, theoretical calculations and synthesis strategy provide a new approach for CO2 conversion.

Porphyromonas gingivalis lipopolysaccharide affects the angiogenic function of endothelial progenitor cells via Akt/FoxO1 signaling.

AIMS: Endothelial progenitor cells (EPCs) function as the angiogenic switch of many physiological and pathological conditions. We aimed to investigate the effects of Porphyromonas gingivalis lipopolysaccharide on the angiogenic capacity of EPCs and delineate the underlying mechanisms. MATERIALS AND METHODS: EPCs were isolated from human umbilical blood. CCK-8 assay was undertaken to analyze the cell viability. The migration and tube formation capacity were assessed by wound healing and tube formation, respectively. The protein expression of Akt/p-Akt, endothelial nitric oxide synthase (eNOS)/p-eNOS, and Forkhead box O1 (FoxO1)/p-FoxO1 was determined by Western blot. The intracellular localization of FoxO1 was evaluated by immunofluorescent staining. RESULTS: P. gingivalis LPS at 10 µg/ml significantly increased the viability (10.9 ± 2.9%), migration (16.3 ± 3.1%), and tube formation (38.6 ± 5.5%) of EPCs, along with increased phosphorylation of Akt, eNOS, and FoxO1. Mechanistically, Akt inhibition by specific inhibitor wortmannin and FoxO1 forced expression by adenovirus transfection in EPCs markedly attenuated the P. gingivalis LPS-induced eNOS activation, tube formation, and migration. Moreover, P. gingivalis LPS-induced phosphorylation and nuclear exclusion of FoxO1 were blunted by Akt inhibition. CONCLUSIONS: The present study suggests that P. gingivalis LPS could affect the angiogenic function of EPCs through the Akt/FoxO1 signaling. The current findings may shed light on the clinical association of periodontitis with aberrant angiogenesis seen in atherosclerotic plaque rupture.

Periodontitis links to concurrent systemic comorbidities among 'self-perceived health' individuals.

BACKGROUND AND OBJECTIVE: Our recent work shows that periodontitis experience reflects host susceptibility to the onset of multiple systemic diseases and conditions. This cross-sectional study further investigated whether and to what extent the existing periodontitis could reflect the concurrent presence of inflammatory comorbidities among 'self-perceived health' individuals. MATERIALS AND METHODS: There were 115 'self-perceived health' adults who completed a questionnaire on demographic characteristics and lifestyles. Twenty medical diagnostic tests were then performed to detect eight common systemic diseases and conditions. Meanwhile, full-mouth periodontal examination was undertaken, and the subjects were classified as two subgroups with or without Generalized Severe Periodontitis (Stages III/IV, generalized). The interlink of periodontal status and concurrent systemic comorbidities was assessed. RESULTS: 98.3% (113/115) of the subjects exhibited at least one undiagnosed systemic disease/disorder. Of them, 52.2% (59/113) and 47.8% (54/113) concurrently presented with 1-5 or ≥6 abnormal test results, respectively. Overall, 96.5% (111/115) had periodontitis. Generalized Severe Periodontitis was present in 43.2% (48/111) of the periodontitis patients, and it was significantly associated with the profiles of abnormal test results after adjusting potential confounders (abnormal test results 1-5 vs ≥6; OR: 3.23, p = .012). CONCLUSIONS: The present study shows that existing severe periodontitis could well reflect the concurrent presence of multiple inflammatory comorbidities. Oral and medical professionals can play proactive roles in enhancing health awareness and healthcare, through strong collaboration and teamwork.

Diagnostic performance of the AAP/EFP classification and the CDC/AAP case definition among pregnant women and a practical screening tool for maternal periodontal diseases.

BACKGROUND AND OBJECTIVE: There is a limited number of studies on the performance assessment of the 2017 AAP/EFP classification and the CDC/AAP case definition among pregnant females. This study evaluated the agreement between these two systems and explored a practical tool for screening maternal periodontal diseases by general dentists. MATERIALS AND METHODS: Totally, 204 systemically healthy females at different phases of pregnancy underwent a full-mouth periodontal examination. Demographic characteristics, lifestyles, and systemic conditions were recorded. Referring to the CDC/AAP definition, the diagnostic performance of the AAP/EFP classification was evaluated by the area under the ROC curve (AUC) and statistical tests (e.g., Youden's index and kappa coefficient). Additionally, a modified scoring system of the FDI Periodontal Diseases Chairside Guide (FDI-CG) was formulated with the addition of pregnancy for testing accordingly. RESULTS: Overall, there were 22.1% of the participants in early phase of pregnancy (7-13 weeks) and 77.9% in late phase (34-36 weeks). The majority of them were below 35 years and non-smokers without gestational diabetes. Notably, 30.9% of subjects presented with Moderate/Severe periodontitis (CDC/AAP), and 35.8% with Stages II-IV periodontitis (AAP/EFP). Referring to the CDC/AAP definition, the AUC, Youden's index, and κ of the AAP/EFP classification were 0.979, 0.890, and 92.9%, respectively. The modified FDI-CG system improved the AUC (0.815), Youden's index (63.0%), and κ (0.544) with reference to the original one. CONCLUSIONS: This study shows that the AAP/EFP classification is in high agreement with the CDC/AAP definition among the pregnant women. The phases of pregnancy-integrated FDI scoring system may serve as a convenient screening tool for maternal periodontal diseases in general dental practice.

