Curcumin alleviated oxidation stress injury by mediating osteopontin in nephrolithiasis rats

Purpose: To explore the role and mechanism of curcumin (Cur) in reducing oxidative stress damage in rats with nephrolithiasis induced by ethylene glycol (EG). Methods: Thirty male rats were divided into normal control, model, positive (10% potassium citrate), Cur-10 (10 mg/kg curcumin) and Cur-20 (20 mg/kg curcumin) groups. Results: The results of kidney tissue section stained by hematoxylin-eosin and von Kossa showed that curcumin treatment can inhibit the formation of kidney stones. The biochemical test results showed that the urea (Ur), creatinine (Cr), uric acid (UA), inorganic phosphorus and Ca2+ concentrations in urine decreased after being treated with curcumin. There were significant differences between different doses of curcumin (P < 0.05). Compared with the Cur-10 group, Cur-20 had a more significant inhibitory effect on malondialdehyde (MDA) (P < 0.05). In addition, reverse transcription polymerase chain reaction (PCR) detection and immunohistochemical results indicated that the osteopontin (OPN) in the kidney was significantly reduced after curcumin treatment. Conclusion: Curcumin could reduce the oxidative stress damage caused by EG-induced kidney stones.

Correlation between serum osteopontin and osteopontin and type 2 diabetes mellitus with coronary heart disease / 中国综合临床

Objective:To study the relationship between serum osteopontin and osteopontin and type 2 diabetes mellitus (T2DM) complicated with coronary heart disease, and to evaluate the correlation between the levels of serum osteopontin and osteopontin with the severity of coronary artery lesions in T2DM patients.Methods:A total of 100 T2DM patients who were suspected to have stable coronary heart disease and underwent coronary angiography from November 2019 to December 2020 were selected from the Affiliated Hospital of Chengde Medical College, according to coronary angiography results, 60 patients with confirmed coronary heart disease were classified as the case group and 40 patients with non-coronary heart disease were classified as the control group for retrospective analysis. The clinical data and biochemical indicators of all patients were recorded, and Gensini score was calculated. The concentration of osteopontin and osteopontin in serum was quantitatively determined by double-antibody enzyme linked immunosorbent assay method. Independent sample t-test was used to compare the mean of normal distribution measurement data between the two groups. The non normal distribution data are represented by M ( Q1, Q3), and Mann Whitney U test is used for comparison between groups. Composition comparison between count data groups χ 2 inspection. Spearman correlation analysis was used to analyze the correlation between serum osteopontin and osteopontin and Gensini score in patients with T2DM. Results:Univariate analysis showed that serum osteopontin and osteopontin were (13.076(8.433, 23.552) μg/L) and (0.437(0.300, 0.630) μg/L) significantly higher in the case group than in the control group (6.367(4.605, 9.048) μg/L) and (0.299(0.196, 0.399) μg/L) respectively, with statistically significant differences ( Z=5.12, 3.28, all P<0.001). Multi-factor logistic regression analysis showed that osteoprotegerin ( OR=2.887, 95% CI:1.850-8.515, P=0.024) and osteopontin ( OR=13.109, 95%CI: 2.557-67.204, P=0.002) were associated with T2DM combined with coronary heart disease, and the risk of T2DM combined with coronary heart disease increased with higher levels of osteoprotegerin and osteopontin. Spearman correlation analysis showed that serum osteopontin and osteoprotegerin were positively correlated with Gensini score in T2DM patients ( r=0.591, 0.467; all P<0.05). Conclusion:Serum osteopontin and osteoprotegerin are associated with T2DM combined with coronary heart disease, and high serum osteopontin and osteoprotegerin are risk factors for T2DM combined with coronary heart disease; serum osteopontin and osteoprotegerin are positively correlated with the degree of coronary artery disease in T2DM patients.

Immunohistochemical Analysis of Alveolar Bone Preserved with Autologous Teeth Graft. Osteopontin Expression and its Regulatory Functions in Preserved Alveolar Ridge

ABSTRACT: The present study investigated the healing response of 12 fresh post-extraction alveolous grafted with particulate autologous teeth to achieve preservation of the post-extraction alveolar ridge. The objective is to elucidate the osteoconductive and osteoinductive properties of the autologous dental graft used as a bone substitute in the alveolar ridge preservation technique. Five patients were included, with at least one hopeless tooth and in need of extraction and preservatio n of the ridge, to receive in the same place a dental implant in prosthetic replacement. In the first surgical stage, dental extractions and preservation of the alveolar ridge were performed, using the teeth extracted and processed with an automatic system as bone substitutes. In the second surgical stage, an incisional bone biopsy was performed in each grafted site, the bone beds were recapitulated in a drilling protocol that allowed the placement of the dental implant, and the harvested bone specimens were prepared for analysis. The histological results of the bone biopsies in all cases showed remnant particles of the dental graft, made up of dentin, partially resorbed, with irregular superficial edges and in close contact with newly forme d bone in transition to mature lamellar bone, in which well differentiated osteocytes were observed. The immunohistochemical results showed a moderate positive expression of osteopontin at the edges of the integrated teeth particles, inside the peritubular dentin space and at the osteodental contact interfaces. In conclusion, the evidence from the study shows that the autologous dental graft is a biocompatible bone substitute, that provides an osteoconductive scaffold that promotes bone cell adhesion and migration for local osteogenesis and that it is associated with moderate in situ expression of osteopontin, which showed a high affinity with mineralized dental tissue, suggesting osteoinductive properties in situ.

