-
International Journal of Molecular... Sep 2022Apical Lesions of Endodontic Origin (ALEO) are initiated by polymicrobial endodontic canal infection. () and () lipopolysaccharides (LPS) can induce a pro-inflammatory...
Apical Lesions of Endodontic Origin (ALEO) are initiated by polymicrobial endodontic canal infection. () and () lipopolysaccharides (LPS) can induce a pro-inflammatory macrophage response through their recognition by TLR2 and TLR4. However, polarization responses induced by and/or LPS in macrophages are not fully understood. We aimed to characterize the polarization profiles of macrophages differentiated from THP-1 cells following and/or LPS stimulation from reference strain and clinical isolates. A modified LPS purification protocol was implemented and the electrophoretic LPS profiles were characterized. THP-1 human monocytes differentiated to macrophages were stimulated with and LPS. Polarization profiles were characterized through cell surface markers and secreted cytokines levels after 24 h of stimulation. TLR2 and TLR4 cell surfaces and transcriptional levels were determined after 24 or 2 h of LPS stimulation, respectively. LPS from induced a predominant M1 profile in macrophages evidenced by changes in the expression of the surface marker CD64 and pro-inflammatory cytokine profiles, TNF-α, IL-1β, IL-6, and IL-12. LPS was unable to induce a significant response. TLR2 and TLR4 expressions were neither modified by or LPS. LPS, but not LPS, induced a macrophage M1 Profile.
Topics: Humans; Lipopolysaccharides; Macrophages; Porphyromonas endodontalis; Porphyromonas gingivalis; Toll-Like Receptor 2; Toll-Like Receptor 4
PubMed: 36077408
DOI: 10.3390/ijms231710011 -
Hua Xi Kou Qiang Yi Xue Za Zhi = Huaxi... Feb 2015This study aims to assess and compare the prevalence of Porphyromonas endodontalis (P. endodontalis) in root canals associated with primary and secondary endodontic...
OBJECTIVE
This study aims to assess and compare the prevalence of Porphyromonas endodontalis (P. endodontalis) in root canals associated with primary and secondary endodontic infections by using 16s rDNA PCR and real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR).
METHODS
A total of 120 adult patients with one radiographically documented periapical lesion were included. Sixty teeth presented with primary endodontic infections and 60 with secondary endodontic infections requiring retreatment. P. endodontalis was identified by using 16s rDNA PCR techniques. The positive DNA expression of P. endodontalis in two types of infected root canals were quantitatively compared by using SYBR GREEN I RTFQ-PCR.
RESULTS
The prevalence of P. endodontalis in the root canals with primary endodontic infections was significantly higher than that in root canals with secondary endodontic infections (P = 0.001). However, RTFQ-PCR results showed no significant difference in DNA expression quantities between the primary and secondary endodontic infections root canals (P = 0.303).
CONCLUSION
P. endodontalis is more highly associated with root canals having primary endodontic infections, although P. endodontalis colonize in both root canals with primary and secondary chronic apical periodontitis.
Topics: Adult; DNA, Bacterial; Dental Pulp Cavity; Gram-Negative Bacterial Infections; Humans; Polymerase Chain Reaction; Porphyromonas endodontalis; Retreatment
PubMed: 25872307
DOI: 10.7518/hxkq.2015.01.020 -
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 -
Frontiers in Cellular and Infection... 2021Apical periodontitis is an inflammatory disease of microbial etiology. It has been suggested that endodontic bacterial DNA might translocate to distant organs blood...
