Espondiloencondrodisplasia con desregulación inmune relacionada a ACP5. Reporte de cuatro casos / Spondyloenchondrodysplasia with immune dysregulation related to ACP5. A report of 4 cases

La espondiloencondrodisplasia con desregulación inmune relacionada a ACP5 (SPENCDI #607944, por la sigla de spondyloenchondrodysplasia with immune dysregulation y el número que le corresponde en OMIM, Online Mendelian Inheritance in Man) es una displasia inmuno-ósea poco frecuente con manifestaciones heterogéneas y gravedad variable. Presenta lesiones espondilometafisarias, disfunción inmune y compromiso neurológico. Se reportan aspectos clínicos, radiológicos y genéticos de cuatro niñas con SPENCDI en un hospital pediátrico. Todas presentaron manifestaciones esqueléticas y tres de ellas enfermedad inmunológica grave. Se encontró en tres pacientes la variante probablemente patogénica c.791T>A; p.Met264Lys en homocigosis, y en una paciente las variantes c.791T>A; p.Met264Lys y c.632T>C; p.lle211Thr (variante de significado incierto con predicción patogénica según algoritmos bioinformáticos) en heterocigosis compuesta en ACP5. La presencia de la variante repetida c.791T>A sugiere la posibilidad de un ancestro en común en nuestra población. El reconocimiento y diagnóstico de esta entidad es importante para lograr un oportuno abordaje, que deberá ser multidisciplinario, orientado hacia la prevención de posibles complicaciones.

Spondyloenchondrodysplasia with immune dysregulation related to ACP5 (SPENCDI, OMIM number 607944) is an uncommon immune-skeletal dysplasia with heterogeneous manifestations and variable severity. It is characterized by spondylar and metaphyseal lesions, immune dysfunction, and neurological involvement. Here we report the clinical, radiological and genetic aspects of 4 girls with SPENCDI treated at a children's hospital. They all had skeletal manifestations and 3 developed severe immune disease. In 3 patients, the likely pathogenic variant c.791T>A; p.Met264Lys (homozygous mutation) was observed, while 1 patient had variants c.791T>A; p.Met264Lys and c.632T>C; p.lle211Thr (variant of uncertain significance with pathogenic prediction based on bioinformatics algorithms) caused by a compound heterozygous mutation in ACP5. The repeated presence of variant c.791T>A suggests the possibility of a common ancestor in our population. The recognition and diagnosis of this disorder is important to achieve a timely approach, which should be multidisciplinary and aimed at preventing possible complications.

Efeito tempo-dose-resposta do montelucaste no processo de reparo alveolar em camundongos 129Sv/Ev / Time-dose-response effect of Montelukast in the process of alveolar repair in 129Sv / Ev mice

Metabólitos do ácido araquidônico são conhecidos por exercerem importante papel nos processos inflamatórios e no metabolismo do tecido ósseo. No entanto, as ações pontuais, especialmente dos leucotrienos derivados da 5-lipoxigenase (5-LO) sobre o processo de reparo ósseo intramembranoso são pouco exploradas. O presente estudo tem como objetivo analisar os efeitos tempo-dose-resposta da droga montelucaste (MTK), potente antagonista dos receptores de cisteinil leucotrienos tipo 1 (CisLT1Rs), no curso do reparo alveolar pós-exodontia em camundongos 129Sv/Ev, bem como nos níveis plasmáticos de marcadores ósseos bioquímicos. Para tanto, foram utilizados 70 camundongos machos jovens divididos em quatro grupos, de acordo com o tratamento: C - Grupo Controle (não tratados); CV - Grupo Controle Veículo, 20 µL de solução fisiológica (SF) 0,9%; MTK2 ­2 mg/kg de MTK e MTK4 ­ 4 mg/kg de MTK. Os animais dos grupos CV, MTK2 e MTK4 foram tratados diariamente por via oral, iniciando 24 horas antes do procedimento cirúrgico, continuando até o final dos períodos experimentais de 7, 14 e 21 dias pós-operatórios. Ao final dos períodos determinados, os animais foram submetidos à eutanásia para coleta de sangue para análise bioquímica dos níveis de cálcio, fosfato, fosfatase ácida resistente ao tartarato (TRAP) total e fosfatase alcalina (FAL), coleta da maxila direita contendo os alvéolos dentários para serem analisados por meio de microtomografia computadorizada (microCT), e análise histopatológica. Os resultados obtidos foram submetidos à testes estatísticos considerando-se nível de confiança de 5%. Observou-se aumento do BV/TV para os animais tratados com MTK em relação aos grupos C e CV, tanto aos 14 dias quanto aos 21 dias, sendo maior no grupo MTK4 aos 14 dias em relação ao grupo MTK2. Do mesmo modo, os animais tratados com MTK em ambas doses apresentaram aumento significativo de Tb.Th em comparação aos grupos C e CV aos 21 dias. Chamou a atenção valores de BV/TV e Tb.Th significativamente reduzidos no grupo CV em comparação ao C, indicando um efeito negativo da manipulação do animal. Na análise histopatológica observou-se reparo ósseo precoce nos animais MTK2 e MTK4 em todos os períodos avaliados, em comparação aos do grupo C, bem como atraso no processo de reparo no grupo CV aos 21 dias. Quanto aos marcadores plasmáticos, observou-se aumento do cálcio no grupo MTK4 em relação ao grupo C aos 7 dias, e aos 21 dias também em relação ao grupo MTK2. Já o fosfato mostrouse significantemente elevado nos períodos de 7 e 21 dias no grupo MTK2 em relação aos demais grupos. FAL e TRAP total não apresentaram níveis plasmáticos significativamente diferentes comparando-se os grupos e períodos. Considerando os resultados obtidos, concluiu-se que o MTK exerceu efeito tempo-dose-dependente, acelerando o processo de reparo ósseo intramembranoso alveolar e interferindo nos níveis plasmáticos de cálcio e fosfato no presente modelo animal(AU)

