Hexavalent chromium exposure alters bone remodeling in the developing tooth alveolus and delays tooth eruption.

Although it has been demonstrated that exposure of lactating rats to CrVI delays tooth eruption, the effects of CrVI exposure on bone remodeling in the developing alveolus during tooth eruption remain unknown. Our purpose was to analyze the effect of CrVI in the alveolus of the first lower molar of rats. Thirty-two suckling Wistar rats were divided into two groups. The experimental group received 12.5 mg/kg body weight/day of potassium dichromate dissolved in saline solution by oral gavage as of day 4 of the experiment; the control group received an equal dose of saline solution. Each group was divided into two sub-sets and euthanized at the ages of 9 and 15 days, respectively. Histochemical and histomorphometric studies of the bone surfaces of the developing tooth alveolus were performed. The percentage of bone formation surfaces was lower in experimental animals than in age-matched controls. The percentage of bone resorption surfaces was significantly lower in 9-day-old experimental rats than in controls and significantly higher in 15-day-old experimental rats than in controls. Exposure to CrVI during lactation alters the sequence of bone resorption and formation in the walls of the developing alveolus, both of which are necessary for tooth eruption, thus causing a delay.

Si bien ya ha sido demostrado que la exposición a CrVI de ratas lactantes retrasa la erupción dentaria, aún se desconocen los efectos de la exposición a CrVI que se producen sobre la modelación y remodelación de las paredes del alvéolo en formación que ocurren en los diferentes estadios de la erupción dentaria. Por tal motivo, el propósito de este trabajo fue estudiar el efecto del CrVI sobre la formación y la reabsorción óseas del alvéolo del primer molar inferior en desarrollo en ratas, a los 9 y a los 15 días de edad, que corresponden a los estadios intraóseo y de penetración mucosa de la erupción dentaria, respectivamente. El grupo experimental recibió una dosis diaria de 12,5 mg/kg de peso corporal de dicromato de potasio disuelto en solución salina por sonda bucal a partir del 4° día; mientras que el grupo control, un volumen equivalente de solución salina. Cada grupo fue dividido en 2 subgrupos de acuerdo al tiempo experimental en el que se llevó a cabo la eutanasia: 9 y 15 días de edad. Se llevaron a cabo estudios histoquímicos e histomorfométricos de las superficies óseas de los alveólos dentarios en formación. Los datos fueron analizados estadísticamente utilizando la prueba t de Student; estableciéndose un valor de p<0,05 como estadísticamente significativo. El porcentaje de superficies en formación fue menor en los animales experimentales de 9 y de 15 días de edad que en los respectivos controles. El porcentaje de superficies en reabsorción en los animales experimentales de 9 días de edad fue significativamente menor y en los animales de 15 días de edad fue significativamente mayor con respecto a sus controles. La exposición al cromo hexavalente durante la lactancia altera la secuencia de la reabsorción y la formación ósea de las paredes del alvéolo en desarrollo necesarias para que la pieza dentaria erupcione, causando su retraso. Los hallazgos obtenidos muestran la importancia del control de sustancias tóxicas en el agua potable, ya que sus efectos pueden alterar la remodelación ósea y por ende, el crecimiento y el desarrollo de los individuos que fueron expuestos durante la infancia temprana.

Hexavalent chromium exposure alters bone remodeling in the developing tooth alveolus and delays tooth eruption / La exposición a cromo hexavalente altera la remodelación ósea del alvéolo dental en desarrollo y retarda la erupción dentaria

ABSTRACT Although it has been demonstrated that exposure of lactating rats to CrVI delays tooth eruption, the effects of CrVI exposure on bone remodeling in the developing alveolus during tooth eruption remain unknown. Our purpose was to analyze the effect of CrVI in the alveolus of the first lower molar of rats. Thirty-two suckling Wistar rats were divided into two groups. The experimental group received 12.5 mg/kg body weight/day of potassium dichromate dissolved in saline solution by oral gavage as of day 4 of the experiment; the control group received an equal dose of saline solution. Each group was divided into two sub-sets and euthanized at the ages of 9 and 15 days, respectively. Histochemical and histomorphometric studies of the bone surfaces of the developing tooth alveolus were performed. The percentage of bone formation surfaces was lower in experimental animals than in age-matched controls. The percentage of bone resorption surfaces was significantly lower in 9-day-old experimental rats than in controls and significantly higher in 15-day-old experimental rats than in controls. Exposure to CrVI during lactation alters the sequence of bone resorption and formation in the walls of the developing alveolus, both of which are necessary for tooth eruption, thus causing a delay.