RESUMEN: El presente estudio investigó el resultado cicatrizal de 12 alvéolos frescos postextracción injertados con dientes autólogos particulados para lograr la preservación del reborde alveolar postextracción. El objetivo es dilucidar las propiedades osteoconductivas y osteoinductivas del injerto dental autólogo utilizado como sustituto óseo en la técnica de preservación de reborde. Se incluyeron 5 pacientes, con al menos un diente sin esperanza y con necesidad de extracción y preservación del reborde, para recibir en el mismo sitio un implante dental en sustitución protésica. En la primera etapa quirúrgica, se realizaron las extracciones dentales y la preservación del reborde alveolar, utilizando como sustituto óseo los dientes extraídos y procesados con un sistema automático. En la segunda etapa quirúrgica, se realizó una biopsia ósea incisional en cada sitio injertado, los lechos óseos fueron recapitulados en un protocolo de fresado que permitió la colocación del implante dental y los especímenes óseos recolectados fueron preparados para su análisis. Los resultados histológicos de las biopsias óseas en todos los casos mostraron partículas remanentes del injerto dental, conformadas por dentina, parcialmente reabsorbidas, con margenes superficiales irregulares y en estrecho contacto con depósitos de hueso de reciente formación en transición hacia hueso laminar maduro, en el cual se observaron osteocitos bien diferenciados. Los resultados inmunohistoquímicos mostraron una expresión positiva moderada de osteopontina en los bordes de las partículas del injerto dental integrado, al interior del espacio peritubular dentinario y en las interfases de contacto osteodental. En conclusión, la evidencia del estudio muestra que el injerto dental autólogo es un sustituto óseo biocompatible, que provee un andamio osteoconductivo promotor de la adhesión y migración de las células óseas para la osteogénesis local y que está asociado a la expresión modera in situ de osteopontina, la cual mostro una alta afinidad con el tejido dental mineralizado, sugiriendo propiedades osteoinductivas in situ.

Assessing the level and diagnostic accuracy of osteopontin and oral health status in periodontitis patients with/without Type-2 Diabetes Mellitus / Avaliação do nível e precisão do diagnóstico de Osteopontina e estado de saúde bucal em pacientes com periodontite com/sem Diabetes Mellitus tipo 2

Objective: Investigating osteopontin (OPN) level in gingival crevicular fluid (GCF) of patients affected by periodontitis with or without Type-2 diabetes mellitus (T2DM). The aim of this study is to explore the possibility of OPN to differentiate between periodontal health and disease. Material and Methods: A total number of 36 participants seeking periodontal treatment were recruited in this pilot study and divided into three study groups. Periodontitis [systemically healthy participants with periodontitis (probing pocket depth) PPD (probing pocket depth) ≥ 4mm], periodontitis and poorly controlled type 2 diabetes mellitus (), and control (systemically and periodontally healthy periodontium) groups. Plaque index (PI), gingival index (GI) and PPD were examined. OPN level was measured in the GCF and analysed, using Enzyme-Linked Immunosorbent assay. Results: PI and GI were significantly higher in T2DM with periodontitis compared to periodontitis and control groups. Both periodontitis and P-T2DM groups showed significant increase in the OPN levels compared to control group (p<0.001). PPD showed the only significant positive association with OPN (p<0.001) compared to other clinical parameters. The receiver operating characteristics curve analysis demonstrated that OPN had higher area under the curve value (AUC: 0.95) in periodontitis compared to P-T2DM patients (AUC: 0.86). Conclusion: In periodontitis groups, clinical parameters were equally deteriorated together with significant increase in the expression of OPN compared to control. Furthermore, GCF levels of OPN were sensitive and specific enough to discriminate between health and periodontitis even with T2DM. This could introduce OPN to be as a candidate diagnostic biomarker of periodontal disease. (AU)

Objetivo: Investigar o nível de osteopontina (OPN) no fluido gengival crevicular (GCF) de pacientes com periodontite com ou sem diabetes mellitus tipo 2 (T2DM). O objetivo deste estudo foi explorar a possibilidade da OPN diferenciar entre saúde e doença periodontal. Material e Métodos: No total, para este estudo piloto foram recrutados 36 participantes que estavam em busca de tratamento periodontal e divididos em três grupos de estudo: grupos periodontite [participantes sistemicamente saudáveis com periodontite (profundidade de sondagem) PPD ≥ 4 mm], grupo periodontite e Diabetes Mellitus tipo 2 mal controlada (P-T2DM) e grupo controle (saudáveis sistemicamente e periodontalmente). Índice de placa (PI), índice gengival (GI) e PPD foram examinados. O nível de OPN foi medido no GCF e analisado usando o ensaio ELISA (Enzyme-Linked Immuno Sorbent Assay). Resultados: PI e GI foram significativamente maiores no T2DM com periodontite em comparação aos grupos com periodontite e controle. Os grupos com periodontite e P-T2DM apresentaram aumento significativo nos níveis de OPN em comparação ao grupo controle (p <0,001). PPD mostrou a única associação positiva significativa com OPN (p <0,001) em comparação com outros parâmetros clínicos. A análise da curva de características operacionais do receptor demonstrou que OPN teve maior área sob o valor da curva (AUC: 0,95) na periodontite em comparação com pacientes com P-T2DM (AUC: 0,86). Conclusão: Nos grupos com periodontite, os parâmetros clínicos foram igualmente deteriorados juntamente com aumento significativo na expressão de OPN em comparação com o grupo controle. Além disso, os níveis de OPN no GCF foram sensíveis e específicos o suficiente para discernir entre saúde e periodontite, mesmo com T2DM. Isso poderia apresentar a OPN como um candidato a biomarcador diagnóstico de doença periodontal.(AU)

Expressions of transforming growth factor-β1, aquaporin and osteopontin in renal injury induced by fluorosis in rats / 中华地方病学杂志

