Usefulness of matrix-assisted laser desorption ionization/time of flight mass spectrometry for the identification of Streptococcus mutans.

This study evaluated the reliability of MALDI-TOF MS coupled with statistical tools for the identification of Streptococcus mutans in comparison with PCR-based techniques. Bacterial isolates were identified and serotyped by conventional PCR, using S. mutans species and serotype-specific primers. For bacterial identification, mass spectra data from S. mutans and other streptococci were compared with Biotyper V 3.1 database and the mass peak lists were examined by cluster and principal component (PCA) analysis. Identification of potential biomarkers was performed using UniProtKB/Swiss-Prot and UniProtKB/TrEMBL databases and BLAST tool of the NCBI database. PCR identified 100% of the isolates as S. mutans. S. mutans strains were typed as serotypes c (85.6%), e (8.6%), k (4.8%), and f (0.9%). Although only the 70% of the strains tested were identified at species level by the Biotyper database, PCA and cluster analysis of mass peaks allowed the identification of 100% S. mutans isolates and its differentiation from the other oral and non-oral streptococci. One mass peak at m/z value of 9572.73 was identified as species-specific biomarker for S. mutans. No biomarkers were identified for S. mutans serotypes. KEY POINTS: • MALDI-TOF MS coupled with statistical tools for the identification of S. mutans. • Detection of species identifying biomarkers by MALDI-TOF MS. • PCR identification and serotyping of S. mutans from saliva samples.

Molecular and serological typing of Streptococcus mutans strains isolated from young Galician population: relationship with the oral health status.

The aim of this study was to determine the prevalence of Streptococcus mutans and its serotypes in samples from oral cavity of young Galician population and their relationship with the oral health state. The variables generally associated with dental caries, such as salivary flow rate, buffering capacity, eating habits, and lifestyle, were also analysed. No relationship was found between the variables studied and the presence of S. mutans in the oral cavity or the existence of dental caries. Presumptive strains of S. mutans were isolated from saliva samples from 48% of the analysed population. The use of conventional microbiological methods, API 20 Strep system, and species-specific polymerase chain reaction (PCR) allowed to substantiate the identity of the strains as S. mutans. Multiplex PCR protocols, developed in this study for the simultaneous detection of S. mutans and serotypes c, e, and f and for detection of S. mutans and serotype k, also confirmed this result and demonstrated that serotype c was predominant in the studied young Galician population (86%). Serotypes e (8%), k (3%), and f (2%) were also detected. Serotype c was detected in carious and caries-free subjects, while the remaining serotypes were only found in subjects with caries.

Effect of antiseptic gels in the microbiologic colonization of the suture threads after oral surgery.

Three different bioadhesive gels were evaluated in a double-blind randomized clinical trial in which microbial growth in the suture thread was assessed following post-surgical application of the aforementioned gels. Also assessed in this trial were, the intensity of post-surgical pain as well as the degree of healing of the patients' surgical wounds. A total of 21 patients (with 42 wisdom teeth) participated in this trial. Chlorhexidine gel, chlorhexidine-chitosan gel, and hyaluronic acid gel were evaluated, with a neutral water-based gel serving as the control agent. The aerobic and facultative anaerobic bacterial recovery on blood agar was lower in the placebo group than in the experimental groups. The most significant difference (p = 0.04) was observed in the chlorhexidine-chitosan group. in which the growth of Blood Agar and Mitis Salivarius Agar was significantly higher than in the placebo group. The intensity of post-surgical pain was very similar among all the groups. Significantly better healing rates were observed in the patients treated with chlorhexidine-chitosan gel when compared with those who used the placebo gel (p = 0.03), and in particular when compared with those patients who used hyaluronic acid gel (p = 0.01). Through our microbiological analyses, we were able to conclude that none of the bioadhesive gels tested resulted in beneficial reductions in the bacterial/fungal populations. However, the healing rates of patients who were treated with chlorhexidine-chitosan were better than those of the patients who used either the placebo gel or the hyaluronic acid gel.

