Evaluación antimicrobiana de rifampicina nanoencapsulada contra aggregatibacter actinomycetemcomitans presente en la periodontitis

Introducción. A pesar de los esfuerzos tanto de la medicina como de la industria farmacéutica, el incremento en la prevalencia de resistencia en bacterias patógenas frente a antibióticos se ha vuelto uno de los mayores problemas en la medicina moderna. El área odontológica tampoco se encuentra exenta, siendo común el uso excesivo de antibióticos lo que contribuye al desarrollo de resistencia antimicrobiana. La primera etapa para el desarrollo de la enfermedad periodontal es la formación de un biofilm de bacterias periodontopatógenas, siendo el Aggregaribacter actinomycetemcomitans (A.a) uno de los más asociados a dicha enfermedad. El tratamiento de esta patología se basa en remover mecánicamente la placa dentobacteriana y, en segunda instancia, en el apoyo de terapia antimicrobiana para coadyuvar la eliminación de las bacterias periodontopatógenas, cuales tienen gran similitud con Mycobacterium tuberculosis. La rifampicina es uno de los antibióticos efectivos contra bacterias multi-resistentes y la primera elección en el tratamiento de tuberculosis activa. Con el fin de mejorar la terapia farmacológica y evadir la resistencia del agente infectivo, se han propuesto nuevas estrategias basadas en sistemas de liberación controlada. Entre los más estudiados en los últimos 10 años se encuentran las nanopartículas poliméricas. El objetivo del presente estudio fue evaluar la actividad antimicrobiana de la rifampicina nanoencapsulada contra el A.a presente en la periodontitis. Materiales y Métodos. Para el estudio, Se tomaron muestras de fluido crevicular en pacientes con bolsas periodontales de 5-10 mm de profundidad. Se inoculo caldo de tripticaseina de soya (TCS) con las muestras tomadas y se incubaron a 37 ° C en condiciones aeróbicas por 7 días. La presencia de Aggregatibacter actinomycetemcomitans (A.a) fue determinado mediante PCR en tiempo real. La Concentración Mínima Inhibitoria (MIC) de rifampicina para interferir con el crecimiento de bacterias orales fue determinada mediante la técnica de dilución de tubos. Posteriormente se prepararon mediante la técnica de nanoprecipitación NP de Eudragit® EPO, L100-55 y PLA entre 100 y 200 nm y su IP con distribución de tamaño homogéneo. Resultados. A.a fue detectado en muestras de fluido crevicular en pacientes con periodontitis, corroborando su asociación con dicha patología. La efectividad de la rifampicina libre contra bacterias orales fue confirmada, obteniéndose una CMI de 1 µg/ml. Las NP con Rifampicina se ajustaron a la misma CMI que la Rif libre. Las NP de Eudragit® EPO cargadas con Rif mostraron que la liberación de la Rif de la NP fue inmediata, mientras que el Eudragit® L100-55 y PLA con Rif no mostró inhibición durante los 5 días de incubación. Esto hace suponer que el fármaco no fue liberado o solo se liberó en una baja proporción que no permitió llegar a la CMI. Conclusión. La rifampicina es una excelente alternativa terapéutica para el tratamiento de la enfermedad periodontal, promoviendo resultados favorables en la evaluación clínica de pacientes. Sería interesante continuar con estudios utilizando otro polímero o mezcla de ellos para favorecer la liberación del fármaco en la NP.

Reabsorción radicular externa apical en incisivos y caninos superiores e inferiores sometidos a tratamiento de Ortodoncia en etapa inicial

La Reabsorción radicular externa apical es un proceso inflamatorio común asociado con el tratamiento de ortodoncia. El objetivo de este estudio fue comparar la magnitud de la RREA de los incisivos y caninos superiores e inferiores en pacientes sometidos a la fase inicial del tratamiento de ortodoncia con tres diferentes tipos de brackets. La muestra consistió en 23 casos de pacientes con edades comprendidas entre 12 y 27 años, fueron divididos aleatoriamente en 3 grupos. Grupo I (n=9; 5 mujeres y 4 varones), utilizando brackets pasivos de autoligado Damon Q Grupo II (n= 8; 4 mujeres y 4 varones) utilizando brackets convencionales prescripción Roth y MBT. Grupo III (n=6; 4 mujeres y 2 varones) utilizando brackets Biofuncional QR. Se analizaron 264 dientes (caninos e incisivos superiores e inferiores) mediante el uso de tomografía computarizada de haz cónico (NewTom VGi Cone Beam 3D Imaging), con el escáner NNT Viewer (versión 4.6 NewTom). Los resultados obtenidos indican que la media de reabsorción se mostró distinta para los diferentes protocolos de tratamiento, así como diferente de acuerdo a la pieza analizada, sin embargo, la prueba de ANOVA no encontró diferencias estadísticamente significativas al comparar la media de reabsorción de cada pieza por tipo de brackets empleados o tratamiento realizado (p >0,05). Se concluye que la RREA se ha producido en todos los dientes evaluados, durante los 6 primeros meses de tratamiento y el diseño del bracket (autoligado pasivo, convencional o QR) no influye en el grado de reabsorción radicular.

