Effects of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans on Modeling Subgingival Microbiome and Impairment of Oral Epithelial Barrier.

Periodontitis is an exemplar of dysbiosis associated with the coordinated action of multiple members within the microbial consortium. The polymicrobial synergy and dysbiosis hypothesis proposes a dynamic host-microbiome balance, with certain modulators capable of disrupting eubiosis and driving shifts towards dysbiosis within the community. However, these factors remain to be explored. We established a Porphyromonas gingivalis- or Aggregatibacter actinomycetemcomitans-modified subgingival microbiome model and 16S rRNA sequencing revealed that P. gingivalis and A. actinomycetemcomitans altered the microbiome structure and composition indicated by α and ß diversity metrics. P. gingivalis increased the subgingival dysbiosis index (SDI), while A. actinomycetemcomitans resulted in a lower SDI. Furthermore, P. gingivalis-stimulated microbiomes compromised epithelium function and reduced expression of tight junction proteins, whereas A. actinomycetemcomitans yielded mild effects. In conclusion, by inoculating P. gingivalis, we created dysbiotic microcosm biofilms in vitro resembling periodontitis-related subgingival microbiota, exhibiting enhanced dysbiosis and impaired epithelium integrity.

Extracellular vesicles from Aggregatibacter actinomycetemcomitans exhibit potential antitumorigenic effects in oral cancer: a comparative in vitro study.

Aggregatibacter actinomycetemcomitans is an opportunistic Gram-negative periodontopathogen strongly associated with periodontitis and infective endocarditis. Recent evidence suggests that periodontopathogens can influence the initiation and progression of oral squamous cell carcinoma (OSCC). Herein we aimed to investigate the effect of A. actinomycetemcomitans-derived extracellular vesicles (EVs) on OSCC cell behavior compared with EVs from periodontopathogens known to associate with carcinogenesis. EVs were isolated from: A. actinomycetemcomitans and its mutant strains lacking the cytolethal distending toxin (CDT) or lipopolysaccharide (LPS) O-antigen; Porphyromonas gingivalis; Fusobacterium nucleatum; and Parvimonas micra. The effect of EVs on primary and metastatic OSCC cells was assessed using cell proliferation, apoptosis, migration, invasion, and tubulogenesis assays. A. actinomycetemcomitans-derived EVs reduced the metastatic cancer cell proliferation, invasion, tubulogenesis, and increased apoptosis, mostly in CDT- and LPS O-antigen-dependent manner. EVs from F. nucleatum impaired the metastatic cancer cell proliferation and induced the apoptosis rates in all OSCC cell lines. EVs enhanced cancer cell migration regardless of bacterial species. In sum, this is the first study demonstrating the influence of A. actinomycetemcomitans-derived EVs on oral cancer in comparison with other periodontopathogens. Our findings revealed a potential antitumorigenic effect of these EVs on metastatic OSCC cells, which warrants further in vivo investigations.

Periodontal pathogen Aggregatibacter actinomycetemcomitans JP2 correlates with colonic leukocytes decrease and gut microbiome imbalance in mice.

AIM: Evidence suggests that translocation of oral pathogens through the oral-gut axis may induce intestinal dysbiosis. This study aimed to evaluate the impact of a highly leukotoxic Aggregatibacter actinomycetemcomitans (Aa) strain on the gut microbiota, intestinal mucosal integrity and immune system in healthy mice. METHODS: Eight-week-old male C57BL6 mice were divided into control (n = 16) and JP2 groups (n = 19), which received intragastric gavage with PBS and with a suspension of Aa JP2 (HK921), respectively, twice a week for 4 weeks. Colonic lamina propria, fecal material, serum, gingival tissues, and mandibles were obtained for analyses of leukocyte populations, inflammatory mediators, mucosal integrity, alveolar bone loss, and gut microbiota. Differences between groups for these parameters were examined by non-parametric tests. RESULTS: The gut microbial richness and the number of colonic macrophages, neutrophils, and monocytes were significantly lower in Aa JP2-infected mice than in controls (p < .05). In contrast, infected animals showed higher abundance of Clostridiaceae, Lactobacillus taiwanensis, Helicobacter rodentium, higher levels of IL-6 expression in colonic tissues, and higher splenic MPO activity than controls (p < .05). No differences in tight junction expression, serum endotoxin levels, and colonic inflammatory cytokines were observed between groups. Infected animals presented also slightly more alveolar bone loss and gingival IL-6 levels than controls (p < .05). CONCLUSION: Based on this model, intragastric administration of Aa JP2 is associated with changes in the gut ecosystem of healthy hosts, characterized by less live/recruited myeloid cells, enrichment of the gut microbiota with pathobionts and decrease in commensals. Negligible levels of colonic pro-inflammatory cytokines, and no signs of mucosal barrier disruption were related to these changes.

