Behavior of Mesenchymal Stem Cells Obtained From Dental Tissues: A Review of the Literature / Comportamiento de células mesenquimales obtenidas de tejidos dentales: Revisión de literatura

Abstract 20. The success of tissue engineering in combination with tissue regeneration depends on the behavior and cellular activity in the biological processes developed within a structure that functions as a support, better known as scaffolds, or directly at the site of the injury. The cell-cell and cell-biomaterial interaction are key factors for the induction of a specific cell behavior, together with the bioactive factors that allow the formation of the desired tissue. Mesenchymal Stem Cells (MSC) can be isolated from the umbilical cord and bone marrow; however, the behavior of Dental Pulp Stem Cells (DPSC) has been shown to have a high potential for the formation of bone tissue, and these cells have even been able to induce the process of angiogenesis. Advances in periodontal regeneration, dentin-pulp complex, and craniofacial bone defects through the induction of MSC obtained from tooth structures in in vitro-in vivo studies have permitted the obtaining of clinical evidence of the achievements obtained to date.

Resumen 24. El éxito de la ingeniería de tejidos en combinación con la regeneración de tejidos depende del comportamiento y la actividad celular en los procesos biológicos desarrollados dentro de una estructura que funciona como soporte mejor conocida como andamio o directamente en el sitio de la lesión. La interación célula-célula y célula-biomaterial son factores claves para la inducción a un comportamiento célular específico junto con factores bioactivos que permitan la formación del tejido deseado. Las células troncales mesenquimales (MSCs) pueden ser aisladas del cordón umbilical y de la medula ósea, sin embargo, el comportamiento de las células troncales de pulpa dental (DPSCs) han demostrado tener un alto potencial para la formación de tejido óseo e incluso han logrado inducir el proceso de angiogénesis. Avances en la regeneración periodontal, complejo dentino-pulpar y defectos óseos craneofaciales a travez de la inducción de MSCs obtenidas de estructuras de dientes en estudios in vitro-in vivo han permitido obtener evidencia clínica de los logros obtenidos hasta el momento.

Trans-Endodontic Zirconium Oxide Implants: A Clinical Alternative / Implantes trans-endodónticos de óxido de zirconia: Una alternativa clínica

Abstract 24. Dental trauma, mainly from the anterior area of the mouth in the incisor teeth at the level of the radicular third, has the extraction of the dental organ as an elective treatment, depending on the severity of the lesion and the site where it presented. This paper reports two cases: one in a young 13-year-old male patient with dental trauma in central incisors, and another in an 18-year-old female patient with radicular reabsorption in central incisors. Both were treated using trans-endodontic implants on teeth 2.1 and 1.1-2.2 each case, respectively. The treatment approach proposed for each case provided good functional and esthetic outcomes. Clinical and radiographic results after 1 year were successful.

Resumen 28. El traumatismo dental del área anterior de la boca en los dientes incisivos a nivel del tercio radicular, indica la extracción del órgano dental como un tratamiento electivo dependiendo de la gravedad de la lesión y el sitio donde se presentó. Este artículo reporta dos casos: uno en un paciente masculino joven de 13 años con trauma dental en los incisivos centrales y otro en una paciente de 18 años con reabsorción radicular en los incisivos centrales. Ambos fueron tratados con implantes trans-endodónticos en los dientes 2.1 y 1.1-2.2 en cada caso, respectivamente. El enfoque de tratamiento propuesto para cada caso proporcionó buenos resultados funcionales y estéticos. Los resultados clínicos y radiográficos después de 1 año fueron exitosos.

Cytotoxicity, Cell Adhesion, and Apoptotic Gene Expression in Periodontal Ligament Fibroblasts treated with Endodontic Sealers.

OBJECTIVE: The present study assessed cytotoxicity, cell adhesion, and apoptotic gene expression in periodontal ligament fibroblasts (PLF) treated with 2 endodontic sealers. METHODS: PLF cells were obtained from nonerupted third molars and cultured. MTS and LIVE/DEAD assays were performed using different treatments and time periods. Cellular adhesion was evaluated using immunocytochemistry for integrin ß1 and vinculin expression, and the gene expressions of nuclear factor kB (NF-кB), P53, and apoptotic protease-activating factor 1 (Apaf-1) were evaluated using PCR. RESULTS: Cell proliferation at 12, 24, and 48 h was statistically significant in the control and PLF groups receiving different treatments; PLF treated with culture medium containing non-hardened (NH) sealers showed a decrease in the number of cells. PLF treated with culture medium containing hardened (H) sealers also exhibited a decreased cell population. Integrin ß1 and vinculin were expressed in both cell cultures treated with Acroseal (NH and H); however, the cell morphology changed and the cell population decreased. The gene expression of NF-kB and that of P53 were significantly different between the control group and the groups treated with the different sealers; mineral trioxide aggregate (MTA) (NH and H) inhibited Apaf-1, and PLF treated with Acroseal H exhibited increased Apaf-1 expression. CONCLUSION: Both sealers showed a certain level of cytotoxicity. The gene expression of NF-κB and P53 in PLF treated with the sealers showed significant changes compared to that of the control group, and MTA inhibited Apaf-1.