Objective:To investigate the expressions of transforming growth factor-β1 (TGF-β1), aquaporin (AQP) and osteopontin (OPN) in renal injury induced by fluorosis in rats.Methods:According to body weight (80 - 100 g), forty-eight SD rats were divided into control group (normal saline), low fluoride group (10 mg/kg) and high fluoride group (20 mg/kg) by random number table, 16 rats in each group (half males and half females). The rats were exposed to fluoride by intraperitoneal injection of sodium fluoride. After the rats were treated with fluoride for 12 weeks, the rats were killed by femoral artery bloodletting and the renal tissue was taken. The contents of TGF-β1, AQP and OPN in serum of rats with fluorosis were analyzed by enzyme-linked immunosorbent assay (ELISA). The expressions of TGF-β1, AQP and OPN in renal tissue of rats with fluorosis were analyzed by quantitative real-time PCR (qRT-PCR) and immunocytochemistry.Results:The serum levels of TGF-β1, AQP and OPN in high fluoride group [(26.42 ± 6.09), (378.60 ± 36.84) μg/L, (603.45 ± 64.32) pg/ml] were higher than those in control group [(2.41 ± 0.42), (157.41 ± 15.26) μg/L, (182.45 ± 30.63) pg/ml] and low fluoride group [(13.15 ± 3.26), (245.65 ± 23.21) μg/L, (359.47 ± 55.26) pg/ml, P < 0.05]. The levels of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). The expressions of TGF-β1, AQP and OPN mRNA in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the expressions of TGF-β1, AQP and OPN mRNA in low fluoride group were higher than those in control group ( P < 0.05). The positive expression scores of TGF-β1, AQP and OPN in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the positive expression scores of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). Conclusion:TGF-β1, AQP and OPN are highly expressed in the renal tissue of rats with fluorosis, which can be used as indicators to judge the damage of renal aggregation system caused by fluorosis.

Role of SPP1 in endogenous protective mechanism underlying neuropathic pain in mice with spinal cord injury: relationship with VEGF-AKT signaling pathway / 中华麻醉学杂志

Objective:To evaluate the role of secreted phosphoprotein 1 (SPP1) in endogenous protective mechanism underlying neuropathic pain (NP) in mice with spinal cord injury and the relationship with the vascular endothelial growth factor (VEGF)/kinase B (Akt) signaling pathway.Methods:Seventy-two clean-grade healthy female Kunming mice, aged 7-8 weeks, weighing 30-35 g, were divided into 4 groups ( n=18 each) using a random number table method: sham group (Sham group), NP caused by spinal cord injury group (NP group), NP caused by spinal cord injury+ SPP1-siRNA group (NS-siRNA group), and NP caused by spinal cord injury+ adeno-associated virus vector group (NP-EV group). A model of NP was established by a semi-transecting of the spinal cord.AAV-SPP1-siRNA-GFP adenovirus and AAV-vector-GFP adenovirus 7 μl were intrathecally injected in NS-siRNA group and NP-EV group, respectively, and 5 days later the model was established.At 1, 2 and 3 weeks after operation, 6 mice in each group were randomly selected to measure paw withdrawal threshold to mechanical stimulation (PWMT) and tail flick latency (TFL) to thermal stimuli.And then the mice were sacrificed and the ipsilateral injured spinal cord tissues were taken for determination of the expression of SPP1 mRNA (by real-time polymerase chain reaction) and expression of SPP1, VEGF, Akt and phosphorylated Akt (p-Akt) (by Western blot). Results:Compared with group Sham, PWMT was significantly decreased, TFL was shortened, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was up-regulated at 1, 2 and 3 weeks after operation in NP, NS-siRNA and NP-EV groups( P<0.05). Compared with group NP, PWMT was significantly decreased, TFL was shortened, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was down-regulated at 1, 2 and 3 weeks after operation in group NS-siRNA( P<0.05). Compared with group NS-siRNA, PWMT was significantly increased, TFL was prolonged, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was up-regulated at 1, 2 and 3 weeks after operation in group NS-siRNA( P<0.05). Conclusion:SPP1 is involved in the endogenous protective mechanism underlying NP in mice with spinal cord injury, which may be related to the activation of the VEGF/AKT signaling pathway.

Graft application and dexamethasone treatment influences new bone formation in rat tibial bone defects / Aplicación de injerto y tratamiento con dexametasona influyen en la formación de hueso nuevo en los defectos en la tibia de rata

SUMMARY: The objective of this study were bone defect complications that occur due to traumas or infections. Bone grafts are required to provide support, fill gaps and improve biological repair in skeletal damage. Dexamethasone plays role in calcium signaling modulation and used in diseases. Aim of this study was to evaluate osteonectin and osteopontin expressions in new bone development after dexamethasone application on tibial bone defects. Rats were divided into defect, defect+graft and defect+graft+dexamethasone treated groups. Tibial bone defect created, and rats were kept immobile for 28 days. Alloplastic material was placed in defect area in second and group third groups. 2.5 mg/kg Dex and normal saline were injected to dexamethasone and defect groups twice a week for 56 days. Inflammation and congestion were increased in defect and defect+graft groups. Defect+graft+dexamethasone group; increased number of osteoblast and osteocyte cells, dense bone matrix, formation of new bone trabeculae was observed. Defect+graft group; osteonectin expression in graft regions, osteoblast cells, some connective tissue cells and fibers were seen whereas in defect+graft+dexamethasone group; osteopontin expression in osteoblast and osteocyte cells of new bone trabeculae were observed. Dexamethasone may lead to formation of new bone trabeculae into the graft material resulting in increased osteoconduction and osteoinductive effect for differentiation of osteon.

