Association of blood heavy metal levels with osteocalcin abnormality and incidence of osteoporosis in Saudi subjects / Associação de anormalidades de osteocalcina nos níveis de metais pesados no sangue e incidência de osteoporose em indivíduos sauditas

Abstract Serum toxic metals have been implicated in development of many diseases. This study investigated the association between blood levels of lead and cadmium with abnormal bone mineral density (BMD) and incidence of osteoporosis. Sixty Saudi male adults age matching were assigned into two groups: A healthy control group (n = 30) and osteoporosis patients diagnosed according to T-score (n = 30). Serum calcium, vitamin D, osteocalcin, lead, cadmium were measured. Osteoporotic group showed a highly significant elevation of blood lead and cadmium levels compared to the control group (p <0.001). BMD was negatively correlated with serum osteocalcin level compared with control. There was a significant negative correlation between the cadmium and lead levels (r=-0.465 and p-value = 0.01) and calcium (p < 0.004). Our findings suggested that high cadmium and lead were negative correlated to BMD and increased the risk factor for osteoporosis.

Resumo Os metais tóxicos do soro têm sido implicados no desenvolvimento de muitas doenças. Este estudo investigou a associação entre os níveis sanguíneos de chumbo e cádmio com densidade mineral óssea anormal (DMO) e incidência de osteoporose. Sessenta adultos sauditas do sexo masculino com idades iguais foram divididos em dois grupos: um grupo de controle saudável (n = 30) e pacientes com osteoporose diagnosticados de acordo com o T-score (n = 30). Cálcio sérico, vitamina D, osteocalcina, chumbo, cádmio foram medidos. O grupo osteoporótico apresentou elevação altamente significativa dos níveis de chumbo e cádmio no sangue em comparação ao grupo controle (p < 0,001). A DMO foi negativamente correlacionada com o nível de osteocalcina sérica em comparação com o controle. Houve correlação negativa significativa entre os níveis de cádmio e chumbo (r = -0,465 ep = 0,01) e cálcio (p < 0,004). Nossos achados sugeriram que cádmio e chumbo elevados foram correlacionados negativamente à DMO e aumentaram o fator de risco para osteoporose.

Mechanism of miR-26a-5p/cAMP response element binding protein 1 molecular axis regulating osteogenic differentiation of adipose-derived mesenchymal stem cells / 中国修复重建外科杂志

OBJECTIVE@#To investigate the regulatory effects of miR-26a-5p on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) by regulating cAMP response element binding protein 1 (CREB1).@*METHODS@#The adipose tissues of four 3-4 weeks old female C57BL/6 mice were collected and the cells were isolated and cultured by digestion separation method. After morphological observation and identification by flow cytometry, the 3rd-generation cells were subjected to osteogenic differentiation induction. At 0, 3, 7, and 14 days after osteogenic differentiation induction, the calcium deposition was observed by alizarin red staining, ALP activity was detected, miR- 26a-5p and CREB1 mRNA expressions were examined by real-time fluorescence quantitative PCR, and CREB1 protein and its phosphorylation (phospho-CREB1, p-CREB1) level were measured by Western blot. After the binding sites between miR-26a-5p and CREB1 was predicted by the starBase database, HEK-293T cells were used to conduct a dual-luciferase reporter gene experiment to verify the targeting relationship (represented as luciferase activity after 48 hours of culture). Finally, miR-26a-p inhibitor (experimental group) and the corresponding negative control (control group) were transfected into ADSCs. Alizarin red staining, ALP activity, real-time fluorescent quantitative PCR (miR-26a-5p) and Western blot [CREB1, p-CREB1, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)] were performed at 7 and 14 days after osteogenic induction culture.@*RESULTS@#The cultured cells were identified as ADSCs. With the prolongation of osteogenic induction culture, the number of calcified nodules and ALP activity significantly increased ( P<0.05). The relative expression of miR-26a-5p in the cells gradually decreased, while the relative expressions of CREB1 mRNA and protein, as well as the relative expression of p-CREB1 protein were increased. The differences were significant between 7, 14 days and 0 day ( P<0.05). There was no significant difference in p-CREB1/CREB1 between different time points ( P>0.05). The starBase database predicted that miR-26a-5p and CREB1 had targeted binding sequences, and the dual-luciferase reporter gene experiment revealed that overexpression of miR-26a-5p significantly suppressed CREB1 wild-type luciferase activity ( P<0.05). After 7 and 14 days of osteogenic induction, compared with the control group, the number of calcified nodules, ALP activity, and relative expressions of CREB1, p-CREB1, OCN, and RUNX2 proteins in the experimental group significantly increased ( P<0.05). There was no significant difference in p-CREB1/CREB1 between the two groups ( P>0.05).@*CONCLUSION@#Knocking down miR-26a-5p promoted the osteogenic differentiation of ADSCs by up-regulating CREB1 and its phosphorylation.