RESUMEN Si bien ya ha sido demostrado que la exposición a CrVI de ratas lactantes retrasa la erupción dentaria, aún se desconocen los efectos de la exposición a CrVI que se producen sobre la modelación y remodelación de las paredes del alvéolo en formación que ocurren en los diferentes estadios de la erupción dentaria. Por tal motivo, el propósito de este trabajo fue estudiar el efecto del CrVI sobre la formación y la reabsorción óseas del alvéolo del primer molar inferior en desarrollo en ratas, a los 9 y a los 15 días de edad, que corresponden a los estadios intraóseo y de penetración mucosa de la erupción dentaria, respectivamente. El grupo experimental recibió una dosis diaria de 12,5 mg/kg de peso corporal de dicromato de potasio disuelto en solución salina por sonda bucal a partir del 4° día; mientras que el grupo control, un volumen equivalente de solución salina. Cada grupo fue dividido en 2 subgrupos de acuerdo al tiempo experimental en el que se llevó a cabo la eutanasia: 9 y 15 días de edad. Se llevaron a cabo estudios histoquímicos e histomorfométricos de las superficies óseas de los alveólos dentarios en formación. Los datos fueron analizados estadísticamente utilizando la prueba t de Student; estableciéndose un valor de p<0,05 como estadísticamente significativo. El porcentaje de superficies en formación fue menor en los animales experimentales de 9 y de 15 días de edad que en los respectivos controles. El porcentaje de superficies en reabsorción en los animales experimentales de 9 días de edad fue significativamente menor y en los animales de 15 días de edad fue significativamente mayor con respecto a sus controles. La exposición al cromo hexavalente durante la lactancia altera la secuencia de la reabsorción y la formación ósea de las paredes del alvéolo en desarrollo necesarias para que la pieza dentaria erupcione, causando su retraso. Los hallazgos obtenidos muestran la importancia del control de sustancias tóxicas en el agua potable, ya que sus efectos pueden alterar la remodelación ósea y por ende, el crecimiento y el desarrollo de los individuos que fueron expuestos durante la infancia temprana.

Evaluation of apoptosis and osteopontin expression in osteocytes exposed to orthodontic forces of different magnitudes.

The in vivo response of osteocytes to different force magnitudes soon after they are applied remains to be elucidated. The aim of this study was to examine the early effects of applying a very light (LF: 0,16 N) and a very strong (SF: 2,26 N) orthodontic force during one hour on apoptosis and osteopontin (OPN) expression on alveolar bone osteocytes, in rats. Results: LF: compared to the control group, they showed a significant increase in OPN expression, and a significant decrease in the number of TUNELpositive osteocytes. SF: compared to the control group, they showed a significant increase in OPN expression and a significant decrease in the number of TUNELpositive osteocytes. Our results show that osteocytes respond very early to the application of tension and pressure forces of different magnitudes, and application of forces decreases the number of apoptotic osteocytes and increases OPN expression. These results allow concluding that osteocytes activate rapidly when subjected to locally applied forces, whether these forces be pressure or tension, light or strong forces.

Hasta el momento no se ha dilucidado la respuesta temprana in vivo de los osteocitos a la aplicación de fuerzas de diferentes magnitudes sobre el hueso. El objetivo de este estudio fue examinar la respuesta temprana de la aplicación de una fuerza ortodóncica muy liviana (FL: 0,16 N) y muy fuerte (FF: 2,26 N) durante una hora sobre la expresión de apoptosis y osteopontina (OPN) en los osteocitos del hueso alveolar, en ratas. Resultados: FL: en comparación con el grupo control, mostraron un aumento significativo en la expresión de OPN y una disminución significativa en el número de osteocitos TUNELpositivos. FF: en comparación con el grupo control, mostraron un aumento significativo en la expresión de OPN y una disminución signi ficativa en el número de osteocitos TUNELpositivos. Nuestros resultados muestran que los osteocitos responden muy temprano a la aplicación de fuerzas de tensión y presión de diferentes magnitudes, y la aplicación de fuerzas disminuye el número de osteocitos apoptóticos y aumenta la expresión de OPN. Estos resultados permiten concluir que los osteocitos se activan rápidamente cuando se los somete a fuerzas aplicadas localmente, ya sean estas fuerzas de presión o tensión, livianas o fuertes.

An experimental model of disuse in the alveolar rat bone: A histomorphometrical study.

Lack of mechanical stimulation is known to cause disuse osteopenia in bones. However, experimental models for disuse osteopenia on bones other than jawbones are not applicable to jawbones. The little available information in this field has been applied to the study of overeruption of teeth lacking antagonists. However, the absence of an antagonist in the opposite jaw means that there is no stimulation by occlusion. Our hypothesis is that the lack of stimulation due to the absence of teeth causes disuse osteopenia in the interradicular bone of the antagonist teeth. Our aim was to develop a model of disuse osteopenia due to the absence of occlusal forces. We used male Wistar rats with 215-230 g body weight, divided into 2 groups: one absolute control group (C) and one experimental group in which the three right lower molars were extracted (E). The left side of the jaw in the experimental group was used as a paired control (PC). The animals were euthanized 7 days after extraction. The jaws were placed in occlusion, fastened and fixed in 10% formalin. The heads were cut in half and radiographs made of both jaws. The upper jaws were processed histologically. After decalcification, bucco-palatine oriented sections were cut at the level of the mesial root and distal roots of the first upper molars. On the radiographs, the distance from the tip of the cusp on the first upper molar to the antagonist edentulous ridge (DA) was measured. On the microphotographs, the following parameters were measured: passive eruption degree (PED), height of periodontal ligament at the level of the furcation (HPL) and interradicular bone volume (BVI). The data were compared statistically using ANOVA and Bonferroni's post-hoc test, considering p<0.05 as statistically significant. DA in experimental animals was 0.34}0.048 mm. PED in experimental animals was significantly greater than in the control groups, both for the buccal plate and for the palatal plate. HPL showed no significant difference between groups. BVI was significantly lower in the experimental group than in the control group. The results showed that the model used produces a condition of disuse osteopenia, shown by the statistically significant reduction in interradicular bone volume. The use of this model at different experimental times will enable the evaluation of cell responses in periodontal tissues, particularly bone tissue, e.g. to compare them to known responses such as the application of orthodontic forces.

