ABSTRACT
A cartilagem de Meckel é uma estrutura transitória embrionária presente durante os estágios iniciais da formação da mandíbula, localizada em toda sua extensão e dividida em três porções, anterior, intermediária e posterior. O enfoque deste trabalho foi direcionado à elucidação do destino final da porção intermediária por meio de um estudo temporal sequenciado. Por isso, foi investigado a presença de células de reabsorção e a presença de fibras colágenas, bem como da proteína óssea osteopontina (OPN) na cartilagem de Meckel na região do germe do 1º molar inferior e no seu entorno. Foram utilizados fetos de ratos Wistar em períodos gestacionais pré-estabelecidos, G18 a G21 (grupos de dias intrauterinos), bem como P0 e P1 (recém-nascidos) para remoção das cabeças. Em sequência, os espécimes foram fixados em solução de formaldeído 4% + glutaraldeído 0,1% com tampão fosfato 0,1M, descalcificados em EDTA 4,13%, desidratados em concentrações crescentes de etanol e incluídos em parafina. As amostras foram coradas em hematoxilina e eosina (HE) e tricrômico de Mallory para análise histológica. Adicionalmente, os grupos G19 a P0 foram submetidos à reação histoquímica de TRAP para determinação da presença de células clásticas. Além disso, os grupos G21 e P0 (dia do nascimento) passaram por reações de imunomarcação para análise da expressão de OPN. Foi observado a degeneração gradual da cartilagem com a observação de mudanças estruturais, a justaposição de células clásticas na superfície da cartilagem por reação histoquímica TRAP a partir do G21, o aparecimento de colágeno tipo I nas fases terminais da degeneração, assim como a marcação positiva para a osteopontina na superfície de G21 e em todo o remanescente da cartilagem de Meckel no grupo P0. O estudo apontou um processo de degeneração da cartilagem com evidências de formação de matriz mineralizada de natureza óssea, a qual foi reabsorvida por células clásticas, sugerindo a ossificação da porção intermediária da cartilagem de Meckel.
Subject(s)
Osteoclasts , Cartilage , Osteopontin , MandibleABSTRACT
Abstract Objective This study aimed to correlate the expression, by immunohistochemistry, of the proteins OPN, ABCB5, and WNT3A from anatomopathological materials obtained from paraffin blocks, slides, or both, from patients with osteosarcoma (OS), analyzing epidemiological characteristics, as well as their presence and influence on the evolution and progression of the disease. Methods After the initial case selection, we searched for the respective paraffin blocks and took only those with sufficient tumor mass to allow additional sections with no complete loss of biological material. The sarcoma area identification in representative paraffin blocks used multisample blocks (tissue microarray [TMA]) created on a BenchMark ULTRA (Roche Diagnostics Corporation, Indianapolis, IN, USA) instrument. Then, we analyzed the association between the expression of ABCB5, WNT3A, and osteopontin (OPN) markers with the variables age, location, and tissue type (Fisher exact test/Chi-squared test). Results The average age of the patients was 23 years, and the rate of males and females was the same. We analyzed 40 slides from 28 OS patients seen from 2005 to 2017. Their follow-up time was 80.0 months, and the 5-year survival rate was 46.7%. Most metastases occurred in lung tissue (92.9%). Proteins ABCB5, OPN, and WNT3A did not present statistical significance when compared with age group, neo-adjuvant, adjuvant, or both, chemotherapy, location, survival, or death. Osteopontin was negative in all samples. WNT3A expression occurred in patients who died early. Conclusion In an immunohistochemical study, ABCB5, OPN, and WNT3A did not have statistical significance. In the parameters analyzed, they did not seem to be a predictive or aggressive factor for OS.
Resumo Objetivo Correlacionar a expressão, por imunoistoquímica, das proteínas OPN, ABCB5 e WNT3A de material anatomopatológico, obtido de blocos de parafina e/ou lâminas, em pacientes com osteossarcoma (OS), analisando as características epidemiológicas, sua presença e influência na evolução e progressão da doença. Métodos Após a seleção inicial dos casos, ocorreu a busca dos respectivos blocos de parafina, dentre os quais foram selecionados somente aqueles que possuíam massa tumoral suficiente para serem realizados cortes adicionais sem que todo o material biológico fosse utilizado. Foram identificadas áreas do sarcoma nos blocos de parafina representativos para a confecção de blocos multiamostrais (microarranjo de tecidos, ou tissue microarray [TMA], em inglês), realizada em instrumento BenchMark ULTRA (Roche Diagnostics Corporation, Indianapolis, IN, USA). Foi então analisada a associação entre a expressão dos marcadores ABCB5, WNT3A e OPN com as variáveis idade, localização e tipo de tecido (teste exato de Fisher/Qui-quadrado). Resultados A média de idade foi de 23 anos, e a incidência de pacientes dos sexos masculino e feminino foi a mesma; foram analisadas 40 lâminas de 28 pacientes com OS, entre 2005 e 2017, com tempo de segmento de 80,0 meses, e o tempo de sobrevida foi de 46,7% em 5 anos. Metástases ocorreram em tecido pulmonar (92,9%). Quanto aos marcadores ABCB5, OPN e WNT3A, não apresentaram significância estatísticas quando comparados com faixa etária, neo-adjuvância e/ou adjuvância quimioterápica, localização, sobrevida ou óbito. O OPN mostrou-se negativo em todas as amostras. E o WNT3A expressou-se nos pacientes com óbitos precoces. Conclusão As proteínas ABCB5, OPN e WNT3A, em estudo imunoistoquímico, não se mostraram presentes com significância estatística. Nos parâmetros analisados, não surgem como sendo fatores preditivos ou de agressividade para o OS.
Subject(s)
Humans , Male , Female , Adult , Immunohistochemistry , Biomarkers, Tumor , Osteosarcoma , Survival Rate , Osteopontin , Neoplasm MetastasisABSTRACT
BACKGROUND@#Congenital scoliosis (CS) is a complex spinal malformation of unknown etiology with abnormal bone metabolism. Fibroblast growth factor 23 (FGF23), secreted by osteoblasts and osteocytes, can inhibit bone formation and mineralization. This research aims to investigate the relationship between CS and FGF23.@*METHODS@#We collected peripheral blood from two pairs of identical twins for methylation sequencing of the target region. FGF23 mRNA levels in the peripheral blood of CS patients and age-matched controls were measured. Receiver operator characteristic (ROC) curve analyses were conducted to evaluate the specificity and sensitivity of FGF23. The expression levels of FGF23 and its downstream factors fibroblast growth factor receptor 3 (FGFr3)/tissue non-specific alkaline phosphatase (TNAP)/osteopontin (OPN) in primary osteoblasts from CS patients (CS-Ob) and controls (CT-Ob) were detected. In addition, the osteogenic abilities of FGF23-knockdown or FGF23-overexpressing Ob were examined.@*RESULTS@#DNA methylation of the FGF23 gene in CS patients was decreased compared to that of their identical twins, accompanied by increased mRNA levels. CS patients had increased peripheral blood FGF23 mRNA levels and decreased computed tomography (CT) values compared with controls. The FGF23 mRNA levels were negatively correlated with the CT value of the spine, and ROCs of FGF23 mRNA levels showed high sensitivity and specificity for CS. Additionally, significantly increased levels of FGF23, FGFr3, OPN, impaired osteogenic mineralization and lower TNAP levels were observed in CS-Ob. Moreover, FGF23 overexpression in CT-Ob increased FGFr3 and OPN levels and decreased TNAP levels, while FGF23 knockdown induced downregulation of FGFr3 and OPN but upregulation of TNAP in CS-Ob. Mineralization of CS-Ob was rescued after FGF23 knockdown.@*CONCLUSIONS@#Our results suggested increased peripheral blood FGF23 levels, decreased bone mineral density in CS patients, and a good predictive ability of CS by peripheral blood FGF23 levels. FGF23 may contribute to osteopenia in CS patients through FGFr3/TNAP / OPN pathway.