Arachidonic acid metabolites are known to play an important role in inflammatory processes and in bone metabolism. However, the role of these products on alveolar bone repair post tooth extraction remains to be explored, especially leukotrienes, derived from 5-lipoxygenase (5-LO). The present study aims to analyze the time-doseresponse effects of the drug montelukast (MTK), a potent type 1 leukotriene cystenyl antagonist (CisLT1Rs), in the alveolar repair process after extraction in male 129Sv/Ev mice. For this purpose, 70 young male mice were used, divided into four groups: C - Control Group (no treatment); VC - Vehicle Control Group, treated with 20 µL of 0.9% SF; MTK2 - treated with 2mg / Kg of MTK and MTK4 - treated with 4mg / Kg of MTK. The animals of the CV, MTK2 and MTK4 groups were treated daily orally (V.O.), starting 24 hours before the surgical procedure, continuing until the end of the experimental periods of 7, 14 and 21 days postoperatively. At the end of the experimental periods, the animals were euthanized for blood collection for serum markers as calcium, phosphate, tartrate-resistant acid phosphate (TRAP) and alkaline phosphatasis (FAL), and to removal of the right maxilla containing the dental socket to be analyzed under computed microtomography (microCT) and histopathology. The results obtained were subjected to statistical tests considering a confidence level of 5%. Results revealed an increase in BV/TV for MTK vs. C and CV groups, in both 14 and 21 days time points. Of note, this increase was higher in MTK4 than in the MTK2 at 14 days. Considering Tb.Th, both MTK2 e MTK4 groups presented positive effects in the BV/TV and Tb.Th increase when compared to controls groups (C and CV) at 21 days. A decrease in BV/TV and Tb.Th was observed in CV compared to C, as a negative effect of animal manipulation. As observed in H&E sections, both MTK2 and MTK4 experimental groups presented an early bone repair in comparison with C group from 7 to 21 days. CV group presented a slight delayed bone healing compared to C. Levels of calcium was increased in MTK4 in comparison to C and MTK2 at 7 and 21 days. Phosphate was significantly elevated at 7 and 21 days in MTK2 in comparison to the other groups. Despite of beneficial effects on observed on morphological levels on sites of healing (microCT and HE), no significant changes were found for bone markers of remodeling in blood plasma (FAL and TRAP). Taken together, these results indicate that MTK induced early bone healing post tooth extraction in 129Sv/Ev mice. Thus, the inhibition of CysLT is suggested to exert a positive influence on intramembranous bone repair post tooth extraction(AU)

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

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)

Infrared light-emitting diode (led) effects on orthodontic tooth movement

Abstract The present study aimed to analyze the effect of LED phototherapy on the presence of hyalinization and root resorption during orthodontic tooth movement (OTM) in rats and to measure the amount of tooth movement. Eighty rats were allocated into two groups: LED and control (CON), where the LED rats were irradiated with infrared LED (850 nm, 30 mW) for 5 min during the first five days of OTM and where controls were not irradiated. Both groups were subdivided into four subgroups (n=10) according to the date of euthanasia (4, 7, 14 and 21 days). Five out of ten LED21 and five of ten CON21 rats were submitted to micro-computed tomography (μCT); μCT scans were taken on days 0, 7, 14 and 21. For histological study, maxillae were processed to light microscopy using Hematoxylin-Eosin (HE) and Tartrate-Resistant Acid Phosphatase (TRAP) histochemistry. The amount of tooth movement did not differ between LED and CON. Hyalinization was observed at the pressure areas in both groups, and it did not show a statistically significant difference between the groups. Root resorption was also observed in both groups after 7 days and it did not represent any differences between the two groups. LED phototherapy was not able to increase the amount of OTM. Similar characteristics of hyalinization and root resorption were observed in both groups.