Anti-biofilm activity of 445 nm and 970 nm diode laser on mixed species colonies of- aggregatibacter actinomycetemcomitans and porphyromonas gingivalis cultured on titanium discs -an in vitro study.

To assess and compare the anti-microbial efficacy of 445 nm and 970 nm diode laser on mixed species biofilm of Aggregatibacter actinomycetemcomitans [A.a] and Porphyromonas gingivalis [P.g] cultured on machined pure titanium discs. A total of 65 machined pure titanium discs with no surface modifications with a 10-mm diameter and a 2-mm height were sterilized by autoclaving at 121 °C for 15 min and incubated with the commercially available bacterial strains ATCC(American Type Culture Collection- P.g 33277 and A.a 29522)mixture of Aggregatibacter actinomycetemcomitans(A.a) and Porphyromonas gingivalis(P.g).After a 2-week incubation period with the mixture of bacteria to develop a mixed species biofilm, the discs were divided into three groups: (1) no treatment (control), (2) 445 nm laser (test), (3) 970 nm laser (test). For each laser wavelength (445 and 970 nm), the discs were exposed to 1.0 W and 2.0 W in continuous wave mode for the times points of 15, 30, and 60 s. The antimicrobial efficacy was assessed by qPCR. A significant reduction in the levels of both species of bacteria was observed between control and the laser intervention groups. A higher efficacy for the 445 nm diode laser against Porphyromonas gingivalis and a similar efficacy against Aggregatibacter actinomycetemcomitans was observed as compared to the 970 nm group. 445 nm wavelength represents a potential and effective laser wavelength which can be used for the management of peri-implant infection. The present study findings also need to be further validated through clinical interventional trials.

The oral microbiome of a family including Papillon-Lefèvre-syndrome patients and clinically healthy members.

AIMS: The oral microbiota composition of patients diagnosed with Papillon-Lefèvre-syndrome and treated for several years were compared to those existing in the oral cavity of the clinically healthy family members and a cohort of patients having various stages of chronic periodontitis. MATERIALS AND METHODS: A family with two sisters affected with severe periodontitis and with the typical skin symptoms of Papillon-Lefèvre-syndrome, and symptomless parents and third sibling were investigated. The Patients received periodontal treatment for several years and their oral microbiome was analysed by amplicon sequencing. Data were evaluated by microbial cluster analysis. RESULTS: The microbiome of the patients with Papillon-Lefèvre-syndrome was predominated with Aggregatibacter actinomycetemcomitans and associated oral periodontopathogens. Although the clinically healthy family members showed no oral disorder, their microbiome resembled that of subjects having mild periodontitis. CONCLUSIONS: Predominance of A. actinomycetemcomitans in the subgingival microbiome of patients with Papillon-Lefèvre-syndrome suggests that specific treatment strategies directed against this pathobiont may improve the oral health status of the affected individuals. TRIAL REGISTRATION: The study was conducted in accordance with the Declaration of Helsinki and the ethical permission has been issued by the Human Investigation Review Board of the University of Szeged, Albert Szent-Györgyi Clinical Centre (Permission No. 63/2017-SZTE). September 19, 2017.  https://u-szeged.hu/klinikaikutatas/rkeb-altal-jovahagyott/rkeb-2017 .

Clinical and microbiological efficacy of intra-pocket application of diode laser in grade C periodontitis: a randomized controlled clinical trial.