Subject(s)
Humans , Osteopontin/genetics , Alkaline Phosphatase/metabolism , Receptor, Fibroblast Growth Factor, Type 3/metabolism , Scoliosis/genetics , Osteoblasts/metabolism , Calcinosis , RNA, Messenger/metabolism , Bone Diseases, Metabolic/metabolism , Fibroblast Growth Factors/geneticsABSTRACT
Purpose: To explore the role and mechanism of curcumin (Cur) in reducing oxidative stress damage in rats with nephrolithiasis induced by ethylene glycol (EG). Methods: Thirty male rats were divided into normal control, model, positive (10% potassium citrate), Cur-10 (10 mg/kg curcumin) and Cur-20 (20 mg/kg curcumin) groups. Results: The results of kidney tissue section stained by hematoxylin-eosin and von Kossa showed that curcumin treatment can inhibit the formation of kidney stones. The biochemical test results showed that the urea (Ur), creatinine (Cr), uric acid (UA), inorganic phosphorus and Ca2+ concentrations in urine decreased after being treated with curcumin. There were significant differences between different doses of curcumin (P < 0.05). Compared with the Cur-10 group, Cur-20 had a more significant inhibitory effect on malondialdehyde (MDA) (P < 0.05). In addition, reverse transcription polymerase chain reaction (PCR) detection and immunohistochemical results indicated that the osteopontin (OPN) in the kidney was significantly reduced after curcumin treatment. Conclusion: Curcumin could reduce the oxidative stress damage caused by EG-induced kidney stones.
Subject(s)
Animals , Male , Rats , Oxidative Stress/drug effects , Ethylene Glycol/analysis , Curcumin/administration & dosage , Osteopontin/analysis , Nephrolithiasis/veterinaryABSTRACT
The present study investigated the healing response of 12 fresh post-extraction alveolous grafted with particulate autologous teeth to achieve preservation of the post-extraction alveolar ridge. The objective is to elucidate the osteoconductive and osteoinductive properties of the autologous dental graft used as a bone substitute in the alveolar ridge preservation technique. Five patients were included, with at least one hopeless tooth and in need of extraction and preservatio n of the ridge, to receive in the same place a dental implant in prosthetic replacement. In the first surgical stage, dental extractions and preservation of the alveolar ridge were performed, using the teeth extracted and processed with an automatic system as bone substitutes. In the second surgical stage, an incisional bone biopsy was performed in each grafted site, the bone beds were recapitulated in a drilling protocol that allowed the placement of the dental implant, and the harvested bone specimens were prepared for analysis. The histological results of the bone biopsies in all cases showed remnant particles of the dental graft, made up of dentin, partially resorbed, with irregular superficial edges and in close contact with newly forme d bone in transition to mature lamellar bone, in which well differentiated osteocytes were observed. The immunohistochemical results showed a moderate positive expression of osteopontin at the edges of the integrated teeth particles, inside the peritubular dentin space and at the osteodental contact interfaces. In conclusion, the evidence from the study shows that the autologous dental graft is a biocompatible bone substitute, that provides an osteoconductive scaffold that promotes bone cell adhesion and migration for local osteogenesis and that it is associated with moderate in situ expression of osteopontin, which showed a high affinity with mineralized dental tissue, suggesting osteoinductive properties in situ.
El presente estudio investigó el resultado cicatrizal de 12 alvéolos frescos postextracción injertados con dientes autólogos particulados para lograr la preservación del reborde alveolar postextracción. El objetivo es dilucidar las propiedades osteoconductivas y osteoinductivas del injerto dental autólogo utilizado como sustituto óseo en la técnica de preservación de reborde. Se incluyeron 5 pacientes, con al menos un diente sin esperanza y con necesidad de extracción y preservación del reborde, para recibir en el mismo sitio un implante dental en sustitución protésica. En la primera etapa quirúrgica, se realizaron las extracciones dentales y la preservación del reborde alveolar, utilizando como sustituto óseo los dientes extraídos y procesados con un sistema automático. En la segunda etapa quirúrgica, se realizó una biopsia ósea incisional en cada sitio injertado, los lechos óseos fueron recapitulados en un protocolo de fresado que permitió la colocación del implante dental y los especímenes óseos recolectados fueron preparados para su análisis. Los resultados histológicos de las biopsias óseas en todos los casos mostraron partículas remanentes del injerto dental, conformadas por dentina, parcialmente reabsorbidas, con margenes superficiales irregulares y en estrecho contacto con depósitos de hueso de reciente formación en transición hacia hueso laminar maduro, en el cual se observaron osteocitos bien diferenciados. Los resultados inmunohistoquímicos mostraron una expresión positiva moderada de osteopontina en los bordes de las partículas del injerto dental integrado, al interior del espacio peritubular dentinario y en las interfases de contacto osteodental. En conclusión, la evidencia del estudio muestra que el injerto dental autólogo es un sustituto óseo biocompatible, que provee un andamio osteoconductivo promotor de la adhesión y migración de las células óseas para la osteogénesis local y que está asociado a la expresión modera in situ de osteopontina, la cual mostro una alta afinidad con el tejido dental mineralizado, sugiriendo propiedades osteoinductivas in situ.