Resumo O presente estudo tem como objetivo analisar o efeito da fototerapia LED na presença da hialinização e reabsorção radicular durante o movimento dentário ortodôntico (MDO) em ratos, e a mensuração da quantidade de movimento dentário. Oitenta ratos foram alocados em dois grupos: LED e Controle (CON), os ratos foram irradiados com um LED infravermelho (850nm, 30mW) por 5 minutos durante os cinco primeiros dias da MDO; e o grupo controle não foi irradiado. Ambos os grupos foram subdivididos em 4 subgrupos (n=10) de acordo com a data da eutanásia (4, 7, 14 e 21 dias). Cinco dos dez ratos LED21 e cinco dos dez ratos CON21foram submetidos a microtomografia computadorizada (μCT); As μCT foram realizadas nos dias 0, 7, 14 e 21. Para o estudo histológico, as maxilas foram processadas para microscopia de luz, usando hematoxilina-eosina (HE) e Fosfatase ácido Tartrate-Resistente (TRAP) para histoquímica. A quantidade de movimento dentário não diferiu entre o LED e o CON. A hialinização foi observada nas áreas de pressão em ambos os grupos e não mostrou diferença estatisticamente significante. Reabsorção radicular também foi observada em ambos os grupos depois de 7 dias e não houve diferença entre os grupos. A fototerapia LED não aumentou a quantidade de MDO. Características similares de hialinização e reabsorção radicular foram observadas em ambos os grupos

Preventive effect and mechanism of puerarin on rat models of disuse osteoporosis / 中国中药杂志

To investigate the preventive effect and possible mechanism of puerarin(Pur) in rat model of disuse osteoporosis(DOP),thirty healthy Wistar female rats of 2 months old were randomly divided into control group(Control), hindlimb suspension group(HLS), and puerarin group(HLS+Pur) in hindlimb suspension, with 10 rats in each group. A disuse osteoporosis model was established by tail suspension method, and 15.4 mg·kg~(-1) puerarin suspension was administered to HLS+Pur group every day, and the same volume of distilled water was administered to Control group and HLS group respectively. After 28 days, the rats were sacrificed by abdominal aorta blood collection, the main organs of the rats were removed, and the bone tissues of the rats were dissected. The organ index of the rats was calculated and the histopathology of the organs was observed under microscope. Bone mineral density test and bone biomechanical experiment were performed. Bone histomorphometry results were observed after bone tissue sectioning, and serum biochemical markers of bone metabolism were determined. There was no significant difference in organ index between the groups. There was no obvious abnormality in the pathological examination of the organs. The results of bone mineral density showed that puerarin could significantly increase the bone density of the tibia and vertebrae caused by hindlimb suspension. The mechanical parameters experiments showed that puerarin could effectively increase the maximum load and elastic modulus of the tibia and vertebrae. Fluorescence labeling showed that the fluorosis interval increased and the bone formation increased during puerarin treatment. The VG staining results showed that compared with the HLS group, in the puerarin group, the number of trabecular bone increased, the thickness of the trabecular bone became thicker, and the bone separation became smaller, which greatly improved the bone microstructure after hindlinb suspension. In addition, serum biochemical indicators showed that puerarin could promote bone formation index bone calcium. The content of osteocalcin(OC) increased and inhibited the formation of tartrate-resistant acid phosphatase 5 b(TRACP 5 b). Puerarin has a preventive effect in the rat model of disuse osteoporosis and its effect is good, and its mechanism may be related to promoting bone formation and inhibiting bone resorption.

Effect of avocado/soybean unsaponifiables on periodontal repair in rats with arthritis and induced periodontitis

Abstract Objective: This study aimed to evaluate the effect of avocado/soybean unsaponifiables (ASU) on periodontal repair in rats with induced periodontitis and arthritis. Methodology: Forty-five rats were submitted to periodontitis induction by insertion of ligatures into the upper second molars, maintained for 15 days. These animals were randomly allocated to 3 groups according to the presence of induced arthritis (ART) and the application of the ASU: Control (CTR) group-healthy animals, where saline solution was administered; ART-animals with induced arthritis, where saline solution was administered; ART/ASU-animals with induced arthritis, where ASU (0.6 mg/ kg) was administered. The drugs were administered daily by gavage and the animals were euthanized after 7, 15 and 30 days of the ligature removal. Bone resorption, inflammatory infiltrate composition and marker proteins expression of the differentiation and formation of osteoclasts (RANKL and TRAP) were assessed. Results: The ART/ASU group presented higher bone volume than the ART group at 7 and 30 days after the ligature removal. Furthermore, the ART group presented higher quantity of inflammatory cells and expression of TRAP and RANKL than the other groups. Conclusion: ASU administration improves the repair of periodontal tissues in an experimental periodontitis model in rats with induced arthritis.

Efeitos do queijo probiótico com adição de Lactobacillus casei-01 na periodontite experimental em ratos / Effects of probiotic cheese supplemented with Lactobacillus casei-01 on experimental periodontitis in rats