Increased local concentrations of growth factors from leucocyte- and platelet-rich fibrin do not translate into improved alveolar ridge preservation: An intra-individual mechanistic randomized controlled trial.

AIM: Leucocyte- and platelet-rich fibrin (L-PRF) has been tested for enhancing alveolar ridge preservation (ARP), but little is known about the local release profile of growth factors (GF), and the clinical equipoise related to its efficacy remains. This study compared the patterns of GF release, early soft tissue healing, and alveolar ridge resorption following unassisted healing and L-PRF application in non-molar extraction sockets. MATERIALS AND METHODS: Atraumatic tooth extraction of two hopeless teeth per patient was followed by unassisted healing or L-PRF placement to fill the socket in 18 systemically healthy, non-smoking subjects. This intra-individual trial was powered to assess changes in horizontal alveolar ridge dimensions 1 mm below the crest of alveolar bone. GF concentrations in wound fluid were assessed with a multiplex assay at 6, 24, 72, and 168 h. Early healing was evaluated with the wound healing index and changes in soft tissue volumes on serial digital scans. Hard tissue changes were measured on superimposed CBCT images after 5 months of healing. RESULTS: L-PRF resulted in higher GF concentrations in wound fluid (WF) than in the control, but no differences in release patterns or time of peak were observed. No inter-group differences in early healing parameters were observed. Alveolar bone resorption was observed in both groups. No significant inter-group differences were observed in hard tissue healing 1, 3, or 5 mm apical to the original bone crest or in the ability to digitally plan a prosthetically guided implant with or without bone augmentation. CONCLUSIONS: L-PRF increased the GF concentrations in WF of extraction sockets without shifting the pattern observed in unassisted healing, while the increased delivery did not translate into clinical benefits in early wound healing or ARP. The current findings question the assumption that increased local concentrations of GF by L-PRF translate into improved clinical outcomes. Additional definitive studies are needed to establish the benefits of L-PRF in ARP (ClinicalTrials.gov NCT03985033).

Lipopolysaccharides affect compressed periodontal ligament cells via Eph-ephrin signaling.

OBJECTIVES: The aim of this study is to investigate the underlying mechanism of the recovery of periodontal ligament cells (PDLCs) sequentially exposed to inflammation and mechanical loading. MATERIALS AND METHODS: We divided PDLCs into four groups: control; compressive force (CF) alone (2.0 g/cm2 ); lipopolysaccharides (LPS) pretreatment (0.1 µg/ml) followed by simultaneous LPS and CF stimulation, simulating uncontrolled periodontitis; and LPS pretreatment followed by CF exposure, simulating controlled periodontitis. The expression of EphB4-ephrinB2 and EphA2-ephrinA2, and the level of osteoclastogenesis and osteogenesis were evaluated. RESULTS: Simultaneous stimulation by LPS and CF, compared with CF alone and sequential LPS and CF exposure, significantly suppressed EphB4 and enhanced ephrinA2 expression. Similarly, the most intense osteoclastic differentiation was observed under simultaneous LPS and CF stimulation, while sequential exposure to LPS and CF only slightly increased osteoclastic cell numbers. Both the activation of EphB4 signaling and ephrinA2 silencing lowered osteoclastic differentiation, which had previously been upregulated by simultaneous LPS and CF stimulation. These treatments also increased osteogenic differentiation. CONCLUSIONS: Simultaneous LPS and CF stimulation critically enhances osteoclastogenesis in PDLCs through the suppression of EphB4 and the induction of ephrinA2 signaling. Sequential LPS and CF exposure partially abolishes the osteolytic effects of simultaneous stimulation.

Periodontitis links to concurrent metabolic disorders and abnormal liver function in pregnant women.