Subject(s)
Humans , Female , Pregnancy , Middle Aged , Alveolar Bone Loss/diagnosis , Alveolar Ridge Augmentation/methods , Transplantation, Autologous , Biopsy , Demography , Alveolar Bone Loss/surgery , Bone Transplantation/methods , Tooth Socket , Osteopontin/metabolism , Alveolar Process/surgery , HistologyABSTRACT
A cartilagem de Meckel é uma estrutura transitória embrionária presente durante os estágios iniciais da formação da mandíbula, localizada em toda sua extensão e dividida em três porções, anterior, intermediária e posterior. O enfoque deste trabalho foi direcionado à elucidação do destino final da porção intermediária por meio de um estudo temporal sequenciado. Por isso, foi investigado a presença de células de reabsorção e a presença de fibras colágenas, bem como da proteína óssea osteopontina (OPN) na cartilagem de Meckel na região do germe do 1º molar inferior e no seu entorno. Foram utilizados fetos de ratos Wistar em períodos gestacionais pré-estabelecidos, G18 a G21 (grupos de dias intrauterinos), bem como P0 e P1 (recém-nascidos) para remoção das cabeças. Em sequência, os espécimes foram fixados em solução de formaldeído 4% + glutaraldeído 0,1% com tampão fosfato 0,1M, descalcificados em EDTA 4,13%, desidratados em concentrações crescentes de etanol e incluídos em parafina. As amostras foram coradas em hematoxilina e eosina (HE) e tricrômico de Mallory para análise histológica. Adicionalmente, os grupos G19 a P0 foram submetidos à reação histoquímica de TRAP para determinação da presença de células clásticas. Além disso, os grupos G21 e P0 (dia do nascimento) passaram por reações de imunomarcação para análise da expressão de OPN. Foi observado a degeneração gradual da cartilagem com a observação de mudanças estruturais, a justaposição de células clásticas na superfície da cartilagem por reação histoquímica TRAP a partir do G21, o aparecimento de colágeno tipo I nas fases terminais da degeneração, assim como a marcação positiva para a osteopontina na superfície de G21 e em todo o remanescente da cartilagem de Meckel no grupo P0. O estudo apontou um processo de degeneração da cartilagem com evidências de formação de matriz mineralizada de natureza óssea, a qual foi reabsorvida por células clásticas, sugerindo a ossificação da porção intermediária da cartilagem de Meckel.
Subject(s)
Osteoclasts , Cartilage , Osteopontin , MandibleABSTRACT
BACKGROUND@#Although osteopontin (OPN) is expressed in the liver and pigment gallstones of patients with hepatolithiasis, its role in pigment gallstone formation remains unclear. This study aimed to explore the function of OPN in pigment gallstone formation.@*METHODS@#Rats were fed a chow diet (CD) or lithogenic diet (LD) for 10 consecutive weeks; blocking tests were then performed using an OPN antibody (OPN-Ab). Incidence of gallstones and levels of several bile components, OPN, tumor necrosis factor alpha (TNF-α), and cholesterol 7 alpha-hydroxylase (CYP7A1) were analyzed. To determine TNF-α expression in hepatic macrophages and both CYP7A1 and bile acid (BA) expression in liver cells, recombinant rat OPN and recombinant rat TNF-α were used to treat rat hepatic macrophages and rat liver cells, respectively. Chi-square or Fisher exact tests were used to analyze qualitative data, Student t-test or one-way analysis of variance were used to analyze qualitative data.@*RESULTS@#Incidence of gallstones was higher in LD-fed rats than in CD-fed rats (80% vs. 10%, P < 0.05). BA content significantly decreased in bile (t = -36.08, P < 0.01) and liver tissue (t = -16.16, P < 0.01) of LD-fed rats. Both hepatic OPN protein expression (t = 9.78, P < 0.01) and TNF-α level (t = 8.83, P < 0.01) distinctly increased in the LD group; what's more, CYP7A1 mRNA and protein levels (t = -12.35, P < 0.01) were markedly down-regulated in the LD group. Following OPN-Ab pretreatment, gallstone formation decreased (85% vs. 25%, χ2 = 14.55, P < 0.01), liver TNF-α expression (F = 20.36, P < 0.01) was down-regulated in the LD group, and CYP7A1 expression (F = 17.51, P < 0.01) was up-regulated. Through CD44 and integrin receptors, OPN promoted TNF-α production in macrophage (F = 1041, P < 0.01), which suppressed CYP7A1 expression (F = 48.08, P < 0.01) and reduced liver BA synthesis (F = 119.4, P < 0.01).@*CONCLUSIONS@#We provide novel evidence of OPN involvement in pigmented gallstone pathogenesis in rats.
Subject(s)
Animals , Rats , Diet/adverse effects , Gallstones/etiology , Lithiasis , Liver , Liver Diseases , Osteopontin/geneticsABSTRACT
Objective: Investigating osteopontin (OPN) level in gingival crevicular fluid (GCF) of patients affected by periodontitis with or without Type-2 diabetes mellitus (T2DM). The aim of this study is to explore the possibility of OPN to differentiate between periodontal health and disease. Material and Methods: A total number of 36 participants seeking periodontal treatment were recruited in this pilot study and divided into three study groups. Periodontitis [systemically healthy participants with periodontitis (probing pocket depth) PPD (probing pocket depth) ≥ 4mm], periodontitis and poorly controlled type 2 diabetes mellitus (), and control (systemically and periodontally healthy periodontium) groups. Plaque index (PI), gingival index (GI) and PPD were examined. OPN level was measured in the GCF and analysed, using Enzyme-Linked Immunosorbent assay. Results: PI and GI were significantly higher in T2DM with periodontitis compared to periodontitis and control groups. Both periodontitis and P-T2DM groups showed significant increase in the OPN levels compared to control group (p<0.001). PPD showed the only significant positive association with OPN (p<0.001) compared to other clinical parameters. The receiver operating characteristics curve analysis demonstrated that OPN had higher area under the curve value (AUC: 0.95) in periodontitis compared to P-T2DM patients (AUC: 0.86). Conclusion: In periodontitis groups, clinical parameters were equally deteriorated together with significant increase in the expression of OPN compared to control. Furthermore, GCF levels of OPN were sensitive and specific enough to discriminate between health and periodontitis even with T2DM. This could introduce OPN to be as a candidate diagnostic biomarker of periodontal disease. (AU)
Objetivo: Investigar o nível de osteopontina (OPN) no fluido gengival crevicular (GCF) de pacientes com periodontite com ou sem diabetes mellitus tipo 2 (T2DM). O objetivo deste estudo foi explorar a possibilidade da OPN diferenciar entre saúde e doença periodontal. Material e Métodos: No total, para este estudo piloto foram recrutados 36 participantes que estavam em busca de tratamento periodontal e divididos em três grupos de estudo: grupos periodontite [participantes sistemicamente saudáveis com periodontite (profundidade de sondagem) PPD ≥ 4 mm], grupo periodontite e Diabetes Mellitus tipo 2 mal controlada (P-T2DM) e grupo controle (saudáveis sistemicamente e periodontalmente). Índice de placa (PI), índice gengival (GI) e PPD foram examinados. O nível de OPN foi medido no GCF e analisado usando o ensaio ELISA (Enzyme-Linked Immuno Sorbent Assay). Resultados: PI e GI foram significativamente maiores no T2DM com periodontite em comparação aos grupos com periodontite e controle. Os grupos com periodontite e P-T2DM apresentaram aumento significativo nos níveis de OPN em comparação ao grupo controle (p <0,001). PPD mostrou a única associação positiva significativa com OPN (p <0,001) em comparação com outros parâmetros clínicos. A análise da curva de características operacionais do receptor demonstrou que OPN teve maior área sob o valor da curva (AUC: 0,95) na periodontite em comparação com pacientes com P-T2DM (AUC: 0,86). Conclusão: Nos grupos com periodontite, os parâmetros clínicos foram igualmente deteriorados juntamente com aumento significativo na expressão de OPN em comparação com o grupo controle. Além disso, os níveis de OPN no GCF foram sensíveis e específicos o suficiente para discernir entre saúde e periodontite, mesmo com T2DM. Isso poderia apresentar a OPN como um candidato a biomarcador diagnóstico de doença periodontal.(AU)
