-
Scientific Reports Nov 2023The objective of this study was to compare and evaluate the changes in periodontal pathogens and periodontal status within 6 months of wearing three orthodontic...
The objective of this study was to compare and evaluate the changes in periodontal pathogens and periodontal status within 6 months of wearing three orthodontic retainers, namely, vacuum-formed retainer (VFR), Hawley retainer (HR), and lingual fixed retainer (LR). In total, 48 patients who underwent orthodontic treatment with ordinary metal brackets were divided into VFR, HR, and LR groups (n = 16 per group). Saliva samples were collected at the time of debonding (T0) and after 1 month (T1), 3 months (T2), and 6 months (T3). Porphyromonas gingivalis (Pg) and Aggregatibacter actinomycetemcomitans (Aa) were quantitatively analyzed using real-time PCR. Gingival index (GI), plaque index (PLI), and probing depth (PD) were measured at the four time points to evaluate changes in periodontal state. SPSS20.0 software was used to analyze the data, and P < 0.05 was considered statistically significant. The trial was registered at the Chinese Clinical Trial Registry (ChiCTR2300073704), the registration was retrospective. Compared to baseline (T0) values, Pg, Aa, GI, PLI, and PD were significantly decreased in all three groups 1 month after wearing the retainer (p < 0.05). Significant differences were observed in Aa at T3 among the three groups, whereby the HR group exhibited significantly better results compared to the VFR and LR groups (p < 0.05). Differences were found among the three groups' Porphyromonas gingivalis at T3, and the HR group was significantly better than the VFR and LR groups (P < 0.05). From T1 to T2, GI, PLI, and PD of the three groups tended to be stable, however differences were observed at T3, with the PLI and PD of the HR group being the lowest among the three groups (p < 0.05). Regardless of the type of retainer used, the periodontal condition of patients was significantly improved after removal of the metal brackets. After 6 months of retainer use, the Hawley retainer was superior to vacuum-formed retainer and lingual fixed retainer with regard to Pg, Aa, and periodontal clinical parameters.
Topics: Humans; Orthodontic Retainers; Retrospective Studies; Porphyromonas gingivalis; Orthodontic Appliances, Fixed; Gingival Diseases
PubMed: 38001102
DOI: 10.1038/s41598-023-46922-2 -
Molecular Oral Microbiology Aug 2023Porphyromonas gingivalis, the causative agent of adult periodontitis, must gain resistance to frequent oxidative and nitric oxide (NO) stress attacks from immune cells...
Porphyromonas gingivalis, the causative agent of adult periodontitis, must gain resistance to frequent oxidative and nitric oxide (NO) stress attacks from immune cells in the periodontal pocket to survive. Previously, we found that, in the wild-type and under NO stress, the expression of PG1237 (CdhR), the gene encoding for a putative LuxR transcriptional regulator previously called community development and hemin regulator (CdhR), was upregulated 7.7-fold, and its adjacent gene PG1236 11.9-fold. Isogenic mutants P. gingivalis FLL457 (ΔCdhR::ermF), FLL458 (ΔPG1236::ermF), and FLL459 (ΔPG1236-CdhR::ermF) were made by allelic exchange mutagenesis to determine the involvement of these genes in P. gingivalis W83 NO stress resistance. The mutants were black pigmented and β hemolytic and their gingipain activities varied with strains. FLL457 and FLL459 mutants were more sensitive to NO compared to the wild type, and complementation restored NO sensitivity to that of the wild type. DNA microarray analysis of FLL457 showed that approximately 2% of the genes were upregulated and over 1% of the genes downregulated under NO stress conditions compared to the wild type. Transcriptome analysis of FLL458 and FLL459 under NO stress showed differences in their modulation patterns. Some similarities were also noticed between all mutants. The PG1236-CdhR gene cluster revealed increased expression under NO stress and may be part of the same transcriptional unit. Recombinant CdhR showed binding activity to the predicted promoter regions of PG1459 and PG0495. Taken together, the data indicate that CdhR may play a role in NO stress resistance and be involved in a regulatory network in P. gingivalis.
Topics: Porphyromonas gingivalis; Nitric Oxide; Hemin; Gingipain Cysteine Endopeptidases; Gene Expression Profiling
PubMed: 37134265
DOI: 10.1111/omi.12414 -
BMC Complementary and Alternative... Aug 2017P-113 (AKRHHGYKRKFH-NH2) is a 12-amino-acid histidine-rich peptide derived from histatin 5 that is highly degradable in high salt concentrations and biological fluids...
