RESUMEN
Abstract The aim of this experimental study was to determine the effect of photobiomodulation therapy on bone repair in a rat tibia osteotomy model at 15 and 30 days. The sample consisted of 36 male Holtzman rats that were randomized into 6 equal groups. Groups A1 and A2: osteotomy + 1 J laser energy. Groups B1 and B2: osteotomy + 3 J laser energy. Groups C1 and C2 (controls): osteotomy only. The bone repair was analyzed by histological evaluation of osteoblasts and osteocytes both at 15 days (groups A1, B1, and C1) and at 30 days (groups A2, B2, and C2). Within the results, in all groups a greater number of osteoblasts was found at 15 days vs 30 days (p<0.05), and a greater number of osteocytes in B1 and C2 vs B2 and C1, respectively (p<0.05). When evaluating the 3 groups worked up to 15 days, more osteoblasts were found in A1 and C1 vs B1 (p<0.001); and osteocytes predominated in A1 and B1 vs C1 (p<0.001). At 30 days there was a greater quantity of osteoblasts in C2 vs A2 and B2 (p<0.05) and of osteocytes in C2 vs B2 (p<0.05). It is concluded that 1 J photobiomodulation therapy improved bone repair at 15 days; however, this improvement was not observed at 30 days because there were no differences between the irradiated groups and the control.
Resumen El objetivo de este estudio experimental fue determinar el efecto de terapia de fotobiomodulación sobre la reparación ósea en un modelo de osteotomía de tibia de rata a los 15 y 30 días. La muestra estuvo compuesta por 36 ratas Holtzman macho que se aleatorizaron en 6 grupos iguales. Grupos A1 y A2: osteotomía + energía láser de 1 Joule. Grupos B1 y B2: osteotomía + energía láser 3 Joule. Grupos C1 y C2 (controles): solo osteotomía. La reparación ósea fue analizada por evaluación histológica de osteoblastos y osteocitos tanto a los 15 días (grupos A1, B1 y C1) como a los 30 días (grupos A2, B2 y C2). Como resultados se encontró que en todos los grupos hubo mayor número de osteoblastos a los 15 días vs. 30 días (p<0,05), y mayor número de osteocitos en B1 y C2 vs B2 y C1, respectivamente (p<0,05). Al evaluar a los animales a los 15 días, se observó mayor número de osteoblastos en A1 y C1 vs B1 (p<0.001); y mayor número de osteocitos en A1 y B1 vs C1 (p<0,001). Al evaluar a los ratones a los 30 días hubo mayor cantidad de osteoblastos en C2 vs A2 y B2 (p<0,05) y de osteocitos en C2 vs B2 (p<0,05). Se concluye que la terapia de fotobiomodulación con 1 Joule mejoró la reparación ósea a los 15 días; sin embargo, dicha mejora no se observó a los 30 días porque no hubo diferencias entre los grupos irradiados y el control.
Asunto(s)
Animales , Ratas , Tibia , Fotobiología , Terapia por Luz de Baja Intensidad , HuesosRESUMEN
Aluminum (Al) is widely used for water purification, cooking pots, cosmetic and pharmaceutical preparations, toothpaste tubes, and food processing industries. Although the transport in the digestive tract is very poor but if the load is high, it can be absorbed and accumulated. About 50-70% of Al accumulates in the bones and can have an impact on human health. Resveratrol (RES), isolated from tempeh as an Indonesian food ingredient, can increase cell viability and has promising cytoprotective effects. RES has the capacity to interact with oxidative stress, so it has the potential as a therapy in bone repair. Therefore, this study aimed to evaluate the effect of RES on the number of osteocytes and bone marrow cells in Al-induced mice. Swiss Webster mice were divided into four groups: (1) untreated groups, (2) AlCl3 -treated groups, (3) Al+Res5 treated groups, and (4) Al+Res10 treated groups. Al dose 200 mg/kg body weight was administered intraperitoneally. RES was given one hour after administration of Al, with doses of 5 and 10 mg/kg Body Weight. Al and RES administration is carried out for one month. All mice were sacrificed, and mouse bones were isolated for histological preparations and a half for genotoxic assays. Bone marrow cells were collected and stained with My Grunwald. The number of micronuclei polychromatic erythrocytes (MNPCE) was examined in 1,000 PCEs per animal. The number of PCEs is counted by at least 200 erythrocytes (PCE + NCE) per animal. The results showed that the administration of Al significantly increased the number of micronuclei (MN) but after administration of RES at doses of 5 and 10 mg/kg Body Weight significantly reduced the number of MN in bone marrow cells. A dose of RES 10 mg/kg BW stimulates proliferation and increases the number of osteocytes in bone significantly. It can be concluded that Al can cause genotoxicity in bone marrow cells and RES is anti-genotoxic and can stimulate osteocyte proliferation.