Subject(s)
Humans , Periodontitis , Bone Resorption , Diabetes Mellitus, Type 2 , OsteopontinABSTRACT
SUMMARY: The objective of this study were bone defect complications that occur due to traumas or infections. Bone grafts are required to provide support, fill gaps and improve biological repair in skeletal damage. Dexamethasone plays role in calcium signaling modulation and used in diseases. Aim of this study was to evaluate osteonectin and osteopontin expressions in new bone development after dexamethasone application on tibial bone defects. Rats were divided into defect, defect+graft and defect+graft+dexamethasone treated groups. Tibial bone defect created, and rats were kept immobile for 28 days. Alloplastic material was placed in defect area in second and group third groups. 2.5 mg/kg Dex and normal saline were injected to dexamethasone and defect groups twice a week for 56 days. Inflammation and congestion were increased in defect and defect+graft groups. Defect+graft+dexamethasone group; increased number of osteoblast and osteocyte cells, dense bone matrix, formation of new bone trabeculae was observed. Defect+graft group; osteonectin expression in graft regions, osteoblast cells, some connective tissue cells and fibers were seen whereas in defect+graft+dexamethasone group; osteopontin expression in osteoblast and osteocyte cells of new bone trabeculae were observed. Dexamethasone may lead to formation of new bone trabeculae into the graft material resulting in increased osteoconduction and osteoinductive effect for differentiation of osteon.
RESUMEN: Los defectos óseos son complicaciones que ocurren debido a traumas o infecciones. Se requieren injertos óseos para proporcionar apoyo, llenar los espacios y mejorar la reparación biológica en el hueso dañado. La dexametasona desempeña un papel importante en la modulación de la señalización del calcio y se usa en enfermedades. El objetivo de este estudio fue evaluar las expresiones de osteonectina y osteopontina en el desarrollo óseo después de la aplicación de dexametasona en defectos óseos tibiales. Las ratas se dividieron en grupos: defecto, defecto + injerto y defecto + injerto + grupos tratados con dexametasona. Se creó un defecto óseo tibial, y las ratas se mantuvieron inmóviles durante 28 días. El material aloplástico se colocó en el área del defecto en el segundo y tercer grupo. Se inyectaron 2,5 mg / kg de dexametasona y solución salina normal a grupos de defectos dos veces por semana durante 56 días. La inflamación y la congestión aumentaron en los grupos de defectos y defectos + injerto; En el grupo defecto + injerto + grupo tratado con dexametasona se observó un aumento en el número de osteoblastos y osteocitos, de matriz ósea densa y en la formación de nuevas trabéculas óseas. En el grupo defecto + grupo de injerto se observó la expresión de osteonectina en las áreas de injerto, osteoblastos, algunas células y fibras de tejido conectivo, mientras que en el grupo defecto + injerto + dexametasona se observó la expresión de osteopontina en osteoblastos y osteocitos y formación de nuevas trabéculas óseas . En conclusión la dexametasona puede conducir a la formación de nuevas trabéculas óseas en el material de injerto, lo que resulta en un aumento de la osteoconducción y un efecto osteoinductivo para la diferenciación del osteón.
Subject(s)
Animals , Male , Rats , Tibia/surgery , Tibia/drug effects , Dexamethasone/administration & dosage , Bone Transplantation , Tibia/pathology , Bone Regeneration , Immunohistochemistry , Osteonectin/physiology , Bone Remodeling , Rats, Wistar , Disease Models, Animal , Osteopontin/physiologyABSTRACT
ABSTRACT Melatonin (MLT) is a potential signaling molecule in the homeostasis of bone metabolism and may be an important mediator of bone formation and stimulation. The aim of this in vitro study was to evaluate the effect of MLT on the viability, mRNA/protein expression and mineralization of pre-osteoblastic cells. The concentrations 5, 2.5, 1, 0.1 and 0.01 mM MLT were tested on pre-osteoblastic cells (MC3T3) compared to control (no MLT), evaluating proliferation and cell viability (C50), gene expression (RT-PCR) and secretion (ELISA) of COL-I and OPN at 24h, 48h and 72h, and the formation of mineral nodules (alizarin red and fast red) after 10 days of treatment. MLT at 5 and 2.5 mM proved to be cytotoxic (C50), so only 0.01, 0.1 and 1 mM were used for the subsequent analyses. OPN mRNA expression increased with MLT at 0.1 mM - 1 mM, which was followed by increased secretion of OPN both at 24h and 72h compared to the remaining groups (p <0.05). COL-I mRNA and COL-1 secretion followed the same pattern as OPN at 0.1 mM MLT at 72h of treatment (p <0.05). Regarding mineralization, all MLT doses (except 1mM) caused an increase (p <0.05) in the formation of mineral nodules compared to the control. Melatonin at 0.01mM - 1mM had a stimulatory effect on osteoblasts by upregulating COL-I and OPN expression/ secretion and mineralization, thereby fostering osteogenesis.
RESUMO A melatonina (MLT) é uma molécula potencial de sinalização na homeostase do metabolismo ósseo e pode ser um importante mediador da formação e estimulação óssea. O objetivo deste estudo in vitro foi avaliar o efeito da MLT na viabilidade, na expressão do mRNA da proteína e mineralização de células préosteoblásticas. As concentrações de MLT 5, 2,5, 1, 0,1 e 0,01 mM foram testadas em células pré-osteoblásticas da linhagem MC3T3 em comparação ao controle (sem MLT), avaliando a proliferação e a viabilidade celular (C50), expressão gênica (rtPCR) e secreção (Elisa) de Colágeno tipo 1 (COL-I) e osteopontina (OPN) às 24, 48 e 72 horas, além da formação de nódulos minerais por meio do teste vermelho de Alizarina fast red após 10 dias de tratamento. MLT a 5 e 2,5 mM provou ser tóxico (C50). Portanto, as concentrações de 0,01, 0,1 e 1 mM foram utilizadas para as análises subsequentes. A expressão do mRNA da OPN aumentou com MLT a 0,1 mM-1mM, seguida pela secreção aumentada de OPN às 24 e 72 horas em comparação aos demais grupos (p<0,05). O mRNA de COL-I e a secreção de COL-I seguiram o mesmo padrão do OPN a 0,1 mM de MLT em 72 horas de tratamento (p<0,05). Em relação à mineralização, todas as doses de MLT (exceto 1mM) causaram aumento (p<0,05) na formação de nódulos minerais em comparação ao controle. A MLT na concentração entre 0,01mM a 1 mM teve um efeito estimulador sobre os osteoblastos, ao regular positivamente a expressão e secreção de COL-I e OPN, além da mineralização, favorecendo a osteogênese.