Impairment of Bony Crypt Development Associated With Hexavalent Chromium Exposure During Tooth Eruption.

Improperly treated hexavalent chromium-containing industrial wastes contaminate drinking water, potentially affecting children taking breast milk or baby bottles prepared with infant formula. Thus, the aim of the present work was to determine the effect of this toxic on bone activity in the developing alveolus during tooth eruption of suckling Wistar rats intoxicated with potassium dichromate. Experimental animals received a daily dose of 12.5mg/kg body weight of potassium dichromate by gavage for 10 days; controls received an equivalent volume of saline solution. Histologic and histomorphometric studies of the mandible were performed. The data were statistically analyzed using Student's t test; statistical significance was set at a value of p <0.05. Experimental animals exhibited delayed tooth eruption, decreased periodontal width and bone volume, a lower percentage of bone formation surfaces, and higher percentage of quiescent surfaces (p<0.05) compared to controls. The delay in tooth eruption observed after exposure to hexavalent chromium is the result of a lower rate of bone remodeling in the developing alveolus. The obtained results show the importance of controlling toxic substances in drinking water, since their effects may alter the growth and development of subjects who were exposed during early infancy.

Desechos industriales que contienen cromo hexavalente inade- cuadamente tratados contaminan el agua de consumo pudiendo afectar a los niños por vía de la leche materna o de la preparación de mamaderas. Por lo tanto, el objetivo del presente trabajo fue determinar el efecto de este tóxico en la actividad del hueso en el alveolo en desarrollo durante la erupción dentaria de ratas Wistar lactantes expuestas a dicromato de potasio. Los animales experimentales recibieron una dosis diaria de 12,5 mg / kg de peso corporal de dicromato de potasio por alimentación forzada durante 10 días; mientras que los controles, un volumen equivalente de solución salina. Se llevaron a cabo estudios histológicos e histomorfométricos de la mandíbula. Los datos fueron analizados estadísticamente utilizando la prueba t de Student; estableciéndose un valor de p<0,05 como estadísticamente significativo. Los animales expuestos a cromo hexavalente mostraron retraso en la erupción dentaria, menor espacio periodontal y volumen óseo; encontrándose disminuidas las superficies en formación y en reabsorción óseas y aumentadas las superficies en reposo (p <0,05) en comparación con los controles. El retraso en la erupción dentaria observado luego de la exposición a cromo hexavalente es el resultado de una menor remodelación ósea en el alveolo en desarrollo. Los resultados obtenidos muestran la importancia del control de sustancias tóxicas en el agua potable, ya que sus efectos pueden alterar el crecimiento y el desarrollo de los individuos que fueron expuestos durante la infancia temprana.

An experimental model of disuse in the alveolar rat bone: A histomorphometrical study

Lack of mechanical stimulation is known to cause disuse osteopenia in bones. However, experimental models for disuse osteopenia on bones other than jawbones are not applicable to jawbones. The little available information in this field has been applied to the study of overeruption of teeth lacking antagonists. However, the absence of an antagonist in the opposite jaw means that there is no stimulation by occlusion. Our hypothesis is that the lack of stimulation due to the absence of teeth causes disuse osteopenia in the interradicular bone of the antagonist teeth. Our aim was to develop a model of disuse osteopenia due to the absence of occlusal forces. We used male Wistar rats with 215-230 g body weight, divided into 2 groups: one absolute control group (C) and one experimental group in which the three right lower molars were extracted (E). The left side of the jaw in the experimental group was used as a paired control (PC). The animals were euthanized 7 days after extraction. The jaws were placed in occlusion, fastened and fixed in 10% formalin. The heads were cut in half and radiographs made of both jaws. The upper jaws were processed histologically. After decalcification, bucco-palatine oriented sections were cut at the level of the mesial root and distal roots of the first upper molars. On the radiographs, the distance from the tip of the cusp on the first upper molar to the antagonist edentulous ridge (DA) was measured. On the microphotographs, the following parameters were measured: passive eruption degree (PED), height of periodontal ligament at the level of the furcation (HPL) and interradicular bone volume (BVI). The data were compared statistically using ANOVA and Bonferroni's post-hoc test, considering p<0.05 as statistically significant. DA in experimental animals was 0.34}0.048 mm. PED in experimental animals was significantly greater than in the control groups, both for the buccal plate and for the palatal plate. HPL showed no significant difference between groups. BVI was significantly lower in the experimental group than in the control group. The results showed that the model used produces a condition of disuse osteopenia, shown by the statistically significant reduction in interradicular bone volume. The use of this model at different experimental times will enable the evaluation of cell responses in periodontal tissues, particularly bone tissue, e.g. to compare them to known responses such as the application of orthodontic forces.