Chemical and Microscopy Characterization of Trans-Endodontic Implants / Caracterización química y microscópica de implantes trans-endodónticos

Abstract Trans-endodontic implants are an artificial extension through root apex anchored in periradicular bone tissue. The aim is to improve the crown-root ratio and to provide stability to dental organ present. Zirconium oxide (ZrO2) is a material of great technological importance, having good natural color, high strength, high toughness, high chemical stability, does not suffer any corrosion, chemical and microbial resistance and excellent esthetic properties. Objective: The aim of this study was to evaluate chemical and microscopy of surface conditions of ZrO2 trans-endodontic implant. Materials and Methods: A blocks of ZrO2 were manufactured for produce trans-endodontic implants and divided in two groups: monoclinic and tetragonal phase. They were evaluated using Scanning Electroning Microscope (SEM), EnergyDispersive X-ray Spectroscopy (EDS), and Atomic Force Microscope (AFM) and Vickers Micro hardness. Results: The Monoclinic phase through AFM analysis showed roughness Ra = 0.320μm, whereas in the Tetragonal phase was 0.126μm, SEM/EDX indicated that the phases are not properly uniform and the addition of the Yttrium to favor the stabilization of the Tetragonal phase. The Vickers hardness analysis showed a value of 1500HV. Conclusion: The characterization of the surface of trans-endodontic zirconium oxide implants provides a guideline to know the surface characteristics of the material, since a greater roughness on the surface of the implant will favor the Osseo-integration capacity.

Resumen Los implantes trans-endodónticos son una extensión artificial a través del ápice radicular anclado en el tejido óseo periradicular. El objetivo es mejorar la relación corona-raíz y proporcionar estabilidad al órgano dental presente. El óxido de zirconio (ZrO2) es un material de gran importancia tecnológica, con buen color natural, alta resistencia, alta tenacidad, alta estabilidad química, no sufre corrosión, resistencia química y microbiana y excelentes propiedades estéticas. Objetivo: El objetivo de este estudio fue evaluar las condiciones superficiales de ZrO2 para su aplicación clínica a los implantes transendodónticos. Materiales y Métodos: se trituraron bloques de ZrO2 en implantes trans-endodónticos y se dividieron en: monoclínico y tetragonal. Luego se evaluaron mediante microscopía electrónica de barrido (SEM), espectroscopia de rayos X de energía dispersiva (EDS) y microscopio de fuerza atómica (AFM) y microdureza vickers. Resultados: La fase monoclínica a través del análisis AFM presenta Ra = 0.320 μm, mientras que en la fase Tetragonal es 0.126 μm, SEM / EDS muestra que las fases no son adecuadamente uniformes y la adición del Ytrio para favorecer la estabilización de la fase tetragonal. El análisis de microdureza mostro un valor de 1500HV. Conclusión: La caracterización de la superficie de los implantes trans-endodónticos de óxido de zirconio, brinda una pauta para conocer las características superficiales del material, ya que al haber una mayor rugosidad en la superficie del implante se verá favorecida la capacidad de oseointegración.

Bioactive Tricalcium Silicate-based Dentin Substitute as an Indirect Pulp Capping Material for Primary Teeth: A 12-month Follow-up.