Apical periodontitis is an inflammatory disease of microbial etiology. It has been suggested that endodontic bacterial DNA might translocate to distant organs blood vessels, but no studies have been conducted. We aimed first to explore overall extraradicular infection, as well as specifically by spp; and their potential to translocate from infected root canals to blood through peripheral blood mononuclear cells. In this cross-sectional study, healthy individuals with and without a diagnosis of apical periodontitis with an associated apical lesion of endodontic origin (both, symptomatic and asymptomatic) were included. Apical lesions (N=64) were collected from volunteers with an indication of tooth extraction. Intracanal samples (N=39) and respective peripheral blood mononuclear cells from apical periodontitis (n=14) individuals with an indication of endodontic treatment, as well as from healthy individuals (n=14) were collected. The detection frequencies and loads (DNA copies/mg or DNA copies/μL) of total bacteria, and were measured by qPCR. In apical lesions, the detection frequencies (%) and median bacterial loads (DNA copies/mg) respectively were 70.8% and 4521.6 for total bacteria; 21.5% and 1789.7 for and 18.4% and 1493.9 for . In intracanal exudates, the detection frequencies and median bacterial loads respectively were 100% and 21089.2 (DNA copies/μL) for total bacteria, 41% and 8263.9 for ; and 20.5%, median 12538.9 for Finally, bacteria were detected in all samples of peripheral blood mononuclear cells including apical periodontitis and healthy groups, though total bacterial loads (median DNA copies/μL) were significantly higher in apical periodontitis (953.6) compared to controls (300.7), p<0.05. was equally detected in both groups (50%), but its bacterial load tended to be higher in apical periodontitis (262.3) than controls (158.8), p>0.05; was not detected. Bacteria and specifically spp. were frequently detected in endodontic canals and apical lesions. Also, total bacteria and DNA were detected in peripheral blood mononuclear cells, supporting their plausible role in bacterial systemic translocation.
Topics: Bacterial Translocation; Cross-Sectional Studies; DNA, Bacterial; Humans; Leukocytes, Mononuclear; Periapical Periodontitis; Porphyromonas endodontalis
PubMed: 33816354
DOI: 10.3389/fcimb.2021.649925 -
Molecular Medicine Reports Feb 2017Periapical lesions are characterized by the destruction of periapical bone, and occur as a result of local inflammatory responses to root canal infection by...
Periapical lesions are characterized by the destruction of periapical bone, and occur as a result of local inflammatory responses to root canal infection by microorganisms including Porphyromonas endodontalis (P. endodontalis). P. endodontalis and its primary virulence factor, lipopolysaccharide (LPS), are associated with the development of periapical lesions and alveolar bone loss. Interleukin‑23 (IL‑23) is critical in the initiation and progression of periodontal disease via effects on peripheral bone metabolism. The present study investigated the expression of IL‑23 in tissue where a periapical lesion was present, and the effect of P. endodontalis LPS on the expression of IL‑23 in periodontal ligament (PDL) cells. Reverse transcription‑ quantitative polymerase chain reaction and immunohistochemistry revealed increased levels of IL‑23 expression in tissue with periapical lesions compared with healthy PDL tissue. Treatment with P. endodontalis LPS increased the expression of IL‑23 in the SH‑9 human PDL cell line. BAY11‑7082, a nuclear factor κB inhibitor, suppressed P. endodontalis LPS‑induced IL‑23 expression in SH‑9 cells. Treatment of RAW264.7 cells with conditioned medium from P. endodontalis LPS‑treated SH‑9 cells promoted osteoclastogenesis. By contrast, RAW264.7 cells treated with conditioned medium from IL‑23‑knockdown SH‑9 cells underwent reduced levels of osteoclastogenesis. The results of the present study indicated that the expression of IL‑23 in PDL cells induced by P. endodontalis LPS treatment may be involved in the progression of periapical lesions via stimulation of the osteoclastogenesis process.
Topics: Animals; Chromones; Humans; Immunohistochemistry; Interleukin-23; Lipopolysaccharides; Mice; Morpholines; NF-kappa B; NFATC Transcription Factors; Nitriles; Osteoblasts; Osteogenesis; Periodontitis; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Porphyromonas endodontalis; Proto-Oncogene Proteins c-fos; RANK Ligand; RAW 264.7 Cells; RNA Interference; RNA, Small Interfering; Sulfones
PubMed: 28000855
DOI: 10.3892/mmr.2016.6041 -
Infection and Drug Resistance 2022Even with the advent of NGS and PCR diagnostic tools, cases of chest infections caused by are still very rare. Such pathogens are less likely to be considered by...
BACKGROUND
Even with the advent of NGS and PCR diagnostic tools, cases of chest infections caused by are still very rare. Such pathogens are less likely to be considered by clinicians. These cases frequently involve the pleura and lead to pneumothorax, hydropneumothorax, or pyopneumothorax, making the disease severe.