Morphogenesis of the Arabidopsis Shoot Apical Meristem

Phyllotaxis patterns in plants, or the arrangement of leaves and flowers radially around the shoot, have fascinated both biologists and mathematicians for centuries. The current model of this process involves the lateral transport of the hormone auxin through the first layer of cells in the shoot apical meristem via the auxin efflux carrier protein PIN1. Locations around the meristem with high auxin concentration are sites of organ formation and differentiation. Many of the molecular players in this process are well known and characterized. Computer models composed of all these components are able to produce many of the observed phyllotaxis patterns. To understand which parts of this model have a large effect on the phenotype I automated parameter testing and tried many different parameter combinations. Results of this showed that cell size and meristem size should have the largest effect on phyllotaxis. This lead to three questions: (1) How is cell geometry regulated? (2) Does cell size affect auxin distribution? (3) Does meristem size affect phyllotaxis? To answer the first question I tracked cell divisions in live meristems and quantified the geometry of the cells and the division planes using advanced image processing techniques. The results show that cell shape is maintained by minimizing the length of the new wall and by minimizing the difference in area of the daughter cells. To answer the second question I observed auxin patterning in the meristem, shoot, leaves, and roots of Arabidopsis mutants with larger and smaller cell sizes. In the meristem and shoot, cell size plays an important role in determining the distribution of auxin. Observations of auxin in the root and leaves are less definitive. To answer the third question I measured meristem sizes and phyllotaxis patterns in mutants with altered meristem sizes. These results show that there is no correlation between meristem size and average divergence angle. But in an extreme case, making the meristem very small does lead to a switch on observed phyllotaxis in accordance with the model.

Microbial Evaluation Of Traumatized Teeth Treated With Triple Antibiotic Paste Or Calcium Hydroxide With 2% Chlorhexidine Gel In Pulp Revascularization.

Revascularization outcome depends on microbial elimination because apical repair will not happen in the presence of infected tissues. This study evaluated the microbial composition of traumatized immature teeth and assessed their reduction during different stages of the revascularization procedures performed with 2 intracanal medicaments. Fifteen patients (7-17 years old) with immature teeth were submitted to the revascularization procedures; they were divided into 2 groups according to the intracanal medicament used: TAP group (n = 7), medicated with a triple antibiotic paste, and CHP group (n = 8), dressed with calcium hydroxide + 2% chlorhexidine gel. Samples were taken before any treatment (S1), after irrigation with 6% NaOCl (S2), after irrigation with 2% chlorhexidine (S3), after intracanal dressing (S4), and after 17% EDTA irrigation (S5). Cultivable bacteria recovered from the 5 stages were counted and identified by means of polymerase chain reaction assay (16S rRNA). Both groups had colony-forming unit counts significantly reduced after S2 (P < .05); however, no significant difference was found between the irrigants (S2 and S3, P = .99). No difference in bacteria counts was found between the intracanal medicaments used (P = .95). The most prevalent bacteria detected were Actinomyces naeslundii (66.67%), followed by Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum, which were detected in 33.34% of the root canals. An average of 2.13 species per canal was found, and no statistical correlation was observed between bacterial species and clinical/radiographic features. The microbial profile of infected immature teeth is similar to that of primarily infected permanent teeth. The greatest bacterial reduction was promoted by the irrigation solutions. The revascularization protocols that used the tested intracanal medicaments were efficient in reducing viable bacteria in necrotic immature teeth.

Efficacy Of Ethylene-diamine-tetra-acetic Acid Associated With Chlorhexidine On Intracanal Medication Removal: A Scanning Electron Microscopy Study.