O alumínio (Al) é amplamente utilizado para purificação de água, panelas, preparações cosméticas e farmacêuticas, tubos de pasta de dente e indústrias de processamento de alimentos. Embora o transporte no trato digestivo seja escasso, se a carga for alta, pode ser, todavia, absorvida e acumulada. Cerca de 50-70% do Al se acumula nos ossos e pode ter impacto na saúde humana. O resveratrol (RES), isolado do tempê indonésio como ingrediente alimentar, pode aumentar a viabilidade celular e tem efeitos citoprotetores promissores. O RES possui a capacidade de interagir com o estresse oxidativo, e por essa razão pode ser utilizado como terapia no reparo ósseo. Portanto, este estudo teve como objetivo avaliar o efeito do RES no número de osteócitos e células da medula óssea em camundongos induzidos por Al. Camundongos Swiss Webster (SW) foram divididos em quatro grupos: (1) grupos não tratados, (2) grupos tratados com AlCl3 , (3) grupos tratados com Al+Res5 e (4) grupos tratados com Al+Res10. Uma dose de 200 mg/kg de peso corporal foi administrada por via intraperitoneal. O RES foi administrado uma hora após a administração do Al, nas doses de 5 e 10 mg/kg de peso corporal. A administração de Al e RES foi realizada por um mês. Todos os camundongos foram sacrificados, e os ossos dos camundongos foram isolados para preparações histológicas e meio para ensaios genotóxicos. As células da medula óssea foram coletadas e coradas com My Grunwald. O número de eritrócitos policromáticos micronúcleos (MNPCE) foi examinado em 1.000 PCEs por animal. O número de PCEs foi contado por pelo menos 200 eritrócitos (PCE + NCE) por animal. Os resultados mostraram que a administração de Al aumentou significativamente o número de micronúcleos (MN), mas após a administração de RES nas doses de 5 e 10 mg/kg de peso corporal reduziu significativamente o número de MN nas células da medula óssea. Uma dose de RES de 10 mg/kg BW estimula a proliferação e aumenta significativamente o número de osteócitos no osso. Dessa forma, pôde-se concluir que o Al pode causar genotoxicidade em células da medula óssea e o RES é antigenotóxico e pode estimular a proliferação de osteócitos.
Asunto(s)
Animales , Ratones , Osteocitos , Células de la Médula Ósea/ultraestructura , Resveratrol , Cloruro de Aluminio/administración & dosificación , Pruebas de MutagenicidadRESUMEN
ABSTRACT Introduction: Induced tooth-bone movement occurs by a synchronicity of dental and bone phenomena, thanks to the osteocytic network, which is a three-dimensional network that controls the bone shape or design. Objective: To describe the tooth-bone movement induced by enhanced anchorage, divided into three distinct moments: zero, start and stop. Question: From this description, the main question arises: with the use of mini-implants/miniplates, what changes in the biology of induced tooth-bone movement? The answer is: nothing changes, either biologically or microscopically. Conclusion: This technique optimizes the treatment time, and the range of therapeutic possibilities is broadened, thanks to the synchronicity of phenomena - which remain the same, in all teeth and bones, yet in a synchronized manner. Bone anchorage represents synchronicity in induced tooth-bone movement.
RESUMO Introdução: A movimentação osseodentária induzida ocorre meio de uma sincronicidade de fenômenos dentários e ósseos, graças à rede osteocítica, uma rede tridimensional de controle do formato ou design ósseo. Objetivo: Descrever a movimentação osseodentária induzida com ancoragem ampliada, dividindo-a em três momentos distintos: zero, start e stop. Questionamento: Dessa descrição origina-se a principal pergunta: com o uso de mini-implantes/miniplacas, o que muda na biologia da movimentação osseodentária induzida? A resposta é: não muda nada, nem biologicamente, nem microscopicamente. Conclusão: O que se otimiza, com essa técnica, é o tempo de tratamento, e se amplia o leque de possibilidades terapêuticas, graças à sincronicidade dos fenômenos - que continuam sendo os mesmos, em todos os dentes e nos ossos, só que de forma sincronizada. A ancoragem óssea representa a sincronicidade na movimentação osseodentária induzida.