Subject(s)
Humans , Osteoblasts/drug effects , Osteogenesis/drug effects , Osteogenesis/genetics , Peptide Fragments/metabolism , Cell Differentiation/drug effects , Cell Differentiation/genetics , Matrix Metalloproteinase 2/metabolism , Osteopontin/metabolism , Melatonin/pharmacology , Osteoblasts/metabolism , Peptide Fragments/genetics , RNA, Messenger/genetics , Enzyme-Linked Immunosorbent Assay , Gene Expression , Gene Expression Regulation, Developmental/drug effects , Matrix Metalloproteinase 2/genetics , Cell Proliferation/drug effects , Cell Proliferation/genetics , Osteopontin/genetics , Real-Time Polymerase Chain ReactionABSTRACT
Osteopontin is a glycophosphoprotein implicated in different physiologic and pathologic processes and is known to be involved in progression and metastasis of various cancers in humans, but this relation is still little explored in the veterinary. The aim was to evaluate the expression of osteopontin in canine mammary carcinomas and its relation with well-established canine mammary tumor biomarkers. For that, expression of OPN, EGFR, HER2, and c-Kit were evaluated along with Ki67 rate in 43 mammary carcinomas. Osteopontin was demonstrated to be expressed by neoplastic epithelial cells in all carcinomas as well as in stromal cells from the tumor microenvironment. Relation between high osteopontin expression and EGFR positivity (P<0.001) and HER2 overexpression (P=0.012) was demonstrated. In conclusion, high OPN expression seems to be related to poor prognosis and MAPK pathway activation, given the association with EGFR and HER2, members of the MAPK signaling pathway.(AU)
A osteopontina é uma glicofosfoproteina implicada em diferentes processos fisiológicos e patológicos, sendo conhecida por estar envolvida na progressão e metástase de vários cânceres nos humanos, no entanto, essa relação é ainda pouco explorada na veterinária. O objetivo deste trabalho foi avaliar a expressão da osteopontina nos carcinomas mamários caninos e sua relação com biomarcadores bem estabelecidos para esta neoplasia. Para isto, foi avaliada a expressão de OPN, EGRH, HER2 e c-Kit juntamente com a taxa de Ki67 em 43 carcinomas mamários. A osteopontina foi expressa pelas células epiteliais neoplásicas em todos os carcinomas, assim como, nas células estromais do microambiente tumoral. Foi demonstrada uma relação entre uma alta expressão de osteopontina e positividade para EGFR (P<0.001) e superexpressão de HER2 (P=0.012). Em conclusão, alta expressão de OPN parece estar relacionada com mau prognóstico e ativação da via MAPK, devido a sua associação com EGRF e HER2, os quais são membros desta via de sinalização.(AU)
Subject(s)
Animals , Female , Dogs , Carcinoma , Biomarkers , Mammary Neoplasms, Animal , Dog Diseases , Osteopontin , ImmunohistochemistryABSTRACT
Objective: To explore the association between serum levels of osteopontin (OPN) and systolic pulmonary artery pressure (sPAP) in healthy men following acute high altitude exposure. Methods: According to the inclusion and exclusion criteria, this observational study included 94 male subjects (aged from 18 to 30 years, dwelling in lowland<500 m) who ascended to Litang (4 100 m) from Chongqing (400 m) by bus with a stair-like journey within 7 days in June 2013. Data including basic information, OPN, superoxide dismutase (SOD), and malondialdehyde (MDA) and echocardiographic derived sPAP were collected within 48 hours before ascent and within 2-7 hours after arrival. Accordingly, subjects were divided into 3 groups based on the tertiles of sPAP after acute high altitude exposure: low sPAP group (26.8-32.3 mmHg (1 mmHg=0.133 kPa)) (n=31), middle sPAP group (32.4-37.4 mmHg) (n=32) and high sPAP group (37.5-55.6 mmHg) (n=31). Associations of serum OPN and SOD levels with sPAP were analysed by univariate and multivariate linear regression analysis. Results: After acute high altitude exposure, the levels of sPAP were significantly increased (P<0.001). There were no differences in age, height, weight, body mass index, percent of Han nationality and smoking among 3 subgroups. However, following acute high altitude exposure, the levels of heart rate, systolic and diastolic blood pressure elevated (all P<0.05), whereas the levels of oxygen saturation were reduced in the total subjects and all subgroups (all P<0.05). Moreover, systolic blood pressure of subjects in the high sPAP group was higher than that in low and middle sPAP groups (both P<0.05), and diastolic blood pressure of subjects in high sPAP group was higher than that in low sPAP group (P<0.05). The serum levels of OPN were increased in total cohort(27.9 (22.5,34.0) μg/L vs. 25.6 (18.4, 33.1) μg/L, P<0.05), and high sPAP group (P<0.05), whereas no differences were found in serum SOD and MDA levels among groups. Furthermore, the serum level of OPN in high sPAP group was higher than that in low sPAP group at high altitude (P<0.05), and there was a trend for decline in SOD level with increasing sPAP (P>0.05). Results from univariable linear regression analysis showed that the serum levels of OPN (r=0.32, P=0.002) and SOD (r=-0.22,P=0.032) were linearly correlated with sPAP in total cohort after high altitude exposure. Multivariate regression analysis showed that the serum levels of OPN(β=0.310,P=0.002) and SOD (β=-0.199,P=0.043) were independently associated with the levels of sPAP at high altitude. Conclusion: After acute high altitude exposure, the serum level of OPN is positively associated with sPAP, suggesting that OPN may be a novel bio-marker for predicting the increase of pulmonary pressure in response to acute high altitude exposure.