PURPOSE: The purpose of this study was to evaluate the clinical and radiographic outcomes of a bioactive tricalcium silicate [Ca3SiO5]-based dentin substitute and a light-activated calcium hydroxide [Ca(OH)2]-based liner as indirect pulp treatment (IPT) interventions for vital primary molars with carious lesions approaching the pulp. METHODS: Eighty children, aged four to eight years old, with 160 bilateral primary teeth without signs or symptoms of irreversibly inflamed or degenerative pulp tissue were treated in a split-mouth design trial comparing IPT using Ca3SiO5 or Ca(OH)2. The teeth were treated and restored with a preformed crown in a single session and assessed clinically and radiographically for one, three, six, and 12 months. RESULTS: Sixty patients with 120 treated molars completed the 12-month evaluations. The combined clinical and radiographic success rates were 98.3 percent (59 out of 60) for Ca3SiO5 and 95 percent for Ca(OH)2 (57 out of 60). No significant differences were found for success rates between the two study groups (P>0.05). The combined success rates for both groups was 96.7 percent. CONCLUSIONS: These results suggest that the indirect pulp treatment procedure with either a bioactive Ca3SiO5-based dentin substitute or a Ca(OH)2-based material may be considered a suitable treatment to achieve acceptable therapeutic results when applied on deeply carious primary teeth without degenerative symptoms.

Expression of enamel proteins in rough endoplasmic reticulum and golgi complex in human dental germs / Expresión de proteínas del esmalte en retículo endoplásmico rugoso y complexo golgiensis en gérmenes dentales humanos

Tooth enamel is the hardest tissue in the body. The organic matrix configuration is provided by the main proteins amelogenin, ameloblastin and enamelysin (MMP20), an enzyme that helps to shape the matrix. The aim of this study was to determine by histochemistry the expression of amelogenin and enamelysin through the rough endoplasmic reticulum in the late stages of amelogenesis, and its expression in the Complexus golgiensis (Golgi complex / Golgi apparatus) in the early stages in human fetuses. In early stages a colocalization of both proteins inside the Golgi apparatus was found, being more evident the relationship between Golgi and amelogenin (99.92 %). In the late stage, a colocalization of both proteins and rugged endoplasmic reticulum was found. With enamelysin being more evident in relation with rough endoplasmic reticulum (99.95 %). Our findings demonstrated the presence of amelogenin and enamelysin in odontoblast and ameloblast. However, the presence of these two proteins in odontoblast remains unknown.

El esmalte dental es el tejido más duro del cuerpo. La configuración de la matriz orgánica es proporcionada por las proteínas principales amelogenina, ameloblastina y enamelisina (MMP20), una enzima que ayuda a dar forma a la matriz. El objetivo de este estudio fue determinar mediante histoquímica la expresión de amelogenina y enamelisina a través del retículo endoplasmático rugoso en las últimas etapas de la amelogénesis , y su expresión en el Complexo golgiensis en las primeras etapas de formación en fetos humanos. En las primeras etapas se observó colocalización de ambas proteínas en el interior del Complexo golgiensis, siendo más evidente la relación entre Golgi y amelogenina (99,92 %). En la última etapa, se identificó una colocalización de ambas proteínas y retículo endoplásmico rugoso. Resulto más evidente la enamelisina en relación con el retículo endoplasmático rugoso (99,95 %). Nuestros resultados demostraron la presencia de amelogenina y enamelisina en odontoblastos y ameloblastos, sin embargo se desconoce la presencia de estas dos proteínas en odontoblastos.

Automatic Electronic Device Used for the Evaluation of Cyclic-Fatigue Resistance of Nickel-Titanium Instruments

he aim of this study was to evaluate the cyclic-fatigue fracture of different Nickel-Titanium motor-driven rotary instruments (ProTaper® Universal, ProFile®, and Mtwo® systems) in artificial canals by means of an Automatic Electronic Device (AED). The study was performed using NickelTitanium instruments 25/0.06 evaluated in canals with a 45-degree curvature and 2-mm radius. The analyses evaluated two parameters: fracture by cyclic fatigue, and time of fracture; in addition, the length of separated fragment was evaluated. Medians and range values were calculated for each group. Data were analyzed by the Kruskall-Wallis and Mann-Whitney U tests to determine statistical difference. The ProFile motor-driven rotator system exhibited highest resistance to fracture due to cyclic fatigue and highest fracture time compared with the ProTaper and Mtwo systems (p <0.05). The equipment proposed in this study (AED) demonstrated efficiency for recording information, automation, scheduled work times and durations, cycle number, time of fracture, pressure changes and, principally control of the human factor.