The aim of this study was to evaluate the effectiveness of 17% ethylene-diamine-tetra-acetic acid (EDTA) used alone or associated with 2% chlorhexidine gel (CHX) on intracanal medications (ICM) removal. Sixty single-rooted human teeth with fully formed apex were selected. The cervical and middle thirds of each canal were prepared with Gates Glidden drills and rotary files. The apical third was shaped with hand files. The specimens were randomly divided into two groups depending on the ICM used after instrumentation: calcium hydroxide Ca(OH)(2) +CHX or Ca(OH)(2) +sterile saline (SS). After seven days, each group was divided into subgroups according to the protocol used for ICM removal: instrumentation and irrigation either with EDTA, CHX+EDTA, or SS (control groups). All specimens were sectioned and processed for observation of the apical thirds by using scanning electron microscopy. Two calibrated evaluators attributed scores to each specimen. The differences between the protocols for ICM removal were analyzed with Kruskal-Wallis and Mann-Whitney U tests. Friedman and Wilcoxon signed rank tests were used for comparison between the score of debris obtained in each root canal third. Remains of Ca(OH)(2) were found in all specimens independently of the protocol and ICM used (P > 0.05). Seventeen percent EDTA showed the best results in removing ICM when used alone (P < 0.05), particularly in those associated with CHX. It was concluded that the chelating agent 17% EDTA significantly improved the removal of ICM when used alone. Furthermore, the type of the vehicle associated with Ca(OH)(2) also plays a role in the ICM removal.

Photoelastic Analysis Of All-on-four Concept Using Different Implants Angulations For Maxilla.

Conventional tilted implants are used in oral rehabilitation for heavily absorbed maxilla to avoid bone grafts; however, few research studies evaluate the biomechanical behavior when different angulations of the implants are used. The aim of this study was evaluate, trough photoelastic method, two different angulations and length of the cantilever in fixed implant-supported maxillary complete dentures. Two groups were evaluated: G15 (distal tilted implants 15°) and G35 (distal tilted implants 35°) n = 6. For each model, 2 distal tilted implants (3.5 x 15 mm long cylindrical cone) and 2 parallel tilted implants in the anterior region (3.5 x 10 mm) were installed. Photoelastic models were submitted to three vertical load tests: in the end of cantilever, in the last pillar and in the all pillars at the same time. We obtained the shear stress by Fringes software and found values for total, cervical and apical stress. The quantitative analysis was performed using the Student tests and Mann-Whitney test; p ≥ 0.05. There is no difference between G15 and G35 for total stress regardless of load type. Analyzing the apical region, G35 reduced strain values considering the distal loads (in the cantilever p = 0.03 and in the last pillar p = 0.02), without increasing the stress level in the cervical region. Considering the load in all pillars, G35 showed higher stress concentration in the cervical region (p = 0.04). For distal loads, G15 showed increase of tension in the apical region, while for load in all pillars, G35 inclination increases stress values in the cervical region.

Tissue culture of Cecropia glaziovii Sneth (urticaceae): vegetative micropropagation and plant regeneration from callus

Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS) medium supplemented with 6-benzylaminopurine alone (BAP) (1.0, 5.0 or 10.0 mg L-1) or combined with -naphthalene acetic acid (NAA) (1.0 or 2.0 mg L-1), after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D) (5.0 mg L-1) combined with BAP (1.0 mg L-1). Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA) (0.1 mg L-1) alone or combined with 2,4-D (1.0 or 5.0 mg L-1) was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA) (1.0 mg L-1). Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.

Influence of different adhesive systems on the pull-out bond strength of glass fiber posts

This in vitro study evaluated the tensile bond strength of glass fiber posts (Reforpost - Angelus-Brazil) cemented to root dentin with a resin cement (RelyX ARC - 3M/ESPE) associated with two different adhesive systems (Adper Single Bond - 3M/ESPE and Adper Scotchbond Multi Purpose (MP) Plus - 3M/ESPE), using the pull-out test. Twenty single-rooted human teeth with standardized root canals were randomly assigned to 2 groups (n=10): G1- etching with 37% phosphoric acid gel (3M/ESPE) + Adper Single Bond + #1 post (Reforpost - Angelus) + four #1 accessory posts (Reforpin - Angelus) + resin cement; G2- etching with 37% phosphoric acid gel + Adper Scotchbond MP Plus + #1 post + four #1 accessory posts + resin cement. The specimens were stored in distilled water at 37°C for 7 days and submitted to the pull-out test in a universal testing machine (EMIC) at a crosshead speed of 0.5 mm/min. The mean values of bond strength (kgf) and standard deviation were: G1- 29.163 ± 7.123; G2- 37.752 ±13.054. Statistical analysis (Student's t-test; a=0.05 showed no statistically significant difference (p<0.05) between the groups. Adhesive bonding failures between resin cement and root canal dentin surface were observed in both groups, with non-polymerized resin cement in the apical portion of the post space when Single Bond was used (G1). The type of adhesive system employed on the fiber post cementation did not influence the pull-out bond strength.