CASE PRESENTATION
A 69-year-old man diagnosed with cerebral infarction a year ago sought medical attention for right-sided pyopneumothorax and respiratory failure. The pathogen found in the pleural fluid was highly suspected to be (). Pleural fluid mNGS confirmed and coinfection. Metronidazole combined with piperacillin tazobactam was administered to counteract infection. Simultaneously, closed chest drainage and thoracoscopic release of pleural adhesions were performed. The patient was cured, discharged from the hospital, and was in good condition after six months of follow-up.
CONCLUSION
When chest infections occur in patients with poor oral hygiene and underlying diseases that may lead to aspiration, the identification of infection should be noted. Early confirmation of the diagnosis often requires mNGS and PCR. Metronidazole is essentially effective against , and medical thoracoscopy can be used to manage pleural conditions if necessary.
PubMed: 36277245
DOI: 10.2147/IDR.S381859 -
Journal of Dental Sciences Apr 2022Because complete microbial elimination of the infected root canal system is nearly impossible to achieve, the use of root canal fillings with antibacterial effects may...
BACKGROUND/PURPOSE
Because complete microbial elimination of the infected root canal system is nearly impossible to achieve, the use of root canal fillings with antibacterial effects may help to minimize intracanal infections. Our previous study, MTA modified by CaF addition, improved the biocompatibility and mineralization potential of human dental pulp cells without adverse effect on the physical properties. In this study, the antibacterial effect of MTA after CaF addition was evaluated for use as a root canal sealer. The objective of this study was to evaluate the antibacterial activity of MTA after the addition of CaF.
MATERIALS AND METHODS
The antibacterial activities of MTA and MTA-CaF mixture against () (), and () were investigated with MTA-CaF powder and eluates. The bacterial growth was measured by optical density using a spectrophotometer and the bacterial colony counting.
RESULTS
MTA inhibited the growth of , and , and the addition of CaF improved this antibacterial effect in a concentration dependent manner ( < 0.05). was more resistant to MTA-CaF than the other groups of bacteria.
CONCLUSION
The addition of more than 5% CaF to MTA can increase the antibacterial activity.
PubMed: 35756781
DOI: 10.1016/j.jds.2021.09.005 -
BioMed Research International 2022This study is aimed at evaluating five mineral oxides (5MO), mineral trioxide aggregate repair high plasticity (MTA HP), and mineral trioxide aggregate (MTA) in relation...
This study is aimed at evaluating five mineral oxides (5MO), mineral trioxide aggregate repair high plasticity (MTA HP), and mineral trioxide aggregate (MTA) in relation to the antimicrobial action over , , , , and ; the genotoxicity over mouse macrophage (RAW 264.7) and osteoblast (Mg-63) cultures; and the morphological analysis using scanning electron microscopy (SEM) analysis (50 k and ×100 k). Sodium hypochlorite (NaOCl), calcium hydroxide, and saline solution were used as control groups in the different analysis. All data were submitted to a normality test and then analyzed with one-way ANOVA, Tukey, and Kruskal-Wallis and Dunn tests, considering ≤ 0.05 significance level. It was found that over and , there was no a significant difference between the calcium silicate-based cements (CSC) and the control group of saline solution, and only 5MO was similar to the NaOCl group. However, over , all groups were effective and showed a statistically significant difference compared to the saline solution group. Conversely, none of the groups were effective over and , except of the NaOCl group. There was a significant difference between 5MO and MTA groups in comparison with NaOCl and MTA HP over osteoblasts and macrophages after 24 hours. SEM images showed small irregular particles interspersed with some elongated needle-like particles and small irregular particles with some larger particles as well as elongated particles. It was concluded that 5MO, MTA, and MTA HP have effective antimicrobial action over . However, only 5MO is effective over and . Besides, 5MO and MTA are not genotoxic over mouse macrophage (RAW 264.7) and osteoblast (Mg-63) cultures.