RESUMEN: Los defectos óseos son complicaciones que ocurren debido a traumas o infecciones. Se requieren injertos óseos para proporcionar apoyo, llenar los espacios y mejorar la reparación biológica en el hueso dañado. La dexametasona desempeña un papel importante en la modulación de la señalización del calcio y se usa en enfermedades. El objetivo de este estudio fue evaluar las expresiones de osteonectina y osteopontina en el desarrollo óseo después de la aplicación de dexametasona en defectos óseos tibiales. Las ratas se dividieron en grupos: defecto, defecto + injerto y defecto + injerto + grupos tratados con dexametasona. Se creó un defecto óseo tibial, y las ratas se mantuvieron inmóviles durante 28 días. El material aloplástico se colocó en el área del defecto en el segundo y tercer grupo. Se inyectaron 2,5 mg / kg de dexametasona y solución salina normal a grupos de defectos dos veces por semana durante 56 días. La inflamación y la congestión aumentaron en los grupos de defectos y defectos + injerto; En el grupo defecto + injerto + grupo tratado con dexametasona se observó un aumento en el número de osteoblastos y osteocitos, de matriz ósea densa y en la formación de nuevas trabéculas óseas. En el grupo defecto + grupo de injerto se observó la expresión de osteonectina en las áreas de injerto, osteoblastos, algunas células y fibras de tejido conectivo, mientras que en el grupo defecto + injerto + dexametasona se observó la expresión de osteopontina en osteoblastos y osteocitos y formación de nuevas trabéculas óseas . En conclusión la dexametasona puede conducir a la formación de nuevas trabéculas óseas en el material de injerto, lo que resulta en un aumento de la osteoconducción y un efecto osteoinductivo para la diferenciación del osteón.

Melatonin has a stimulatory effect on osteoblasts by upregulating COL-I and OPN expression/secretion / Melatonina tem efeito estimulador em osteoblastos, pela regulação positiva da expressão/secreção de COL-I e OPN

ABSTRACT Melatonin (MLT) is a potential signaling molecule in the homeostasis of bone metabolism and may be an important mediator of bone formation and stimulation. The aim of this in vitro study was to evaluate the effect of MLT on the viability, mRNA/protein expression and mineralization of pre-osteoblastic cells. The concentrations 5, 2.5, 1, 0.1 and 0.01 mM MLT were tested on pre-osteoblastic cells (MC3T3) compared to control (no MLT), evaluating proliferation and cell viability (C50), gene expression (RT-PCR) and secretion (ELISA) of COL-I and OPN at 24h, 48h and 72h, and the formation of mineral nodules (alizarin red and fast red) after 10 days of treatment. MLT at 5 and 2.5 mM proved to be cytotoxic (C50), so only 0.01, 0.1 and 1 mM were used for the subsequent analyses. OPN mRNA expression increased with MLT at 0.1 mM - 1 mM, which was followed by increased secretion of OPN both at 24h and 72h compared to the remaining groups (p <0.05). COL-I mRNA and COL-1 secretion followed the same pattern as OPN at 0.1 mM MLT at 72h of treatment (p <0.05). Regarding mineralization, all MLT doses (except 1mM) caused an increase (p <0.05) in the formation of mineral nodules compared to the control. Melatonin at 0.01mM - 1mM had a stimulatory effect on osteoblasts by upregulating COL-I and OPN expression/ secretion and mineralization, thereby fostering osteogenesis.

RESUMO A melatonina (MLT) é uma molécula potencial de sinalização na homeostase do metabolismo ósseo e pode ser um importante mediador da formação e estimulação óssea. O objetivo deste estudo in vitro foi avaliar o efeito da MLT na viabilidade, na expressão do mRNA da proteína e mineralização de células préosteoblásticas. As concentrações de MLT 5, 2,5, 1, 0,1 e 0,01 mM foram testadas em células pré-osteoblásticas da linhagem MC3T3 em comparação ao controle (sem MLT), avaliando a proliferação e a viabilidade celular (C50), expressão gênica (rtPCR) e secreção (Elisa) de Colágeno tipo 1 (COL-I) e osteopontina (OPN) às 24, 48 e 72 horas, além da formação de nódulos minerais por meio do teste vermelho de Alizarina fast red após 10 dias de tratamento. MLT a 5 e 2,5 mM provou ser tóxico (C50). Portanto, as concentrações de 0,01, 0,1 e 1 mM foram utilizadas para as análises subsequentes. A expressão do mRNA da OPN aumentou com MLT a 0,1 mM-1mM, seguida pela secreção aumentada de OPN às 24 e 72 horas em comparação aos demais grupos (p<0,05). O mRNA de COL-I e a secreção de COL-I seguiram o mesmo padrão do OPN a 0,1 mM de MLT em 72 horas de tratamento (p<0,05). Em relação à mineralização, todas as doses de MLT (exceto 1mM) causaram aumento (p<0,05) na formação de nódulos minerais em comparação ao controle. A MLT na concentração entre 0,01mM a 1 mM teve um efeito estimulador sobre os osteoblastos, ao regular positivamente a expressão e secreção de COL-I e OPN, além da mineralização, favorecendo a osteogênese.

Serum levels of bone sialoprotein, osteopontin, and β2-microglobulin in stage I of multiple myeloma

Context: The fluctuations of proteins in multiple myeloma (MM) are well-known markers for checking the status of the patients. Aims: The objective of this study was to examine three proteins that have an important role in disease progression. Subjects and Methods: The study was performed with two groups: 30 MM stage I patients' (14 females/16 males; aged 60.83 ± 12.38 years) as case group and 40 healthy individuals (18 females/22 males; aged 57.65 ± 6.43 years) as control group. Both groups have been matched in gender and age. Bone sialoprotein (BSP), osteopontin (OPN), and β2-microglobulin (β2M) were measured with an enzyme-linked immunosorbent assay. Results: Serum BSP levels of MM-I patients was significantly higher than that of healthy controls (29.24 ± 5.57 vs. 20.89 ± 3.67, P = 0.001). OPN levels of MM-I patients were significantly lower than that of healthy individuals (12.03 ± 3.45 vs. 19.35 ± 4.67, P = 0.001). β2M levels of patients and controls were similar (1.49 ± 0.67 vs. 1.29 ± 0.55, P = 0.193). Conclusions: The results suggested that myeloma cells may affect the production of BSP and OPN, which possibly contributes to osteoclastic bone resorption in MM-I patients. Their levels may be a useful biomarker for assessing bone destruction in MM-I patients and distinguishing MM-I from healthy individuals

Osteopontin expression and its relationship with prognostic biomarkers in canine mammary carcinomas / Expressão de osteopontina e sua relação com biomarcadores prognóstico nos carcinomas mamários caninos