Expression Changes of Hypoxia-Inducible Factor-1α in G-CSF Induced Hematopoietic Stem Cell Mobilization / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression and its relative mechanism of hypoxia-inducible factor-1α(HIF-1α) in bone marrow(BM) of mice during G-CSF mobilization of hematopoietic stem cells (HSC) .@*METHODS@#Flow cytometry was used to detect the proportion of Lin-Sca-1+ c-kit+ (LSK) cells in peripheral blood of C57BL/6J mice before and after G-CSF mobilization. And the expression of HIF-1α and osteocalcin (OCN) mRNA and protein were detected by RQ-PCR and immunohistochemistry. The number of osteoblasts in bone marrow specimens of mice was counted under the microscope.@*RESULTS@#The proportion of LSK cells in peripheral blood began to increase at day 4 of G-CSF mobilization, and reached the peak at day 5, which was significantly higher than that of control group (P<0.05). There was no distinct difference in the expression of HIF-1α mRNA between bone marrow nucleated cells and osteoblasts of steady-state mice (P=0.073), while OCN mRNA was mainly expressed in osteoblasts, which was higher than that in bone marrow nucleated cells (P=0.034). After mobilization, the expression level of HIF-1α increased, but OCN decreased, and the number of endosteum osteoblasts decreased. The change of HIF-1α expression was later than that of OCN and was consistent with the proportion of LSK cells in peripheral blood.@*CONCLUSION@#The expression of HIF-1α in bone marrow was increased during the mobilization of HSC mediated by G-CSF, and one of the mechanisms may be related to the peripheral migration of HSC induced by osteoblasts inhibition.

Association of blood heavy metal levels with osteocalcin abnormality and incidence of osteoporosis in Saudi subjects / Associação de anormalidades de osteocalcina nos níveis de metais pesados no sangue e incidência de osteoporose em indivíduos sauditas

Serum toxic metals have been implicated in development of many diseases. This study investigated the association between blood levels of lead and cadmium with abnormal bone mineral density (BMD) and incidence of osteoporosis. Sixty Saudi male adults age matching were assigned into two groups: A healthy control group (n = 30) and osteoporosis patients diagnosed according to T-score (n = 30). Serum calcium, vitamin D, osteocalcin, lead, cadmium were measured. Osteoporotic group showed a highly significant elevation of blood lead and cadmium levels compared to the control group (p <0.001). BMD was negatively correlated with serum osteocalcin level compared with control. There was a significant negative correlation between the cadmium and lead levels (r=-0.465 and p-value = 0.01) and calcium (p < 0.004). Our findings suggested that high cadmium and lead were negative correlated to BMD and increased the risk factor for osteoporosis.

Os metais tóxicos do soro têm sido implicados no desenvolvimento de muitas doenças. Este estudo investigou a associação entre os níveis sanguíneos de chumbo e cádmio com densidade mineral óssea anormal (DMO) e incidência de osteoporose. Sessenta adultos sauditas do sexo masculino com idades iguais foram divididos em dois grupos: um grupo de controle saudável (n = 30) e pacientes com osteoporose diagnosticados de acordo com o T-score (n = 30). Cálcio sérico, vitamina D, osteocalcina, chumbo, cádmio foram medidos. O grupo osteoporótico apresentou elevação altamente significativa dos níveis de chumbo e cádmio no sangue em comparação ao grupo controle (p < 0,001). A DMO foi negativamente correlacionada com o nível de osteocalcina sérica em comparação com o controle. Houve correlação negativa significativa entre os níveis de cádmio e chumbo (r = -0,465 ep = 0,01) e cálcio (p < 0,004). Nossos achados sugeriram que cádmio e chumbo elevados foram correlacionados negativamente à DMO e aumentaram o fator de risco para osteoporose.

Effects of 275 nm and 310 nm ultraviolet irradiation on bone metabolism in ovariectomized osteoporotic rats / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effect of 275 nm and 310 nm ultraviolet irradiation on ovariectomized rats' bone metabolism.@*METHODS@#Twenty four 3-month-old female Sprague-Dawley (SD) rat were randomly divided into control group, sham operated group, 275 nm ultraviolet (UV) irradiation group and 310 nm UV irradiation group. Each group contained 6 rats. The rats in the two irradiation groups were treated with bilateral ovariectomy. The rats in sham operated group received sham operation (They were given the same back incision and a bit of par-ovarian fat were removed). Control group received no disposition. About 24 weeks after operation, all the rats received detailed bone mineral density (BMD) detection again. Detection regions include cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur. Next, osteopenia rats in 275 nm irradiation group were UV irradiated 275 nm with fixed illumination intensity (15 μW/cm2) everyday for 16 weeks. The osteopenia rats in 310 nm irradiation group were UV irradiated 310 nm with fixed illumination intensity (15 μW/cm2) everyday for 16 weeks. The backs of the rats were shaved regularly as irradiation area (6 cm×8 cm). After 16-week irradiation, all the rats' BMD of cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur were measured. At the end of the trial, all the rats' blood specimens were obtained and serum 25(OH)D, procollagen type Ⅰ N-peptide (PINP) and osteocalcin (OC) were measured.@*RESULTS@#Compared with control group [(238.78±26.74) mg/cm3], the BMD of the whole body were significantly lower in 275 nm [(193.34±13.28) mg/cm3] and 310 nm [(191.19±18.48) mg/cm3] irradiation groups (P=0.002, P=0.001). There were no significant difference between sham operated group [(227.20±14.32) mg/cm3] and control group. After 16-week ultraviolet irradiation, the BMD of the whole body were significantly increased in 275 nm [(193.34±13.28) mg/cm3 vs. (221.68±25.52) mg/cm3, P=0.005] and 310 nm groups [(191.19±18.48) mg/cm3 vs. (267.48±20.54) mg/cm3, P < 0.001] after corresponding irradiation. The BMD of the four body regions (lumbar vertebra, proximal femur, mid femur and distal femur) had significantly increased after irradiation in 275 nm irradiation group. For 310 nm irradiation group, the BMD in cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur also had increased significantly after 310 nm ultraviolet irradiation. The concentration of serum 25(OH)D and OC was higher in 275 nm irradiation group than in control group [(46.78±5.59) μg/L vs. (21.32±6.65) μg/L, P=0.002;(2.05±0.53) U/L vs. (1.32±0.07) U/L, P=0.022]. Compared with the control, the concentration of serum 25(OH)D [(58.05±12.74) μg/L], OC [(2.04±0.53) U/L] and PINP [(176.16±24.18) U/L] was significantly higher (P < 0.001, P=0.015, P=0.005) in 310 nm irradiation group. However, there were no significantly difference between sham operated group and the control.@*CONCLUSION@#Both 275 nm and 310 nm ultraviolet could improve rats' vitamin D synthesis. Both 275 nm and 310 nm ultraviolet could improve osteopenia rats' bone condition. The irradiation of 310 nm might be more effective on bone condition improvement.