Se sabe que la falta de estimulo mecanico produce un cuadro de osteopenia por desuso en huesos de la economia. Los modelos experimentales de osteopenia por desuso utilizados en otros huesos no son aplicables a los huesos maxilares. La escasa informacion que existe en este campo se aplico al estudio de la sobreerupcion de dientes sin antagonista. Sin embargo, la ausencia de antagonista en el maxilar opuesto hace que falte el estimulo de la oclusion. Por tal razon, nuestra hipotesis es que la falta de estimulo por ausencia de piezas dentarias provoca un cuadro de osteopenia por desuso en el hueso interradicular de los dientes antagonistas. Nuestro objetivo fue poner a punto un modelo de osteopenia por desuso debido a la ausencia de fuerzas oclusales. Se emplearon ratas Wistar machos de entre 215-230 g de peso corporal divididas en 2 grupos, un grupo control absoluto (C) y un grupo experimental al que se le extrajeron los tres molares inferiores derechos (E). El lado izquierdo del maxilar del grupo experimental, fue utilizado como control apareado (CA). A los 7 dias de efectuadas las extracciones se realizo la eutanasia de los animales. Los maxilares se colocaron en oclusion, se precintaron y asi ubicados se fijaron en formol 10%. Se dividieron las hemicabezas y tomaron radiografias de ambos maxilares siendo procesados histologicamente los maxilares superiores. Luego de la descalcificacion se obtuvieron cortes orientados en sentido buco-palatino a nivel de la raiz mesial y de las raices distales de los 1o molares superiores. Sobre las radiografias se midio la distancia desde el vertice cuspideo del 1o molar superior al reborde antagonista desdentado (DA), y sobre microfotografias se midieron los siguientes parametros: grado de erupcion pasiva (GE), altura del ligamento periodontal a nivel de la furcacion (ALP) y volumen oseo interradicular (VOI). Los datos se compararon estadisticamente mediante el test ANOVA y prueba post hoc de Bonferroni, considerando una p<0,05 como estadisticamente significativa. La DA en los animales experimentales fue de 0,34}0,048 mm. La GE en los animales experimentales fue significativamente mayor que en los grupos control, tanto a nivel de la tabla vestibular como de la tabla palatina. La ALP no mostro diferencias significativas entre los grupos. El VOi fue significativamente menor en el grupo experimental con respecto a los controles. Los resultados mostraron que el modelo empleado logra una condicion de osteopenia por desuso manifestada por la disminucion del volumen oseo interradicular. La utilizacion de este modelo de desuso a diferentes tiempos experimentales permitira evaluar las respuestas celulares de los tejidos periodontales, especial - mente del tejido oseo, permitiendo por ejemplo, compararlas con respuestas conocidas como la aplicacion de fuerzas ortodoncicas.

Impairment of bony crypt development associated with hexavalent chromium exposure during tooth eruption

Improperly treated hexavalent chromium-containing industrial wastes contaminate drinking water, potentially affecting children taking breast milk or baby bottles prepared with infant formula. Thus, the aim of the present work was to determine the effect of this toxic on bone activity in the developing alveolus during tooth eruption of suckling Wistar rats intoxicated with potassium dichromate. Experimental animals received a daily dose of 12.5mg/kg body weight of potassium dichromate by gavage for 10 days; controls received an equivalent volume of saline solution. Histologic and histomorphometric studies of the mandible were performed. The data were statistically analyzed using Student's t test; statistical significance was set at a value of p <0.05. Experimental animals exhibited delayed tooth eruption, decreased periodontal width and bone volume, a lower percentage of bone formation surfaces, and higher percentage of quiescent surfaces (p<0.05) compared to controls. The delay in tooth eruption observed after exposure to hexavalent chromium is the result of a lower rate of bone remodeling in the developing alveolus. The obtained results show the importance of controlling toxic substances in drinking water, since their effects may alter the growth and development of subjects who were exposed during early infancy.