Preventive effects of the novel antimicrobial peptide Nal-P-113 in a rat Periodontitis model by limiting the growth of Porphyromonas gingivalis and modulating IL-1β and TNF-α production.
BACKGROUND
P-113 (AKRHHGYKRKFH-NH2) is a 12-amino-acid histidine-rich peptide derived from histatin 5 that is highly degradable in high salt concentrations and biological fluids such as serum, plasma and saliva. Nal-P-113, a novel antimicrobial peptide whose histidine residues are replaced by the bulky amino acids β-naphthylalanine, causes the antimicrobial peptide to retain its bactericidal activity even in physiological environments. This study evaluated the effect of the novel antimicrobial peptide Nal-P-113 in a rat periodontitis model and the mechanisms of action of Nal-P-113 for suppressing periodontitis.
METHODS
Periodontitis was induced in mandibular first molars in rats receiving a ligature and infected with Porphyromonas gingivalis. Animals were randomly divided into six groups: a, P. gingivalis W83 alone; b, P. gingivalis W83 with 6.25 μg/mL of Nal-P-113; c, P. gingivalis W83 with 25 μg/mL of Nal-P-113; d, P. gingivalis W83 with 100 μg/mL of Nal-P-113; e, P. gingivalis W83 with 400 μg/mL of Nal-P-113; and f, control without P. gingivalis W83 or Nal-P-113. Morphometric analysis was used to evaluate alveolar bone loss. Microbiological assessment of the presence of Porphyromonas gingivalis and total bacteria was performed using absolute quantitative real-time PCR and scanning electron microscopy. Gingival tissue was collected for western blot and immunohistochemical assays of IL-1β and TNF-α levels.
RESULTS
Alveolar bone loss was inhibited by 100 μg/mL or 400 μg/mL of Nal-P-113 compared to the control group (P < 0.05). Lower amounts of P. gingivalis and total bacteria were found in groups d and e compared with group a (P < 0.05). A decrease in the levels of IL-1β and TNF-α was detected in group d and group e compared to the control group (P < 0.05). The amount of P. gingivalis was positively correlated with IL-1β and TNF-α expression in periodontal tissue (P < 0.05).
CONCLUSIONS
Nal-P-113 exhibited protective effects on Porphyromonas gingivalis-induced periodontitis in rats by limiting the amount of bacteria and modulating IL-1β and TNF-α production. The use of Nal-P-113 in vivo might serve as a beneficial preventive or therapeutic approach for periodontitis.
Topics: Animals; Disease Models, Animal; Humans; Interleukin-1beta; Male; Peptides; Periodontitis; Porphyromonas gingivalis; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha
PubMed: 28851350
DOI: 10.1186/s12906-017-1931-9 -
BMC Microbiology Sep 2020Dental implants have become well-established in oral rehabilitation for fully or partially edentulous patients. However, peri-implantitis often leads to the failure of...
BACKGROUND
Dental implants have become well-established in oral rehabilitation for fully or partially edentulous patients. However, peri-implantitis often leads to the failure of dental implants. The aim of this study was to understand the core microbiome associated with peri-implantitis and evaluate potential peri-implantitis pathogens based on canine peri-implantitis model.
RESULTS
In this study, three beagle dogs were used to build peri-implantitis models with ligature-induced strategy. The peri-implant sulcular fluids were collected at four different phases based on disease severity during the peri-implantitis development. Microbial compositions during peri-implantitis development were monitored and evaluated. The microbes were presented with operational taxonomic unit (OTU) classified at 97% identity of the high-throughput 16S rRNA gene fragments. Microbial diversity and richness varied during peri-implantitis. At the phylum-level, Firmicutes decreased and Bacteroides increased during peri-implantitis development. At the genus-level, Peptostreptococcus decreased and Porphyromonas increased, suggesting peri-implantitis pathogens might be assigned to these two genera. Further species-level and co-occurrence network analyses identified several potential keystone species during peri-implantitis development, and some OTUs were potential peri-implantitis pathogens.
CONCLUSION
In summary, canine peri-implantitis models help to identify several potential keystone peri-implantitis associated species. The canine model can give insight into human peri-implantitis associated microbiota.