Subject(s)
Adolescent , Adult , Humans , Male , Young Adult , Altitude , Blood Pressure Determination , Osteopontin , Pulmonary Artery , SystoleABSTRACT
A regeneração óssea guiada (RGO) tornou-se uma prática comum e importante na odontologia, sendo necessário o uso de membranas para sua realização, uma vez que são barreiras que evitam o crescimento de tecido mole nas áreas de defeitos ósseos. Entre as características mais relevantes das membranas absorvíveis estão: o suporte sanguíneo (diretamente relacionado com a porosidade do material) e suporte mecânico ósseo que depende do tempo de reabsorção da membrana. O objetivo desse estudo foi avaliar e comparar duas membranas de colágeno por meio de estudo histológico, histomorfométrico, imunoistoquímico e por contagem de células inflamatórias o processo de regeneração óssea guiada utilizando a membrana de colágeno derivada de pericárdio porcino (Jason®-Instituto Straumann AG, Suíça) em defeitos críticos de 7 mm de diâmetro criados em setenta e duas calvárias de ratos (Rattus Albinus, variedade Wistar). Esses animais foram divididos em 3 grupos: grupo membrana de colágeno porcino (BioGide® - Geistlich Wohlhusen, Suíça), grupo membrana de colágeno de pericárdio porcino (Jason®-Instituto Straumann AG, Suíça) e grupo coágulo, sendo este preenchidos somente com coágulo sem membrana. Esses 3 grupos forma subdivididos em quatro subgrupos de acordo com os tempos avaliados: 7, 15, 30 e 60 dias. Como resultado tivemos na análise histológica e histométrica maior neoformação óssea com o grupo de membrana pericárdio porcino nos períodos de 7 dias (199 pontos) e não foi significante, com 15 dias (494 pontos) estatisticamente significativo, com 30 dias (979 pontos) não significante. Esses valores se modificam 60 dias, mostrando maior superioridade a membrana de colágeno porcino (BioGide®- Geistlich Wohlhusen, Suíça) (1151 pontos) estatisticamente significativo. No analises global a membrana de colágeno porcino (Jason®-Instituto Straumann AG, Suíça) foi superior que a membrana de pericárdio porcino (BioGide®- Geistlich Wohlhusen, Suíça) (p= 0,021) estatisticamente significativo. A análise imunoistoquímica confirmou os achados histométricos, demostrando maior presença da osteocalcina no grupo de pericárdio porcino aos 7 e 15 dias e presença da osteopontina pouco evidente. Já com a membrana de colágeno porcino a osteopontina foi mais imunomarcada aos períodos de 7 e 15 dias, e a presencia de osteocalcina mais evidente aos 30 e 60 dias, corroborando com os resultados iniciais. Na contagem de células inflamatórias para o tempo de 7 dias não houve diferenças estatísticas, já na contagem de vasos sanguíneos houve diferença significativa no período de 15 dias com maior quantidade de vasos para membrana de colágeno porcino, com estes achados concluímos que tanto a membrana de colágeno de pericárdio porcino (Jason® -Instituto Straumann AG, Suíça) quanto a membrana de colágeno porcino (BioGide®-Geistlich Wohlhusen, Suíça) podem ser consideradas como material de escolha apropriada para regeneração óssea guiada, com maior proporção de osso neoformado com a membrana de colágeno porcino(AU)
Guided bone regeneration (RGO) has become a common and important practice in dentistry, requiring the use of membranes to perform it, since they are barriers that prevent the growth of soft tissue in areas of bone defects. Among the most relevant characteristics of absorbable membranes are: blood support (directly related to the porosity of the material) and mechanical bone support that depends on the time of membrane resorption. The goal of this study was to evaluate and compare two collagen membranes by means of histological, histomorphometric, immunohistochemical study and by inflammatory cell counting the guided bone regeneration process using the collagen membrane derived from porcine pericardium (Jason®-Instituto Straumann AG, Switzerland) in critical defects of 7 mm in diameter created in seventy-two calvaria of rats (Rattus Albinus, Wistar variety). These animals were divided into 3 groups: porcine collagen membrane group (BioGide® - Geistlich Wohlhusen, Switzerland), porcine pericardium collagen group (Jason®-Instituto Straumann AG, Switzerland) and clot group, which were filled with only a clot without membrane. These 3 groups were subdivided into four subgroups according to the evaluated times: 7, 15, 30 and 60 days. As a result, we had a greater bone neoformation in the histological and histometric analysis with the porcine pericardial membrane group in the periods of 7 days (199 points) and it was not statistically significant with 15 days (494 points), with 30 days (979 points) not significant. These values change 60 days, showing greater superiority to the porcine collagen membrane (BioGide®- Geistlich Wohlhusen, Switzerland) (1151 points) statistically significant. In the global analysis, the porcine collagen membrane (Jason®Instituto Straumann AG, Switzerland) was superior than the porcine pericardium membrane (BioGide®- Geistlich Wohlhusen, Switzerland) (p = 0.021) statistically significant. The immunohistochemical analysis confirmed the histometric findings, showing a greater presence of osteocalcin in the porcine pericardium group at 7 and 15 days and the presence of osteopontin little evident. As for the porcine collagen membrane, osteopontin was more immunostained at 7 and 15 days, and the presence of osteocalcin was more evident at 30 and 60 days, corroborating the initial results. In the inflammatory cell count for the 7-day period, there were no statistical differences, whereas in the blood vessel count, there was a significant difference in the 15-day period with a greater number of vessels for porcine collagen membrane, with these findings we conclude that both the porcine pericardial collagen (Jason® - Straumann AG Institute, Switzerland) and porcine collagen membrane (BioGide®-Geistlich Wohlhusen, Switzerland) can be considered as the material of choice for guided bone regeneration, with a higher proportion of neoformed bone according to porcine collagen membrane(AU)
Subject(s)
Animals , Rats , Skull , Bone Regeneration , Collagen , Inflammation , Membranes , Bone and Bones , Immunohistochemistry , Osteocalcin , Cell Count , Rats, Wistar , Guided Tissue Regeneration , OsteopontinABSTRACT
Abstract Introduction: Oral peripheral and central giant cell granulomas are lesions with little-known etiology and pathogenesis. Objective: The aim of this study was to compare matrix metalloproteinases-2 and osteopontin protein expression in the multinucleated giant cells and mononuclear cells of the peripheral and central giant cell granuloma lesions. Methods: In this retrospective study, the presence of matrix metalloproteinases-2 and osteopontin in 37 cases of central giant cell granuloma and 37 cases of peripheral giant cell granuloma paraffin blocks were assessed by streptavidin-biotin immunohistochemistry. Independent sample t-test, Chi-square, Mann-Whitney tests and Spearman's rank correlation coefficient were used. Results: The osteopontin was expressed in both multinucleated giant cells and mononuclear cells in all cases of peripheral and central giant cells granulomas. However, the matrix metalloproteinases-2 expression was positive in 86.5% of giant cells and it was positive in all of mononuclear cells in peripheral giant cells granuloma. In central giant cells granulomas, 91.8% of giant cells and all mononuclear cells were positive for matrix metalloproteinases-2 marker. Percentage and Intensity of staining were significantly higher in central than peripheral giant cells lesions, for both markers (p ˂ 0.05). Conclusion: This study showed that the expression of osteopontin in giant cells supports the theory of osteolcastic nature of these cells. Also, the presence of osteopontin and matrix metalloproteinases-2 in mononuclear cells may indicate the monocyte-macrophage origin of these cells, as the differentiation of the precursors of the mononuclear stromal monocyte/macrophage to osteoclasts is possibly affected by the expression of osteolytic factors. Also, may be differences in biological behaviors of these lesions are associated with the level of osteopontin and matrix metalloproteinases-2 expression.
