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1.
J Clin Periodontol ; 2024 Aug 05.
Artículo en Inglés | MEDLINE | ID: mdl-39104016

RESUMEN

AIM: To identify serum- and salivary-derived inflammatory biomarkers of periodontitis progression and determine their response to non-surgical treatment. MATERIALS AND METHODS: Periodontally healthy (H; n = 113) and periodontitis patients (P; n = 302) were monitored bi-monthly for 1 year without therapy. Periodontitis patients were re-examined 6 months after non-surgical periodontal therapy (NSPT). Participants were classified according to disease progression: P0 (no sites progressed; P1: 1-2 sites progressed; P2: 3 or more sites progressed). Ten salivary and five serum biomarkers were measured using Luminex. Log-transformed levels were compared over time according to baseline diagnosis, progression trajectory and after NSPT. Significant differences were sought using linear mixed models. RESULTS: P2 presented higher levels (p < .05) of salivary IFNγ, IL-6, VEGF, IL-1ß, MMP-8, IL-10 and OPG over time. Serum analytes were not associated with progression. NSPT led to clinical improvement and significant reduction of IFNγ, IL-6, IL-8, IL-1ß, MMP-8, IL-10, OPG and MMP-9 in saliva and of CRP, MMP-8, MMP-9 and MPO in serum. CONCLUSIONS: Periodontitis progression results from a sustained pro-inflammatory milieu that is reflected in salivary biomarkers, but less so in serum, likely because of the limited amount of progression per patient. NSPT can significantly decrease the levels of several salivary analytes.

2.
Arch Oral Biol ; 152: 105721, 2023 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-37196563

RESUMEN

OBJECTIVE: The aim of this study was to optimize the sensitivity, specificity and cost-effectiveness of the RNA-Oligonucleotide Quantification Technique (ROQT) in order to identify periodontal pathogens that remain unrecognized or uncultured in the oral microbiome. DESIGN: Total nucleic acids (TNA) were extracted from subgingival biofilm samples using an automated process. RNA, DNA and Locked Nucleic Acid (LNA) digoxigenin-labeled oligonucleotide probes targeting 5 cultivated/named species and 16 uncultivated or unnamed bacterial taxa were synthesized. Probe specificity was determined by targeting 96 oral bacterial species; sensitivity was assessed using serial dilutions of reference bacterial strains. Different stringency temperatures were compared and new standards were tested. The tested conditions were evaluated analyzing samples from periodontally healthy individuals, and patients with moderate or severe periodontitis. RESULTS: The automated extraction method at 63°C along with LNA-oligunucleotides probes, and use of reverse RNA sequences for standards yielded stronger signals without cross-reactions. In the pilot clinical study, the most commonly detected uncultivated/unrecognized species were Selenomonas sp. HMT 134, Prevotella sp. HMT 306, Desulfobulbus sp. HMT 041, Synergistetes sp. HMT 360 and Bacteroidetes HMT 274. In the cultivated segment of the microbiota, the most abundant taxa were T. forsythia HMT 613 and Fretibacterium fastidiosum (formerly Synergistetes) HMT 363. CONCLUSIONS: In general, samples from severe patients had the greatest levels of organisms. Classic (T. forsythia, P. gingivalis) and newly proposed (F. alocis and Desulfobulbus sp. HMT 041) pathogens were present in greater amounts in samples from severe periodontitis sites, followed by moderate periodontitis sites.


Asunto(s)
Placa Dental , Periodontitis , Humanos , Placa Dental/microbiología , ARN , Periodontitis/microbiología , Oligonucleótidos , ADN Bacteriano , Porphyromonas gingivalis/genética
3.
J Periodontol ; 92(9): 1222-1231, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-33866555

RESUMEN

BACKGROUND: Despite widespread use, the impact of minocycline hydrochloride microspheres on the shifts of oral bacterial species resistant to minocycline remains unknown. This study aimed at examining the percentage and taxonomy of minocycline-resistant isolates in saliva and subgingival plaque samples before and after minocycline microspheres application in periodontitis patients during maintenance. METHODS: Patients received supra- and sub-gingival debridement with (test) or without (control) minocycline microspheres application to sites with probing depth >4 mm and were clinically monitored at baseline, 1, 3, and 6 months. Samples were collected at baseline, 1 and 6 months and analyzed via cultivation with or without 4 µg/mL minocycline. Percentage of resistant strains was determined by colony counting and taxonomy by checkerboard DNA-DNA hybridization. Significant clinical changes were sought with the Mann-Whitney test and differences in percentage of resistant isolates with the Friedman and Mann-Whitney tests. RESULTS: Groups showed similar clinical improvements. Mean percentage of resistant isolates rose at 1 month and decreased at 6 months in saliva and plaque samples in test group (P <0.05) but remained unchanged in control group. Percentage of resistant isolates of Gemella morbillorum and Eubacterium saburreum increased significantly at 6 months in both groups. Antibiotic resistance by Aggregatibacter actinomycetemcomitans, Tannerella forsythia, and Porphyromonas gingivalis was either absent or infrequent. CONCLUSION: Minocycline microspheres result in transient selection of minocycline resistant species in saliva and subgingival plaque samples.