The key research of sports in maintaining human health / A pesquisa chave do esporte na manutenção da saúde humana / La investigación clave del deporte para mantener la salud humana

ABSTRACT Introduction One of the evaluation factors of human health is bone health, and an evaluation index of bone health is osteoporosis. Sports are an effective way to improve the human body. Objective The paper discusses the effects of different exercise intensities on human bone health. Methods The thesis selected 51 female college students, designed different exercise intensities of fitness running intervention programs, and conducted a 12-month exercise intervention. We divide female college students into three groups. The subjects' bone mineral density (BMD), serum alkaline phosphatase (ALP), and serum osteocalcin (BGP) were tested before and after the experiment. Results The differences in femoral BMD, serum ALP, serum BGP, and lumbar spine BMD of the three groups of volunteers were significant (P<0.05), while the differences in ulna and radius BMD were not significant. Conclusions Sports can promote human bone health. At the same time, the effect of fitness running on human BMD is site-specific. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução Um dos fatores de avaliação da saúde humana é a saúde óssea, e um índice de avaliação da saúde óssea é a osteoporose. Os esportes são uma forma eficaz de melhorar o corpo humano. Objetivo o artigo discute os efeitos de diferentes intensidades de exercício na saúde óssea humana. Métodos A tese selecionou 51 universitárias, elaborou diferentes intensidades de exercícios em programas de intervenção de corrida de aptidão e conduziu uma intervenção de exercícios de 12 meses. Dividimos as universitárias em três grupos. A densidade mineral óssea (BMD), fosfatase alcalina sérica (ALP) e osteocalcina sérica (BGP) dos indivíduos foram testadas antes e depois do experimento. Resultados As diferenças na DMO femoral, ALP sérica, BGP sérica e DMO da coluna lombar dos três grupos de voluntários foram significativas (P <0,05), enquanto as diferenças na DMO da ulna e rádio não foram significativas. Conclusão O esporte pode promover a saúde óssea humana. Ao mesmo tempo, o efeito da corrida adaptativa na DMO humana é específico do local. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción Uno de los factores de evaluación de la salud humana es la salud ósea y un índice de evaluación de la salud ósea es la osteoporosis. Los deportes son una forma eficaz de mejorar el cuerpo humano. Objetivo El artículo analiza los efectos de diferentes intensidades de ejercicio en la salud ósea humana. Métodos La tesis seleccionó a 51 estudiantes universitarias, diseñó diferentes intensidades de ejercicio de programas de intervención para correr y realizó una intervención de ejercicio de 12 meses. Dividimos a las estudiantes universitarias en tres grupos. La densidad mineral ósea (DMO), la fosfatasa alcalina sérica (ALP) y la osteocalcina sérica (BGP) de los sujetos se analizaron antes y después del experimento. Resultados Las diferencias en la DMO femoral, la ALP sérica, la BGP sérica y la DMO de la columna lumbar de los tres grupos de voluntarios fueron significativas (P <0,05), mientras que las diferencias en la DMO del cúbito y del radio no fueron significativas. Conclusión Los deportes pueden promover la salud ósea humana. Al mismo tiempo, el efecto de la actividad física en la DMO humana es específico del sitio. Nivel de evidencia II; Estudios terapéuticos: investigación de los resultados del tratamiento.

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin

ABSTRACT Purpose Herein we evaluated the effects of platelet concentrate (PC) and platelet-poor plasma (PPP) on bone repair using noncritical defects in the calvaria of rabbits and compared them to the presence of TGF-β1 and osteocalcin on reparative sites. Methods Five noncritical defects of 8.7 mm in diameter were created on the calvaria of 15 animals. Each defect was treated differently, using autograft (ABG), ABG associated with PC (ABG + PC), ABG with PPP (ABG + PPP), isolated PPP, and blood clot (control). The animals were submitted to euthanasia on the second, fourth and sixth week post-surgery. Results The defects that received ABG+PC or PPP demonstrated lower bone formation when compared to specimens that received ABG in the same period. These results coincided to significant higher immunopositivity for TGF-β1 for specimens that received PC, and lower presence of cytokine in the group PPP. However, either higher or lower presence of TGF-β1 were also correlated to lower presence of osteocalcin. Likewise, these results were similar to findings in specimens treated only with PPP when compared to control. Conclusions PC and PPP were not effective when applied in association with ABG. Similarly, isolated use of PPP was not beneficial in optimizing the bone repair.