Desechos industriales que contienen cromo hexavalente inade - cua damente tratados contaminan el agua de consumo pudiendo afectar a los ninos por via de la leche materna o de la preparacion de mamaderas. Por lo tanto, el objetivo del presente trabajo fue determinar el efecto de este toxico en la actividad del hueso en el alveolo en desarrollo durante la erupcion dentaria de ratas Wistar lactantes expuestas a dicromato de potasio. Los animales experimentales recibieron una dosis diaria de 12,5 mg / kg de peso corporal de dicromato de potasio por alimentacion forzada durante 10 dias; mientras que los controles, un volumen equivalente de solucion salina. Se llevaron a cabo estudios histologicos e histomorfometricos de la mandibula. Los datos fueron analizados estadisticamente utilizando la prueba t de Student; estableciendose un valor de p<0,05 como esta - disticamente significativo. Los animales expuestos a cromo hexavalente mostraron retraso en la erupcion dentaria, menor espacio periodontal y volumen oseo; encontrandose disminuidas las superficies en formacion y en reabsorcion oseas y aumentadas las superficies en reposo (p <0,05) en comparacion con los controles. El retraso en la erupcion dentaria observado luego de la exposicion a cromo hexavalente es el resultado de una menor remodelacion osea en el alveolo en desarrollo. Los resultados obtenidos muestran la importancia del control de sustancias toxicas en el agua potable, ya que sus efectos pueden alterar el crecimiento y el desarrollo de los individuos que fueron expuestos durante la infancia temprana.

Histomorphometric study and three-dimensional reconstruction of the osteocyte lacuno-canalicular network one hour after applying tensile and compressive forces.

The occurrence of very early morphological changes in the osteocyte lacuno-canalicular network following application of tensile and/or compressive forces remains unknown to date. Thus, the aim of this study was to perform a morphological and morphometric evaluation of the changes in the three-dimensional structure of the lacuno-canalicular network and the osteocyte network of alveolar bone that take place very early after applying tensile and compressive forces in vivo, conducting static histomorphometry on bright-field microscopy and confocal laser scanning microscopy images. Our results showed that both the tensile and compressive forces induced early changes in osteocytes and their lacunae, which manifested as an increase in lacunar volume and changes in lacunar shape and orientation. An increase in canalicular width and a decrease in the width and an increase in the length of cytoplasmic processes were also observed. The morphological changes in the lacuno-canalicular and osteocyte networks that occur in vivo very early after application of tensile and compressive forces would be an indication of an increase in permeability within the system. Thus, both compressive and tensile forces would cause fluid displacement very soon after being applied; the latter would in turn rapidly activate alveolar bone osteocytes, enhancing transmission of the signals to the entire osteocyte network and the effector cells located at the bone surface.

The lacuno-canalicular system (LCS) and osteocyte network of alveolar bone by confocal laser scanning microscopy (CLSM).

The osteocyte lacuno-canalicular system (OLCS) is a large network intercommunicating the lacunae and canaliculi which contain the osteocytes and their cytoplasmic processes within the mineralized bone matrix. The vitality and functioning of the osteocytes and cytoplasmic processes depend upon this intercommunication. To date, the 3-dimensional features of OLCS in the alveolar bone have not been studied; therefore the aim of this study was to use confocal scanning microscopy to do so. Samples of alveolar bone from male Wistar rats were fixed in buffer formalin and stained with basic fuchsin to visualize the lacuno-canalicular system. In decalcified samples of the same bone, the actin was labeled using fluorescent phallotoxin to visualize the osteocyte network. The samples were observed at the level of the mesial root of the first upper molar in bucco-palatal direction using a confocal laser scanning microscope. The results showed that in the area near the inner aspect (bundle bone) of the buccal plate, the osteocyte lacunae are oval-shaped and relatively uniform in size, aligned parallel to each other and with their major axes parallel to the periodontal bone surface, and the osteocytes are oval-shaped, with their main axes perpendicular to the periodontal bone surface, and the cytoplasmic processes irradiate in all directions. In the area near the inner aspect (bundle bone) of the palatal plate, the osteocyte lacunae are rounded, have different sizes and their orientation does not follow any specific pattern, and the osteocyte bodies have major axes parallel to the periodontal surface, a larger number of cytoplasmic processes, and run in a straighter direction than in the buccal plate. These results will contribute to the understanding of the changes that may occur in OLCS microarchitecture as a result of a pathological process, surgical technique or force applied to the alveolar bone.

The lacuno-canalicular system (LCS) and osteocyte network of alveolar bone by confocal laser scanning microscopy (CLSM) / Características del sistema lacuno-canalicular (lcs) y la red osteocitaria del hueso alveolar observadas por microscopía láser de barrido confocal (clsm)