Identificación de microorganismos por reacción en cadena de la polimerasa en necrosis pulpar y periodontitis apical / Identification of microorganisms by polymerase chain reaction in pulp necrosis and apical periodontitis

Objetivos: identificar, mediante la técnica de reacción en cadena de la polimerasa, diez especies bacterianas procedentes de conductos radiculares necróticos, así como analizar su asociación con signos y síntomas de la periodontitis apical (dolor, exudado y movilidad), dentro de un área geográfica específica, Santiago de Compostela (noroeste de España). Métodos: se realizó un estudio descriptivo, en un plazo de recogida de muestras de un año. Se extrajeron 43 muestras de los conductos radiculares necróticos, a razón de una muestra por pacientes, quienes fueron examinados previamente para determinar la presencia de periodontitis apical radiográfica, de la cual fueron descritos sus signos y síntomas. Se utilizaron puntas de papel absorbentes estériles y se procedió a extraer de dichas muestras el ADN por medio de la técnica del fenol-cloroformo. El producto se amplificó por medio de la reacción en cadena de la polimerasa, usando cebadores específicos para diez microorganismos. El resultado se visualizó por medio de electroforesis, utilizando un transiluminador UV. Los resultados se analizaron estadísticamente, para así establecer qué microorganismos estaban presentes de forma individual y cuáles se presentaron formando asociaciones como el llamado red complex, en relación con los signos y síntomas presentes en la periodontitis apical. Resultados: Fusobacterium nucleatum fue el microorganismo con presencia de forma individual más frecuente (83,72 por ciento de las muestras). En cuanto al estudio de la asociación de los microorganismos con los signos y síntomas, Porphyromonas endodontalis presentó una asociación significativa con respecto al exudado (p< 0,05) y movilidad (p< 0,05). En cambio, Enterococcus spp. y Treponema denticola (p< 0,01) presentaron ambos una asociación significativa con respecto al dolor. En cuanto a la incidencia del red complex fue de 6 casos en total. Conclusiones: Fusobacterium nucleatum y Streptococcus spp. son los microorganismos identificados con mayor porcentaje en comparación a los restantes estudiados. Porphyromonas endodontalis es el microorganismo con mayor asociación estadística con respecto a los signos y síntomas de la periodontitis apical(AU)

Objectives: identify by polymerase chain reaction technique ten bacterial species obtained from necrotic root canals, and analyze their association with signs and symptoms of apical periodontitis (pain, exudate and mobility) in a specific geographic area: Santiago de Compostela (northwestern Spain). Methods: a descriptive study was conducted based on a one-year sample collection period. Forty-three samples were taken from necrotic root canals, at a rate of one sample per patient, who had been previously examined for radiographic apical periodontitis, of which the signs and symptoms were described. Sterile absorbent paper points were used to extract the DNA samples, applying the phenol-chloroform technique. The product was amplified by polymerase chain reaction, using specific primers for ten microorganisms. The result was visualized by electrophoresis using a UV transilluminator. Outcomes were analyzed statistically to determine which microorganisms were present individually and which formed associations such as the so-called red complex, according to the signs and symptoms present in the apical periodontitis. Results: Fusobacterium nucleatum was the most common individual microorganism (83.72 percent of the samples). As to the association of microorganisms with signs and symptoms, Porphyromonas endodontalis was found to be significantly associated with exudate (p< 0.05) and mobility (p< 0.05), while both Enterococcus spp. and Treponema denticola (p< 0.01) had a significant association with pain. Incidence of the red complex was 6 cases in all. Conclusions: Fusobacterium nucleatum and Streptococcus spp. were the microorganisms identified as displaying the highest percentages. Porphyromonas endodontalis showed the greatest statistical association with signs and symptoms of apical periodontitis(AU)

Identificación de microorganismos por reacción en cadena de la polimerasa en necrosis pulpar y periodontitis apical / Identification of microorganisms by polymerase chain reaction in pulp necrosis and apical periodontitis