BACKGROUND: Periodontitis is a microbially induced disease destroying structures anchoring teeth to jaw bones. Although metronidazole in combination with spiramycin is the effective conventional treatment of stage III grade C periodontitis, it has several systemic side effects. Laser therapy is widely used nowadays as an adjunct to scaling and root planing (SRP) to modulate inflammatory host response and eradicate microbes, due to bactericidal and detoxifying effects. Since microbiological analysis is one of the diagnostic methods identifying periodontal risk; our research aimed to investigate the efficacy of intra-pocket application of diode laser (980 nm) versus antibiotic therapy in enhancing clinical and microbiological parameters in stage III grade C periodontitis. METHODS: A randomized controlled clinical trial was conducted on fifty patients with stage III grade C periodontitis, divided equally into two groups. We managed test group by SRP with intra-pocket application of diode laser (980 nm) and the control group by SRP with systemic antibiotic administration (spiramycin and metronidazole). Then, we measured periodontal pocket depth (PPD) and clinical attachment loss (CAL) for both groups, before treatment (baseline), four and twelve weeks after. Moreover, we collected gingival crevicular fluid from both groups at baseline, four and twelve weeks after treatment and analyzed by real-time polymerase chain reaction to detect the relative count of Aggregatibacter actinomycetemcomitans and Porhyromonas gingivalis. RESULTS: Compared to baseline, all assessed clinical and microbiological parameters attested improvement at the end of the study period in each group individually with no significant difference between the two studied groups. Although, at twelve weeks, flare up of bacterial levels was detected with systemic antibiotic administration. CONCLUSION: Laser therapy can be considered as an effective treatment modality in stage III grade C periodontitis, avoiding the systemic antibiotic side effects and solving the recurrence problems due to bacterial resistance by long term usage. TRIAL REGISTRATION: NCT05222737 retrospectively on 03/02/2022, Clinicaltrial.gov.

The role of NLRP3 in regulating gingival epithelial cell responses evoked by Aggregatibacter actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) has myriads of virulence factors among which leukotoxin provides A. actinomycetemcomitans with the advantage to thrive in the surrounding hostile environment and evade host immune defences. The NLRP3 inflammasome has been associated with periodontal disease development. However, our understanding of the involvement of caspase-1, caspase-4, and NLRP3 in the release of IL-1ß and other inflammatory mediators from gingival epithelial cells during a A. actinomycetemcomitans infection is limited. The aim of this study was to investigate how the inflammasome-associated proteins caspase-1, caspase-4 and NLRP3 regulate the immune response of gingival epithelial cells during a A. actinomycetemcomitans infection. Human gingival epithelial cells (Ca9-22) deficient in NLRP3, caspase-1 or caspase-4 were created using CRISPR/Cas9. Gingival epithelial cells were stimulated with the A. actinomycetemcomitans low-leukotoxic strain NCTC9710 or the highly leukotoxic JP2 strain HK 165 for 6, 12 and 24 h. The results showed that the JP2 strain HK1651 induced higher IL-1ß and IL-1RA release and mediated more epithelial cell death compared to the NCTC9710 strain. These findings were found to be capsase-1, caspase-4 and NLRP3-dependant. A targeted protein analysis of inflammation-related proteins showed that the expression of 37 proteins were identified as being significantly altered after HK1651 infection compared to unstimulated Cas9 and NLRP3-deficient cells. Of the 37 proteins, 23 of these inflammation-related proteins released by NLRP3-deficient cells differed significantly compared to Cas9 cells after infection. This suggests that NLRP3 has a broad effect on the inflammatory response in gingival epithelial cells.

SOCS2 regulates alveolar bone loss in Aggregatibacter actinomycetemcomitans-induced periodontal disease.