Osteopontin is a glycophosphoprotein implicated in different physiologic and pathologic processes and is known to be involved in progression and metastasis of various cancers in humans, but this relation is still little explored in the veterinary. The aim was to evaluate the expression of osteopontin in canine mammary carcinomas and its relation with well-established canine mammary tumor biomarkers. For that, expression of OPN, EGFR, HER2, and c-Kit were evaluated along with Ki67 rate in 43 mammary carcinomas. Osteopontin was demonstrated to be expressed by neoplastic epithelial cells in all carcinomas as well as in stromal cells from the tumor microenvironment. Relation between high osteopontin expression and EGFR positivity (P<0.001) and HER2 overexpression (P=0.012) was demonstrated. In conclusion, high OPN expression seems to be related to poor prognosis and MAPK pathway activation, given the association with EGFR and HER2, members of the MAPK signaling pathway.(AU)

A osteopontina é uma glicofosfoproteina implicada em diferentes processos fisiológicos e patológicos, sendo conhecida por estar envolvida na progressão e metástase de vários cânceres nos humanos, no entanto, essa relação é ainda pouco explorada na veterinária. O objetivo deste trabalho foi avaliar a expressão da osteopontina nos carcinomas mamários caninos e sua relação com biomarcadores bem estabelecidos para esta neoplasia. Para isto, foi avaliada a expressão de OPN, EGRH, HER2 e c-Kit juntamente com a taxa de Ki67 em 43 carcinomas mamários. A osteopontina foi expressa pelas células epiteliais neoplásicas em todos os carcinomas, assim como, nas células estromais do microambiente tumoral. Foi demonstrada uma relação entre uma alta expressão de osteopontina e positividade para EGFR (P<0.001) e superexpressão de HER2 (P=0.012). Em conclusão, alta expressão de OPN parece estar relacionada com mau prognóstico e ativação da via MAPK, devido a sua associação com EGRF e HER2, os quais são membros desta via de sinalização.(AU)

Is there a clinical pathological correlation of colorectal adenocarcinoma with the immunohistochemical expression of opn and abcb5? / Há correlação clínicopatológica do adenocarcinoma colorretal com a expressão imunoistoquímica do opn e abcb5?

ABSTRACT Background: Studies with biomarkers in TMA (tissue microarray) have been showing important results regarding its expression in colon cancer. Aim: Correlate the expression profile of the OPN and ABCB5 biomarkers with the epidemiological and clinicopathological characteristics of the patients, the impact on the progression of the disease and the death. Method: A total of 122 CRC patients who underwent surgical resection, immunomarking and their relationship with progression and death events were evaluated. Result: The average age was 61.9 (±13.4) years. The cases were distributed in 42 (35.9%) in the ascending/transverse colon, 31 (26.5%) in the sigmoid, 27 in the rectum (23.1%), 17 (14.5%) in the descending colon. Most patients had advanced disease (stages III and IV) in 74 cases (60.9%). There was a predominance of moderately differentiated tumors in 101 samples (82.8%); despite this, the poorly differentiated subtype proved to be an independent risk factor for death in 70%. Metastasis to the liver proved to be an independent risk factor for death in 75% (18/24), as well as patients with primary rectal tumors in 81.5% (22/27). Conclusion: The immunohistochemical expression of the OPN and ABCB5 markers was not associated with epidemiological and clinicopathological characteristics. Regarding the progression of disease and death, it was not possible to observe a correspondence relationship with the evaluated markers.

RESUMO Racional: Estudos com biomarcadores com TMA (tissue microarray) vêm demostrando resultados importantes em relação à expressão de biomarcadores em câncer de cólon. Objetivo: Correlacionar o perfil de expressão dos biomarcadores OPN e ABCB5 com as características epidemiológicas e clinicopatológicas dos pacientes, o impacto na progressão de doença e no evento óbito. Método: Foram avaliados 122 pacientes de CCR submetidos à ressecção cirúrgica e à imunomarcação e relação com os eventos progressão e óbito. Resultado: A média de idade encontrada foi de 61,9 (±13,4) anos. Os casos distribuíram-se em 42 (35,9%) no cólon ascendente/transverso, 31 (26,5%) no sigmoide, 27 no reto (23,1%), 17 (14,5%) no cólon descendente. A maioria dos pacientes apresentou doença avançada (estadio III e IV) em 74 casos (60,9%). Houve predomínio de tumor moderadamente diferenciado em 101 amostras (82,8%); apesar disso, o subtipo pouco diferenciado mostrou-se como fator de risco independente para óbito em 70% dos casos. Metástase para o fígado mostrou-se fator de risco independente para óbito em 75% dos casos (18/24), assim como pacientes com tumores primários de reto em 81,5% (22/27). Conclusão: A expressão imunoistoquímica dos marcadores OPN e ABCB5 não apresentou associação com as características epidemiológicas e clinicopatológicas. Em relação à progressão de doença e evento óbito, não se conseguiu observar relação de correspondência com os marcadores avaliados.