Objetivos: identificar, mediante la técnica de reacción en cadena de la polimerasa, diez especies bacterianas procedentes de conductos radiculares necróticos, así como analizar su asociación con signos y síntomas de la periodontitis apical (dolor, exudado y movilidad), dentro de un área geográfica específica, Santiago de Compostela (noroeste de España). Métodos: se realizó un estudio descriptivo, en un plazo de recogida de muestras de un año. Se extrajeron 43 muestras de los conductos radiculares necróticos, a razón de una muestra por pacientes, quienes fueron examinados previamente para determinar la presencia de periodontitis apical radiográfica, de la cual fueron descritos sus signos y síntomas. Se utilizaron puntas de papel absorbentes estériles y se procedió a extraer de dichas muestras el ADN por medio de la técnica del fenol-cloroformo. El producto se amplificó por medio de la reacción en cadena de la polimerasa, usando cebadores específicos para diez microorganismos. El resultado se visualizó por medio de electroforesis, utilizando un transiluminador UV. Los resultados se analizaron estadísticamente, para así establecer qué microorganismos estaban presentes de forma individual y cuáles se presentaron formando asociaciones como el llamado red complex, en relación con los signos y síntomas presentes en la periodontitis apical. Resultados: Fusobacterium nucleatum fue el microorganismo con presencia de forma individual más frecuente (83,72 por ciento de las muestras). En cuanto al estudio de la asociación de los microorganismos con los signos y síntomas, Porphyromonas endodontalis presentó una asociación significativa con respecto al exudado (p< 0,05) y movilidad (p< 0,05). En cambio, Enterococcus spp. y Treponema denticola (p< 0,01) presentaron ambos una asociación significativa con respecto al dolor. En cuanto a la incidencia del red complex fue de 6 casos en total. Conclusiones: Fusobacterium nucleatum y Streptococcus spp. son los microorganismos identificados con mayor porcentaje en comparación a los restantes estudiados. Porphyromonas endodontalis es el microorganismo con mayor asociación estadística con respecto a los signos y síntomas de la periodontitis apical(AU)

Objectives: identify by polymerase chain reaction technique ten bacterial species obtained from necrotic root canals, and analyze their association with signs and symptoms of apical periodontitis (pain, exudate and mobility) in a specific geographic area: Santiago de Compostela (northwestern Spain). Methods: a descriptive study was conducted based on a one-year sample collection period. Forty-three samples were taken from necrotic root canals, at a rate of one sample per patient, who had been previously examined for radiographic apical periodontitis, of which the signs and symptoms were described. Sterile absorbent paper points were used to extract the DNA samples, applying the phenol-chloroform technique. The product was amplified by polymerase chain reaction, using specific primers for ten microorganisms. The result was visualized by electrophoresis using a UV transilluminator. Outcomes were analyzed statistically to determine which microorganisms were present individually and which formed associations such as the so-called red complex, according to the signs and symptoms present in the apical periodontitis. Results: Fusobacterium nucleatum was the most common individual microorganism (83.72 percent of the samples). As to the association of microorganisms with signs and symptoms, Porphyromonas endodontalis was found to be significantly associated with exudate (p< 0.05) and mobility (p< 0.05), while both Enterococcus spp. and Treponema denticola (p< 0.01) had a significant association with pain. Incidence of the red complex was 6 cases in all. Conclusions: Fusobacterium nucleatum and Streptococcus spp. were the microorganisms identified as displaying the highest percentages. Porphyromonas endodontalis showed the greatest statistical association with signs and symptoms of apical periodontitis(AU)

Effect of zein on biodegradable inserts for the delivery of tetracycline within periodontal pockets.

Treatment with antibiotics within the periodontal pocket against bacterial infections represents a useful and adjunctive tool to conventional therapy for healing and teeth preservation. With this function in view, an implantable, tetracycline delivery device for the treatment of periodontal disease was developed. The aim of this study was to develop biodegradable, tetracycline-loaded microparticles made of two polymers: PLGA and zein which were compressed into monolithic devices. In this polymer delivery system, the encapsulation efficiency, release characteristics, drug-polymer interaction, and antibacterial activity of loaded drug were investigated. The interaction of tetracycline with the corn protein zein was studied by nuclear magnetic resonance (NMR), Fourier transform infrared, and X-ray diffraction. The hydrophobic interaction of tetracycline with zein in the formulations was deduced from the NMR studies, whereas X-ray diffraction studies showed a new crystalline state of the drug in the presence of the protein. Zein was not denatured by preparation of inserts. Sustained release of tetracycline was obtained, and the proportion of zein in the inserts had a great impact on the drug release. Finally, an effective tetracycline release from inserts against Staphylococcus aureus was achieved over 30 days. In conclusion, the PLGA:zein delivery system described in this study was found to be effective in controlled delivery of tetracycline, and hence may be suitable for intra-pocket delivery of antimicrobial agents in the treatment of periodontitis.