INTRODUCTION: The role of suppressor of cytokine signaling 2 (SOCS2) in Aggregatibacter actinomycetemcomitans (Aa)-induced alveolar bone loss is unknown; thus, it was investigated in this study. METHODS: Alveolar bone loss was induced by infecting C57BL/6 wild-type (WT) and Socs2-knockout (Socs2-/-) mice with Aa. Bone parameters, bone loss, bone cell counts, the expression of bone remodeling markers, and cytokine profile were evaluated by microtomography, histology, qPCR, and/or ELISA. Bone marrow cells (BMC) from WT and Socs2-/- mice were differentiated in osteoblasts or osteoclasts for analysis of the expression of specific markers. RESULTS: Socs2-/- mice intrinsically exhibited irregular phenotypes in the maxillary bone and an increased number of osteoclasts. Upon Aa infection, SOCS2 deficiency resulted in the increased alveolar bone loss, despite decreased proinflammatory cytokine production, in comparison to the WT mice. In vitro, SOCS2 deficiency resulted in the increased osteoclasts formation, decreased expression of bone remodeling markers, and proinflammatory cytokines after Aa-LPS stimulus. CONCLUSIONS: Collectively, data suggest that SOCS2 is a regulator of Aa-induced alveolar bone loss by controlling the differentiation and activity of bone cells, and proinflammatory cytokines availability in the periodontal microenvironment and an important target for new therapeutic strategies. Thus, it can be helpful in preventing alveolar bone loss in periodontal inflammatory conditions.

Efficacy of hypochlorous acid as an alternative oral antimicrobial agent on human gingival fibroblasts, Aggregatibacter actinomycetemcomitans, and Candida albicans biofilms in vitro.

This study compared the cytotoxicity and antimicrobial activity of hypochlorous acid (HOCl) at 50 ppm and 200 ppm and 0.2% chlorhexidine (CHX) at various time intervals, in vitro. Cell viability and cytotoxicity of human gingival fibroblasts (HGF) were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and the lactate dehydrogenase assay. Antimicrobial effects on Aggregatibacter actinomycetemcomitans and Candida albicans were determined using the time-kill method. All solutions exhibited a significant impact on HGFs in a dose- and time-dependent manner. 50 ppm HOCl demonstrated the highest cell viability, followed by 200 ppm HOCl. Both HOCl solutions were less cytotoxic to HGFs than 0.2% CHX. 50 ppm and 200 ppm HOCl demonstrated stronger efficiencies than CHX against A. actinomycetemcomitans and C. albicans. The data suggest that HOCl solutions have potential as an alternative antiseptic to CHX due to their lower cytotoxicity and superior antimicrobial activity, but optimal dosage of HOCl requires further investigations.

The Roles of Periodontal Bacteria in Atherosclerosis.

Atherosclerosis (AS) is an inflammatory vascular disease that constitutes a major underlying cause of cardiovascular diseases (CVD) and stroke. Infection is a contributing risk factor for AS. Epidemiological evidence has implicated individuals afflicted by periodontitis displaying an increased susceptibility to AS and CVD. This review concisely outlines several prevalent periodontal pathogens identified within atherosclerotic plaques, including Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum. We review the existing epidemiological evidence elucidating the association between these pathogens and AS-related diseases, and the diverse mechanisms for which these pathogens may engage in AS, such as endothelial barrier disruption, immune system activation, facilitation of monocyte adhesion and aggregation, and promotion of foam cell formation, all of which contribute to the progression and destabilization of atherosclerotic plaques. Notably, the intricate interplay among bacteria underscores the complex impact of periodontitis on AS. In conclusion, advancing our understanding of the relationship between periodontal pathogens and AS will undoubtedly offer invaluable insights and potential therapeutic avenues for the prevention and management of AS.

Effectiveness of Diode (810 nm) Laser in Periodontal Parameters and Reduction of Subgingival Bacterial Load in Periodontitis Patients.