Influência do LED Violeta associado ou não ao gel clareador na inflamação, mineralização e maturação colágena do tecido dentinário e pulpar / Influence of violet LED associated or not with peroxide gel on inflammation, mineralization, and collagen fiber maturation in dentin and pulp tissue

Objetivo: Avaliar a influência do LED Violeta, associado ou não ao gel clareador a base de peróxido de hidrogênio (PH) a 17,5% no complexo dentino-pulpar de ratos. Materiais e métodos: Molares superiores de 80 ratos foram distribuídos nos grupos (n = 10): CONT ­ sem tratamento, PH ­ 1 aplicação de 30 minutos de PH a 17,5%, LED ­ 1 aplicação de 20 minutos do LED Violeta, e PH+LED - aplicação do PH e LED Violeta. Imediatamente (T0), e aos 7 (T1), 15 (T2) e 30 dias (T3) após o tratamento, os ratos foram eutanasiados e as maxilas processadas para avaliação histológica, imunoistoquímica (IL-17, IL-23 e osteocalcina), e de picrosírius red, sendo realizados os testes de Wilcoxon e Mann-Whitney e Teste-T pareado e Teste-T, respectivamente. (α = 0,05). Resultados: Necrose e infiltrado inflamatório severo foram observados nos grupos PH e PH+LED. Apenas o grupo PH+LED manteve a imunomarcação severa para IL-17 e IL-23, diferindo do grupo LED e PH que apresentaram moderada imunomarcação em T0. Os grupos PH e PH+LED apresentaram severa imunomarcação de OCN em T2 e moderada imunomarcação em T3. O grupo LED apresentou menor quantidade de fibras imaturas em T2 e T3 que o grupo CONT. Conclusão: A terapia com LED violeta não induziu inflamação e fibrose no tecido pulpar, apesar de acelerar a maturação das fibras de colágeno da dentina e, quando associada ao peróxido de hidrogênio, pode tornar os dentes mais sensíveis(AU)

Objective: Was to evaluated the influence of the Violet LED, associated or not with a 17.5% hydrogen peroxide (HP) bleaching gel in the dentin-pulp complex of rats. Materials and methods: Upper molars of eighty Wistar rats were distributed in the groups (n = 10): CONT - without treatment, HP - 1 application of 30 minutes of 17.5% HP, LED - 1 application of 20 minutes of the Violet LED, and HP+LED - application of HP and LED Violet. Immediately (T0), and at 7 (T1), 15 (T2) and 30 days (T3) after treatment, the rats were euthanized and the jaws were processed for histological, immunohistochemical evaluation (IL-17, IL-23 and osteocalcin), and picrosirius red, with Wilcoxon and Mann-Whitney tests and paired T-test and T-test, respectively (α = 0.05). Results: Necrosis and severe inflammatory infiltrate were observed in the PH and PH+LED groups. Only the PH+LED group maintained severe immunostaining for IL-17 and IL-23, differing from the LED and PH group which presented moderate T0 immunostaining. The PH and PH+LED groups presented severe immunostaining of OCN in T2 and moderate immunostaining in T3. The LED group had a lower amount of immature fibers in T2 and T3 than the CONT group. Conclusion: Violet LED therapy induced no inflammation and fibrosis in the pulp tissue, however accelerating the maturation of dentin collagen fibers and, when associated with hydrogen peroxide, can make the teeth more sensitive(AU)

Correlation between genotypes with metabolic markers and microstructure of bones in children with Gitelman syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the correlation between the genotypes and metabolic markers and microstructure of bones in children with Gitelman syndrome (GS).@*METHODS@#For 15 children with GS and 10 healthy individuals, baseline data and bone metabolic markers including parathyroid hormone, alkaline phosphatase, osteocalcin, N-terminal propeptide of type I procollagen, beta isomer of the C-terminal telopeptide of type I collagen and 25-hydroxyvitamin D, high-resolution peripheral quantitative computed tomography indicators (volumetric bone mineral density, bone microstructure indicators) were collected. Genetic testing was carried out to determine their genotypes.@*RESULTS@#The volumetric bone mineral density, bone geometry and bone microstructure parameters of the GS group were better than those of the healthy controls (P<0.05). Variants of the SLC12A3 gene were identified in 9 of the 15 patients but none of the 10 healthy controls.@*CONCLUSION@#The phenotype of GS children is influenced by the interaction of genetic variants, though the phenotype associated with high frequency mutations showed no specificity. There is also a correlation between their genotype and the bone microstructure.