RESUMEN
OBJECTIVES: This animal study was conducted to evaluate the osteocyte index in the peri-implant bone around immediately restored implants under static lateral overload. MATERIAL AND METHODS: Seven mongrel dogs received three implants on each side of the mandible. Forty-two implants were distributed into three groups (14 implants per group); each animal received two implants connected to a 4.5-mm opened expansion device (experimental group); in the other mandible side, two implants were connected into an expansion device without activation (control group); one implant each side of the mandible was left submerged (unload group). After 4 months under daily mechanical and chemical plaque control, the animals were euthanized; dental implants and surrounding bone were removed and processed to obtain thin ground sections. Histomorphometry was used to evaluate the osteocyte index in the peri-implant bone contact to implant. RESULTS: A higher, statistically significant mean number of osteocytes × 10-5 µm2 (54.74 ± 23.91) was found in the control group compared with the test group (22.57 ± 22.55) (p = 0.0221). The correlation between percentage of bone-implant contact and osteocyte index for submerged implants was not statistically significant (p = 0.2667), whereas the value for immediately loaded implants was statistically significant (p = 0.0480). CONCLUSION: The lower number of osteocytes in the peri-implant bone around overloaded implants could be related to the need for functional adaptation of the bone tissue to overloading and to the hypothesized involvement of the osteocytes in the maintenance of the bone matrix in the control group. CLINICAL RELEVANCE: Osteocytes play a pivotal role in bone adaptation to mechanical loading, and the osteocyte network has been regarded as being the main mechanosensory mechanism.
Asunto(s)
Implantes Dentales , Placa Dental , Animales , Implantación Dental Endoósea , Perros , Mandíbula/cirugía , Oseointegración , OsteocitosRESUMEN
Following radiotherapy, patients have decreased bone mass and increased risk of fragility fractures. Diabetes mellitus (DM) is also reported to have detrimental effects on bone architecture and quality. However, no clinical or experimental study has systematically characterized the bone phenotype of the diabetic patients following radiotherapy. After one month of streptozotocin injection, three-month-old male rats were subjected to focal radiotherapy (8 Gy, twice, at days 1 and 3), and then bone mass, microarchitecture, and turnover as well as bone cell activities were evaluated at 2 months post-irradiation. Micro-computed tomography results demonstrated that DM rats exhibited greater deterioration in trabecular bone mass and microarchitecture following irradiation compared with the damage to bone structure induced by DM or radiotherapy. The serum biochemical, bone histomorphometric, and gene expression assays revealed that DM combined with radiotherapy showed lower bone formation rate, osteoblast number on bone surface, and expression of osteoblast-related markers (ALP, Runx2, Osx, and Col-1) compared with DM or irradiation alone. DM plus irradiation also caused higher bone resorption rate, osteoclast number on bone surface, and expression of osteoclast-specific markers (TRAP, cathepsin K, and calcitonin receptor) than DM or irradiation treatment alone. Moreover, lower osteocyte survival and higher expression of Sost and DKK1 genes (two negative modulators of Wnt signaling) were observed in rats with combined DM and radiotherapy. Together, these findings revealed a higher deterioration of the diabetic skeleton following radiotherapy, and emphasized the clinical importance of health maintenance.
RESUMEN
Objetive: To determine the expressions of the bone surface marker CD44 in samples of alveolar bone previously regenerated with allograft, xenograft, and mixed, using the technique of guided bone regeneration. Material and Methods: This exploratory study was approved by the institutional research and ethics committee. By means of intentional sampling and after obtaining informed consent for tissue donation, 20 samples of alveolar bone previously regenerated with guided bone regeneration therapy with particulate bone graft and membrane were taken during implant placement. The samples were stained with hematoxylin-eosin for histological analysis, and by immunohistochemistry for the detection of CD44. Results: Sections with hematoxylin-eosin showed bone tissue with the presence of osteoid matrix and mature bone matrix of usual appearance. Of the CD44+ samples, 80% were allograft and 20% xenograft. The samples with allograft-xenograft were negative. There were no differences in the intensity of CD44 expression between the positive samples. The marker was expressed in osteocytes, stromal cells, mononuclear infiltrate, and some histiocytes. Eighty percent of the CD44+ samples and 100% of the samples in which 60 or more cells were labelled corresponded to allografts (p=0.000). A total of 67% of the samples from the anterior sector, and 40% from the posterior sector were CD44+ (p=0.689). Conclusion: This study shows for the first time that guided bone regeneration using allografts is more efficient for the generation of mature bone determined by the expression of CD44, compared to the use of xenografts and mixed allograft-xenograft, regardless of the regenerated anatomical area.