AIM: This split-mouth randomized trial (RCT) aimed to assess the effect of diode laser on the clinical parameters in patients with periodontitis, compare the results with scaling and root planing (SRP) alone, and assess the implications of diode laser (DL) on plaque bacteria. MATERIALS AND METHODS: Seventeen periodontitis patients were randomly assigned into two equal groups based on the therapy delivered. Group I (control site) received just SRP at baseline, while group II (test site) received both SRP and DL irradiation. For both groups, the clinical periodontal parameters probing pocket depth (PPD), and clinical attachment level (CAL) were measured at baseline, 30 days, and 90 days. Microbiological amount was also measured at baseline, 30, and 90 days after periodontal treatment. The amounts of Aggregatibacter actinomycetemcomitans (A.a), Prevotella intermedia (Pr. intermedia), and Porphyromonas gingivalis (P. gingivalis) were determined using real-time PCR probing with specific bacterial primers. RESULTS: In both groups, PPD and CAL showed statistically significant reductions at different time intervals (p < 0.05). No significant difference were observed in CAL values after 1 and 3 months in both test and control groups (p > 0.05). The mean values of the concentration of A.a, Pr. intermedia and P. gingivalis were lower in the case group as compared to the control group and the difference was statistically significant after 1 month (*p = 0.001). CLINICAL SIGNIFICANCE: According to this study, non-invasive laser treatment has the potential to improve clinical outcomes by lowering the quantity of A.a, Pr. intermedia and P. gingivalis. CONCLUSION: In both groups, a considerable decrease in the periodontal pathogens A.a, Pr. intermedia and P. gingivalis were discovered; however, the intergroup comparison was insignificant in relation to PD and CAL. The adjunctive treatment with diode laser showed better efficacy in ensuring a better periodontal treatment than SRP alone. How to cite this article: Abdullah LA, Hashim N, Rehman MM, et al. Effectiveness of Diode (810 nm) Laser in Periodontal Parameters and Reduction of Subgingival Bacterial Load in Periodontitis Patients. J Contemp Dent Pract 2023;24(12):1008-1015.

Virulence traits and plasmid-mediated quinolone resistance among Aggregatibacter actinomycetemcomitans from Iraq: Low rate of highly virulent JP2 genotype.

Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) is an important causative agent of periodontitis acting by employment of a series of virulence factors. Our aim was to evaluate the virulence traits and plasmid-mediated quinolone resistance of A. actinomycetemcomitans isolates in Iraq. A total of 1580 samples were collected from dental caries (n = 1190) and periodontitis (n = 390) among which 200 samples were positive. The antibiotic susceptibility pattern and biofilm formation were performed. The antibiotic resistance and virulence determinants were screened using polymerase chain reaction (PCR) technique. The ltx3 and ltx4 primers were used for identification of highly virulent JP2 type. A. actinomycetemcomitans was identified among dental caries (n = 114) and periodontitis (n = 86) samples. The JP2 type was identified in six periodontitis samples. Sixty (30% of) isolates were multidrug-resistant (MDR). Eighty-four (42% of) A. actinomycetemcomitans isolates formed strong biofilms and 44% of them had moderate-level biofilms. The detected virulence genes included ltxA (96%), cdtB (64%), qseB (62%), qseC (58%) and rcpA (58%). There was a significant relation between the existence of ltxA (42%, p = 0.041) and rcpA (64%, p = 0.022) genes and the biofilm formation. The JP2 genotype was identified in six adolescents with periodontitis. The rate of qnrA, qnrB, qnrC, qnrD, qnrS and aac(6')-Ib-cr plasmid-mediated quinolone resistance genes included 22%, 18%, 16%, 16%, 14% and 0%, respectively. The qnrA (66.7%) and qnrB (53.4%) genes were significantly detected higher in MDR strains. Herein, A. actinomycetemcomitans from dental caries and periodontitis had relatively high rate of resistance to ß-lactams but low resistance levels to quinolones. Moreover, most of the resistant isolates carried the qnrA-S genes. A majority of them had ltxA gene, half of them contained all the virulence genes and JP2 genotype was found in six isolates from periodontitis. Furthermore, half of the isolates produced biofilms which was significantly related to the ltxA and rcpA genes. Screening of virulence genes and antibiotic resistance pattern determination contribute to the control, diagnosis and eradication of these isolates.

Pilus PilA of the naturally competent HACEK group pathogen Aggregatibacter actinomycetemcomitans stimulates human leukocytes and interacts with both DNA and proinflammatory cytokines.