El sistema lacuno-canalicularosteocitario (OLCS) comprende una amplia red de intercomunicación entre las lagunas y loscanalículos que contienen a los osteocitos y sus procesos citoplasmáticos dentro de la matriz ósea mineralizada, de lo que depende su vitalidad y funcionamiento. Hasta el momento no se han estudiado las características tridimensionales delOLCS en el hueso alveolar por lo que el objetivo del presente trabajo fue de eterminarla por microscopía de barrido co nfocal. Muestras de hueso alveolar de ratas Wistar machos, luego fijadas en formol buffer fueron teñidas con fucsina básicapar a visualizar el sistema lacuno-canalicular y a muestrasdescalcificadas del mismo hueso mediante una falotoxinafluorescente se les marcó la actina para visualizar la redosteocitaria. Las muestras se observaron a nivel de la raízmesial del primer molar superior en sentido buco-palatinocon un microscopio láser de barrido confocal. Los resultadosmostraron que en la zona aledaña a la cortical periodontal dela tabla ósea vestibular las lagunas osteocitarias presentanforma ovalada y tamaño relativamente uniforme, alinéandoseparalelas entre sí y con su eje mayor paralelo a la superficieósea periodontal en tanto que los osteocitos, de forma ovalada,presentan su eje mayor orientado perpendicularmente a dicha superficie y con los procesos citoplasmáticos irradiándose en todos los sentidos del espacio. En la zona aledaña ala cortical periodontal de la tabla ósea palatina las lagunas osteocitarias se presentan de forma redondeada y tienen distintos tamaños, sin seguir un patrón de orientación específico, y los cuerpos osteocitarios presentan su eje mayor paralelo a una dirección más recta que en la tabla vestibular.

The lacuno-canalicular system (LCS) and osteocyte network of alveolar bone by confocal laser scanning microscopy (CLSM) / Características del sistema lacuno-canalicular (lcs) y la red osteocitaria del hueso alveolar observadas por microscopía láser de barrido confocal (clsm)

El sistema lacuno-canalicularosteocitario (OLCS) comprende una amplia red de intercomunicación entre las lagunas y loscanalículos que contienen a los osteocitos y sus procesos citoplasmáticos dentro de la matriz ósea mineralizada, de lo que depende su vitalidad y funcionamiento. Hasta el momento no se han estudiado las características tridimensionales delOLCS en el hueso alveolar por lo que el objetivo del presente trabajo fue de eterminarla por microscopía de barrido co nfocal. Muestras de hueso alveolar de ratas Wistar machos, luego fijadas en formol buffer fueron teñidas con fucsina básicapar a visualizar el sistema lacuno-canalicular y a muestrasdescalcificadas del mismo hueso mediante una falotoxinafluorescente se les marcó la actina para visualizar la redosteocitaria. Las muestras se observaron a nivel de la raízmesial del primer molar superior en sentido buco-palatinocon un microscopio láser de barrido confocal. Los resultadosmostraron que en la zona aledaña a la cortical periodontal dela tabla ósea vestibular las lagunas osteocitarias presentanforma ovalada y tamaño relativamente uniforme, alinéandoseparalelas entre sí y con su eje mayor paralelo a la superficieósea periodontal en tanto que los osteocitos, de forma ovalada,presentan su eje mayor orientado perpendicularmente a dicha superficie y con los procesos citoplasmáticos irradiándose en todos los sentidos del espacio. En la zona aledaña ala cortical periodontal de la tabla ósea palatina las lagunas osteocitarias se presentan de forma redondeada y tienen distintos tamaños, sin seguir un patrón de orientación específico, y los cuerpos osteocitarios presentan su eje mayor paralelo a una dirección más recta que en la tabla vestibular.(AU)

Evaluation of vascular endothelial growth factor (VEGF) in interradicular bone marrow in olpadronate treated animals.

UNLABELLED: Vascular endothelial growth factor (VEGF) is a protein that increases vascular permeability and induces the proliferation, migration and survival of endothelial cells. Bisphosphonates (BPs) are antiresorptive drugs that are widely used in the treatment of bone metabolism diseases and bone metastases. Since 2003, cases of bisphosphonate-related osteonecrosis of the jaw (BRONJ) have been reported. Few papers explain the mechanisms that induce BRONJ; some authors mention alterations in bone remodelling and a certain antiangiogenic effect of BPs. The aim of this study is to evaluate the expression of VEGF in bone marrow cells and the number of blood vessels and area occupied by them in animals treated with the BP sodium olpadronate (OPD). We used 16 Wistar rats, 60 days old, divided into two groups, experimental (OPD) and control. The OPD group received 0.3 mg/kg/week intraperitoneal OPD for 5 weeks. The control group received an equivalent intraperitoneal volume of physiological saline solution. After euthanasia, hemimandibles were processed and mesio-distal histological sections of the first molar were prepared. Sections were stained with hematoxylin-eosin (HE), immunohistochemical detection of VEGF was performed (sc-7269) and the following histomorphometric parameters were evaluated: In HE-stained sections--number of blood vessels per sq. mm. and percentage (%) of area occupied by blood vessels in relation to total area evaluated; in sections with immunohistochemical detection of VEGF--number of VEGF+ bone marrow cells per sq. mm. Data underwent statistical analysis. Number of blood vessels/mm2 was significantly lower in the OPD group (OPD: 92 +/- 16; CONTROL: 140 +/- 31; p < 0.05) and % vascular area/total area evaluated showed no significant difference (OPD: 15.6 +/- 6.1; CONTROL: 10.2 +/- 4.2). Number of VEGF+ cells/mm2 was lower in the OPD group than in the control group, with statistically significant differences (OPD: 7804.8 +/- 597; CONTROL: 13187.6 +/- 894; p < .001). The results of this study suggest that monosodium olpadronate has an antiangiogenic effect. Further studies are needed to reveal its potential as an antitumor agent and its connection with the onset of BRONJ.