OBJECTIVE: This cross-sectional study investigated the association of periodontitis with the metabolic status and hepatic function in pregnant women. MATERIALS AND METHODS: Full-mouth periodontal conditions, metabolic profiles, and hepatic function were assessed in 219 self-reported healthy pregnant females. The association of periodontal status with the systemic parameters was evaluated by parametric and non-parametric tests, and multivariate logistic regression analysis. RESULTS: Overall, periodontal status was positively associated with the metabolic profiles and hepatic function test results. The subjects with periodontitis exhibited higher levels of body mass index (BMI) (p < 0.01) and serum aspartate transaminase (AST) (p < 0.05), elevated diastolic blood pressure (DBP) (p < 0.05), and lower levels of high-density lipoprotein cholesterol (p < 0.05) than those of the counterparts. The periodontitis severity was strongly correlated with BMI and AST levels, and the extent of periodontal inflammation was related to DBP (p < 0.01). The periodontitis patients at 34-36 gestational weeks showed higher blood pressure and AST levels than those of non-periodontitis subjects (p < 0.05). CONCLUSION: Our findings on the notable links of periodontitis to concurrent metabolic disorders and abnormal liver function in pregnant women highlight the need of proactive integration of regular periodontal screening and healthcare in maternal programs for promoting optimal health and wellbeing of mothers-to-be and newborns.

RNA Sequencing Reveals the Upregulation of FOXO Signaling Pathway in Porphyromonas gingivalis Persister-Treated Human Gingival Epithelial Cells.

Porphyromonas gingivalis as the keystone periodontopathogen plays a critical role in the pathogenesis of periodontitis, and crucially accounts for inflammatory comorbidities such as cardiovascular disease and Alzheimer's disease. We recently identified the existence of P. gingivalis persisters and revealed the unforeseen perturbation of innate response in human gingival epithelial cells (HGECs) due to these noxious persisters. Herein, RNA sequencing revealed how P. gingivalis persisters affected the expression profile of cytokine genes and related signaling pathways in HGECs. Results showed that metronidazole-treated P. gingivalis persisters (M-PgPs) impaired the innate host defense of HGECs, in a similar fashion to P. gingivalis. Notably, over one thousand differentially expressed genes were identified in HGECs treated with M-PgPs or P. gingivalis with reference to the controls. Gene Ontology and KEGG pathway analysis demonstrated significantly enriched signaling pathways, such as FOXO. Importantly, the FOXO1 inhibitor rescued the M-PgP-induced disruption of cytokine expression. This study suggests that P. gingivalis persisters may perturb innate host defense, through the upregulation of the FOXO signaling pathway. Thus, the current findings could contribute to developing new approaches to tackling P. gingivalis persisters for the effective control of periodontitis and P. gingivalis-related inflammatory comorbidities.

Periodontal disease increases the host susceptibility to COVID-19 and its severity: a Mendelian randomization study.

BACKGROUND: Emerging evidence shows that periodontal disease (PD) may increase the risk of Coronavirus disease 2019 (COVID-19) complications. Here, we undertook a two-sample Mendelian randomization (MR) study, and investigated for the first time the possible causal impact of PD on host susceptibility to COVID-19 and its severity. METHODS: Summary statistics of COVID-19 susceptibility and severity were retrieved from the COVID-19 Host Genetics Initiative and used as outcomes. Single nucleotide polymorphisms associated with PD in Genome-wide association study were included as exposure. Inverse-variance weighted (IVW) method was employed as the main approach to analyze the causal relationships between PD and COVID-19. Three additional methods were adopted, allowing the existence of horizontal pleiotropy, including MR-Egger regression, weighted median and weighted mode methods. Comprehensive sensitivity analyses were also conducted for estimating the robustness of the identified associations. RESULTS: The MR estimates showed that PD was significantly associated with significantly higher susceptibility to COVID-19 using IVW (OR = 1.024, P = 0.017, 95% CI 1.004-1.045) and weighted median method (OR = 1.029, P = 0.024, 95% CI 1.003-1.055). Furthermore, it revealed that PD was significantly linked to COVID-19 severity based on the comparison of hospitalization versus population controls (IVW, OR = 1.025, P = 0.039, 95% CI 1.001-1.049; weighted median, OR = 1.030, P = 0.027, 95% CI 1.003-1.058). No such association was observed in the cohort of highly severe cases confirmed versus those not hospitalized due to COVID-19. CONCLUSIONS: We provide evidence on the possible causality of PD accounting for the susceptibility and severity of COVID-19, highlighting the importance of oral/periodontal healthcare for general wellbeing during the pandemic and beyond.

Study of the Cycloaddition of CO2 with Styrene Oxide Over Six-Connected spn Topology MOFs (Zr, Hf) at Room Temperature.