Objetivo: Este estudo avaliou os efeitos da ingestão de alimento funcional, queijo Prato contendo Lactobacillus casei-01, no desenvolvimento da periodontite experimental (PE) em ratos. Materiais e Métodos: 44 ratos machos Wistar foram divididos em 4 grupos (n = 11): C (controle) - animais sem indução de PE e alimentados com ração convencional (RC) + queijo convencional (CONV); PROB - animais sem indução de PE e alimentados com RC + queijo probiótico (PROB); PE - animais com indução de PE e alimentados com RC + queijo CONV e PE-PROB - animais com indução de PE e alimentados com RC + queijo PROB. O queijo Prato foi fabricado por método tradicional com bactérias lácticas, suplementado ou não com a cepa probiótica Lactobacillus casei-01. No dia 0 do experimento, queijo CONV ou PROB, de acordo com o grupo experimental, foi administrado oralmente para todos os animais 1 x / dia até o final do experimento. No dia 28, a PE foi induzida nos primeiros molares inferiores direito e esquerdo dos animais dos grupos PE e PE-PROB. No dia 42, todos os animais foram submetidos à eutanásia. Foram realizadas análises microtomográfica na região da bifurcação e nos sítios vestibular, lingual e interproximais [volume ósseo (BV), número de trabéculas (Tb.N), espessura das trabéculas (Tb.Th), perda óssea alveolar (ABL)] e análises histológica (parâmetros dos tecidos moles e duros), histométrica [porcentagem de osso na furca (PBF)] e imunoistoquímica [fator de necrose tumoral-α (TNFα), interleucina (IL)-1ß, IL-10, fator de crescimento transformador-ß1 (TGFß1) e fosfatase ácida resistente ao tartarato (TRAP)] na região da furca. Foram realizadas análises histológica (parâmetros de inflamação, celularidade e estruturação tecidual) e histométrica [altura de vilosidades (VH) e profundidade de cripta (CD)] nos tecidos intestinais. Os dados foram submetidos à análise estatística. Resultados: Os valores de BV foram similares em todos os grupos experimentais. No entanto, os grupos PE e PE-PROB apresentaram Tb.N significativamente maior e Tb.Th significativamente menor que os grupos C e PROB. O grupo PE apresentou PBF significativamente menor que os grupos C e PROB, e similar ao grupo PE-PROB. Os grupos PE e PE-PROB apresentaram maior expressão de citocinas pró-inflamatórias (TNFα e IL-1ß) e TRAP que os grupos C e PROB. O grupo PE-PROB apresentou maior expressão de citocinas anti-inflamatórias (TGFß1 e IL-10) que o grupo PE. Não houve diferenças significativas para os valores de VH e CD entre os gru pos experimentais. Conclusão: O consumo de queijo Prato, tanto CONV como PROB, teve efeito protetor nos tecidos periodontais durante o desenvolvimento da PE em ratos. Contudo, o queijo PROB parece ter estimulado principalmente a expressão de citocinas antiinflamatórias, quando comparado ao queijo CONV, favorecendo o processo de reparo tecidual(AU)

Objective: This study evaluated the effects of a functional food (Prato cheese) intake supplemented with Lactobacillus casei-01 in experimental periodontitis (EP) in rats. Materials and Methods: 44 Wistar male rats were divided into 4 groups (n = 11): C (control) - animals without EP induction and fed with conventional feed (CF) + conventional cheese (CONV); PROB - animals without EP induction and fed with CF + probiotic cheese (PROB); EP - animals with EP induction and fed with CF + CONV cheese; and EP-PROB - animals with EP induction and fed with CF + PROB cheese. Prato cheese was produced by traditional lactic bacteria manufacturing method, supplemented or not with the probiotic strain Lactobacillus casei-01. On day 0, either CONV or PROB cheese, according to the experimental group, was orally administered to all animals once a day until the end of the experiment. On day 28, EP was induced on the right and left mandibular first molars of the animals of groups EP and EP-PROB. On day 42, all animals were euthanized. Microtomographic analysis was performed in the furcation region and at the buccal, lingual and interproximal sites [bone volume (BV), trabeculae number (Tb.N), trabeculae thickness (Tb.Th), alveolar bone loss (ABL)]. Histologic (hard and soft tissue parameters), histometric [bone percentage in the furcation area (PBF)] and immunohistochemical [transforming growth factor-α (TNF-α), interleukin (IL)-1ß, IL-10, transforming growth factor-ß1 (TGF- ß1) and tartrate resistant acid phosphatase (TRAP)] analyses were performed in the furcation region. Histologic (inflammatory, cellularity and tissue structure parameters) and histometric [villi height (VH) and crypt depth (CD)] analyses were performed in the intestinal tissues. Data were statistically analyzed. Results: BV values were similar in all experimental groups. However, groups EP and EP-PROB presented significantly greater Tb.N and significantly lower Tb.Th than groups C and PROB. Group EP presented significantly lower PBF than groups C and PROB, and similar to group EP-PROB. Groups EP and EP-PROB presented higher expression of pro-inflammatory cytokines (TNF-α and IL-1ß) and TRAP than groups C and PROB. Group EP-PROB presented higher expression of anti-inflammatory cytokines (TGFß1 e IL-10) than group EP. No significant differences were observed among groups for VH and CD values. Conclusion: CONV as well as PROB Prato cheese intake promoted a protective effect on periodontal tissues during EP induction in rats. However, PROB cheese seems to mainly stimulate the e xpression of antiinflammatory cytokines when compared with CONV cheese, favoring tissue repair process(AU)