Resumo Introdução: Os granulomas periféricos e centrais de células gigantes são lesões com etiologia e patogênese pouco conhecidas. Objetivo: Comparar a expressão das proteínas metaloproteinases da matriz-2 e osteopontina nas células gigantes multinucleadas e células mononucleares no granuloma periférico e central de células gigantes. Método: Neste estudo retrospectivo, a presença de metaloproteinases da matriz-2 e osteopontina em 37 casos de granuloma central de células gigantes e 37 casos de granuloma periférico de células gigantes em blocos de parafina foi avaliada por imuno-histoquímica pela estreptavidina-biotina. Foram usados teste t para amostra independente, teste de qui-quadrado, Mann-Whitney e coeficiente de correlação de Spearman. Resultados: A osteopontina foi expressa em células gigantes multinucleadas e células mononucleares em todos os casos de granuloma periférico de células gigantes e granuloma central de células gigantes. No entanto, a expressão de metaloproteinases da matriz-2 foi positiva em 86,5% de células gigantes e foi positiva em todas as células mononucleares em granuloma periférico de células gigantes. Em granuloma central de células gigantes, 91,8% das células gigantes e todas as células mononucleares foram positivas para o marcador metaloproteinases da matriz-2. A porcentagem e intensidade de coloração em granuloma central de células gigantes foram significantemente maiores do que em granuloma periférico de células gigantes para ambos os marcadores (p ˂ 0,05). Conclusão: Este estudo mostrou que a expressão de osteopontina em células gigantes apoia a teoria da natureza osteoclástica dessas células. Além disso, a presença de osteopontina e metaloproteinases da matriz-2 em células mononucleares pode indicar a origem dos monócitos-macrófagos dessas células, uma vez que a diferenciação dos precursores do monócito/macrófago estromal mononuclear em osteoclastos é possivelmente afetada pela expressão de fatores osteolíticos. Além disso, as diferenças nos comportamentos biológicos dessas lesões estão associadas ao nível de expressão de osteopontina e metaloproteinases da matriz-2.
Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Young Adult , Granuloma, Giant Cell/pathology , Jaw Diseases/pathology , Matrix Metalloproteinase 2/analysis , Osteopontin/analysis , Reference Values , Severity of Illness Index , Immunohistochemistry , Sex Factors , Retrospective Studies , Age Factors , Statistics, Nonparametric , StreptavidinABSTRACT
Objective: To analyze osteopontin mRNA expression levels in subjects with periodontitis prior to (baseline) and 7, 14, and 28 days following scaling and root planing (SRP). Material and Methods: Gingival crevicular fluid was collected as clinical samples from four subjects with periodontitis (pocket depth, 4-5 mm) aged 35-54 years old as well as from three healthy subjects (controls). The osteopontin mRNA expression levels were measured by quantitative real-time polymerase chain reaction. Spearman's rank correlation between osteopontin levels in gingival crevicular fluid and the modified gingival index (MGI) was also performed. Results: The Wilcoxon signed-rank test showed no significant difference in osteopontin mRNA expression levels between baseline and 28 days following SRP (p=0.068). The Friedman test showed no significant difference in osteopontin mRNA expression levels between baseline and following SRP (7, 14, or 28 days) (p>0.05). Spearman's rank correlation showed no significant correlation between osteopontin mRNA expression levels and MGI (r=0.087; p=0.749). Conclusion: Following SRP of periodontal tissue, there was a decreasing trend in osteopontin mRNA expression; however, this finding was not statistically significant. Nevertheless, osteopontin can be used as a biomarker to monitor the healing process; however, further studies are required to clarify our results.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Periodontitis , RNA, Messenger , Root Planing/methods , Osteopontin , Case-Control Studies , Statistics, Nonparametric , IndonesiaABSTRACT
Biomarkers for hepatocellular carcinoma (HCC) following curative resection are not currently sufficient for prognostic indication of overall survival (OS) and disease-free survival (DFS). The aim of this study was to investigate the prognostic performance of osteopontin (OPN), matrix metalloproteinase 7 (MMP7), and pregnancy specific glycoprotein 9 (PSG9) in patients with HCC. A total of 179 prospective patients with HCC provided plasma before hepatectomy. Plasma OPN, MMP7, and PSG9 levels were determined by enzyme-linked immunosorbent assay. Correlations between plasma levels, clinical parameters, and outcomes (OS and DFS) were overall analyzed. High OPN ( ⩾ 149.97 ng/mL), MMP7 ( ⩾ 2.28 ng/mL), and PSG9 ( ⩾ 45.59 ng/mL) were prognostic indicators of reduced OS (P < 0.001, P < 0.001, and P = 0.007, respectively). Plasma PSG9 protein level was an independent factor in predicting OS (P = 0.008) and DFS (P = 0.038). Plasma OPN + MMP7 + PSG9 elevation in combination was a prognostic factor for OS (P < 0.001). OPN was demonstrated to be a risk factorassociated OS in stage I patients with HCC and patients with low α-fetoprotein levels ( < 20 ng/mL). These findings suggested that OPN, MMP7, PSG9 and their combined panels may be useful for aiding in tumor recurrence and mortality risk prediction of patients with HCC, particularly in the early stage of HCC carcinogenesis.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Blood , Carcinoma, Hepatocellular , Blood , Mortality , Enzyme-Linked Immunosorbent Assay , Hepatectomy , Liver Neoplasms , Blood , Mortality , Matrix Metalloproteinase 7 , Blood , Osteopontin , Blood , Pregnancy-Specific beta 1-Glycoproteins , Prognosis , Prospective Studies , Risk Assessment , Survival AnalysisABSTRACT
Abstract Stanozolol (ST) is a synthetic androgen with high anabolic potential. Although it is known that androgens play a positive role in bone metabolism, ST action on bone cells has not been sufficiently tested to support its clinical use for bone augmentation procedures. Objective: This study aimed to assess the effects of ST on osteogenic activity and gene expression in SaOS-2 cells. Material and Methods: SaOS-2 deposition of mineralizing matrix in response to increasing doses of ST (0-1000 nM) was evaluated through Alizarin Red S and Calcein Green staining techniques at 6, 12 and 24 days. Gene expression of runt-related transcription factor 2 (RUNX2), vitamin D receptor (VDR), osteopontin (SPP1) and osteonectin (ON) was analyzed by RT-PCR. Results: ST significantly influenced SaOS-2 osteogenic activity: stainings showed the presence of rounded calcified nodules, which increased both in number and in size over time and depending on ST dose. RT-PCR highlighted ST modulation of genes related to osteogenic differentiation. Conclusions: This study provided encouraging results, showing ST promoted the osteogenic commitment of SaOS-2 cells. Further studies are required to validate these data in primary osteoblasts and to investigate ST molecular pathway of action.