Asunto(s)
Antibacterianos , Farmacorresistencia Bacteriana , Minociclina , Periodontitis/terapia , Aggregatibacter actinomycetemcomitans , Antibacterianos/uso terapéutico , Clostridiales , Gemella , Humanos , Microesferas , Minociclina/uso terapéutico , Porphyromonas gingivalis
4.
Compend Contin Educ Dent ; 38(8 Suppl): 22-25, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-29227113

RESUMEN

A better characterization of the peri-implant microbiome can improve the understanding of the etiology of peri-implant diseases. Ultimately, more detailed information about the peri-implantitis microbiome will lead to better strategies for prevention, supportive therapy, and risk assessment, as well as early diagnosis of peri-implantitis and timely intervention, all of which are critical for the long-term retention of implants.


Asunto(s)
Microbiota , Periimplantitis/microbiología , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Periimplantitis/historia , Periodontitis/microbiología
5.
J Clin Periodontol ; 44(12): 1274-1284, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28766745

RESUMEN

AIM: To compare the microbiome of healthy (H) and diseased (P) peri-implant sites and determine the core peri-implant microbiome. MATERIALS AND METHODS: Submucosal biofilms from 32 H and 35 P sites were analysed using 16S rRNA sequencing (MiSeq, Illumina), QIIME and HOMINGS. Differences between groups were determined using principal coordinate analysis (PCoA), t tests and Wilcoxon rank sum test and FDR-adjusted. The peri-implant core microbiome was determined. RESULTS: PCoA showed partitioning between H and P at all taxonomic levels. Bacteroidetes, Spirochetes and Synergistetes were higher in P, while Actinobacteria prevailed in H (p < .05). Porphyromonas and Treponema were more abundant in P while Rothia and Neisseria were higher in H (p < .05). The core peri-implant microbiome contained Fusobacterium, Parvimonas and Campylobacter sp. T. denticola, and P. gingivalis levels were higher in P, as well as F. alocis, F. fastidiosum and T. maltophilum (p < .05). CONCLUSION: The peri-implantitis microbiome is commensal-depleted and pathogen-enriched, harbouring traditional and new pathogens. The core peri-implant microbiome harbours taxa from genera often associated with periodontal inflammation.


Asunto(s)
Bacterias/clasificación , Bacterias/aislamiento & purificación , Implantes Dentales/microbiología , Microbiota/genética , Periimplantitis/microbiología , Adulto , Anciano , Pérdida de Hueso Alveolar/microbiología , Bacterias/genética , Carga Bacteriana , Secuencia de Bases , Biopelículas/crecimiento & desarrollo , Estudios de Casos y Controles , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Femenino , Humanos , Masculino , Consorcios Microbianos/genética , Persona de Mediana Edad , Bolsa Periodontal/microbiología , ARN Ribosómico 16S/genética
6.
J Clin Periodontol ; 37(4): 313-23, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-20447254

RESUMEN

AIM: To examine relationships between subgingival biofilm composition and levels of gingival crevicular fluid (GCF) cytokines in periodontal health and generalized aggressive periodontitis (GAP). MATERIALS AND METHODS: Periodontal parameters were measured in 25 periodontally healthy and 31 GAP subjects. Subgingival plaque and GCF samples were obtained from 14 sites from each subject. Forty subgingival taxa were quantified using checkerboard DNA-DNA hybridization and the concentrations of eight GCF cytokines were measured using Luminex. Cluster analysis was used to define sites with similar subgingival microbiotas in each clinical group. Significance of differences in clinical, microbiological and immunological parameters among clusters was determined using the Kruskal-Wallis test. RESULTS: GAP subjects had statistically significantly higher GCF levels of interleukin-1beta (IL-1beta) (p<0.001), granulocyte-macrophage colony-stimulating factor (GM-CSF) (p<0.01) and IL-1beta/IL-10 ratio (p<0.001) and higher proportions of Red and Orange complex species than periodontally healthy subjects. There were no statistically significant differences in the mean proportion of cytokines among clusters in the periodontally healthy subjects, while the ratio IL-1beta/IL-10 (p<0.05) differed significantly among clusters in the aggressive periodontitis group. CONCLUSIONS: Different subgingival biofilm profiles are associated with distinct patterns of GCF cytokine expression. Aggressive periodontitis subjects were characterized by a higher IL-1beta/IL-10 ratio than periodontally healthy subjects, suggesting an imbalance between pro- and anti-inflammatory cytokines in aggressive periodontitis.


Asunto(s)
Periodontitis Agresiva/inmunología , Placa Dental/microbiología , Líquido del Surco Gingival/inmunología , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interacciones Microbianas/inmunología , Adulto , Periodontitis Agresiva/metabolismo , Periodontitis Agresiva/microbiología , Bacterias/clasificación , Bacterias/genética , Biopelículas , Biomarcadores/análisis , Estudios de Casos y Controles , Análisis por Conglomerados , ADN Bacteriano/análisis , Placa Dental/inmunología , Femenino , Líquido del Surco Gingival/metabolismo , Líquido del Surco Gingival/microbiología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/análisis , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Interleucina-10/análisis , Interleucina-1beta/análisis , Masculino , Valores de Referencia , Curetaje Subgingival
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