The role of autophagy in ossification of the human ligamentum flavum / 中国组织工程研究

BACKGROUND: Pathological mechanism of ossification of the ligamentum flavum is unclear. There is no effective drug or non-surgical treatment in clinical practice. Current studies have found that osteopontin and autophagy play an important role in the process of osteogenesis, but their role in ossification of the ligamentum flavum has not been elucidated. OBJECTIVE: To seek for the potential target of drug therapy by exploring the mechanism of ossification of the ligamentum flavum. METHODS: (1) Surgical specimens of the ligamentum flavum were taken from patients with ossification of the ligamentum flavum, thoracic vertebrae or simple lumbar disc herniation undergoing posterior total laminectomy and decompression. These specimens were divided into two groups: An ossification group and a non-ossification group. Eight specimens from each group were collected. Osteopontin, osteocalcin and autophagy indexes Beclin-1, LC3 and P62 were stained by immunohistochemistry. (2) The ligamentum flavum cells were isolated and cultured by adherence method. The third generation cells were treated with osteopontin at different concentrations for different time to construct an in vitro model of ligamentum flavum ossification. (3) Autophagy inhibitor 3-methyladenine with different concentrations was used to intervene with non-ossified ligamentum flavum cells, followed by induction with 100 μg/L osteopontin. Western blot assay was used to detect the expression of alkaline phosphatase, osteocalcin. (4) Non-ossified ligamentum flavum cells were induced with 100 μg/L osteopontin, and the induction was terminated at 0, 15, 30, 60, and 120 minutes, respectively. The phosphorylation of ERK1/2, JNK and P38, which are important molecules in the MAPK signaling pathway, was detected by western blot. (5) Finally, after inhibition by ERK1/2 phosphorylation blocker U0126, the expression of alkaline phosphatase and osteocalcin was detected by western blot after induction with 100 μg/L steopontin. RESULTS AND CONCLUSION: (1) Immunohistochemical staining of osteopontin and osteocalcin in ossified and non-ossified ligamentum flavum was positive. In the ossified ligamentum flavum, Beclin-1 was positive, but LC3 and P62 were not. Beclin-1, LC3 and P62 were all positive in the non-ossified ligamentum flavum. (2) The expression of alkaline phosphatase and osteocalcin in the ossified ligamentum flavum cells was higher than that in the non-ossified ligamentum flavum cells. Osteopontin could induce ossification of the ligamentum flavum in a concentration- and time-dependent manner. (3) The degree of ossification was negatively correlated with the degree of autophagy, that is, the more obvious autophagy was, the weaker ossification was. (4) Osteopontin could phosphorylate the MAPK signaling pathway in a time-dependent manner. After inhibiting the phosphorylation of MAPK, osteopontin could still induce the ossification of ligamentum flavum cells. To conclude, in the process of ligamentum flavum ossification, the upstream and downstream relationships of ERK1/2, osteopontin, alkaline phosphatase and osteocalcin molecules in signaling pathway are ERK1/2→osteopontin→osteocalcin/alkaline phosphatase.

Beta-ecdysterone promotes in vitro proliferation and osteogenic differentiation of MC3T3-E1 cells / 中国组织工程研究

BACKGROUND: β-ecdysterone as a “phytoestrogen” has the ability to stimulate protein synthesis, promote carbohydrate and lipid metabolism, relieve hyperglycemia and hyperlipidemia, protect endothelial cells from apoptosis and induce their proliferation. Some scholars have reported that it also plays an important role in the treatment of osteoporosis, fractures and other bone inflammatory diseases. OBJECTIVE: To observe the effect of β-ecdystrone on the proliferation of mouse pre-osteoblasts(MC3T3-E1 cells) in vitro, and to explore whether β-ecdysterone can induce osteogenic differentiation of MC3T3-E1 at a safe dose. METHODS: The fourth generation MC3T3-E1 cells were cultured in the osteogenic induction medium for 7, 10, 14, 21, and 28 days. The osteogenic differentiation proteins(alkaline phosphatase, type I collagen, osteopontin, and calcified nodules) were detected at different time points, to identify whether the cells have the ability of osteogenic differentiation. MC3T3-E1 cells were then seeded into the induction medium containing different final concentrations of β-ecdysterone(0.01, 0.1, 1, 10, 100 µmol/L). The proliferation activity of the cells was detected by cell counting kit-8 method at days 1, 2, 3, 4, 5, 6, and 7 after induction. The control group(general induction medium group) and the experimental group(general induction medium + β-ecdysterone) were cultured under the same conditions, and the expression levels of osteogenic marker proteins in each group of cells at different time periods were determined. RESULTS AND CONCLUSION: In the MC3T3-E1 cells stimulated by the osteogenic induction medium, alkaline phosphatase staining and type I collagen florescence staining showed higher expression at day 10 of induction, and this was also confirmed by detection of alkaline phosphatase activity(P 0.05). The expression of alkaline phosphatase and type I collagen was higher in the experimental group than in the control group at day 10 of induction. The expression of osteopontin and osteocalcin in the cells was higher at day 14 of induction, and there was no significant difference in the calcified nodule staining between the two groups at day 28 of induction. These findings indicate that β-ecdysterone can promote the proliferation of MC3T3-E1 cells in vitro and induce MC3T3-E1 cells to differentiate into osteoblasts at a safe dose.

Mechanism of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma Extracts in Delaying High Glucose-induced Vascular Calcification in Mice / 中国实验方剂学杂志

Objective::To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on vascular calcification induced by high glucose in mice by observing the expression of osteopontin (OPN) and smooth muscle 22α (SM22α) as well as vascular calcium deposition in the common carotid artery and thoracic aorta of mice. Method::Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin(STZ), and fed on a high-fat diet for 7 months. Then, the mice were randomly divided into model group, low-dose and high-dose Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). Each group was intragastrically administered once a day for 9 weeks. The changes in blood glucose were measured. Seven days before the end of the administration, a group of 4-week old male C57BL/6 mice were purchased and fed normally for one week as a youth group. At the end of the administration, the common carotid artery and thoracic aorta tissues of the mice were collected. Von Kossa staining was used to determine the degree of calcium deposition in the common carotid artery and thoracic aorta. The expression levels of OPN and SM22α protein in the common carotid artery and thoracic aorta were detected by immunohistochemistry. The expression of OPN and SM22α protein in the common carotid artery of mice was determined by Western blot. Result::As compared with the young group, the blood glucose of the normal control group was slightly increased without statistical difference, the common carotid artery and thoracic aorta were uniformly stained, and no black granular precipitate was observed. As compared with the normal control group, the blood glucose of the model group was increased (P<0.01), with a large amount of brown-black particles deposited in the intimal elastic fibers, showing obvious calcium salt deposition. As compared with the model group, blood glucose was significantly decreased in each administration group (P<0.05, P<0.01), and the degree of vascular calcium salt deposition was significantly reduced. There were no significant changes in expression levels of OPN protein and SM22α protein in the common carotid artery and thoracic aorta between the youth group and normal control group. As compared with the normal control group, the expression of intimal OPN protein in the common carotid artery and thoracic aorta of the model group was positive, SM22α protein expression was weakly positive, and the gray value of OPN protein expression in the common carotid artery was significantly increased (P<0.01), while the gray value of SM22α protein was decreased significantly (P<0.01). As compared with the model group, the expression levels of intimal OPN protein and SM22α protein in the common carotid artery and thoracic aorta of each administration group were significantly improved, and the gray value of OPN protein expression in the common carotid artery was reduced (P<0.05, P<0.01), while SM22α protein expression was significantly increased (P<0.01). Conclusion::High glucose can induce calcification of common carotid artery and thoracic aorta in mice and accelerate vascular aging. This formation process may be related to the expression of OPN and SM22α. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can reduce vascular calcification and delay vascular aging by regulating the expression of OPN and SM22α.