Short-Term effect of COX-2 selective inhibitor as an adjunct for the treatment of periodontal disease: a clinical double-blind study in humans

Adjunctive therapeutic strategies that modulate the inflammatory mediators can play a significant role in periodontal therapy. In this double-blind, placebo-controlled study, 60 subjects diagnosed as periodontitis patients were evaluated for 28 days after periodontal treatment combined with selective cyclooxygenase-2 (COX-2) inhibitor. The experimental group received scaling and root planning (SRP) combined with the Loxoprofen antiinflammatory drug (SRP+Loxoprofen). The control group received SRP combined with placebo (SRP+placebo). Plaque index (PI), probing pocket depth (PD) and bleeding on probing (BOP) were monitored with an electronic probe at baseline and after 14 and 28 days. Both groups displayed clinical improvement in PD, PI and BOP. They also showed statistically similar values (p>0.05) of PD reduction on day 14 (0.4 mm) and on day 28 (0.6 mm). At the baseline, few deeper sites (>7 mm) from SRP+Loxoprofen group were responsible and most PD reduction was observed after 14 days (p<0.05). The percentage of remaining deep pockets (>7 mm) after 14 days in the SRP+Loxoprofen group was significantly lower (p<0.05) than in the SRP+placebo group. Loxoprofen presents potential effect as an adjunct of periodontal disease treatment, but long-term clinical trials are necessary to confirm its efficacy.

Histopathological evaluation of root canal filling materials for primary teeth

This study aimed to assess the response of apical and periapical tissues of dogs' teeth after root canal filling with different materials. Forty roots from dogs' premolars were prepared biomechanically and assigned to 4 groups filled with: Group I: commercial calcium hydroxide and polyethylene glycol-based paste (Calen®) thickened with zinc oxide; Group II: paste composed of iodoform, Rifocort® and camphorated paramonochlorophenol; Group III: zinc oxide-eugenol cement; Group IV: sterile saline. After 30 days, the samples were subjected to histological processing. The histopathological findings revealed that in Groups I and IV the apical and periapical regions exhibited normal appearance, with large number of fibers and cells and no resorption of mineralized tissues. In Group II, mild inflammatory infiltrate and mild edema were observed, with discrete fibrogenesis and bone resorption. Group III showed altered periapical region and thickened periodontal ligament with presence of inflammatory cells and edema. It may be concluded that the Calen paste thickened with zinc oxide yielded the best tissue response, being the most indicated material for root canal filling of primary teeth with pulp vitality.

Periapical repair after root canal filling with different root canal sealers

The aim of this study was to evaluate periapical repair after root canal filling with different endodontic sealers. Sixty-four root canals from dog´s teeth were filled, divided into 4 groups (n=16). Root canals were instrumented with K-type files and irrigated with 1% sodium hypochlorite solution. Root canals were filled in the same session by active lateral condensation of the cones and sealers: Intrafill, AH Plus, Roeko Seal and Resilon/Epiphany System. After 90 days, the animals were euthanized and the tissues to be evaluated were processed and stained with hematoxylin and eosin. For histopathological analysis, the following parameters were evaluated: inflammatory process, mineralized tissue resorption, and apical mineralized tissue deposition. Histopathological analysis demonstrated that Intrafill had less favorable results in terms of apical and periapical repair, compared to the other sealers (p<0.05). AH Plus, Roeko Seal, and Epiphany sealers had similar and satisfactory results (p>0.05). In conclusion, AH Plus and the materials Roeko Seal and Epiphany are good options for clinical use in Endodontics.