Each HACEK group pathogen, which can cause infective endocarditis, expresses type IVa pili. The type IVa major pilin PilA plays a role in bacterial colonization, virulence, twitching motility, and the uptake of extracellular DNA. The type IV prepilin homolog PilA of the periodontal pathogen A. actinomycetemcomitans (AaPilA) is linked to DNA uptake and natural competence. Our aim was to investigate the virulence properties and immunogenic potential of AaPilA. Since Neisseria meningitidis PilE, which shares sequence similarity with AaPilA, participates in sequestering host cytokines, we examined the ability of AaPilA to interact with various cytokines. Moreover, we investigated the structural characteristics of AaPilA with molecular modeling. AaPilA was conserved among A. actinomycetemcomitans strains. One of the 18 different natural variants, PilAD7S, is present in naturally competent strains. This variant interacted with DNA and bound interleukin (IL)-8 and tumor necrosis factor (TNF)-α. Specific anti-AaPilA antibodies were present in A. actinomycetemcomitans-positive periodontitis patient sera, and the production of reactive oxygen species from human neutrophils was less effectively induced by the ΔpilA mutant than by the wild-type strains. However, AaPilA did not stimulate human macrophages to produce proinflammatory cytokines, nor was it cytotoxic. The results strengthen our earlier hypothesis that the DNA uptake machinery of A. actinomycetemcomitans is involved in the sequestration of inflammatory cytokines. Furthermore, AaPilA stimulates host immune cells, such as B cells and neutrophils, making it a potential virulence factor.

Aggregatibacter actinomycetemcomitans: From Basic to Advanced Research.

Aggregatibacter actinomycetemcomitans is a major periodontal pathogen that was identified firstly in actinomycotic lesions and later in advanced forms of periodontal diseases as well as in oral cavity of healthy subjects. The particular pathogenicity of this specie makes it a target for extensive studies both at fundamental and practical scales. The current advances in experimental and clinical research related to this bacterium focus the light on epidemiologic features, virulence, and invasiveness aspects as well as on identification challenges, bacterial susceptibility, and anti-virulence strategies. The present chapter provide to scientists and periodontal researchers a comprehensive overview on the main advances made in this field with a special focus on epidemiologic dissemination, microbial diagnosis, virulence factors and clinical implementations of such progress.

Aggregatibacter actinomycetemcomitans colonization and persistence in a primate model.

Aggregatibacter actinomycetemcomitans is associated with aggressive periodontitis resulting in premature tooth loss in adolescents. Tooth adherence and biofilm persistence are prerequisites for survival in the oral domain. Here, using a rhesus monkey model, 16S rRNA sequencing, and weighted network analysis, we assessed colonization of A. actinomycetemcomitans variants and ascertained microbial interactions in biofilm communities. Variants in A. actinomycetemcomitans leukotoxin (ltx) were created, labeled, inoculated, and compared with their progenitor strain for in vivo colonization. Samples of tooth-related plaque were assessed for colonization at baseline and after debridement and inoculation of labeled strains. Null, minimal, and hyper-Ltx-producing strains were created and assessed for hydroxyapatite binding and biofilm formation in vitro. Ltx-hyperproducing strains colonized with greater prevalence and at higher levels than wild type or ltx mutants (P = 0.05). Indigenous and inoculated A. actinomycetemcomitans strains that attached were associated with lactate-producing species (i.e., Leptotrichia, Abiotrophia, and Streptoccocci). A. actinomycetemcomitans was found at 0.13% of the total flora at baseline and at 0.05% 4 wk after inoculation. In vivo data were supported by in vitro results. We conclude that hyper-Ltx production affords these strains with an attachment advantage providing a foothold for competition with members of the indigenous microbiota. Increased attachment can be linked to ltx gene expression and up-regulation of adherence-associated genes. Growth of attached A. actinomycetemcomitans in vivo was enhanced by lactate availability due to consorting species. These associations provide A. actinomycetemcomitans with the constituents required for its colonization and survival in the complex and competitive oral environment.

In vitro evaluation of physical and chemical parameters involved in aPDT of Aggregatibacter actinomycetemcomitans.