Subject(s)
Humans , Osteogenesis/drug effects , Stanozolol/pharmacology , Gene Expression/drug effects , Anabolic Agents/pharmacology , Osteoblasts/drug effects , Time Factors , Calcification, Physiologic/drug effects , Linear Models , Osteonectin/analysis , Osteonectin/drug effects , Reproducibility of Results , Analysis of Variance , Receptors, Calcitriol/analysis , Receptors, Calcitriol/drug effects , Cell Line, Tumor/drug effects , Core Binding Factor Alpha 1 Subunit/analysis , Core Binding Factor Alpha 1 Subunit/drug effects , Osteopontin/analysis , Osteopontin/drug effects , Real-Time Polymerase Chain ReactionABSTRACT
Abstract Objective This study aimed to evaluate the inflammatory effect and bone formation in sterile surgical failures after implantation of a collagen sponge with mesenchymal stem cells from human dental pulp (hDPSCs) and Aloe vera. Material and Methods Rattus norvegicus (n=75) were divided into five experimental groups according to treatment: G1) control (blood clot); G2) Hemospon®; G3) Hemospon® in a culture medium enriched with 8% Aloe vera; G4) Hemospon® in a culture medium containing hDPSCs and G5) Hemospon® in a culture medium enriched with 8% Aloe vera and hDPSCs. On days 7, 15 and 30, the animals were euthanized, and the tibia was dissected for histological, immunohistochemistry and immunofluorescence analyses. The results were analyzed using nonparametric Kruskal-Wallis test and Dunn's post-test. Results On days 7 and 15, the groups with Aloe vera had less average acute inflammatory infiltrate compared to the control group and the group with Hemospon® (p<0.05). No statistically significant difference was found between the groups regarding bone formation at the three experimental points in time. Osteopontin expression corroborated the intensity of bone formation. Fluorescence microscopy revealed positive labeling with Q-Tracker® in hDPSCs before transplantation and tissue repair. Conclusion The results suggest that the combination of Hemospon®, Aloe vera and hDPSCs is a form of clinical treatment for the repair of non-critical bone defects that reduces the inflammatory cascade's effects.
Subject(s)
Humans , Animals , Male , Rats , Bone Regeneration/drug effects , Bone Regeneration/physiology , Plant Extracts/pharmacology , Dental Pulp/cytology , Mesenchymal Stem Cell Transplantation/methods , Aloe/chemistry , Osteogenesis/drug effects , Osteogenesis/physiology , Tibia/drug effects , Tibia/physiology , Tibia/pathology , Time Factors , Immunohistochemistry , Hemostatics/pharmacology , Cell Survival/drug effects , Cell Survival/physiology , Cells, Cultured , Reproducibility of Results , Collagen/pharmacology , Treatment Outcome , Osteopontin/analysis , Flow Cytometry , Microscopy, FluorescenceABSTRACT
Abstract Purpose: The effects of resveratrol administration on calvarial bone defects with alloplastic graft material was investigated for osteoinductive reaction and bone development in rats. Methods: Healthy male rats were randomly divided into 3 groups consisting of 10 rats. Groups were as follows: control (defect) group, defect + graft group, and defect + graft + resveratrol group. A calvarial bone defect was created in all groups, alloplastic bone grafts were applied to the defect in the 2nd and 3rd group, resveratrol (5 mg/kg/day) was added to the drinking water of the animals following graft application for 28 days in the 3rd group. Results: Increase in osteoclasts and necrotic changes were observed histopathologically in the control group. In the 2nd group, reduction of inflammation, congestion of blood vessels, increased osteblastic activity, osteoinductive effect, progression of osteocyte development and increased collagen fibers in connective tissue were observed. In the 3rd group, osteoblasts seemed to secrete bone matrix and accelerate osteoinductive effect with increased osteopregenitor activity and positive osteopontin and osteonectin expressions. Conclusion: Resveratrol treatment was thought to be an alternative and supportive drug for implant application by inducing new bone formation in the calvaral defect region as a result of short-term treatment.
Subject(s)
Animals , Male , Rats , Skull/surgery , Bone Regeneration/drug effects , Bone Transplantation/methods , Bone Substitutes/administration & dosage , Resveratrol/administration & dosage , Osteoblasts/drug effects , Osteogenesis/drug effects , Skull/drug effects , Drug Administration Schedule , Osteonectin/administration & dosage , Osseointegration/drug effects , Bone Substitutes/therapeutic use , Disease Models, Animal , Osteopontin/administration & dosageABSTRACT
SUMMARY: The purpose of this study was to investigate effects of diabetes mellitus (DM) on the alveolar bone with histopathological and immunohistochemical methods. Wistar rats were divided into two groups, control and diabetes group. Control group was fed standard rat chow and drinking water for 8 weeks. Single dose (Streptozotocin) STZ (55 mg/kg), was dissolved in sodium citrate buffer and introduced intraperitoneal injection. Diabetes group and control group were compared in terms of glucose values. The blood glucose concentration in diabetic rats was significantly high (p <0.05). In diabetes group; periodontal membrane and the dilation of blood vessels, hemorrhage has also been a significant increase in inflammatory cells. In the diabetes group, osteonectin showed positive expression in periodontal membrane and showed negative expression in osteocytes of alveolar bone. Osteopontin expression in fibroblast cells and periodontal membrane collagen fibrils was positive, alveolar cells, osteocytes and bone matrix bone was found positive. Diabetes results showed that there formed periodontitis; due to the increase in inflammation inhibiting bone formation delaying the development of early bone cells.
RESUMEN: El objetivo de este estudio fue investigar los efectos de la diabetes mellitus (DM) sobre el hueso alveolar con métodos histopatológicos e inmunohistoquímicos. Las ratas Wistar se dividieron en dos grupos, grupo control y grupo de diabetes. El grupo control fue alimentado con comida estándar y agua potable durante 8 semanas. La dosis única Streptozotocina (STZ) (55 mg/ kg), se disolvió en tampón de citrato de sodio y se introdujo mediante inyección intraperitoneal. El grupo diabetes y el grupo control se compararon en términos de valores de glucosa. La concentración de glucosa en sangre en ratas diabéticas fue significativamente alta (p <0,05). En el grupo diabetes hubo un aumento significativo de la membrana periodontal y dilatación de los vasos sanguíneos y hemorragia, con un aumento significativo de células inflamatorias. En el grupo diabetes, la osteonectina mostró una expresión positiva en la membrana periodontal además se observó expresión negativa en los osteocitos del hueso alveolar. La expresión de osteopontina en fibroblastos y fibrillas de colágeno en membrana periodontal fue positiva, las células alveolares, osteocitos y hueso de la matriz ósea dio positivo. Los resultados de la diabetes mostraron que existía periodontitis, debido al aumento de la inflamación que inhibió la formación ósea retardando el desarrollo de células óseas tempranas.