Experimental acute sepsis reduced number of osteocalcin immunolabeled cells in periodontal ligament

Abstract The aim of this study was to evaluate the effect of acute sepsis in the periodontal ligament, alveolar and furcation bone in absence of periodontitis induction through histological and immunohistochemical analyses. A septic rat model was established by cecal ligation and puncture (CLP). Twelve rats were randomly divided into CLP (n=6) and Sham (n=6) groups. The animals were euthanized at 24 h and hemimandibles were submitted to histomorfometric (bone matrix, collagenous fibers, fibroblasts, osteocytes, inflammatory cells, and blood vessels) and immunohistochemical (BMP-2/4, RANKL and osteocalcin) evaluation in alveolar bone, furcation bone and periodontal ligament. Our results demonstrated that histomorphometric parameters were similar in alveolar bone, furcation bone and periodontal ligament of Sham and CLP rats. Regarding to immunohistochemical analyses, the number of BMP-2/4 and RANKL immunolabeled cells was also similar in both groups. Furthermore, it was detected a reduction in the osteocalcin immunolabeled cells in periodontal ligaments of CLP compared to Sham rats (p=0.0014). In conclusion, the acute sepsis induction resulted in reduced number of osteocalcin labelled cells in periodontal ligament region. Moreover, no significant histological differences were observed in the periodontium of rats under acute sepsis. Considering the role of osteocalcin in bone remodeling, the study contributes to revealing the importance of careful periodontal evaluation in the presence of sepsis.

Resumo O objetivo deste estudo foi avaliar o efeito da sepse aguda no ligamento periodontal, osso alveolar e osso da furca por meio de análise histológica e imunohistoquímica. O modelo de sepse em ratos foi estabelecido pelo procedimento de ligação e perfuração do ceco (CLP). Doze ratos foram divididos de forma randomizada em ratos sépticos (n=6) e controle - grupo Sham (n=6). Os animais foram eutanasiados após 24 horas e suas hemimandíbulas foram submetidas aos procedimentos histotécnicos para análise histomorfométricos (matriz óssea, fibras colágenas, fibroblastos, osteócitos, células inflamatórias e vasos sanguíneos) e imunohistoquímicos (BMP-2/4, RANKL e osteocalcina) no osso alveolar, osso de furca e ligamento periodontal. Nossos resultados demonstraram que os parâmetros histomorfométricos foram similares no osso alveolar, osso de furca e ligamento periodontal dos animais do grupo sepse e do grupo Sham. Em relação à análise por imunohistoquímica, o número de células imunomarcadas para BMP-2/4 e RANKL também foi similar em ambos os grupos. Entretanto, houve redução (p=0.0014) no número de células imunomarcadas para osteocalcina no ligamento periodontal de ratos sépticos em relação ao grupo Sham. Como conclusão, o estabelecimento de sepse aguda resultou em um número reduzido de células imunomarcadas para osteocalcina na região do ligamento periodontal (p=0,0014). Além disso, não foram observadas diferenças histológicas significativas no periodonto de ratos na presença de sepse aguda. Considerando o papel da osteocalcina na remodelação óssea, este estudo contribui para revelar a importância da avaliação periodontal cuidadosa na presença de sepse.

A influência da ocitocina e do antagonista do receptor de ocitocina no metabolismo ósseo, estresse oxidativo, padrões da marcha e análise do tipo ansioso de ratas senescentes / The influence of oxytocin and the oxytocin receptor antagonist on bone metabolism, oxidative stress, gait and analysis of the anxious type of senescent rats

O objetivo deste estudo foi avaliar a ação da ocitocina (OT) endógena, bem como o efeito potencializador da OT exógena sobre o metabolismo ósseo, estresse oxidativo, marcha e análise do tipo ansioso de ratas na periestropausa. Ao completar 19 meses, os animais receberam injeções de solução salina (0,15M/ip), Atosiban (AT) (At; 300 µg/Kg/ip), OT (Ot; 134 µg/Kg/ip) ou At+Ot (injeções de OT 5 minutos após AT), sendo duas injeções de cada substância por dia, com intervalos de 12 horas entre elas, a cada 30 dias até a idade de 21 meses. Após trinta dias sem tratamentos, foi realizada a coleta de amostras biológicas. Aspartato aminotransferase (AST), marcador de dano hepático, foi menor em Ot e At+Ot. Substância ácida reativa ao ácido tiobarbitúrico (TBARs É¥mol/L), marcador do dano oxidativo lipídico, foi maior no grupo Ot comparado ao At (p = 0,0093), e menor no At+Ot em relação ao Ot (p = 0,0040). Houve maior defesa antioxidante enzimática avaliada por meio da superóxido dismutase (SOD) no grupo Ot em comparação ao Veh (p < 0,0312). Por sua vez, no grupo At houve maior atividade enzimática da fosfatase alcalina (FAL) em relação ao Veh e Ot (p < 0,0001; At+Ot: p = 0,0015). A espessura do tecido ósseo compacto foi menor no grupo At em relação ao Veh (p = 0,0228), no entanto, foi maior no grupo Ot em relação ao Veh e At (p = 0,0132, p < 0,0001); no grupo At+Ot foi menor quando comparado ao grupo Ot (p = 0,0003). O número de trabéculas ósseas foi menor no grupo At comparado ao Veh (p = 0,0240), e maior em Ot em relação ao At (p = 0,0084). Quanto a análise imunoistoquímica realizada no osso cortical do colo do fêmur, o grupo Ot apresentou maior expressão de osteocalcina (OCN) em comparação aos grupos Veh e At (p = 0,05 e 0,0033), e menor expressão no grupo At+Ot em relação ao grupo Ot (p = 0,05). A expressão de fosfatase ácida resistente ao tartarato (TRAP) foi menor no grupo Ot comparado aos grupos Veh e At (p = 0,05 e 0,0033), contudo foi maior no grupo At+Ot comparado ao Ot (p = 0,05). A densidade mineral óssea areal (DMO) foi significativamente maior nos grupos Ot e At+Ot em relação à Veh (p < 0,0001) e grupo At (p = 0,0231, p = 0,0418). Por sua vez, a relação mineral-matriz (vPO4/Proline) foi maior e a substituição de carbonato tipo B (CO3/vPO4) foi menor no grupo Veh. O teste de deambulação por comprimento (cm) usado para avaliar função musculoesquelética, aumentou em última análise no grupo Ot em relação ao grupo Veh - F (p = 0,0078), At - F (p = 0,0023), bem como aumentou sobre Ot - I (p = 0,0094). O teste do labirinto, usado para estudar o comportamento chamado "tipo ansioso", demonstrou que a OT inverte a redução nas entradas dos braços fechados, reduz o tempo gasto no centro causado pelo At. Os resultados obtidos neste estudo demonstram que a OT ajuda a modular o ciclo de remodelação óssea de ratas senescentes, melhorando os parâmetros de densitometria óssea e os parâmetros funcionais musculoesquelético(AU)