The effect of colchicine on alveolar bone loss in ligature-induced periodontitis

Abstract Colchicine is widely used in the treatment of several inflammatory diseases due to its anti-inflammatory effect, but effects on bone metabolism are unclear. The aim of this study was to evaluate the effects of systemically-administered colchicine on healthy periodontium and experimentally-induced periodontitis. In total, 42 male Wistar rats were included in this study. A non-ligated group constituting the negative control group (Control, C, n = 6) and a ligature-only group forming the positive control group (LO, n = 12) were created separately. Twelve rats were treated with 0.4 mg/kg colchicine and another 12 with 1 mg/kg colchicine. In the colchicine-administered groups, right mandibles constituted the ligated groups (1 mgC-L or 0.4 mgC-L) and left mandibles formed the corresponding non-ligated controls (1mgC or 0.4mgC). Silk ligatures were placed at the gingival margin of the lower first molars. The animals were euthanized at different time-points of healing (11 or 30 days). Alveolar bone loss was clinically measured and TRAP+ osteoclasts, osteoblastic activity, and MMP-1 expression were examined histologically. There was no increase in alveolar bone loss with either colchicine dose in healthy periodontium (p > 0.05) and the highest level of alveolar bone loss, TRAP+ osteoclast number, and MMP-1 expression were measured in the LO group (p < 0.05). The 0.4 mgC-L group showed less alveolar bone loss at 11 days (p < 0.05), but greater loss at 30 days. The 1 mgC-L group showed higher osteoblast number than the other ligated groups (p < 0.05) at both time-points. In summary, colchicine did not increase alveolar bone loss in healthy periodontium and also may tend to reduce periodontitis progression. However, further extensive study is necessary to understand the mechanism of colchicine action on alveolar bone loss in periodontitis.

Effect of ProRoot MTA® and Biodentine® on osteoclastic differentiation and activity of mouse bone marrow macrophages

Abstract Objectives This investigation aimed to assess the differentiation inhibitory effects of ProRoot MTA® (PMTA) and Biodentine® (BIOD) on osteoclasts originated from murine bone marrow macrophages (BMMs) and compare these effects with those of alendronate (ALD). Materials and Methods Mouse BMMs were cultured to differentiate into osteoclasts with macrophage colony-stimulating factor and receptor activator of NF-κB (RANKL), treated with lipopolysaccharide. After application with PMTA, BIOD, or ALD, cell toxicities were examined using WST-1 assay kit, and RANKL-induced osteoclast differentiation and activities were determined by resorption pit formation assay and tartrate-resistant acid phosphate (TRAP) staining. The mRNA levels of osteoclast activity-related genes were detected with quantitative real time polymerase chain reaction. Expressions of molecular signaling pathways were assessed by western blot. All data were statistically analyzed with one-way ANOVA and Tukey's post-hoc test (p<0.05). Results Mouse BMMs applied with PMTA, BIOD, or ALD showed highly reduced levels of TRAP-positive osteoclasts. The BIOD treated specimens suppressed mRNA expressions of cathepsin K, TRAP, and c-Fos. Nonetheless, it showed a lower effect than PMTA or ALD applications. Compared with ALD, PMTA and BIOD decreased RANKL-mediated phosphorylation of ERK1/2 and IκBα. Conclusions PMTA and BIOD showed the inhibitory effect on osteoclast differentiation and activities similar to that of ALD through IκB phosphorylation and suppression of ERK signaling pathways.

Effect of minimally invasive anterior lateral approach for total hip arthroplasty on serum type I collagen propeptide, tartrate-resistant acid phosphatase 5b and hip joint function in patients with femoral neck fracture / 中国骨伤

OBJECTIVE@#To analyze the level of serum N-terminal propeptide of type I precollagen (PINP) and tartrate resistant acid phosphatase 5b(TRACP-5b) in patients with femoral neck fracture(FNF) after minimally invasive anterior lateral approach with total hip arthroplasty and the effects of hip function.@*METHODS@#From September 2016 to May 2017, 98 cases of femoral neck fracture were divided into control group and observation group, 49 cases in each group. There were 49 patients in control group, including 30 males and 19 females, ranging in age from 63 to 72 years old, who underwent minimally invasive anterolateral total hip arthroplasty. There were 49 patients in observation group, including 29 males and 20 females, ranging in age from 62 to 73 years old, who underwent minimally invasive anterolateral total hip arthroplasty. The perioperative conditions(operation time, bleeding volume, incision length, hospitalization time), bone metabolism indexes including PINP, TRACP-5b, fibroblast growth factor(FGF), bone gla-protein(BGP), propetide carboxy-terminal procollagen (PICP), bone-specific alkaline phosphatase(BAP), and pain mediators such as prostaglandin E2 (PGE2) and 5-hydroxytrytamine (5-HT) levels were compared between the two groups. The hip joint function and complications were evaluated.@*RESULTS@#The operation time of the observation group was longer than that of the control group(0.05). PINP, fibroblast growth factor, BGP, PICP and BAP in observation group were higher than those in control group 1 month after operation, and TRACP-5b was lower than those in control group(0.05).@*CONCLUSIONS@#Minimally invasive anterolateral approach total hip arthroplasty is safe and reliable, and can improve hip function, improve bone metabolism, promote fracture healing, alleviate pain in patients with femoral neck fracture, which is worthy of promotion.