Topics: Animals; Bacterial Typing Techniques; Bacteroides; Bone-Implant Interface; Dental Implants; Disease Models, Animal; Dogs; Firmicutes; Genetic Variation; Humans; Ligation; Male; Microbiota; Peptostreptococcus; Peri-Implantitis; Phylogeny; Porphyromonas; RNA, Ribosomal, 16S; Spirochaeta
PubMed: 32993514
DOI: 10.1186/s12866-020-01982-6 -
The Type IX Secretion System (T9SS): Highlights and Recent Insights into Its Structure and Function.Frontiers in Cellular and Infection... 2017Protein secretion systems are vital for prokaryotic life, as they enable bacteria to acquire nutrients, communicate with other species, defend against biological and... (Review)
Review
Protein secretion systems are vital for prokaryotic life, as they enable bacteria to acquire nutrients, communicate with other species, defend against biological and chemical agents, and facilitate disease through the delivery of virulence factors. In this review, we will focus on the recently discovered type IX secretion system (T9SS), a complex translocon found only in some species of the phylum. T9SS plays two roles, depending on the lifestyle of the bacteria. It provides either a means of movement (called gliding motility) for peace-loving environmental bacteria or a weapon for pathogens. The best-studied members of these two groups are , a commensal microorganism often found in water and soil, and , a human oral pathogen that is a major causative agent of periodontitis. In and some other periodontopathogens, T9SS translocates proteins, especially virulence factors, across the outer membrane (OM). Proteins destined for secretion bear a conserved C-terminal domain (CTD) that directs the cargo to the OM translocon. At least 18 proteins are involved in this still enigmatic process, with some engaged in the post-translational modification of T9SS cargo proteins. Upon translocation across the OM, the CTD is removed by a protease with sortase-like activity and an anionic LPS is attached to the newly formed C-terminus. As a result, a cargo protein could be secreted into the extracellular milieu or covalently attached to the bacterial surface. T9SS is regulated by a two-component system; however, the precise environmental signal that triggers it has not been identified. Exploring unknown systems contributing to bacterial virulence is exciting, as it may eventually lead to new therapeutic strategies. During the past decade, the major components of T9SS were identified, as well as hints suggesting the possible mechanism of action. In addition, the list of characterized cargo proteins is constantly growing. The actual structure of the translocon, situated in the OM of bacteria, remains the least explored area; however, new technical approaches and increasing scientific attention have resulted in a growing body of data. Therefore, we present a compact up-to-date review of this topic.
Topics: Bacterial Proteins; Bacterial Secretion Systems; Bacteroidetes; Flavobacterium; Humans; Porphyromonas gingivalis; Protein Processing, Post-Translational; Protein Transport; Virulence Factors
PubMed: 28603700
DOI: 10.3389/fcimb.2017.00215 -
Molecular Oral Microbiology Apr 2021Porphyromonas gingivalis, a pathogen of chronic periodontitis, is an asaccharolytic microorganism that solely utilizes nutritional amino acids as its energy source and... (Review)
Review
Porphyromonas gingivalis, a pathogen of chronic periodontitis, is an asaccharolytic microorganism that solely utilizes nutritional amino acids as its energy source and cellular constituents. The bacterium is considered to incorporate proteinaceous nutrients mainly as dipeptides, thus exopeptidases that produce dipeptides from polypeptides are critical for survival and proliferation. We present here an overview of dipeptide production by P. gingivalis mediated by dipeptidyl-peptidases (DPPs), e.g., DPP4, DPP5, DPP7, and DPP11, serine exopeptidases localized in periplasm, which release dipeptides from the N-terminus of polypeptides. Additionally, two other exopeptidases, acylpeptidyl-oligopeptidase (AOP) and prolyl tripeptidyl-peptidase A (PTP-A), which liberate N-terminal acylated di-/tri-peptides and tripeptides with Pro at the third position, respectively, provide polypeptides in an acceptable form for DPPs. Hence, a large fraction of dipeptides is produced from nutritional polypeptides by DPPs with differential specificities in combination with AOP and PTP-A. The resultant dipeptides are then incorporated across the inner membrane mainly via a proton-dependent oligopeptide transporter (POT), a member of the major facilitator superfamily. Recent studies also indicate that DPP4 and DPP7 directly link between periodontal and systemic diseases, such as type 2 diabetes mellitus and coagulation abnormality, respectively. Therefore, these dipeptide-producing and incorporation molecules are considered to be potent targets for prevention and treatment of periodontal and related systemic diseases.