l objetivo del presente estudio fue evaluar la fractura a la fatiga cíclica de diferentes instrumentos rotatorios de Niquel-Titanio (sistemas ProTaper® Universal, ProFile® y Mtwo®) en conductos artificiales por medio de un Dispositivo Electrónico Automático (DEA). Se usaron instrumentos de Niquel-Titanio 25/0.06, los cuales se evaluaron en conductos con una curvatura de 45 grados y 2 milímetros de radio. Se analizaron dos parámetros: Fractura a la fatiga cíclica y tiempo a la fractura; además se evaluó la longitud del fragmento separado. Medianas y rangos fueron calculados para cada uno de los grupos. Los resultados fueron analizados por las pruebas de Kruskall-Wallis y U de Mann-Whitney para determinar diferencias estadísticas. El Sistema ProFile mostró una mayor resistencia y tiempo a la fractura en comparación con los sistemas ProTaper y Mtwo (p <0.05). El equipo propuesto en este estudio (DEA) demostró eficiencia para el registro de la información, tiempos de trabajo y duración, número de ciclos, tiempo a la fractura, cambios en la presión y principalmente control del factor humano.

Sustained release of calcium hydroxide from poly(DL-lactide-co-glycolide) acid microspheres for apexification.

Calcium hydroxide (CH) loaded poly(DL-lactide-co-glycolide) acid (PLGA) microspheres (MS) might be used for apexification requiring a sustained release of Ca(2+). The aim of this study was to formulate and characterize CH-PLGA-MS. The CH-loaded MS were prepared by either oil-in-water (O/W) or water-in-oil/in-water (W/O/W) emulsion solvent evaporation technique. MS produced by the O/W technique exhibited a larger diameter (18.63 ± 7.23 µm) than the MS produced by the W/O/W technique (15.25 ± 7.37 µm) (Mann-Whitney U test P < 0.001). The CH encapsulation efficiency (E e) and Ca(2+) release were calculated from data obtained by absorption techniques. Ca(2+) release profile was evaluated for 30 days. To know the E e, the CH-loaded MS were dissolved in 1 M NaOH to release all its content and a Ca(2+) colorimetric marker was added to this solution. The reagent marked the Ca(2+) in blue color, which was then measured by a UV-Vis system (650 nm). The percentage of E e was calculated on the basis of the theoretical loading. The E e of the O/W-produced MS was higher (24 %) than the corresponding percentage of the W/O/W-produced MS (11 %). O/W- and W/O/W-produced MS released slower and lower Ca(2+) than a control CH paste with polyethylene glycol 400 (Kruskal-Wallis test). O/W-produced MS released higher Ca(2+) than W/O/W-produced MS (statistically significant differences; P < 0.05). In conclusion, the CH-PLGA-MS were successfully formulated; the technique of formulation influenced the size, encapsulation efficiency and release profile. The MS were better sustained release system than the CH paste.

Sensory Neuropeptides and Endogenous Opioids Expression in Human Dental Pulp with Asymptomatic Inflammation: In Vivo Study.

PURPOSE: This study quantified the expression of substance P (SP), calcitonin gene-related peptide (CGRP), ß-endorphins (ß-End), and methionine-enkephalin (Met-Enk) in human dental pulp following orthodontic intrusion. METHODS: Eight patients were selected according to preestablished inclusion criteria. From each patient, two premolars (indicated for extraction due to orthodontic reasons) were randomly assigned to two different groups: the asymptomatic inflammation group (EXPg), which would undergo controlled intrusive force for seven days, and the control group (CTRg), which was used to determine the basal levels of each substance. Once extracted, dental pulp tissue was prepared to determine the expression levels of both neuropeptides and endogenous opioids by radioimmunoassay (RIA). RESULTS: All samples from the CTRg exhibited basal levels of both neuropeptides and endogenous opioids. By day seven, all patients were asymptomatic, even when all orthodontic-intrusive devices were still active. In the EXPg, the SP and CGRP exhibited statistically significant different levels. Although none of the endogenous opioids showed statistically significant differences, they all expressed increasing trends in the EXPg. CONCLUSIONS: SP and CGRP were identified in dental pulp after seven days of controlled orthodontic intrusion movement, even in the absence of pain.

Effectiveness of three electronic apex locators to determine root canal working length / Efectividad de tres localizadores apicales electrónicos para determinar la longitud radicular de trabajo

This study was to evaluate in vivo the accuracy of three electronic apex locators (EALs) in determining working length (WL) using hand files and a wear technique. Thirty two premolars that were completely formed apically and that were scheduled for extraction for orthodontic reasons from patients between ages of 15 and 20 years old were included. Electronic measurement of WL was performed using the EAL according to the manufacturer’s instructions. The following three EAL were used: A. Root ZX II; B. Raypex 5, and C. Propex II. There were significant difference (p=0.0002) when comparing median differences among the three EAL. Statistical analysis revealed significant differences between Root ZX II vs. Raypex 5 and Root ZX II vs. Propex II (p=0.0044; p=0.0002), while between aypex 5 and Propex II, there were no statistically significant differences with respect to the accuracy of the EAL in determining WL (p=0.1087). The present findings suggest that Root ZX II presented the highest agreement rate for determining the final WL...