Effect of acute administration of nicotine and ethanol on tooth movement in rats

Abstract The aim of this study was to evaluate the effect of acute administration of nicotine and ethanol on tooth movement in rats. Two hundred rats were divided into eight groups: S: saline; N: nicotine; E: ethanol; NE: nicotine and ethanol; SM: saline with tooth movement; NM: nicotine with tooth movement; EM: ethanol with tooth movement; and NEM: nicotine and ethanol with tooth movement. All the solutions were applied for 32, 44, or 58 days, according to the subgroup. Orthodontic movement (25 cN) was initiated 30 days after solution administration in the groups with tooth movement. The rats were euthanized 2, 14, or 28 days after initiation of tooth movement. Tooth sections were stained using picrosirius and tartrate-resistant acid phosphatase (TRAP). The data were compared by ANOVA using Tukey's HSD and Games-Howell. On day 28 of tooth movement, the NEM group had a lower percentage of type I collagen compared to the SM group (p = 0.0448), and the S group had a higher number of osteoclasts/μm2 compared to the N group (p = 0.0405). Nicotine and ethanol did not affect the tooth movement rate, regardless of induction of orthodontic movement. Nicotine influenced the number of osteoclasts by decreasing their quantity when dental movement was not induced. When nicotine was associated with ethanol, it interfered in the maturation of collagen fibers during orthodontic movement.

Alveolar bone healing in rats: micro-CT, immunohistochemical and molecular analysis

Abstract Alveolar bone healing after upper incisor extraction in rats is a classical model of preclinical studies. The underlying morphometric, cellular and molecular mechanism, however, remains imprecise in a unique study. Objectives The aim of this study was therefore to characterize the alveolar bone healing after upper incisor extraction in rats by micro computed tomographic (Micro-CT), immunohistochemical and real-time polymerase chain reaction (RT-PCR) analysis. Material and Methods Thirty animals (Rattus norvegicus, Albinus Wistar) were divided into three groups after upper incisors extraction at 7, 14, and 28 days. Micro-CT was evaluated based on the morphometric parameters. Subsequently, the histological analyses and immunostaining of osteoprotegerin (OPG), receptor activator of nuclear kappa B ligand (RANKL) and tartrate resistant acid phosphate (TRAP) was performed. In addition, RT-PCR analyses of OPG, RANKL, the runt-related transcription factor 2 (RUNX2), osteocalcin (OC), osteopontin (OPN), osterix (OST) and receptor activator of nuclear kappa B (RANK) were performed to determine the expression of these proteins in the alveolar bone healing. Results Micro-CT: The morphometric parameters of bone volume and trabecular thickness progressively increased over time. Consequently, a gradual decrease in trabecular separation, trabecular space and total bone porosity was observed. Immunohistochemical: There were no differences statistically significant between the positive labeling for OPG, RANKL and TRAP in the different periods. RT-PCR: At 28 days, there was a significant increase in OPG expression, while RANKL expression and the RANKL/OPG ratio both decreased over time. Conclusion Micro-CT showed the newly formed bone had favorable morphometric characteristics of quality and quantity. Beyond the RUNX2, OC, OPN, OST, and RANK proteins expressed in the alveolar bone healing, OPG and RANKL activity showed to be essential for activation of basic multicellular units during the alveolar bone healing.

Bone repair access of BoneCeramic™ in 5-mm defects: study on rat calvaria

Abstract Objective: The aim of this study was to evaluate the osteoconductive potential of BoneCeramic™ on bone healing in rat calvaria 5-mm defects. Material and Methods: A 5-mm calvaria bone defect was induced in three groups and the defect was not filled with biomaterial [Clot Group (CG)], autogenous bone (AG), or Bone Ceramic Group (BCG). Animals were euthanized after 14 or 28 days and the bone tissue within the central area of the bone defect was evaluated. Results were compared using ANOVA and Tukey test (p<0.05). Immunohistochemistry was performed using primary antibodies against osteocalcin, RUNX-2, TRAP, VEGF proteins, and 3-dimensional images of the defects in μCT were obtained to calculate bone mineral density (BMD). Results: In BCG, the defect was completely filled with biomaterial and new bone formation, which was statistically superior to that in the GC group, at both time-points (p<0.001 for 14 days; p=0.002 for 28 days). TRAP protein showed weak, RUNX-2 showed a greater immunolabeling when compared with other groups, VEGF showed moderate immunostaining, while osteocalcin was present at all time-points analyzed. The μCT images showed filling defect by BCG (BMD= 1337 HU at 28 days). Conclusion: Therefore, the biomaterial tested was found to be favorable to fill bone defects for the reporting period analyzed.