A series of MOFs with a 6-connected spn topology were synthesized (MOF-808-(Zr, Hf), PCN-777-(Zr, Hf), MOF-818-(Zr, Hf)). Through the in situ DRIFTS of NH3 adsorption-desorption, we found that the activated catalyst mainly contains Lewis acid sites. The effects of different organic ligands on the Lewis acid of the Zr6 cluster were analyzed by XPS and NH3 -TPD, and the relative Lewis acidity of the same metal was obtained: PCN-777>MOF-808>MOF-818. In the Py-FTIR results, we confirmed that MOF-818 has a higher acid site density. In the activity test, MOFs with mesoporous structure showed better catalytic activity under normal temperature and pressure. Among them, MOF-818 can still maintain a high degree of crystallinity after catalysis. Finally, we use density functional theory to propose the mechanism of the cycloaddition reaction of carbon dioxide and styrene oxide. The results show that the metal is coordinated with styrene oxide and halogens attack the ß carbon of the epoxide.

Identification and integrated analysis of differentially expressed long non-coding RNAs associated with periodontitis in humans.

BACKGROUND AND OBJECTIVE: Long non-coding RNAs (lncRNAs) can act as competing endogenous RNAs (ceRNAs) to compete for micro-RNAs (miRNAs) in regulation of downstream genes, various biological functions and diseases. Yet, the expression and regulation of lncRNAs in periodontitis are not fully understood. The objective of the study was to identify potential genes (lncRNA, messenger RNA [mRNA] and miRNA) involved in periodontitis, construct lncRNA-miRNA-mRNA ceRNA networks, explore gene functions and validate gene expressions. MATERIAL AND METHODS: The data sets for the lncRNA, mRNA and miRNA expression profiles in gingival samples from periodontally healthy subjects and chronic periodontitis patients were obtained from the Gene Expression Omnibus. The differentially expressed lncRNAs (DElncRNAs), mRNAs (DEmRNAs) and miRNAs (DEmiRNAs) were identified, and ceRNA networks were then constructed. The expression of DElncRNAs and DEmRNAs was examined by quantitative real-time polymerase chain reaction (qPCR). Moreover, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed for exploring the potential functions and biological pathways. RESULTS: The GSE80715 and GSE54710 data sets were retrieved. Subsequently, 26 DElncRNAs, 436 DEmRNAs and 12 DEmiRNAs were identified (|fold change| ≥2, adjusted p < 0.05). Further bioinformatics analysis contributed to establishment of the ceRNA networks, which consisted of 10 DElncRNAs, 11 DEmiRNAs and 83 DEmRNAs. Notably, the qPCR results showed a marked decrease in the expression of lncRNA H19 and two mRNAs (NOS1 and MAPT) which further supported the identified ceRNA network. The GO results revealed that the up-regulated mRNAs were significantly enriched in inflammatory processes, whilst the down-regulated mRNAs were enriched in cellular potentials. CONCLUSION: Non-coding RNAs are critically involved in the regulatory mechanisms in the pathogenesis of periodontitis. Further study is warranted to investigate the specific underlying genetic traits and networks.

Gingival bleeding on brushing as a sentinel sign of gingival inflammation: A diagnostic accuracy trial for the discrimination of periodontal health and disease.

AIM: To assess the accuracy of self-reported gingival bleeding on brushing (GBoB) for differentiating between periodontal health and disease and explore the optimal haemoglobin concentration that enables visual detection of GBoB. MATERIALS AND METHODS: Self-assessment of GBoB was conducted in supervised sessions for 408 consecutive adults. The haemoglobin levels in saliva/toothpaste slurry (TPS) were analysed, followed by a full-mouth periodontal examination. Periodontal diagnoses were made based on the 2017 classification of periodontal diseases. Gingival inflammation was defined as presence of at least 10% of sites with bleeding on probing (BOP). Logistic regression and area under the receiver operating characteristic curve (AUROC) analyses were applied to assess the accuracy of GBoB. RESULTS: Overall, 37.1% of the subjects claimed self-reported GBoB, and they had higher values of BOP (median: 25.0%; interquartile range (IQR): 16.0%-37.5%) than those without GBoB (median: 13.5%; IQR: 8.0%-24.8%, p < .001). The concentration/total amount of haemoglobin in TPS was positively correlated with the number of bleeding sites (r = .409/r = .520, p < .001). Haemoglobin concentration of 90.58 µg/ml or 0.51 µl blood volume enabled visual detection of GBoB with an AUROC of 0.848. Self-reported GBoB exhibited significantly increased values of diagnostic odds ratios (3-8) for varying degrees of gingival inflammation and periodontal disease (gingivitis and periodontitis). It showed low to moderate accuracy for discriminating periodontitis and gingivitis from periodontal health, with a sensitivity of 37.1% and 61.3% and a specificity of 84.8% and 84.4%, respectively. Absence of self-reported GBoB and low levels of haemoglobin had 93%-98% predictive values for periodontal health. CONCLUSIONS: Despite its low sensitivity for the discrimination of periodontitis, self-reported GBoB is a promising sentinel sign for periodontal health and disease, and gingival inflammation in particular. It is visually detectable after minor blood loss. After validation in an independent population, identification of GBoB may promote earlier detection and better prevention and treatment of periodontal disease, thereby eventually reducing the global burden of the disease.