Effect of different doses of hexavalent chromium on mandibular growth and tooth eruption in juvenile Wistar rats.

Not only workers employed at industrial plants are exposed to intoxication with the element they manipulate, the population at large is also at risk of suffering health problems caused by contaminating wastes inadequately treated for their safe disposal. As a result certain toxic substances, such as hexavalent chromium,has reached the general population including children. The present study sought to evaluate the effect of intoxication with hexavalent chromium on body and mandibular growth and tooth eruption in suckling Wistar rats. Potassium dichromate was administered by gavage in a dose of 6.25 or 12.5mg/kg body weight (b.w.) to one of the two groups of 4-day-old Wistar rats during 10 days. Our results showed that the effects of chromium are dose-dependent. Morphometric studies of body growth showed lower body weight in both experimental groups and shorter tail length in animals receiving 12.5mg/kg b.w. dose of chromium, compared with controls. All parameters of mandibular growth were lower in the experimental group receiving 12.5mg/kg b.w. of chromium. Differences in tooth eruption were observed at the level of the first molar in animals receiving 12.5mg/kg and of the second molar in those receiving 6.25mg/kg b.w. of chromium. Chromium was found to affect all the studied parameters.

Estandarización del método para el estudio de fuerzas ortodóncicas de magnitud conocida en maxilares de rata / Standardization of the method for the study of forces orthodontics of magnitude known in maxillary of rats

La ortodoncia se basa en la aplicación de fuerzas sobre las piezas dentarias en las que toma anclaje, provocando su movilización dentaria debido a la amplificación de los procesos de reabsorción y formación óseas en los alvéolos en los que se alojan dichas piezas. El objetivo de nuestro trabajo fue estandarizar la metodología utilizada para obtener la mayor precisión en los procedimientos, para poder evaluar correctamente el efecto de fuerzas ortodóncicas de magnitud conocida sobre piezas de muy pequeñas dimensiones como son los maxilares de rata. Tomando como base el modelo de ortodoncia experimental propuesto por otros autores para la rata se modificaron algunos aspectos de la aparatología y dispositivos utilizados.La precisión del sistema utilizado se evaluó sobre maxilares de ratas a las que se les instaló la aparatología con distintos tiempos de aplicación de las fuerzas. La aplicación de la fuerza pudo medirse apropiadamente en función del desplazamiento dentario sobre modelos cuando el tiempo de aplicación de la fuerza fue mayor a 48 horas. Sin embargo, cuando el tiempo de aplicación fue inferior, la fuerza sólo fue factible de medición en función de la compresión / ensanchamiento del espacio periodontal sobre los preparados histológicos de los maxilares. Los resultados obtenidos indican que la metodología propuesta, habiendo sido estandarizada, es útil para evaluar apropiadamente los efectos de fuerzas ortodóncicas experimentales sobre maxilares de rata (AU)

Estandarización del método para el estudio de fuerzas ortodóncicas de magnitud conocida en maxilares de rata / Standardization of the method for the study of forces orthodontics of magnitude known in maxillary of rats

La ortodoncia se basa en la aplicación de fuerzas sobre las piezas dentarias en las que toma anclaje, provocando su movilización dentaria debido a la amplificación de los procesos de reabsorción y formación óseas en los alvéolos en los que se alojan dichas piezas. El objetivo de nuestro trabajo fue estandarizar la metodología utilizada para obtener la mayor precisión en los procedimientos, para poder evaluar correctamente el efecto de fuerzas ortodóncicas de magnitud conocida sobre piezas de muy pequeñas dimensiones como son los maxilares de rata. Tomando como base el modelo de ortodoncia experimental propuesto por otros autores para la rata se modificaron algunos aspectos de la aparatología y dispositivos utilizados.La precisión del sistema utilizado se evaluó sobre maxilares de ratas a las que se les instaló la aparatología con distintos tiempos de aplicación de las fuerzas. La aplicación de la fuerza pudo medirse apropiadamente en función del desplazamiento dentario sobre modelos cuando el tiempo de aplicación de la fuerza fue mayor a 48 horas. Sin embargo, cuando el tiempo de aplicación fue inferior, la fuerza sólo fue factible de medición en función de la compresión / ensanchamiento del espacio periodontal sobre los preparados histológicos de los maxilares. Los resultados obtenidos indican que la metodología propuesta, habiendo sido estandarizada, es útil para evaluar apropiadamente los efectos de fuerzas ortodóncicas experimentales sobre maxilares de rata

Influence of short-term diabetes on osteocytic lacunae of alveolar bone. A histomorphometric study.