Topics: Animals; Mice; Aluminum Compounds; Anti-Infective Agents; Calcium; Calcium Compounds; Drug Combinations; Materials Testing; Oxides; Root Canal Filling Materials; Saline Solution; Silicates
PubMed: 35592523
DOI: 10.1155/2022/2155226 -
Journal of Endodontics Sep 2014Periapical infections secondary to pulpal necrosis are associated with bacterial contamination of the pulp. Porphyromonas endodontalis, a gram-negative organism, is... (Comparative Study)
Comparative Study
INTRODUCTION
Periapical infections secondary to pulpal necrosis are associated with bacterial contamination of the pulp. Porphyromonas endodontalis, a gram-negative organism, is considered to be a pulpal pathogen. P. gingivalis is phylogenetically related to P. endodontalis and synthesizes several classes of novel complex lipids that possess biological activity, including the capacity to promote osteoclastogenesis and osteoclast activation. The purpose of this study was to extract and characterize constituent lipids of P. endodontalis and evaluate their capacity to promote proinflammatory secretory responses in the macrophage cell line, RAW 264.7, as well as their capacity to promote osteoclastogenesis and inhibit osteoblast activity.
METHODS
Constituent lipids of both organisms were fractionated by high-performance liquid chromatography and were structurally characterized using electrospray mass spectrometry or electrospray-mass spectrometry/mass spectrometry. The virulence potential of P. endodontalis lipids was then compared with known biologically active lipids isolated from P. gingivalis.
RESULTS
P. endodontalis total lipids were shown to promote tumor necrosis factor alpha secretion from RAW 264.7 cells, and the serine lipid fraction appeared to account for the majority of this effect. P. endodontalis lipid preparations also increased osteoclast formation from RAW 264.7 cells, but osteoblast differentiation in culture was inhibited and appeared to be dependent on Toll-like receptor 2 expression.
CONCLUSIONS
These effects underscore the importance of P. endodontalis lipids in promoting inflammatory and bone cell activation processes that could lead to periapical pathology.
Topics: Animals; Cell Differentiation; Cell Line; Cells, Cultured; Ceramides; Chromatography, High Pressure Liquid; Female; Inflammation Mediators; Lipids; Macrophages; Mice; Mice, Inbred C57BL; Mice, Knockout; Osteoblasts; Osteoclasts; Phosphatidylethanolamines; Porphyromonas endodontalis; Porphyromonas gingivalis; Serine; Spectrometry, Mass, Electrospray Ionization; Sphingomyelins; Tandem Mass Spectrometry; Toll-Like Receptor 2; Tumor Necrosis Factor-alpha; Virulence Factors
PubMed: 25146013
DOI: 10.1016/j.joen.2014.02.017 -
Veterinary Microbiology May 2017Periodontitis is a polymicrobial infectious disease that causes occlusion change, tooth loss, difficulty in rumination, and premature culling of animals. This study...
Periodontitis is a polymicrobial infectious disease that causes occlusion change, tooth loss, difficulty in rumination, and premature culling of animals. This study aimed to detect species of the genera Porphyromonas and Prevotella present in the periodontal pocket of sheep with lesions deeper than 5mm (n=14) and in the gingival sulcus of animals considered periodontally healthy (n=20). The presence of microorganisms was evaluated by polymerase chain reaction (PCR) using specific primers for Porphyromonas asaccharolytica, Porphyromonas endodontalis, Porphyromonas gingivalis, Porphyromonas gulae, Prevotella buccae, Prevotella intermedia, Prevotella loescheii, Prevotella melaninogenica, Prevotella nigrescens, Prevotella oralis, and Prevotella tannerae. Prevalence and risk analysis were performed using Student's t-test and Spearman's correlation. Among the Prevotella and Porphyromonas species detected in the periodontal lesions of sheep, P. melaninogenica (85.7%), P. buccae (64.3%), P. gingivalis (50%), and P. endodontalis (50%) were most prevalent. P. gingivalis (15%) and P. oralis (10%) prevailed in the gingival sulcus. P. gulae and P. tannerae were not detected in the 34 samples studied. Data evaluation by t-test verified that occurrence of P. asaccharolytica, P. endodontalis, P. gingivalis, P. buccae, P. intermedia, P. melalinogenica, and P. nigrescens correlated with sheep periodontitis. The findings of this study will be an important contribution to research on pathogenesis of sheep periodontitis and development of its control measures.
Topics: Animals; Bacteria, Anaerobic; Biofilms; Periodontal Pocket; Periodontitis; Polymerase Chain Reaction; Porphyromonas; Prevotella; Sheep; Sheep Diseases
PubMed: 28619155
DOI: 10.1016/j.vetmic.2017.03.032