Estudio fue evaluar in vivo la exactitud de tres localizadores apicales electrónicos(LAEs) para determinar la longitud de trabajo (LT) usando instrumentos manuales y una técnica de desgaste. Treinta y dos premolares con formación apical completa e indicados para extracción por razones ortodóncicas de pacientes de edad entre 15 y 20 años fueron incluidos en el estudio. Seusaron tres LAE; A. Root ZX II; B. Raypex 5, y C. Propex II. Se encontraron diferencias significativas (p=0.0002) cuando se compararon las medianas entre los tres LAE. El análisismostró diferencias entre Root ZX II vs. Raypex 5 y Root ZX II vs. Propex II (p=0.0044; p=0,0002), mientras queentre Raypex 5 y Propex II, no se encontraron diferencias estadísticamente significativas en la determinación de la LT (p=0.1087). Los presentes hallazgos sugieren que Root ZX II mostró la mayor exactitud para determinar la LT final...

Expression of substance P, calcitonin gene-related peptide, ß-endorphin and methionine-enkephalin in human dental pulp tissue after orthodontic intrusion: a pilot study.

OBJECTIVE: To determine the levels of two sensory neuropeptides (substance P [SP] and calcitonin gene-related peptide [CGRP]) and two endogenous opioids (methionine-enkephalin [Met-Enk] and ß-endorphin [ß-End]) in dental pulp tissue samples subjected to controlled orthodontic intrusive forces. MATERIALS AND METHODS: Sixteen healthy premolars were selected from eight patients who were undergoing extraction for orthodontic purposes. Eight were randomly used as controls, and the other eight were assigned to an experimental group (controlled orthodontic intrusive forces applied for 24 hours). After this period, teeth were extracted, and pulp samples were obtained. All samples were processed to quantify the expression levels of SP, CGRP, Met-Enk, and ß-End using commercial radioimmunoassay kits. RESULTS: All samples exhibited basal levels of both neuropeptides and endogenous opioids. After 24 hours of the intrusive stimulus, all patients reported a tolerable discomfort localized at the involved premolar. Only SP was significantly increased (P<.05). For the other molecules, no statistically significant differences were observed (P>.05); however, they expressed important increasing trends. CONCLUSIONS: The expression levels of SP and CGRP in dental pulp samples from the experimental group support the positive correlation between the symptomatic clinical scenario and increased expression levels of neuropeptides, clarifying the role of neurogenic inflammation in early injury response.

Comparison between rotary and manual techniques on duration of instrumentation and obturation times in primary teeth.

OBJECTIVE: The aim of this study was to compare the duration of instrumentation and obturation times and quality of root canal filling between rotary and manual instrumentation techniques in primary teeth. STUDY DESIGN: A randomized, controlled clinical trial was performed that included deciduous teeth with pulp necrotic. Forty necrotic teeth were included; 20 were instrumented with a rotary technique (experimental group) and 20 with a manual technique (control group). The time taken for instrumentation and for obturation were recorded in minutes, and the quality of the root canal filling was recorded as optimal, under-filled, or overfilled. RESULTS: The use of the rotary technique diminished the time of instrumentation to 63% and time of obturation to 68%, and it improved the quality of the root canalfilling. CONCLUSION: The use of rotary instruments in the pulpectomy of primary molars represents a promising technique; the time is significantly reduced.

Design of a controlled release system of OP-1 and TGF-ß1 based in microparticles of sodium alginate and release characterization by HPLC-UV.