Objetivo: Determinar la expresión del marcador de membrana óseo CD44 en muestras de hueso alveolar previamente regenerado con aloinjerto, xenoinjerto y mezcla mediante la técnica de regeneración ósea guiada. Material y Métodos: Con aval del Comité de Investigación y Ética, se realizó un estudio exploratorio. Por muestreo intencional y firma de consentimiento informado de donación, se tomaron durante la colocación del implante, 20 muestras de hueso alveolar previamente regenerado con terapia de regeneración ósea guiada con injerto óseo particulado y membrana. Las muestras fueron teñidas con hematoxilina-eosina para el análisis histológico y por inmunohistoquímica para la detección del CD44. Resultados: : Los cortes con hematoxilina-eosina mostraron tejido óseo con presencia de matriz osteoide y matriz ósea madura de aspecto usual. De las muestras CD44+, 80% fueron de aloinjerto y 20% de xenoinjerto. Las muestras con aloinjerto-xeoninjerto fueron negativas. No hubo diferencias en la intensidad de la expresión del CD44 entre las muestras positivas. El marcador se expresó en osteocitos, células estromales, infiltrado mononuclear y algunos histiocitos. El 80% de las muestras CD44+ y el 100% de las muestras con marcación de 60 o más células correspondían a aloinjertos (p=0,000). El 67% de las muestras del sector anterior y el 40% del sector posterior fueron CD44+ (p=0,689). Conclusión: Este estudio muestra por primera vez que la regeneración ósea guiada usando aloinjertos, es más eficiente para la generación de hueso maduro determinado por la expresión de CD44, comparado con el uso de xenoinjertos y mezcla de aloinjerto-xenoinjerto, independientemente del sector anatómico regenerado.
Asunto(s)
Humanos , Masculino , Femenino , Receptores de Hialuranos/metabolismo , Injerto de Hueso Alveolar , Osteocitos , Regeneración Ósea , Implantes Dentales , Receptores de Hialuranos/genética , Aloinjertos , XenoinjertosRESUMEN
Osteoarticular brucellosis is the most frequent complication of active disease. A large amount of cells in bone are osteocytes. Since bone remodeling process is regulated by hormones we sought to study the effect of cortisol and DHEA in Brucella abortus-infected osteocytes. Cortisol treatment inhibited the expression of IL-6, TNF-α, MMP-2 and RANKL in B. abortus-infected osteocytes. DHEA could reverse the inhibitory effect of cortisol on MMP-2 production. B. abortus infection inhibited connexin 43 (Cx43) expression in osteocytes. This expression was increased when cortisol was incorporated during the infection and DHEA treatment partially reversed the effect of cortisol. Osteocytes-infected with B. abortus induced osteoclast's differentiation. Yet, the presence of cortisol, but not DHEA, during osteocyte infection inhibited osteoclastogenesis. Glucocorticoid receptor (GR) is implicated in the signaling of cortisol. Infection with B. abortus was able to increase GRα/ß ratio. Levels of intracellular cortisol are not only dependent on GR expression but also a result of the activity of the isoenzymes 11ß-hydroxysteroid dehydrogenase (11ß-HSD)-1 (cortisone to cortisol conversion), 11ß-HSD2 (cortisol to cortisone conversion). B. abortus infection increased 11ß-HSD 1/2 ratio and cortisone mimicked the effect of cortisol. Our results indicated that cortisol and DHEA could modulate osteocyte responses during B. abortus infection.
Asunto(s)
Brucella abortus/fisiología , Brucelosis/patología , Osteocitos/microbiología , Osteoprotegerina/metabolismo , Ligando RANK/metabolismo , 11-beta-Hidroxiesteroide Deshidrogenasas/genética , Animales , Brucella abortus/crecimiento & desarrollo , Brucella abortus/metabolismo , Brucelosis/metabolismo , Células Cultivadas , Conexina 43/metabolismo , Cortisona/farmacología , Medios de Cultivo Condicionados/farmacología , Citocinas/metabolismo , Deshidroepiandrosterona/farmacología , Hidrocortisona/metabolismo , Hidrocortisona/farmacología , Metaloproteinasa 2 de la Matriz/metabolismo , Ratones , Viabilidad Microbiana , Osteocitos/citología , Osteocitos/efectos de los fármacos , Osteocitos/metabolismo , Osteogénesis/efectos de los fármacos , Osteoprotegerina/antagonistas & inhibidores , Receptores de Glucocorticoides/genética , Transducción de SeñalRESUMEN
Secondary hyperparathyroidism is a complication of chronic kidney disease that compromises skeletal integrity. In patients with secondary hyperparathyroidism undergoing parathyroidectomy, parathyroid hormone levels dramatically decrease. The effects of parathyroidectomy on bone tissue are poorly understood, especially regarding the proteins expressed by osteocytes, such as fibroblast growth factor 23, dentin matrix protein 1, matrix extracellular phosphoglycoprotein, sclerostin, receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin, which regulate bone turnover. The objective of this study was to characterize the bone expression of these proteins by immunohistochemistry and correlate these results with those of bone histomorphometry before and after parathyroidectomy. We studied bone biopsies that were obtained from 23 patients before and 12â¯months after parathyroidectomy. We observed an improvement in bone microarchitecture, but impaired mineralization after parathyroidectomy. We found significant increases in sclerostin and osteoprotegerin expression and a decrease in the RANKL/osteoprotegerin ratio after parathyroidectomy, suggesting that their expression is regulated by parathormone. These proteins correlated with structural and bone formation parameters. We conclude that after parathyroidectomy, significant changes occur in the bone expression of osteocyte proteins and that these proteins potentially regulate bone remodeling.