The objective of this study was to evaluate the endogenous oxytocin (OT) action, as well as the potentiating effect of exogenous OT on the bone metabolism, oxidative stress, gait and analysis of the anxious type of rats in periestropause. Upon completing 19 months, the animals received injections of saline solution (0.15M/ip), Atosiban (AT) (At; 300 µg/Kg/ip), OT (Ot; 134 µg/Kg/ip) or At+Ot (OT injections 5 minutes after AT), being two injections of each substance per day, with intervals of 12 hours between them, every 30 days until the age of 21 months. After thirty days without treatment, biological samples were collected. Aspartate aminotransferase (AST), a marker of liver damage, was lower after Ot and At+Ot. Acid reactive substance to thiobarbituric acid (TBARs µmol/L), marker of lipid oxidative damage, was higher in the Ot group compared to At (p = 0.0093), and lower in At+Ot compared to Ot (p = 0.0040). There was a higher antioxidant enzymatic defense evaluated by means of superoxide dismutase (SOD) in the Ot group compared to Veh (p < 0.0312). In turn, in the At group there was greater alkaline phosphatase (FAL) enzymatic activity in relation to Veh and Ot (p < 0.0001; At+Ot: p = 0.0015). The thickness of the compact bone tissue was smaller in the At group in relation to Veh (p = 0.0228), however, it was greater in the Ot group in relation to Veh and At (p = 0.0132, p < 0.0001); in the At+Ot group it was smaller when compared to Ot (p = 0.0003). The number of bone trabecules was smaller in the At group compared to the Veh (p = 0.0240), and greater in Ot in relation to the At (p = 0.0084). As for the immunohistochemical analysis performed on the cortical bone of the femoral neck, the Ot group presented a higher expression of osteocalcin (OCN) compared to the Veh and At groups (p = 0.05 and 0.0033), and lower expression in the At+Ot group compared to the Ot group (p = 0.05). The tartrate-resistant acid phosphatase (TRAP) expression was lower in the Ot group compared to the Veh and At groups (p = 0.05 and 0.0033), however it was higher in the At+Ot group compared to Ot (p = 0.05). The sandal mineral density (BMD) was significantly higher in the Ot and At+Ot groups compared to Veh (p < 0.0001) and At group (p = 0.0231, p = 0.0418). In turn, the parent mineral ratio (vPO4/Proline) was higher and the replacement of carbonate type B (CO3/vPO4) was lower in the Veh group. The walking test per length (cm) used to evaluate musculoskeletal function was ultimately increased in group Ot in relation to group Veh - F (p = 0.0078), At - F (p = 0.0023), as well as increased over Ot - I (p = 0.0094). The labyrinth test, used to study the so-called "anxious type" behavior, demonstrated that the OT reverses the reduction in the entries of the closed arms, reducing the time spent in the center caused by At. The results obtained in this study show that the OT helps to modulate the cycle of bone remodeling of senescent rats, improving the parameters of bone densitometry and the musculoskeletal functional parameters(AU)

Avaliação da regeneração óssea guiada por meio histológico, histométrico, imunoistoquímico e perfil inflamatório de membranas reabsorvíveis de colágeno porcino em defeitos críticos em calotas de ratos / Evaluation histological, histometric, immunohistochemistry analysis and inflammatory profile of bone-guided regeneration with membranes of reabsorbable porcine collagen in critical defects in rodent skullcaps

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)

Cyclosporine A impairs bone repair in critical defects filled with different osteoconductive bone substitutes