Association between serum levels of osteopontin and systolic pulmonary artery pressure among healthy adults post acute high altitude exposure / 中华心血管病杂志

Objective: To explore the association between serum levels of osteopontin (OPN) and systolic pulmonary artery pressure (sPAP) in healthy men following acute high altitude exposure. Methods: According to the inclusion and exclusion criteria, this observational study included 94 male subjects (aged from 18 to 30 years, dwelling in lowland<500 m) who ascended to Litang (4 100 m) from Chongqing (400 m) by bus with a stair-like journey within 7 days in June 2013. Data including basic information, OPN, superoxide dismutase (SOD), and malondialdehyde (MDA) and echocardiographic derived sPAP were collected within 48 hours before ascent and within 2-7 hours after arrival. Accordingly, subjects were divided into 3 groups based on the tertiles of sPAP after acute high altitude exposure: low sPAP group (26.8-32.3 mmHg (1 mmHg=0.133 kPa)) (n=31), middle sPAP group (32.4-37.4 mmHg) (n=32) and high sPAP group (37.5-55.6 mmHg) (n=31). Associations of serum OPN and SOD levels with sPAP were analysed by univariate and multivariate linear regression analysis. Results: After acute high altitude exposure, the levels of sPAP were significantly increased (P<0.001). There were no differences in age, height, weight, body mass index, percent of Han nationality and smoking among 3 subgroups. However, following acute high altitude exposure, the levels of heart rate, systolic and diastolic blood pressure elevated (all P<0.05), whereas the levels of oxygen saturation were reduced in the total subjects and all subgroups (all P<0.05). Moreover, systolic blood pressure of subjects in the high sPAP group was higher than that in low and middle sPAP groups (both P<0.05), and diastolic blood pressure of subjects in high sPAP group was higher than that in low sPAP group (P<0.05). The serum levels of OPN were increased in total cohort(27.9 (22.5,34.0) μg/L vs. 25.6 (18.4, 33.1) μg/L, P<0.05)， and high sPAP group (P<0.05), whereas no differences were found in serum SOD and MDA levels among groups. Furthermore, the serum level of OPN in high sPAP group was higher than that in low sPAP group at high altitude (P<0.05), and there was a trend for decline in SOD level with increasing sPAP (P>0.05). Results from univariable linear regression analysis showed that the serum levels of OPN (r=0.32, P=0.002) and SOD (r=-0.22，P=0.032) were linearly correlated with sPAP in total cohort after high altitude exposure. Multivariate regression analysis showed that the serum levels of OPN(β=0.310，P=0.002) and SOD (β=-0.199，P=0.043) were independently associated with the levels of sPAP at high altitude. Conclusion: After acute high altitude exposure, the serum level of OPN is positively associated with sPAP, suggesting that OPN may be a novel bio-marker for predicting the increase of pulmonary pressure in response to acute high altitude exposure.

Immunohistochemical analysis of osteopontin expression in oral squamous cell carcinoma

Background: Oral squamous cell carcinoma (OSCC) is the most common malignant neoplasm of the oral cavity. Osteopontin (OPN) has been proved as a biomarker in varying malignant tumors. Only limited studies detail the role of OPN in OSCC. Aims: This study aims to demonstrate the expression of OPN in OSCC and to correlate the expression of OPN with the histologic grades of OSCC. Settings and Design: This is a retrospective immunohistochemical study in Dravidian population (linguistically Malayalam). Materials and Methods: Thirty diagnosed cases of OSCC were subjected to immunohistochemistry using OPN antibody for detection of OPN expression. Ten normal oral mucosal specimens were also stained as controls. Statistical Analysis Used: Chi-square test and ANOVA followed by Bonferroni test. Results: OPN expression was significantly higher in OSCC patients than in controls. In normal oral mucosal specimens, none of them showed OPN immunoreactivity. A significant difference was observed between total scores and intensities of normal and varying grades of OSCC. A significant difference was also observed between the percentage of positive cells for OPN expression of normal and varying grades of OSCC. However, no significant difference was observed between the percentage of positive cells for OPN expression of well-, moderate-, poorly-differentiated carcinomas. Correlation of OPN expression with lymph node status, site, and sex was found to be statistically insignificant. Conclusion: Insights gained from this study may lead to research targeted at the treatment of OSCC.

