Person: Sanz Alonso, Mariano
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First Name
Mariano
Last Name
Sanz Alonso
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Odontología
Department
Especialidades Clínicas Odontológicas
Area
Estomatología
Identifiers
3 results
Search Results
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Item Detection and quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis in blood samples with different microbiological identification methods: An in vitro study(Archives of Oral Biology, 2017) Marín Cuenda, María José; Ambrosio Elejalde, Nagore; Virto Ruiz, Leire; Diz, Pedro; Álvarez, Maximiliano; Herrera, David; Sanz Alonso, Mariano; Figuero Ruiz, ElenaBackground: Culture-based methods (culture broth bottles or lysis methods) have been the standard for detecting bacteremia. More recently, quantitative polymerase chain reaction (qPCR) was proposed as a more sensitive and specific test although none of them has been validated for the identification of periodontal pathogens (fastidious growing bacteria) in blood samples. Objective: To compare the ability to detect and quantify Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis (alone or in combination) in blood samples with three culture techniques [direct anaerobic culturing (DAC), haemo-culture (BACTEC), and lysis-centrifugation (LC)] and a non-culture dependent approach (qPCR) in an in vitro study. Material and methods: Blood samples from 12 periodontally healthy volunteers were contaminated with three concentrations [104,102 and 101 colony forming units (CFU)/mL] of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination. Samples were analysed by DAC, BACTEC, LC and qPCR. Sensitivity, specificity, predictive values, kappa index and Lińs correlation coefficients were calculated. Results: DAC, LC and qPCR were able to detect the three target species at all concentrations. An excellent concordance (correlation coefficient r: 0.92-1) was observed between DAC and the reference standard (sensitivity raging 93.33-100% and specificity 88.89-100%) values. BACTEC was not able to identify P. gingivalis in any of the performed experiments. qPCR provided false negative results for S.oralis. Conclusions: DAC showed the best results for the proper identification and quantification of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination, in blood samples.Item An in vitro biofilm model associated to dental implants: Structural and quantitative analysis of in vitro biofilm formation on different dental implant surfaces(Dental Materials, 2014) Sánchez Beltrán, María Del Carmen; Llama-Palacios, Arancha; Fernández, Eva; Figuero Ruiz, Elena; Marín Cuenda, María José; León, Rubén; Blanc, Vanesa; Herrera González, David; Sanz Alonso, MarianoObjectives: The impact of implant surfaces in dental biofilm development is presently unknown. The aim of this investigation was to assess in vitro the development of a complex biofilm model on titanium and zirconium implant surfaces, and to compare it with the same biofilm formed on hydroxyapatite surface. Methods: Six standard reference strains were used to develop an in vitro biofilm over sterile titanium, zirconium and hydroxyapatite discs, coated with saliva within the wells of pre-sterilized polystyrene tissue culture plates. The selected species used represent initial (Streptococcus oralis and Actinomyces naeslundii), early (Veillonella parvula), secondary (Fusobacterium nucleatum) and late colonizers (Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans). The developed biofilms (growth time 1 to 120h) were studied with confocal laser scanning microscopy using a vital fluorescence technique and with low-temperature scanning electron microscopy. The number (colony forming units/biofilm) and kinetics of the bacteria within the biofilm were studied with quantitative PCR (qPCR). As outcome variables, the biofilm thickness, the percentage of cell vitality and the number of bacteria were compared using the analysis of variance. Results: The bacteria adhered and matured within the biofilm over the three surfaces with similar dynamics. Different surfaces, however, demonstrated differences both in the thickness, deposition of the extracellular polysaccharide matrix as well as in the organization of the bacterial cells. Significance: While the formation and dynamics of an in vitro biofilm model was similar irrespective of the surface of inoculation (hydroxyapatite, titanium or zirconium), there were significant differences in regards to the biofilm thickness and three-dimensional structure.Item Antibacterial effects of polymeric PolymP-n Active nanoparticles. An in vitro biofilm study(Dental Materials, 2019) Sánchez Beltrán, María Del Carmen; Toledano-Osorio, Manuel; Bueno, Jaime; Figuero Ruiz, Elena; Toledano, Manuel; Medina-Castillo, A.L.; Osorio, Raquel; Herrera González, David; Sanz Alonso, MarianoObjective: to study the antibacterial effect of polymeric PolymP-n Active nanoparticles using an in vitro subgingival biofilm model. Methods: Hydroxyapatite discs coated with five modalities of nanoparticles (NPs): NPs, NPs doped with zinc, calcium, silver and doxycycline, PBS as control, and Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were studied in a static in vitro biofilm model (12, 24, 48, and 72h). Nano-roughness of the different disc surfaces (SRa, in nm) and morphological characteristic of the biofilms (thickness (μm) and bacterial viability) were studied by different microscopy modalities. Quantitative Polymerase Chain Reaction was used to assess the effect of the nanoparticles on the bacterial load (colony forming unit per milliliter) (CFUmL-1). Analysis of variance and post-hoc testing with T3 Dunnett́s, and Student Newman Keuls correction was used. Results were considered statistically significant at p<0.05. Results: Surfaces containing the different nanoparticles showed significant increments in roughness when compared to controls (p<0.05). A similar biofilm formation and dynamics was observed, although reductions in bacterial viability were detected in biofilms in contact with the different nanoparticles, more pronounced with silver and doxycycline NPs. Doxycycline-NPs biofilms resulted in unstructured biofilm formation and significantly lower number of the six species when compared with the other nanoparticles specimens and controls (p<0.001 in all cases). Significance: Polymeric PolymP-n Active nanoparticles when combined with silver and doxycycline showed a significant antibacterial effect when tested in an in vitro subgingival biofilm model.