Effects of triethylene glycol dimethacrylate (TEGDMA) on the odontoclastic differentiation ability of human dental pulp cells

Abstract Objectives: The primary purpose of this study was to examine the effects of triethylene glycol dimethacrylate (TEGDMA) on odontoclastic differentiation in the dental pulp tissue. Material and Methods: The effects of different TEGDMA dosages on the odontoclastic differentiation capability of dental pulp cells were analyzed in vitro using the following methodologies: i) flow cytometry and tartrate-resistant acid phosphatase (TRAP) staining; ii) apoptotic effects using Annexin V staining; iii) mRNA expression of osteoprotegerin (OPG) and receptor activator of nuclear factor (NF)-kB ligand (RANKL) genes by quantitative Real-time PCR (qRT-PCR); and iv) OPG and RANKL protein expression by enzyme-linked immunosorbent assay (ELISA). Results: TEGDMA caused relatively less odontoclastic differentiation in comparison with the control group; however, odontoclastic differentiation augmented with increasing doses of TEGDMA (p<0.05). The mRNA and protein expression of OPG was lower in TEGDMA treated pulp cells than in the control group (p<0.05). While the mRNA expression of RANKL remained unchanged compared to the control group (p>0.05), its protein expression was higher than the control group (p<0.05). In addition, TEGDMA increased the apoptosis of dental pulp cells dose dependently. Conclusions: TEGDMA reduced the odontoclastic differentiation ability of human dental pulp cells. However, odontoclastic differentiation ratios increased proportionally with the increasing dose of TEGDMA.

Hepatic Osteodystrophy: The Mechanism of Bone Loss in Hepatocellular Disease and the Effects of Pamidronate Treatment

OBJECTIVES: The present study was designed to evaluate the bone phenotypes and mechanisms involved in bone disorders associated with hepatic osteodystrophy. Hepatocellular disease was induced by carbon tetrachloride (CCl4). In addition, the effects of disodium pamidronate on bone tissue were evaluated. METHODS: The study included 4 groups of 15 mice: a) C = mice subjected to vehicle injections; b) C+P = mice subjected to vehicle and pamidronate injections; c) CCl4+V = mice subjected to CCl4 and vehicle injections; and d) CCl4+P = mice subjected to CCl4 and pamidronate injections. CCl4 or vehicle was administered for 8 weeks, while pamidronate or vehicle was injected at the end of the fourth week. Bone histomorphometry and biomechanical analysis were performed in tibiae, while femora were used for micro-computed tomography and gene expression. RESULTS: CCl4 mice exhibited decreased bone volume/trabecular volume and trabecular numbers, as well as increased trabecular separation, as determined by bone histomorphometry and micro-computed tomography, but these changes were not detected in the group treated with pamidronate. CCl4 mice showed increased numbers of osteoclasts and resorption surface. High serum levels of receptor activator of nuclear factor-κB ligand and the increased expression of tartrate-resistant acid phosphatase in the bones of CCl4 mice supported the enhancement of bone resorption in these mice. CONCLUSION: Taken together, these results suggest that bone resorption is the main mechanism of bone loss in chronic hepatocellular disease in mice.

Effect of resveratrol on peak bone mass in growing rats / 浙江大学学报·医学版

Objective: To investigate the effect of resveratrol on peak bone mineral density and bone mass in growing rats. Methods: Thirty-six female healthy Wistar rats were randomly divided into control group, icariin group and resveratrol group with 12 rats in each group. Icariin (25 mg·kg-1·d-1), resveratrol (8.4 mg·kg-1·d-1) or equal volume of distilled water were given by gavage to icariin group, resveratrol group and control group, respectively. The rats were sacrificed after 12 weeks. The organ indexes were calculated and pathology sections were observed; the bone mineral density (BMD), bone biomechanics, serum bone metabolism index, and results of micro-CT scan were analyzed. Results: During the experiment, the body weight of rats showed an increasing trend and there was no significant difference among three groups (P0.05). There were no significant differences in organ index of vital organs and pathological changes among the groups (all P0.05). Compared with the control group, the whole body BMD, and the BMDs of femur and vertebrae in icariin and resveratrol groups were significantly increased after 12 weeks (all PPPPPPPConclusion: Resveratrol can inhibit bone resorption and enhance bone formation, so as to improve the peak bone mass and bone density, enhance bone strength and improve the microstructure of bone tissue in young rats.

Effects of avocado/soybean unsaponifiables (ASU) on the treatment of ligature-induced periodontitis in rats

Abstract The purpose of this study is to evaluate the effect of the avocado/soybean unsaponifiables (ASU) on the treatment of induced periodontitis in rats. Periodontitis was induced in 84 rats via ligature placement around the second upper molar, which was removed after 7 days, and scaling and root planning (SRP) was performed at this time. Subsequently, the rats were randomly allocated to four groups with 21 animals each: One SRP group in which saline solution was administered (SS), and three groups in which ASU was administered (0.6 g/kg/day), beginning either 7 days before the induction of periodontitis (SRP/ASU-7), on the day of periodontitis induction (SRP/ASU0), or on the day of treatment (SRP/ASU+7). ASU and SS were administered daily by gavage until the sacrifice of the animals (7, 15, and 30 days after SRP). The % bone in the furcation area was evaluated by histomorphometry and micro-CT. The expression of proteins (TRAP, RANKL, and alkaline phosphatase) and mRNA (IL-1β, TNF-α, IL-6, RANKL, and alkaline phosphatase) were evaluated by immunohistochemistry and qPCR. The SRP/ASU+7 group presented a higher percentage of bone fill in the furcation area and higher expression of alkaline phosphatase than in the SRP group (at 7 and 30 days, respectively). The SRP/ASU0 and SRP/ASU+7 groups presented lower expression levels of RANKL mRNA than the SRP and SRP/ASU-7 groups at 15 days. ASU administration on the day of the SRP treatment of the ligature-induced periodontitis promoted subtle beneficial effects on periodontal repair following the treatment of induced periodontitis within the experimental period of 7 days.