The aim of this study was to evaluate, for the first time, the histomorphometry of the cellular and lacunar features of the osteocytes of alveolar bone in acute streptozotocin-induced diabetic insulin-treated or untreated rats. Eighteen male Wistar rats weighing 200 to 260 g were assigned to one of the following groups: I) control group (C), II) diabetic group (DBT), and III) insulin treated diabetic group (DBT+INS). Experimental diabetes was induced by a single intraperitoneal injection of 60 mg/kg of body weight of streptozotocin. Insulin treatment began 24 h after the streptozotocin injection in animals of group DBT+INS in a dose of 4-6 IU of Humulin NPH insulin given as a single daily subcutaneous (s.c.) injection each morning between 07.00 and 10.00 h. The animals were euthanized on the 8th day. The upper maxillae were removed and fixed in buffered formalin, decalcified in EDTA, embedded in paraffin and stained with H-E for histologic and histomorphometric evaluation. Bone activity and lacunar density, osteocyte and empty lacunar densities, lacunar volume and lacunar shape were evaluated. Differences between variables were assessed by one-way ANOVA. Surface bone activity values revealed that bone resorption was significantly greater in the DBT group than in the C group (p < 0.05). Total lacunar density was significantly reduced in the DBT and DBT+INS groups as compared to control (p < 0.05). Concomitantly, a statistically significant reduction in osteocyte density and an increase, albeit not statistically significant, in empty lacunar density was observed in DBT and DBT+INS groups versus control. Lacunar volume did not exhibit statistically significant differences. The osteocyte lacunae in the DBT group lost their rounded shape and acquired intermediate shapes. This study reveals an early response of osteocytes to hyperglycemia, before systemic compensatory mechanisms are turned on. The effects are not always compensated by insulin treatment.

Alterations of tooth eruption and growth in pups suckling from diabetic dams.

Several studies have confirmed a decrease in the quality and quantity of milk of mothers with diabetes during lactation. However, it remains unclear how maternal diabetes affects the offspring specifically during lactation. The aim of this study was to evaluate body and mandibular growth and tooth eruption in pups suckling from diabetic dams. The study was performed on 13 Wistar rat pups that were born to dams that were subjected to experimental diabetes on the day of parturition. Body weight and body size were recorded regularly throughout the study. The experimental pups and a group of eight age-matched pups suckling from nondiabetic dams were killed at weaning. Both hemimandibles were excised and fixed. Right hemimandibles were radiographed to assess mandibular growth and tooth eruption. The left hemimandibles were processed to obtain buccolingually oriented sections at the level of the first mesial root of the first lower molar. Histologic and histomorphometric studies were performed. Results showed that body weight and body size were significantly lower in experimental animals at weaning compared with their age-matched controls. Smaller mandible size and reduced tooth eruption in experimental animals compared with controls were observed. The length, width, and bone volume of the developing alveolus were reduced in experimental animals compared with controls. The results obtained in this study allow the conclusion that suckling from diabetic dams results in reduced body, mandible size, and tooth eruption of the pups at weaning.

Effect of orthodontic forces on root resorption in molars submitted to experimental periodontitis.

The present experimental study addresses the issue of the development and size of root resorption areas in molars of rats submitted to orthodontic forces of 51 or 75 g and the induction of periodontal disease by placing a cotton ligature around the cervix of the first upper molars for 48 hours. Immediately on removal of the ligature or 48 hs later we put an orthodontic device in place. The device comprised two steel bands. The arms of a helicoidal spring that exerted force towards palatine passed through the palatine tubes welded to the bands. The number of odontoclasts and the percentage of root resorption areas were determined histomorphometrically on bucco-palatine sections obtained at the level of the central roots. The data showed an increase in root resorption areas when the orthodontic forces were applied to molars of rats following the induction of periodontitis. The magnitude of the force was proportional to the size of the resorption areas and to the number of odontoclasts. Both these end-points exhibited smaller values when the forces were applied once the inflammatory reaction had subsided. The present study shows that the risk of development of root resorption areas in patients with periodontal disease submitted to orthodontic treatment would be lower if lighter forces were applied and treatment were delayed until the inflammatory signs have subsided.

Effect of orthodontic forces on root resorption in molars submitted to experimental periodontitis.

The present experimental study addresses the issue of the development and size of root resorption areas in molars of rats submitted to orthodontic forces of 51 or 75 g and the induction of periodontal disease by placing a cotton ligature around the cervix of the first upper molars for 48 hours. Immediately on removal of the ligature or 48 hs later we put an orthodontic device in place. The device comprised two steel bands. The arms of a helicoidal spring that exerted force towards palatine passed through the palatine tubes welded to the bands. The number of odontoclasts and the percentage of root resorption areas were determined histomorphometrically on bucco-palatine sections obtained at the level of the central roots. The data showed an increase in root resorption areas when the orthodontic forces were applied to molars of rats following the induction of periodontitis. The magnitude of the force was proportional to the size of the resorption areas and to the number of odontoclasts. Both these end-points exhibited smaller values when the forces were applied once the inflammatory reaction had subsided. The present study shows that the risk of development of root resorption areas in patients with periodontal disease submitted to orthodontic treatment would be lower if lighter forces were applied and treatment were delayed until the inflammatory signs have subsided.