Diagnosis and root canal treatment in a mandibular premolar with three canals

This paper presents a case report of a left mandibular second premolar with three canals and three different apical foramina. A 39-year-old male patient presented to our clinic with pain in the mandibular left second premolar. Initially, pain was caused by cold stimulus and later was spontaneously. The intraoral clinical examination revealed a fractured amalgam restoration with occlusal caries. Percussion and cold (Endo-Frost) tests were positive. The radiographic examination showed the presence of two roots. The probable diagnosis was an acute pulpitis. After access cavity, it was observed remaining roof of the pulp chamber and mild bleeding in the tooth lingual area, indicating the possible presence of a third canal. The endodontic treatment was completed in a single session using Root ZX apex locator and K3 NiTi rotary system with surgical diameter corresponding to a .02/45 file in the three canals and irrigation with 1% sodium hypochlorite. The canals were obtured with gutta-percha cones and Sealer 26 using the lateral condensation technique. After 1 year of follow-up, the tooth was asymptomatic and periapical repair was observed radiographically. Internal alterations should be considered during the endodontic treatment of mandibular second premolars. The correct diagnosis of these alterations by the analysis of preoperative radiographs can help the location of two or more canals, thereby avoiding root therapy failure.

Immediate implants placed into infected sockets: a case report with 3-year follow-up

The esthetics and functional integrity of the periodontal tissue may be compromised by dental loss. Immediate implants became a viable option to maintain the periodontal architecture because of their anatomic compatibility with the dental socket and the possibility of eliminating local contamination. This article describes the procedure of immediate implant placement in the anterior maxilla replacing teeth with chronic periapical lesions, which were condemned due to endodontic lesions persisting after failed endodontic treatment and endodontic surgery, and discusses the relationship between the procedure and periapical lesions. Surgical removal of hopeless teeth 11, 12 and 21 was performed conservatively in such a way to preserve the anatomy and gingival esthetics. A second surgical access was gained at the apical level, allowing the debridement of the surgical chamber for elimination of the periapical lesion, visual orientation for setting of the implants and filling of the surgical chamber with xenogenous bovine bone graft. After this procedure, the bone chamber was covered with an absorbent membrane and the healing screws were positioned on the implants. Later, a provisional partial removable denture was installed and the implants were inserted after 6 months. After 3 years of rehabilitation, the implants present satisfactory functional and esthetic conditions, suggesting that immediate implant placement combined with guided bone regeneration may be indicated for replacing teeth lost due to chronic periapical lesions with endodontic failure history in the anterior maxilla.

Effectiveness of different final irrigation protocols in removing debris in flattened root canals

This study evaluated in vitro the capacity of debris removal from the apical third of flattened root canals, using different final irrigation protocols. Thirty human mandibular central incisors with a mesiodistal flattened root were prepared using rotary instrumentation by Endo-Flare 25.12 and Hero 642 30.06, 35.02, 40.02 files, irrigated with 2 mL of 1% NaOCl after each file. The specimens were randomly distributed into 5 groups according to the final irrigation of root canals: Group I: 10 mL of distilled water (control), Group II: 10 mL of 1% NaOCl for 8 min, Group III: 2 mL of 1% NaOCl for 2 min (repeated 4 times), Group IV: 10 mL of 2.5% NaOCl for 8 min, and Group V: 10 mL of 2.5% NaOCl for 2 min (repeated 4 times). The apical thirds of the specimens were subjected to histological processing and 6-μm cross-sections were obtained and stained with hematoxylin-eosin. The specimens were examined under optical microscopy at ×40 magnification and the images were subjected to morphometric analysis using the Scion image-analysis software. The total area of root canal and the area with debris were measured in square millimeters. Analysis of variance showed no statistically significant difference (p>0.05) among the groups GI (2.39 ± 3.59), GII (2.91 ± 2.21), GIII (0.73 ± 1.36), GIV (0.95 ± 0.84) and GV (0.51 ± 0.22). In conclusion, the final irrigation protocols evaluated in this study using the Luer syringe presented similar performance in the removal of debris from the apical third of flattened root canals.

Method for determination of root curvature radius using cone-beam computed tomography images

This article describes and discusses a method to determine root curvature radius by using cone-beam computed tomography (CBCT). The severity of root canal curvature is essential to select instrument and instrumentation technique. The diagnosis and planning of root canal treatment have traditionally been made based on periapical radiography. However, the higher accuracy of CBCT images to identify anatomic and pathologic alterations compared to panoramic and periapical radiographs has been shown to reduce the incidence of false-negative results. In high-resolution images, the measurement of root curvature radius can be obtained by circumcenter. Based on 3 mathematical points determined with the working tools of Planimp® software, it is possible to calculate root curvature radius in both apical and coronal directions. The CBCT-aided method for determination of root curvature radius presented in this article is easy to perform, reproducible and allows a more reliable and predictable endodontic planning, which reflects directly on a more efficacious preparation of curved root canals.