Abstract The aim of this study was to assess the influence of cyclosporine administration on the repair of critical-sized calvaria defects (CSDs) in rat calvaria filled with diverse biomaterials. Sixty animals were divided into two groups: the control (CTR) group (saline solution) and the cyclosporine (CCP) group (cyclosporine, 10 mg/kg/day). These medications were administered daily by gavage, beginning 15 days before the surgical procedure and lasting until the day the animals were euthanized. A CSD (5 mm Ø) was made in the calvaria of each animal, which was allocated to one of 3 subgroups, according to the biomaterial used to fill the defect: coagulum (COA), deproteinized bovine bone (DBB), or biphasic calcium phosphate ceramics of hydroxyapatite and β-phosphate tricalcium (HA/TCP). Euthanasia of the animals was performed 15 and 60 days after the surgical procedure (n = 5 animals/period/subgroup). Bone repair (formation) assessment was performed through microtomography and histometry, while the analyses of the expression of the BMP2, Osteocalcin, and TGFβ1 proteins were performed using immunohistochemistry. The CSDs not filled with biomaterials demonstrated lower bone formation in the CCP group. At 15 days, less bone formation was observed in the CSDs filled with DBB, a smaller volume of mineralized tissue was observed in the CSDs filled with HA/TCP, and the expression levels of BMP2 and osteocalcin were lower in the CCP group compared to the CTR group. The use of cyclosporine impaired bone repair in CSD, and this effect can be partially explained by the suppression of BMP2 and osteocalcin expression.

Upper Zone of Growth Plate and Cartilage Matrix (UCMA) Levels in Patients with Chronic Kidney Disease

Abstract Chronic kidney disease (CKD) is an important health problem across the world affecting the adult population with an enormous social and economic burden. Calcium regulation is also affected in patients with CKD, and related to several disorders including vascular calcifications, mineral bone disorders, and cardiovascular diseases (CVD). Upper zone of growth plate and cartilage matrix (UCMA) is vitamin K-dependent protein (VKDP) and acts as a calcification inhibitor in the cardiovascular system. The molecular mechanism of UCMA action remains unclear in CKD. In the current study, we aimed to investigate serum total UCMA levels and its association with calcium metabolism parameters in CKD patients including hemodialysis (HD) patients. Thirty-seven patients with CKD stage 3-5, 41 HD patients, and 34 healthy individuals were enrolled in this cross-sectional study. Serum UCMA and calcification related protein levels (Matrix Gla Protein (MGP), Osteocalcin (OC), and Fetuin-A) were analyzed with enzyme-linked immunosorbent assay (ELISA). Calcium mineral disorder parameters (Serum Ca, P, iPTH) were quantified with routine techniques. We, for the first time, report the potential biomarker role of UCMA in CKD including HD. Serum total UCMA levels were significantly higher in patients with CKD including HD patients than the healthy controls. Also, serum UCMA levels showed negative correlations with serum calcium, and eGFR, while showed positive relationships with P, iPTH, MGP, OC. Increased total UCMA levels may have a role in the Ca metabolism disorder and related to the pathogenesis of Vascular Calcification in patients with CKD.

Effect of enhanced masticatory force on OPG, RANKL and MGF in alveolar bone of ovariectomized rats

Abstract Menopause induces oral bone loss, leading to various oral diseases. Mastication importantly affects bone metabolism in the jawbone. Objective: To analyze the effect of enhanced masticatory force on osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), and mechano-growth factor (MGF) in alveolar bone of ovariectomized rats and to study the mechanics mechanism of the alveolar bone of ovariectomized rats response to enhanced masticatory force. Methodology: Thirty Sprague Dawley rats were randomly divided into three groups: sham-operation group (fat around the removed ovary + normal hard diet), model group (ovariectomy + normal hard diet), and experimental group (ovariectomy + high hard diet). It was a 2-month experiment. Enzyme-linked immunosorbent assay (ELISA) detected serum estradiol (E2), osteocalcin (BGP) and alkaline phosphatase (ALP) in rats. Bone histomorphometric indices in the third molar region of maxilla were detected by micro-CT; protein expressions of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Western blot; and gene expression of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Quantitative Real-Time PCR. Results: Comparing with model group, serum E2 in experimental group increased but not significantly, serum BGP and serum ALP in experimental group decreased but not significantly, OPG in experimental group in alveolar bone increased significantly, RANKL in experimental group in alveolar bone decreased significantly, RANKL/OPG ratio in experimental group decreased significantly, MGF in experimental group in alveolar bone increased significantly, bone volume to total volume fraction increased significantly in experimental group, trabecular thickness increased significantly in experimental group, and trabecular separation decreased significantly in experimental group. Conclusion: Enhanced masticatory force affected the expression of OPG, RANKL, and MGF in alveolar bone of ovariectomized rats, improved the quality of jaw bone of ovariectomized rats, and delayed oral bone loss by ovariectomy.

Impact of history of periodontitis on gene expression of bone-related factors in young patients

Abstract Although dental implants and bone regenerative procedures are important approaches for the reestablishment of esthetics and function in young patients with a history of generalized aggressive periodontitis (GAP), no predictable outcomes have been reported, and the host osteo-immunoinflammatory response may play a relevant role in this context. In view of the lack of molecular investigations into the bone tissue condition of young patients with periodontitis, the aim of this study was to evaluate the gene expression of bone-related factors in this population. Bone biopsies were obtained from the posterior mandible in 16 individuals previously diagnosed with GAP and on periodontal support therapy and from 17 periodontally healthy (PH) patients. The gene expression of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, receptor activator of the NF-κB ligand (RANKL), osteoprotegerin (OPG), osteocalcin (OC), bone sialoprotein (BSP), and type I collagen (COL-I), important biomarkers of bone turnover, was evaluated by qRT-PCR. Lower TGF-β and OPG mRNA levels were observed in GAP patients compared to PH individuals (p ≤ 0.05). There were no between-group differences in levels of TNF-α, BSP, RANKL, OC, or COL-I mRNA (p>0.05). In young adults, a history of periodontal disease can negatively modulate the gene expression of important bone-related factors in alveolar bone tissue. These molecular outcomes may contribute to the future development of therapeutic approaches to benefit bone healing in young patients with history of periodontitis via modulation of osteo-immuno-inflammatory biomarkers.