Asunto(s)
Huesos/metabolismo , Hiperparatiroidismo Secundario/metabolismo , Paratiroidectomía , Insuficiencia Renal Crónica/metabolismo , Insuficiencia Renal Crónica/cirugía , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Humanos , Osteocitos/fisiología , Osteoprotegerina/metabolismoRESUMEN
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.
Asunto(s)
Apoptosis , Osteocitos/fisiología , Osteopontina/biosíntesis , Estrés Mecánico , Técnicas de Movimiento Dental , Proceso Alveolar/citología , Animales , Masculino , Aparatos Ortodóncicos , Osteocitos/metabolismo , Ratas , Ratas WistarRESUMEN
Estrogen maintains osteocyte viability, whereas its deficiency induces osteocyte apoptosis. As autophagy is important for osteocyte viability, we hypothesized whether the anti-apoptotic effect of estrogen is related to autophagy in osteocytes. Thirty adult female rats were sham-operated (SHAM) or ovariectomized (OVX). After three weeks, twelve rats of SHAM and OVX groups were killed before treatment (basal period), whereas the remaining rats received estrogen (OVXE) or vehicle (OVX) for 45 days. Fragments of maxilla containing alveolar process of the first molars were embedded in paraffin or Araldite. Paraffin-sections were stained with hematoxylin/eosin for histomorphometry, or subjected to the silver impregnation method for morphological analysis of osteocyte cytoplasmic processes. Autophagy was analyzed by immunohistochemical detections of beclin-1, MAP-LC3α and p62, whereas apoptosis was evaluated by immunohistochemical detections of cleaved caspase-3 and BAX, TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) method and by ultrastructural analysis. Araldite-semithin sections were subjected to the Sudan-black method for detection of lipids. OVX-basal group showed high frequency of caspase-3-, TUNEL- and p62-positive osteocytes accompanied with low frequency of beclin-1- and MAP-LC3α-positive osteocytes. At 45 days, OVXE group exhibited higher number of osteocytes, higher frequency of beclin-1- and MAP-LC3α-positive osteocytes, and lower frequency of caspase-3, BAX-, TUNEL- and p62-positive osteocytes than OVX group. Significant reduction in bone area was observed in the OVX compared to OVXE and SHAM groups. The highest frequency of Sudan-Black-positive osteocytes and osteocytes with scarce cytoplasmic processes, or showing apoptotic features were mainly observed in OVX groups. Our results indicate that estrogen deficiency decreases autophagy and increases apoptosis, whereas estrogen replacement enhances osteocyte viability by inhibiting apoptosis and maintaining autophagy in alveolar process osteocytes. These results suggest that the anti-apoptotic effect of estrogen may be, at least in part, related to autophagy regulation in osteocytes.
Asunto(s)
Autofagia/fisiología , Supervivencia Celular/fisiología , Estrógenos/metabolismo , Osteocitos/metabolismo , Proceso Alveolar/metabolismo , Animales , Apoptosis/fisiología , Proteínas Reguladoras de la Apoptosis/metabolismo , Caspasa 3/metabolismo , Femenino , Etiquetado Corte-Fin in Situ/métodos , Ovariectomía/métodos , Ratas , Ratas WistarRESUMEN
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. Grants: UBACyT 200201301002270 BA and School of Dentistry, University of Buenos Aires (AU)
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 (AU)
Asunto(s)
Animales , Ratas , Osteocitos , Estrés Mecánico , Técnicas de Movimiento Dental , Apoptosis , Osteopontina , Inmunohistoquímica , Interpretación Estadística de Datos , Etiquetado Corte-Fin in Situ , Mecanotransducción Celular , Proceso AlveolarRESUMEN
The selection of an appropriate demineralizing solution in pathology laboratories depends on several factors such as the preservation of cellularity, urgency of diagnostic and financial costs. The aim of this study was to test different decalcification bone procedures in order to establish the best value of these in formalin-fixed and paraffin-embedded samples. Femurs were removed from 13 adult male Wistar rats to obtain 130 bone disks randomly divided into five groups that were demineralized in different concentrations of nitric acid (Group I); formic acid (Group II); acetic acid (Group III); EDTA, pH7.4 (Group IV) and Morsés solution (Group V). Serial, 3-µm-thick sections were obtained and stained with hematoxylin-eosin to calculate the percentage of osteocyte-occupied lacunae. The sections were also stained with Masson's trichrome in conjunction with picrosirius red under polarized light followed by a semi-quantitative analysis to verify the adjacent muscle-to-bone integrity and preservation of collagen fibres. The highest percentage of osteocyte-occupied lacunae was found with 10% acetic acid solution (95.64 ± 0.95%) and Group I (nitric acid) demanded the shorter time (0.8-5.7days). Of all solutions, 5% nitric acid incurred the lowest cost to achieve complete demineralization compared with other solutions (p < .001). Group IV (EDTA) had the highest integrity of muscle and collagen type I and III (P < 0.01). Demineralization with 10% acetic acid was the most effective at preserving bone tissue, while 5% EDTA was the best at maintaining collagen and adjacent muscle to bone. In conclusion, nitric acid at 5% showed the most efficient result as it balanced both time and cost as a demineralizing solution.