Topics: Diabetes Mellitus, Type 2; Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; Humans; Periplasm; Porphyromonas gingivalis; Proteins
PubMed: 33006264
DOI: 10.1111/omi.12317 -
The Chinese Journal of Dental Research 2019To investigate the immunoinflammatory response in the crosstalk of human oral keratinocytes (HOKs) with selected periodontal commensals and pathogens.
OBJECTIVE
To investigate the immunoinflammatory response in the crosstalk of human oral keratinocytes (HOKs) with selected periodontal commensals and pathogens.
METHODS
Four representative viable oral bacteria, including periodontal commensals (Streptococcus mutans, Sm; and Actinomyces israelii, Ai) and pathogens (Aggregatibacter actinomycetemcomitans, Aa; and Porphyromonas gingivalis, Pg), were selected. A viable bacteria-HOKs interactive model was tested under various conditions of oxygen, antibiotics, duration and multiplicity of infection (MOI). The expression of IL-6 and IL-8 in HOKs was assessed by real-time qPCR and ELISA.
RESULTS
An MOI of 1 was determined to be the appropriate ratio of bacteria and HOKs with substantial amounts of viable bacterial cells and HOKs in an antibiotic-free medium under aerobic conditions for 2 h. Sm and Pg significantly upregulated the expression of IL-6 and IL-8 (P < 0.05), while Ai and Aa could not induce significant levels of these cytokines with reference to the control.
CONCLUSION
Within the limitations of this study, the current findings suggest that periodontal commensals and pathogens may differentially modulate immunoinflammatory response in human oral keratinocytes.
Topics: Aggregatibacter actinomycetemcomitans; Cytokines; Humans; Keratinocytes; Porphyromonas gingivalis; Streptococcus mutans
PubMed: 31172138
DOI: 10.3290/j.cjdr.a42514 -
Nan Fang Yi Ke Da Xue Xue Bao = Journal... Jul 2023To investigate the effect of (Pg) infection on IFNGR1 palmitoylation and biological behaviors of esophageal squamous cell carcinoma (ESCC) cells and the clinical...
OBJECTIVE
To investigate the effect of (Pg) infection on IFNGR1 palmitoylation and biological behaviors of esophageal squamous cell carcinoma (ESCC) cells and the clinical implications.
METHODS
The expression levels of IFNGR1 protein in ESCC cell lines KYSE30 and KYSE70 were detected using Western blotting at 24 and 48 h after Pg infection, and 2-BP was used to detect IFNGR1 palmitoylation in the cells. KYSE70 cells with wild-type IFNGR1 (IFNGR1-WT cells) and with IFNGR1-C122A palmitoylation site mutation induced by site-specific mutagenesis (IFNGR1-C122A cells) were both infected with Pg, and the changes in palmitoylation of IFNGR1-C122A were analyzed using immunofluorescence and Click-iT assays. The changes in proliferation, migration and invasion ability of the infected cells were evaluated using plate cloning assay, scratch assay and Transwell assay, and IFNGR1 co-localization with lysosomal marker LAMP2 was dected using immunofluorescence assay. Immunohistochemistry was used to detect Pg infection and IFNGR1 protein expression in 50 ESCC tissues, and their correlation with the clinicopathological characteristics and survival outcomes of the patients was analyzed.
RESULTS
Pg infection down-regulated the protein expression of IFNGR1 in ESCC and promoted IFNGR1 palmitoylation at site 122. In IFNGR1-WT cells, Pg infection significantly enhanced cell proliferation, migration and invasion ( < 0.05). Similarly, Pg also significantly promoted proliferation, migration and invasion of IFNGR1-C122A cells, but to a lesser extent as compared with the wild-type cells ( < 0.05). Immunofluorescence assay showed that Pg and ZDHHC3 promoted IFNGR1 degradation within the lysosome. Immunohistochemical studies of the ESCC tissue samples showed a negative correlation between IFNGR1 and Pg expression, and a reduced IFNGR1 expression was correlated with a poorer survival outcome of the patient.
CONCLUSION
Pg infection enhances IFNGR1 palmitoylation to promote progression of ESCC, and elimination of Pg and inhibiting IFNGR1 palmitoylation may effectively control ESCC progression.