Cyclosporine A impairs bone repair in critical defects filled with different osteoconductive bone substitutes

Abstract The aim of this study was to assess the influence of cyclosporine administration on the repair of critical-sized calvaria defects (CSDs) in rat calvaria filled with diverse biomaterials. Sixty animals were divided into two groups: the control (CTR) group (saline solution) and the cyclosporine (CCP) group (cyclosporine, 10 mg/kg/day). These medications were administered daily by gavage, beginning 15 days before the surgical procedure and lasting until the day the animals were euthanized. A CSD (5 mm Ø) was made in the calvaria of each animal, which was allocated to one of 3 subgroups, according to the biomaterial used to fill the defect: coagulum (COA), deproteinized bovine bone (DBB), or biphasic calcium phosphate ceramics of hydroxyapatite and β-phosphate tricalcium (HA/TCP). Euthanasia of the animals was performed 15 and 60 days after the surgical procedure (n = 5 animals/period/subgroup). Bone repair (formation) assessment was performed through microtomography and histometry, while the analyses of the expression of the BMP2, Osteocalcin, and TGFβ1 proteins were performed using immunohistochemistry. The CSDs not filled with biomaterials demonstrated lower bone formation in the CCP group. At 15 days, less bone formation was observed in the CSDs filled with DBB, a smaller volume of mineralized tissue was observed in the CSDs filled with HA/TCP, and the expression levels of BMP2 and osteocalcin were lower in the CCP group compared to the CTR group. The use of cyclosporine impaired bone repair in CSD, and this effect can be partially explained by the suppression of BMP2 and osteocalcin expression.

Reference Intervals for Bone Turnover Markers in Korean Healthy Women

BACKGROUND: Adequate suppression of bone turnover rate is important to decrease fracture risk without mineralization defect due to oversuppression. This study was performed to determine reference intervals (RIs) for 2 bone turnover markers, serum C-terminal telopeptide of type I collagen (CTX) and osteocalcin, in Korean women.METHODS: A total of 461 Korean women (287 premenopausal and 174 postmenopausal) without any disease or drug history affecting bone metabolism was included. Serum CTX and osteocalcin were measured after overnight fasting. Bone mineral density (BMD) was measured at the 1st to 4th lumbar vertebra using dual energy X-ray absorptiometry. Subjects with normal spinal BMD (T-score ≥−1.0) were included in this study.RESULTS: After stable concentrations were maintained, both CTX and osteocalcin were abruptly increased in 50 to 59 years, and then decreased with increasing age. Median levels and interquartile range of serum CTX and osteocalcin in all subjects were 0.322 (0.212–0.461) ng/mL and 15.68 (11.38–19.91) ng/mL. RIs for serum CTX and osteocalcin in all subjects were 0.115 to 0.861 ng/mL and 6.46 to 36.76 ng/mL. Those were higher in postmenopausal women (CTX, 0.124–1.020 ng/mL, osteocalcin, 5.42–41.57 ng/mL) than in premenopausal women (CTX, 0.101–0.632 ng/mL, osteocalcin, 6.73–24.27 ng/mL). If we use target reference levels as lower half of premenopausal 30 to 45 years in patients with antiresorptive drugs, those were 0.101 to 0.251 ng/mL and 6.40 to 13.36 ng/mL.CONCLUSIONS: We established RIs for serum CTX and osteocalcin in healthy Korean women with normal lumbar spine BMD. Premenopausal RIs for serum CTX and osteocalcin would be useful to monitor patients with low bone mass using osteoporosis drugs.

Paget's Disease of Bone Affecting Peripheral Limb: Difficulties in Diagnosis: A Case Report

In terms of management of Paget's disease of bone (PDB), early diagnosis and proper management achieving remission is essential with lifelong specialist follow-up. We present the case of a 40-year-old woman with PDB affecting mainly the distal extremities (ankle and wrist). The patient visited our hospital in 2012 with heel pain. Plain radiography revealed osteoporosis, and a bone scan revealed hot uptake. Initial laboratory investigations showed normal serum calcium, 25-hydroxy-vitamin D, and parathyroid hormone levels; however, osteocalcin, C-terminal telopeptide of type I collagen, and bone alkaline phosphatase levels were elevated. A bone mineral density scan showed T- and Z-scores of −2.5 and −2.7, respectively, and bisphosphonate treatment was initiated. Biopsy performed on the calcaneal lateral wall revealed inconclusive findings. Follow-up biopsy on the left distal radius was performed 7 years later to investigate wrist pain, and this examination led to a final diagnosis as PDB. We suggest inconclusive biopsy result during the early phase of PDB and highly recommend follow-up evaluation in osteoporosis with atypical behavior.