Asunto(s)
Técnica de Desmineralización de Huesos/economía , Técnica de Desmineralización de Huesos/métodos , Técnica de Descalcificación/economía , Técnica de Descalcificación/métodos , Fémur/química , Estudios de Tiempo y Movimiento , Ácido Acético/química , Animales , Huesos/química , Colágeno Tipo I/análisis , Colágeno Tipo III/análisis , Ácido Edético/química , Formiatos/química , Masculino , Músculos/fisiología , Ácido Nítrico/química , Ratas , Ratas Wistar , Coloración y Etiquetado/economía , Coloración y Etiquetado/métodosRESUMEN
Actualmente, los mecanismos biológicos que subyacen a la estimulación ortopédica funcional están en proceso de entendimiento; sin embargo, se sabe que el osteocito juega un rol esencial, al recibir y transformar dicho estímulo funcional hacia una señal bioquímica, lo que da como consecuencia la secreción de diversas moléculas. Estas se movilizan entre los osteocitos, gracias a su extensa red de uniones comunicantes, y llegan en última instancia a activar a las células efectoras del tejido óseo: osteoblastos y osteoclastos. El objetivo de la revisión es actualizar y compendiar algunos de los más importantes mecanismos celulares y moleculares subyacentes a la terapia ortopédica funcional de los maxilares.
Currently, the biological mechanisms underlying functional orthopedic stimulation are in process of understanding. However, it is known that osteocyte plays an essential role, to receive and process the functional stimulus to biochemical signals giving as result the secretion of various molecules. Such molecules are mobilized between the osteocytes, thanks to its extensive network of gap junctions, ultimately coming to activate effector cells of bone tissue: osteoblasts and osteoclasts. The aim of the review is to update some of the cellular and molecular mechanisms underlying functional orthopedic therapy of the maxillary.
RESUMEN
Introducción: Estudio experimental donde se procuró determinar el efecto osteoinductor del mineral trióxido agregado (MTA) versus el cemento Portland tipo I sobre lesiones óseas mandibulares. Metodología: Se emplearon 12 conejos machos de la raza New Zealand de 3 meses de edad, los cuales fueron divididos en 4 grupos iguales. Todos los conejos fueron anestesiados utilizando pentobarbital sódico. Se procedió a la incisión en la piel mandibular para exponer el hueso sobre el que se realizó 3 cavidades de 3mm cada una. En una cavidad se colocó MTA, en otra cemento Portland y en la tercena ninguna pasta. Se procedió al sacrificio de los grupos experimentales a la 1era, 2da, 3era y 4ta semana respectiva y se evaluó las muestras obtenidas de las áreas quirúrgicas mediante conteo de osteocitos y osteoblastos. Resultados: Tanto el MTA como el cemento Portland poseen la misma capacidad osteoinductiva en la 1era, 2da y 3era semana (p>0,05). Sin embargo, en la 4ta semana el MTA tuvo mayor capacidad osteoinductora al estimular mayor número de osteoblastos que el cemento Portland (p=0,024). Conclusiones: El MTA y el cemento Portland tipo I mostraron similar efecto osteoinductor durante las 3 primeras semanas de evaluación. El MTA demostró mayor efecto osteoinductor durante la cuarta semana de valoración.
Introduction: An experimental study was carried out to determine the osteoinductive effect of Mineral Trioxide Aggregate (MTA) versus Portland Cement type I on mandibular bone lesions. Methodology: Twelve 3-month-old male New Zealand rabbits were divided into 4 equal groups. All rabbits were anesthetized using Pentobarbital. An incision of the mandibular skin was performed to expose the bone on which 3 cavities of 2mm each one were made. In one cavity MTA was placed, in another Portland Cement type I and the third remained empty. The experimental groups were sacrificed at the 1st, 2nd, 3rd and 4th respective weeks and evaluated histologically by counting osteocytes and osteoblasts. Results: Both MTA and Portland cement have the same osteoinductive capacity in the 1st, 2nd and 3rd week (0.05
RESUMEN
The functional demand imposed on bone promotes changes in the spatial properties of osteocytes as well as in their extensions uniformly distributed throughout the mineralized surface. Once spatial deformation is established, osteocytes create the need for structural adaptations that result in bone formation and resorption that happen to meet the functional demands. The endosteum and the periosteum are the effectors responsible for stimulating adaptive osteocytes in the inner and outer surfaces. Changes in shape, volume and position of the jaws as a result of skeletal correction of the maxilla and mandible require anchorage to allow bone remodeling to redefine morphology, esthetics and function as a result of spatial deformation conducted by orthodontic appliances. Examining the degree of changes in shape, volume and structural relationship of areas where mini-implants and miniplates are placed allows us to classify mini-implants as devices of subabsolute anchorage and miniplates as devices of absolute anchorage.