Periodontitis is an infectious disease characterized by the destruction of supporting tissues. Antimicrobial photodynamic therapy (aPDT) has been proposed as an improved method for eliminating microorganisms. Its efficiency depends on the correct use of physical and chemical parameters. Thus, these parameters and their relations were evaluated in this study with the purpose of establishing lethal conditions for combating bacterial agents. Diode lasers and light-emitting diodes (LEDs) were characterized to evaluate the absorption profile and resonance of methylene blue (MB) and toluidine blue O (TBO). The relations between light energy density and photosensitizer absorption were determined. Two methodologies were used to evaluate the effects of aPDT against Aggregatibacter actinomycetemcomitans. LED light exhibited a broad emission spectrum with a peak light wavelength of 637 nm and 99% purity. The resonance intensity of MB was higher with diode laser irradiation, and TBO showed higher resonance intensity with LED irradiation. There was no difference in the absorption profile of photosensitizers using diode lasers or LEDs, and variations in power density did not result in an increasing or decrease in light absorption. A. actinomycetemcomitans was susceptible to photodynamic processes. Emission spectra and peak light wavelengths of light sources combined with the absorption profiles of photosensitizers were the main parameters involved in determining the efficiency of photodynamic effects. Power density did not alter the light absorption of photosensitizers. The association between adequate irradiation characteristics and photosensitizer absorption results in complete inactivation of A. actinomycetemcomitans. In addition, the bactericidal effect was not altered by an increase in energy densities.

The effect of Er,Cr:YSGG laser on periodontopathic bacteria elimination: an in vitro study.

In vitro bacterial elimination using the erbium, chromium: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser against periodontopathic bacteria was investigated. Bacterial suspensions of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were spread on agar plates and the Er,Cr:YSGG laser was applied at 40 mJ pulse energy for durations of 30 s, 60 s, and 90 s. The agar plates were incubated, and growth inhibition zones were assessed. Optimal laser irradiation durations to achieve maximal bacterial elimination were evaluated using laser ablation on the bacterial colonies. The remaining viable bacteria were determined by the colony-forming unit (CFU) counting method. Growth inhibition zones were observed at all irradiation durations for both A. actinomycetemcomitans and P. gingivalis. Mean logarithmic values of CFU/ml after bacterial colony irradiation for 0 s (control), 12 s × 1 lap, 24 s × 1 lap, 48 s × 1 lap, and 24 s × 2 laps were 8.82 ± 0.35, 7.31 ± 0.94, 6.32 ± 0.61, 3.17 ± 2.90, and 0.00, respectively, for A. actinomycetemcomitans and 9.83 ± 0.50, 9.42 ± 0.11, 6.90 ± 1.60, 2.33 ± 3.19, and 0.00 for P. gingivalis. Significant differences were found between the control group and the two irradiated groups 48 s × 1 lap and 24 s × 2 laps (p < 0.05), and also between irradiated groups 12 s × 1 lap and 24 s × 2 laps (p < 0.05). An Er,Cr:YSGG laser with power setting 1.5 W and 30 Hz frequency showed potential for bacterial elimination against A. actinomycetemcomitans and P. gingivalis in vitro. Significant bacterial elimination (> 99.99%) was observed after 48 s of irradiation.

Prevalence of the JP2 genotype of Aggregatibacter actinomycetemcomitans in the world population: a systematic review.

PURPOSE: To investigate the global prevalence of the JP2 genotype of Aggregatibacter actinomycetemcomitans (Aa). METHODS: A comprehensive electronic search of databases, PUBMED, MEDLINE, EMBASE, BIOSIS, and SCOPUS, was conducted up to August 2021. All published articles and studies were considered, excluding animal studies, editorials, personal opinions, letters to editor, conference abstracts, posters, and those studies without full text. The primary objective of this systematic review was to determine the presence of the JP2 genotype of Aa in the world population. RESULTS: A total of 295 articles were identified, of which 62 were preselected, and 51 were finally included in this review. Due to variable study designs and high heterogeneity, a meta-analysis was not conducted. A total of 9744 subjects were screened for the presence of the JP2 genotype of Aa worldwide, and only 621 cases were found positive. CONCLUSIONS: A relatively high presence of JP2 genotype of Aa was found in subjects from South America, North America, and Africa. There were no studies estimating the presence of the JP2 genotype of Aa in the Oceania region. The heterogeneity and quality of the included publications suggest that caution should be exercised when interpreting the data and that there remains an important need for additional evidence. CLINICAL RELEVANCE: Periodontitis is a highly prevalent inflammatory oral disorder with substantial aesthetic, functional, and psychological implications for patients. The JP2 genotype of Aa is implicated in the pathogenesis of periodontitis. To the best of our knowledge, there is a lack of systematic reviews estimating the presence of the JP2 genotype of Aa in the global population. We identified a relatively high presence of the JP2 genotype of Aa in specific geographic areas of the world, and we propose that cross-sectional and longitudinal studies are lacking in the Oceania region and need to be conducted to estimate the presence of the JP2 genotype of Aa in this region.