Topics: Humans; Esophageal Neoplasms; Porphyromonas gingivalis; Lipoylation; Esophageal Squamous Cell Carcinoma; Lysosomes
PubMed: 37488798
DOI: 10.12122/j.issn.1673-4254.2023.07.12 -
Journal of Oral Science 2020Porphyromonas gingivalis (P. gingivalis) is one of the major pathogenic bacteria of periodontitis or peri-implantitis. P. gingivalis tends to attach to the implant's...
Porphyromonas gingivalis (P. gingivalis) is one of the major pathogenic bacteria of periodontitis or peri-implantitis. P. gingivalis tends to attach to the implant's neck with the formation of biofilm, leading to peri-implantitis. d-arginine has been shown to have a potential antimicrobial role. In this study, P. gingivalis was cultured in Brain Heart Infusion broth together with d-arginine. After 3 days (inhibition) or 6 days (dissociation), these were characterized using crystal violet (CV) staining for the biofilm, extracellular polysaccharide (EPS) production from the biofilm, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay for biofilm activation. Furthermore, the P. gingivalis biofilm was observed by scanning electron microscopy (SEM). d-arginine effectively reduced biomass accumulation and promoted dissociation at concentrations of ≥50 mM and 100 mM, respectively. Through CV staining, d-arginine concentrations of EPS production from the biofilm for inhibition and dissociation effects was ≥50 mM and 100 mM, respectively. In addition, d-arginine affected biofilm activation for the corresponding concentrations: ≥60 mM for inhibition and ≥90 mM for dispersal. Under SEM observation, d-arginine changed the P. gingivalis biofilm structure in relatively high concentrations for inhibition or dissociation, respectively. The authors concluded that d-arginine could inhibit the formation of P. gingivalis biofilm and promote the dissociation of P. gingivalis biofilm.
Topics: Arginine; Biofilms; Humans; Microscopy, Electron, Scanning; Peri-Implantitis; Porphyromonas gingivalis
PubMed: 31996524
DOI: 10.2334/josnusd.19-0075 -
PloS One 2014Dipeptidyl peptidases (DPPs) that liberate dipeptides from the N-terminal end of oligopeptides are crucial for the growth of Porphyromonas species, anaerobic...
Dipeptidyl peptidases (DPPs) that liberate dipeptides from the N-terminal end of oligopeptides are crucial for the growth of Porphyromonas species, anaerobic asaccharolytic gram negative rods that utilize amino acids as energy sources. Porphyromonas endodontalis is a causative agent of periapical lesions with acute symptoms and Asp/Glu-specific DPP11 has been solely characterized in this organism. In this study, we identified and characterized two P. endodontalis DPPs, DPP5 and DPP7. Cell-associated DPP activity toward Lys-Ala-4-methylcoumaryl-7-amide (MCA) was prominent in P. endodontalis ATCC 35406 as compared with the Porphyromonas gingivalis strains ATCC 33277, 16-1, HW24D1, ATCC 49417, W83, W50, and HNA99. The level of hydrolysis of Leu-Asp-MCA by DPP11, Gly-Pro-MCA by DPP4, and Met-Leu-MCA was also higher than in the P. gingivalis strains. MER236725 and MER278904 are P. endodontalis proteins belong to the S9- and S46-family peptidases, respectively. Recombinant MER236725 exhibited enzymatic properties including substrate specificity, and salt- and pH-dependence similar to P. gingivalis DPP5 belonging to the S9 family. However, the kcat/Km figure (194 µM-1·sec-1) for the most potent substrate (Lys-Ala-MCA) was 18.4-fold higher as compared to the P. gingivalis entity (10.5 µM-1·sec-1). In addition, P. endodontalis DPP5 mRNA and protein contents were increased several fold as compared with those in P. gingivalis. Recombinant MER278904 preferentially hydrolyzed Met-Leu-MCA and exhibited a substrate specificity similar to P. gingivalis DPP7 belonging to the S46 family. In accord with the deduced molecular mass of 818 amino acids, a 105-kDa band was immunologically detected, indicating that P. endodontalis DPP7 is an exceptionally large molecule in the DPP7/DPP11/S46 peptidase family. The enhancement of four DPP activities was conclusively demonstrated in P. endodontalis, and remarkable Lys-Ala-MCA-hydrolysis was achieved by qualitative and quantitative potentiation of the DPP5 molecule.
Topics: Amino Acid Sequence; Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; Enzyme-Linked Immunosorbent Assay; Isoenzymes; Porphyromonas; Sequence Homology, Amino Acid; Species Specificity
PubMed: 25494328
DOI: 10.1371/journal.pone.0114221