Asunto(s)
Placas Óseas , Implantes Dentales , Métodos de Anclaje en Ortodoncia/instrumentación , Matriz Ósea/fisiología , Remodelación Ósea/fisiología , Resorción Ósea/fisiopatología , Humanos , Mandíbula/citología , Maxilar/citología , Mecanotransducción Celular/fisiología , Miniaturización , Diseño de Aparato Ortodóncico , Osteoblastos/fisiología , Osteoclastos/fisiología , Osteocitos/fisiología , Osteogénesis/fisiología , Periostio/fisiología , Técnicas de Movimiento Dental/instrumentaciónRESUMEN
The functional demand imposed on bone promotes changes in the spatial properties of osteocytes as well as in their extensions uniformly distributed throughout the mineralized surface. Once spatial deformation is established, osteocytes create the need for structural adaptations that result in bone formation and resorption that happen to meet the functional demands. The endosteum and the periosteum are the effectors responsible for stimulating adaptive osteocytes in the inner and outer surfaces.Changes in shape, volume and position of the jaws as a result of skeletal correction of the maxilla and mandible require anchorage to allow bone remodeling to redefine morphology, esthetics and function as a result of spatial deformation conducted by orthodontic appliances. Examining the degree of changes in shape, volume and structural relationship of areas where mini-implants and miniplates are placed allows us to classify mini-implants as devices of subabsolute anchorage and miniplates as devices of absolute anchorage.
Uma demanda funcional sobre o osso promove alterações na forma espacial da rede de osteócitos e seus prolongamentos, distribuídos uniformemente na estrutura mineralizada. A partir da deformação espacial captada, os osteócitos comandam a necessidade de adaptações estruturais, formando osso em novas áreas e reabsorvendo em outras, para que sejam atendidas as demandas funcionais. O endósteo e o periósteo são os verdadeiros efetores desses estímulos osteocíticos adaptativos, nas superfícies internas e externas. As alterações de forma, volume e posição dos ossos maxilares, nas correções esqueléticas da maxila e mandíbula, requerem uma ancoragem para que a remodelação óssea redefina a morfologia, a estética e as funções, a partir de deformações espaciais dirigidas por aparelhos. Verificar o grau de alterações na forma, volume e relações estruturais das áreas onde se fixaram os mini-implantes e as miniplacas poderá levar à classificação dos mini-implantes como dispositivos de ancoragem subabsoluta e as miniplacas, como de ancoragem absoluta.
Asunto(s)
Humanos , Placas Óseas , Implantes Dentales , Métodos de Anclaje en Ortodoncia/instrumentación , Matriz Ósea/fisiología , Remodelación Ósea/fisiología , Resorción Ósea/fisiopatología , Miniaturización , Mandíbula/citología , Maxilar/citología , Mecanotransducción Celular/fisiología , Diseño de Aparato Ortodóncico , Osteoblastos/fisiología , Osteoclastos/fisiología , Osteocitos/fisiología , Osteogénesis/fisiología , Periostio/fisiología , Técnicas de Movimiento Dental/instrumentaciónRESUMEN
Introducción: La hipoxia caracteriza a los tejidos inicialmente injuriados; la variación en la disposición del oxígeno tisular en el proceso de reparación ósea determina la aparición de diversas moléculas implicadas en la sanación del tejido. Objetivos: Determinar si la variación de la presión de oxígeno ambiental influirá en la formación ósea posterior a una osteotomía en cuyes nativos del nivel del mar. Diseño: Experimental. Lugar: Facultad de Odontología de la Universidad Nacional Mayor de San Marcos, Lima, Perú, e Instituto Veterinario de Investigaciones Tropicales y de Altura, Mantaro, Perú. Material biológico: Cuyes. Intervenciones: Se utilizó 5 grupos de 10 cuyes cada uno, uno sin inducción de osteotomía (grupo control) y cuatro grupos experimentales: mar 15 días y mar 30 días (expuestos a PO2 ambiental de 157 mmHg), altura 15 días y altura 30 días (expuestos a PO2 ambiental de 107 mmHg). Principales medidas de resultados: Conteo de osteocitos. Resultados: El grupo mar 15 días presentó menor número de osteocitos comparado con el grupo altura 15 días (63 180 vs. 80 310, p<0,05). A su vez, el grupo mar 30 días presentó también menor número de osteocitos comparado con el grupo altura 30 días (160 640 vs. 167 370, p<0,05). Conclusión: La menor presión de oxígeno ambiental favoreció una mayor formación ósea en cuyes nativos del nivel mar...