Enhancing production of herpes simplex virus type 1 in oral epithelial cells by co-infection with Aggregatibacter actinomycetemcomitans.

BACKGROUND/PURPOSE: The association between herpetic/bacterial co-infection and periodontal diseases has been reported. However, how interactions between herpesviruses and periodontal bacteria dampen periodontal inflammation is still unclear. This study determined effects of co-infection with oral bacteria, including Streptococcus sanguinis, Fusobacterium nucleatum or Aggregatibacter actinomycetemcomitans, in herpes simplex virus type 1 (HSV-1)-infected oral epithelial cells. METHODS: Cell viability was determined by detection the activity of mitochondrial dehydrogenase. Viral production was measured using the plaque assay. Levels of bacterial and viral DNA were determined by real-time polymerase chain reaction. Secretion of interleukin (IL)-6 and IL-8 was measured using the enzyme-linked immunosorbent assay. RESULTS: Viability was not further reduced by bacterial co-infection in HSV-1-infected cells. Co-infection with HSV-1 and S. sanguinis or F. nucleatum reduced the viral yield whereas co-infection with HSV-1 and A. actinomycetemcomitans significantly enhanced the viral yield in oral epithelial cells. The enhancing effect of A. actinomycetemcomitans was not affected by bacterial heat-inactivation. Co-infection with HSV-1/A. actinomycetemcomitans increased intracellular levels of both viral and bacterial DNA. Secretion of IL-6 and IL-8 stimulated by A. actinomycetemcomitans infection was partly reduced by co-infection with HSV-1 in oral epithelial cells. CONCLUSION: In contrast to S. sanguinis and F. nucleatum, A. actinomycetemcomitans enhanced the yield of HSV-1. Either HSV-1 or A. actinomycetemcomitans may be benefited from co-infection, in aspects of increases in production of viral and bacterial DNA as well as reductions in cytokine secretion. These findings echoed with previous clinical studies showing co-infection of HSV and A. actinomycetemcomitans in patients with aggressive periodontitis.

In Vitro Selection of Lactobacillus and Bifidobacterium Probiotic Strains for the Management of Oral Pathobiont Infections Associated to Systemic Diseases.

The human oral pathobionts Aggregatibacter actinomycetemcomitans, Streptococcus mitis and Streptococcus mutans, in dysbiosis-promoting conditions, lead to oral infections, which also represent a threat to human systemic health. This scenario may be worsened by antibiotic misuse, which favours multi-drug resistance, making the research on pathogen containment strategies more than crucial. Therefore, we aimed to in vitro select the most promising probiotic strains against oral pathogen growth, viability, biofilm formation, and co-aggregation capacity, employing both the viable probiotics and their cell-free supernatants (CFSs). Interestingly, we also assessed probiotic efficacy against the three-pathogen co-culture, mimicking an environment similar to that in vivo. Overall, the results showed that Lactobacillus CFSs performed better than the Bifidobacterium, highlighting Limosilactobacillus reuteri LRE11, Lacticaseibacillus rhamnosus LR04, Lacticaseibacillus casei LC04, and Limosilactobacillus fermentum LF26 as the most effective strains, opening the chance to deeper investigation of their action and CFS composition. Altogether, the methodologies presented in this study can be used for probiotic efficacy screenings, in order to better focus the research on a viable probiotic, or on its postbiotics, suitable in case of infections.