Diagnostic accuracy of self-reported measures of periodontal disease: A clinical validation study using the 2017 case definitions.

AIM: To clinically validate a self-reported questionnaire for periodontal disease and assess its accuracy for differentiating periodontal health and different stages of periodontitis. METHODS: A Chinese (Cantonese) version of a self-reported questionnaire was prepared by translating and validating the original English questions proposed by the Centers for Disease Control and Prevention/American Academy of Periodontology (CDC/AAP). The utility of the CDC/AAP questionnaire and its individual questions was assessed against a full-mouth periodontal examination. Periodontal case definition was based on the 2017 World Workshop classification of periodontal diseases. Multivariable logistic regression and the area under the receiver operating characteristic curve (AUROC) analysis were performed to assess the accuracy of the questionnaire. RESULTS: 408 subjects enrolled in this study, including those with periodontal health (16.2%), gingivitis (15.2%), Stages I/II periodontitis (31.8%) and Stages III/IV periodontitis (36.8%). Overall, the questionnaire had poor accuracy in detecting the presence of Stages I/II periodontitis with an AUROC 0.608. While it showed moderate to high accuracy for identifying periodontal disease (gingivitis and periodontitis), periodontitis and Stages III/IV periodontitis with an AUROC of 0.837, 0.803 and 0.870, respectively. Self-reported measures in combination with age and tobacco smoking showed excellent performance for identifying Stages III/IV periodontitis with a high AUROC of 0.953, a sensitivity of 95.7%, and a specificity of 89.0%. Specific questions and combinations provided greater utility to discriminate the various periodontal case definitions. CONCLUSIONS: The self-reported CDC/AAP questionnaire may be a feasible tool for detecting periodontitis, and its combination with demographic and lifestyle factors is useful for the identification of individuals with Stages III/IV periodontitis. This questionnaire seems less helpful in screening of Stages I/II periodontitis. Further studies are needed to test the validity in larger community-based populations.

Diagnostic accuracy of a point-of-care aMMP-8 test in the discrimination of periodontal health and disease.

AIM: To assess the diagnostic utility of an oral rinse active matrix metalloproteinase-8 (aMMP-8) point-of-care test (POCT) for differentiating periodontal health, gingivitis, as well as different stages and grades of periodontitis. MATERIALS & METHODS: The aMMP-8 index test was undertaken in 408 consecutive adults, followed by a full-mouth periodontal examination. The reference standard was the 2017 World Workshop classification of periodontal diseases. Sensitivity, specificity, and the area under the receiver operating characteristic curve (AUROC) were assessed. RESULTS: 68.6% of the participants were diagnosed with periodontitis, including Stages I (15.9%), II (15.9%), III (29.7%) and IV (7.1%). A positive aMMP-8 POCT was associated with periodontitis after adjusting for age, gender, tobacco smoking and systemic diseases, while it was unable to differentiate among the stages/grades of periodontitis and between gingivitis/periodontal health. This test showed a sensitivity of 33.2% and a specificity of 93.0% for detecting periodontitis (threshold level >10 ng/ml). The levels of aMMP-8 adjusted by the number of teeth present (aMMP-8/NTP) performed better for periodontitis (sensitivity: 67.1%; specificity: 68.8%). Notably, aMMP-8/NTP were strongly predictive for Stage IV periodontitis (threshold level =0.4312 ng/ml) (sensitivity: 89.7%; specificity: 73.6%; and AUROC: 0.856). The test performance greatly improved in combination with age and smoking, with a sensitivity of 82.5%, a specificity of 84.4%, and an AUROC of 0.883. CONCLUSION: This aMMP-8 POCT is able to detect periodontitis with better specificity than sensitivity across the spectrum of its severity. This test may be useful for periodontal screening in conjunction with subject characteristics and/or other sensitive screening tools. Further validation studies are needed.