A new system for sustained release of growth factors, such as osteogenic protein 1 (OP-1) and transforming growth factor ß1 (TGF-ß1), intended to repair and promote dental tissue regeneration in rats was designed and characterized in this work. The release system was made with microparticles of sodium alginate, produced by ionic gelling dripping technique. The release profiles of OP-1 and TGF-ß1 from biopolymer matrix were determined by high-performance liquid chromatography (HPLC), and with this purpose, an HPLC-UV method was developed. About 80% of each growth factor was released in the first 24 h, reaching almost 100% in 168 h. The system was tested during the tissue repair in rat molars in comparison with calcium hydroxide and both growth factors not encapsulated. The dentin sialoprotein (DSP) was used as a repair marker. It was detected by immunohistochemistry, after 14- and 28-d post-treatment. X (2) test (p ≤ 0.001) and Fisher exact test (p ≤ 0.05) were applied for assessment of the amount of immunostaining. The treatment with encapsulated OP-1 showed an increased DSP immunostaining after 14 d and did not find any significant difference with the immunostaining observed for calcium hydroxide treatment. Treatment with TGF-ß1 did not show significant difference with calcium hydroxide. Treatment with both factors OP-1 and TGF-ß1 showed higher DSP immunostaining in comparison with calcium hydroxide treatment. In conclusion, the combination of both growth factors encapsulated showed more DSP immunostaining in comparison with each one separated, either encapsulated or not.

Amelogenin and enamelysin localization in human dental germs.

Odontogenesis is extensively studied in animal models but less understood in human. In early amelogenesis, amelogenin constitutes 90% of enamel organic matrix, which is degraded by enamelysin and replaced by hydroxyapatite crystals. Here, amelogenin and enamelysin distribution changes during amelogenesis were shown by co-localization experiments by confocal microscopy. Early bell stage showed more amelogenin labeling than enamelysin, as free immune-reactive granular patches towards basal membrane between ameloblast and odontoblast. Increased amelogenin expression and secretion towards extracellular matrix formation region was found. Enamelysin distribution was perinuclear in early bell stage. During late bell stage, a decreasing amelogenin labeling in contrast with enamelysin increasing along the cells was found, suggesting specific temporal amelogenin degradation. Enamelysin was located initially around nuclei and later was found in all the ameloblast and stellate reticulum cytoplasm. Amelogenin was observed inside ameloblast, stellate reticulum, and intermediate stratum cells in the enamel as well as in the newly formed dentin extracellular matrix. In contrast, in dentin more amelogenin than enamelysin was found located close to the periphery.

Identification of cultivable microorganisms from primary teeth with necrotic pulps.

The objective of this study was to identify cultivable microorganisms from primary teeth with necrotic pulps. This experimental study included 21 patients of both sexes between 4 and 7 years of age with necrotic pulps in primary teeth. Twenty-one maxillary and mandibular molars containing at least 1 necrotic canal, an abscess or sinus tract, one or more radiolucent areas in the furcation or periapical region, teeth having at least two thirds of root length, and carious lesions directly exposed to the oral environment were included. After antisepsis of the oral cavity, anesthesia of the affected tooth, and isolation and disinfection of the operative field, 3 sterile absorbent paper points were sequentially placed for 30 seconds for the collection of samples. The samples were immediately processed in an anaerobic chamber, and all isolated microorganisms were identified. Anaerobic species (anaerobic facultative and moderate anaerobes) were isolated in all root canals; 68.4% of root canal samples studied showed a polymicrobial nature. Most of the isolate consisted of Bifidobacterium Spp2 and Streptococcus intermedius. Other less frequently encountered species were Actinomyces israelii, Bifidobacterium spp 1, Clostridium spp, and Candida albicans. Results indicate the existence of combinations of bacterial species in root canal infections of the primary dentition with necrotic pulps, anaerobic bacteria predominating.

Clinical efficacy of 5% sodium hypochlorite for removal of stains caused by dental fluorosis.

The objective of this study was to evaluate the clinical efficacy of 5% sodium hypochlorite solution for removal of stains caused by dental fluorosis in young patients. A clinical trial involved 33 patients with diffuse opacities on the enamel surfaces of maxillary incisors due to effects of dental fluorosis. The protocol of treatment 3 steps: (1) cleaning and enamel etching with 37% phosphoric acid in order to eliminate the layer that covers the fluorotic enamel surface and allow better penetration of the bleaching agent, (2) application of 5% sodium hypochlorite to remove stains caused by organic material, and (3) filling the opened micro-cavities with a light-cured, composite surface sealant to prevent restaining. The whiteness of the enamel lesions before and after treatment were expressed in L*, a*, and b* color space measurements using a Minolta Chroma Meter CR300. Analysis of parameters of [symbol:see text]E (L*, a*, b*) showed that changes were observed in the L* (brightness) and a* (redness), which paralleled the [symbol: see text]E differences. There was no significant difference in the b* (yellow) parameter. The technique described in this study appears to have advantages over other methods for improving the appearance of fluorotic lesions. It is simple, low cost, non invasive so the enamel keeps its structure, relatively rapid, and safe; it requires no special materials, and it can be used with safety on young permanent teeth.