Introduction: Hypoxia characterizes initially injured tissue; variation in oxygen available determines the appearance of molecules involved in bone repair. Objectives: To determine if environmental oxygen pressure variation influences bone formation following osteotomy in sea level native guinea pigs. Design: Experimental. Setting: Faculty of Dentistry, Universidad Nacional Mayor de San Marcos, Lima, Peru, and Tropical and High Altitude Research Veterinary Institute, Mantaro, Peru. Biological material: Guinea pigs. Interventions: Five groups of 10 guinea pigs each included one without osteotomy (control group) and four experimental groups: sea 15 days and sea 30 days (exposed to environmental PO2 157 mmHg), high altitude 15 days and high altitude 30 days (exposed to environmental PO2 107 mmHg). Main outcome measures: Osteocytes count. Results: The 15 days sea group had lower number of osteocytes compared with the 15 days high altitude group (63 180 vs. 80 310, p<0.05). The 30 days sea group had also lower number of osteocytes compared with the 30 days high altitude group (160 640 vs. 167 370, p<0.05). Conclusions: The lower environmental oxygen pressure favored higher bone formation in sea level native guinea pigs...
Asunto(s)
Masculino , Animales , Cobayas , Hipoxia , Consumo de Oxígeno , Experimentación Animal , Osteocitos , Osteogénesis , Ensayo ClínicoRESUMEN
Os alvos principais na compreensão da biopatologia óssea centravam-se nos osteoblastos e clastos, mas nos últimos anos têm se deslocado para os osteócitos como mecanotransdutores do tecido ósseo, a partir da rede tridimensional, pelo entrelaçamento e contato de seus prolongamentos interligando uma célula a outras 20 a 40, tal qual uma rede neural. Pela mecanotransdução e a partir de mediadores como a esclerostina e o RANKL, os osteócitos podem influenciar na biopatologia óssea por interferirem na atividade dos osteoblastos e clastos. Quando necessário mais osso, os osteócitos liberam menos esclerostina; quando é necessário inibir a formação óssea, os osteócitos liberam mais esclerostina. O RANKL está ligado à osteoclastogênese local para que se tenha mais células capazes de reabsorver a matriz mineralizada. Algumas terapêuticas inovadoras das doenças ósseas metabólicas têm tido como alvo esses mediadores e os osteócitos. Estudar a presença e os efeitos específicos da esclerostina e do RANKL na osseointegração pode levar a um maior detalhamento de seus fenômenos biológicos.
The main targets for the comprehension of bone pathobiology were focused in osteoblasts and clasts, but in recent years it has shifted to the osteocytes as mechanotransductors of the bone tissue, from the three-dimensional network, by interconnecting its extensions linking a cell to other 20 to 40, like a neural network. By mechanotransduction and from mediators as sclerostin and RANKL, the osteocytes may influence bone pathobiology by interfering with the activity of osteoblasts and clasts. When more bone is necessary, osteocytes release less sclerostin, when it is necessary to inhibit bone formation, osteocytes release more sclerostin. RANKLis connected to local osteoclastogenesis in order to have more cells capable of reabsorbing the mineralized matrix. New therapeutic ways of controlling the metabolic bone diseases have been targeted at these mediators. Studying the presence and the specific effects of sclerostin and RANKL in osseointegration can lead to greater detailing of their biological phenomena.
Asunto(s)
Humanos , Mecanotransducción Celular , Huesos/citología , Osteocitos/citología , Ligando RANK , Matriz Ósea , Remodelación Ósea , Resorción Ósea , Implantación Dental , Enfermedades Óseas Metabólicas/terapia , Oseointegración , OsteogénesisRESUMEN
Osteoblasts and clasts were primary targets for the understanding of bone biopathology. In recent years, evidence has shifted attention to the osteocytes. The biology of induced tooth movement and jaw orthopedics should research the role of osteocytes and the specific effects of mediators such as RANKL and sclerostin. The sclerostin represents a regulatory molecule: When more bone is necessary, osteocytes release less sclerostin, when it is necessary to inhibit bone formation, osteocytes release more sclerostin. RANKL is connected to local osteoclastogenesis in order to have more cells capable of reabsorbing the mineralized matrix. New therapeutic ways of controlling the metabolic bone diseases have been targeted at these mediators.
RESUMEN
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.