Imunomarcação para TRAP em tecido de reparação óssea de ratos espontaneamente hipertensos (SHR) tratados com Atenolol / Immunostaining for TRAP in bone healing tissue of Spontaneously Hypertensive Rats (SHR) treated with Atenolol

Introdução: A hipertensão arterial tem sido um dos maiores problemas de saúde no mundo, com grandes alterações para as doenças cardiovasculares e renais. O tecido ósseo tem função importante no suporte, proteção e locomoção e está sob o controle de fatores sistêmicos como hormônios e fatores locais, entre eles os fatores de crescimento e citocinas. A Fosfatase Ácida Tartarato Resistente (TRAP) é uma enzima que faz parte da família das fosfatases ácidas e apresenta localização intracelular; mais especificamente dentro do compartimento lisossomal de osteoclasto, macrófagos e células dendríticas, tem sido utilizada como um marcador histoquímico da atividade osteoclástica. Objetivos: Avaliar a expressão da proteína TRAP em alvéolos dentários de ratos hipertensos (SHR) e normotensos tratados ou não com atenolol. Métodos: Neste estudo foram utilizados 4 grupos de ratos sendo: 1) W (wistar sem tratamento), 2) WT (wistar tratado com atenolol), 3) S (SHR sem tratamento) e 4) ST (SHR tratado com atenolol), submetidos a exodontia do incisivo superior direito, com eutanásia no 7º, 14º, 21 e 28º dia pós-operatório. A análise dos mecanismos biológicos envolvidos no processo de reparo alveolar foi obtida pela análise da expressão de proteínas TRAP por meio da técnica de imunoistoquímica. Os resultados foram analisados pela média e erro padrão da média e aplicado o teste paramétrico ANOVA, com pos-test de Tukey para avaliar os períodos dentro de cada grupo e entre os grupos, sendo consideradas as diferenças significativas quando p<0,05. Resultados: Os resultados mostraram que a marcação TRAP aumenta em alvéolo dentais de ratos Wistar durante todos os períodos pós ­ operatórios. A marcação TRAP aumenta apenas ao 14o nos dias de reparação alveolar em alvéolo dental de SHR não tratados. O atenolol não altera o processo de reparo alveolar em ratos Wistar, porém o atenolol promoveu a redução da marcação de TRAP em SHR ao 14º dia. Conclusão: A hipertensão aumenta a expressão da proteína TRAP no 14o dia pós-cirúrgico de reparação alveolar e o atenolol promove redução da marcação aumentada de TRAP ao 14º dia pós-cirúrgico em alvéolos de SHR(AU)

Introduction: Arterial hypertension has been one of the world's biggest health problems, with considerable alterations for cardiovascular and renal diseases. The bone tissue has an important role in support, protection and locomotion and is controlled by systemic factors like hormones and local factors, such as growth factors and cytokines. The Tartrate-resistant Acid Phosphatase (TRAP) is an enzyme that belongs to the Acid Phosphatases family and has an intracellular location, more specifically inside the lysosomal compartment of osteoclasts, macrophages and dendritic cells. It has been used as a histochemical marker of the osteoclast activity. Objectives: Evaluate TRAP protein's expression in the dental alveoli of normotensive and hypertensive rats (SHR) treated or not treated with Atenolol. Methods: In this study, four groups of rats were used: 1) W (with no treatment), 2) WT (wistar treated with Atenolol), 3) S (SHR without treatment) and 4) ST (SHR treated with Atenolol), all of which underwent exodontia of the upper right incisor with euthanasia on the 7th, 14th, 21st and 28th day after the operation. The analysis of the biological mechanisms involved in the process of alveolar repair was obtained by the expression of TRAP proteins in the alveolar process through an immunohistochemistry technique. The results were analyzed through the average and its standard error. The parametric test ANOVA was applied with Tukey's posttest were applied to evaluate the periods within each group and between the groups, considering the significant differences when p< 0,05. Results: The results demonstrated that TRAP staining increases in the dental alveoli of Wistar rats during all the post-surgical periods. TRAP staining increases only on the 14th day of alveolar recovery in the dental alveoli of non-treated SHR. Atenolol does not change the process of alveolar repair in Wistar rats, but Atenolol promoted the reduction of TRAP staining among SHR on the 14th day. Conclusion: Hypertension increases the expression of TRAP proteins on the 14th alveolar recovery postsurgical day and Atenolol promotes the reduction of the increased TRAP staining on the 14th postsurgical day in SHR's alveoli(AU)