A comparative study of osteopontin and MMP-2 protein expression in peripheral and central giant cell granuloma of the jaw / Estudo comparativo da expressão das proteínas osteopontina e MMP-2 no granuloma central e periférico de células gigantes de mandíbula

Abstract Introduction: Oral peripheral and central giant cell granulomas are lesions with little-known etiology and pathogenesis. Objective: The aim of this study was to compare matrix metalloproteinases-2 and osteopontin protein expression in the multinucleated giant cells and mononuclear cells of the peripheral and central giant cell granuloma lesions. Methods: In this retrospective study, the presence of matrix metalloproteinases-2 and osteopontin in 37 cases of central giant cell granuloma and 37 cases of peripheral giant cell granuloma paraffin blocks were assessed by streptavidin-biotin immunohistochemistry. Independent sample t-test, Chi-square, Mann-Whitney tests and Spearman's rank correlation coefficient were used. Results: The osteopontin was expressed in both multinucleated giant cells and mononuclear cells in all cases of peripheral and central giant cells granulomas. However, the matrix metalloproteinases-2 expression was positive in 86.5% of giant cells and it was positive in all of mononuclear cells in peripheral giant cells granuloma. In central giant cells granulomas, 91.8% of giant cells and all mononuclear cells were positive for matrix metalloproteinases-2 marker. Percentage and Intensity of staining were significantly higher in central than peripheral giant cells lesions, for both markers (p ˂ 0.05). Conclusion: This study showed that the expression of osteopontin in giant cells supports the theory of osteolcastic nature of these cells. Also, the presence of osteopontin and matrix metalloproteinases-2 in mononuclear cells may indicate the monocyte-macrophage origin of these cells, as the differentiation of the precursors of the mononuclear stromal monocyte/macrophage to osteoclasts is possibly affected by the expression of osteolytic factors. Also, may be differences in biological behaviors of these lesions are associated with the level of osteopontin and matrix metalloproteinases-2 expression.

Resumo Introdução: Os granulomas periféricos e centrais de células gigantes são lesões com etiologia e patogênese pouco conhecidas. Objetivo: Comparar a expressão das proteínas metaloproteinases da matriz-2 e osteopontina nas células gigantes multinucleadas e células mononucleares no granuloma periférico e central de células gigantes. Método: Neste estudo retrospectivo, a presença de metaloproteinases da matriz-2 e osteopontina em 37 casos de granuloma central de células gigantes e 37 casos de granuloma periférico de células gigantes em blocos de parafina foi avaliada por imuno-histoquímica pela estreptavidina-biotina. Foram usados teste t para amostra independente, teste de qui-quadrado, Mann-Whitney e coeficiente de correlação de Spearman. Resultados: A osteopontina foi expressa em células gigantes multinucleadas e células mononucleares em todos os casos de granuloma periférico de células gigantes e granuloma central de células gigantes. No entanto, a expressão de metaloproteinases da matriz-2 foi positiva em 86,5% de células gigantes e foi positiva em todas as células mononucleares em granuloma periférico de células gigantes. Em granuloma central de células gigantes, 91,8% das células gigantes e todas as células mononucleares foram positivas para o marcador metaloproteinases da matriz-2. A porcentagem e intensidade de coloração em granuloma central de células gigantes foram significantemente maiores do que em granuloma periférico de células gigantes para ambos os marcadores (p ˂ 0,05). Conclusão: Este estudo mostrou que a expressão de osteopontina em células gigantes apoia a teoria da natureza osteoclástica dessas células. Além disso, a presença de osteopontina e metaloproteinases da matriz-2 em células mononucleares pode indicar a origem dos monócitos-macrófagos dessas células, uma vez que a diferenciação dos precursores do monócito/macrófago estromal mononuclear em osteoclastos é possivelmente afetada pela expressão de fatores osteolíticos. Além disso, as diferenças nos comportamentos biológicos dessas lesões estão associadas ao nível de expressão de osteopontina e metaloproteinases da matriz-2.

Expression of Osteopontin mRNA During Periodontal Healing Following Scaling and Root Planing

Objective: To analyze osteopontin mRNA expression levels in subjects with periodontitis prior to (baseline) and 7, 14, and 28 days following scaling and root planing (SRP). Material and Methods: Gingival crevicular fluid was collected as clinical samples from four subjects with periodontitis (pocket depth, 4-5 mm) aged 35-54 years old as well as from three healthy subjects (controls). The osteopontin mRNA expression levels were measured by quantitative real-time polymerase chain reaction. Spearman's rank correlation between osteopontin levels in gingival crevicular fluid and the modified gingival index (MGI) was also performed. Results: The Wilcoxon signed-rank test showed no significant difference in osteopontin mRNA expression levels between baseline and 28 days following SRP (p=0.068). The Friedman test showed no significant difference in osteopontin mRNA expression levels between baseline and following SRP (7, 14, or 28 days) (p>0.05). Spearman's rank correlation showed no significant correlation between osteopontin mRNA expression levels and MGI (r=0.087; p=0.749). Conclusion: Following SRP of periodontal tissue, there was a decreasing trend in osteopontin mRNA expression; however, this finding was not statistically significant. Nevertheless, osteopontin can be used as a biomarker to monitor the healing process; however, further studies are required to clarify our results.

Association between neuroimmunological indicators and autism spectrum disorders / 中国学校卫生

Objective@#To explore the relationship among osteopontin(OPN), Interleukin-17A(IL-17A), anti-MBP auto-antibody and autism spectrum disorder(ASD), and to provide the theoretical basis for the etiology and pathogenesis of ASD.@*Methods@#Forty autistic children and forty matched healthy children were enrolled in this case-control study. The levels of OPN, IL-17A, anti-MBP autoantibody in serum were measured by enzyme-linked immunosorbent assay (ELISA). The associations between those metabolic levels and the severity and intelligence of ASD children were performed by Pearson or Spearman correlation.@*Results@#Children with ASD had higher serum levels of OPN, IL-17A [(296.89±162.95),0.93] pg/mL compared to healthy control[(217.98±113.39), 0.62] pg/mL(P<0.05). Serum OPN, IL-17A, and anti-MBP auto-antibody levels in ASD group were not correlated with the scores of ABC, CARS, and PPVT(P>0.05). However, anti-MBP auto-antibodies level in children with ASD were positively correlated with OPN and IL-17A levels, respectively(r=0.35, 0.34, P<0.05).@*Conclusion@#It was obvious that the ASD children were found with neuroimmunologic abnormality, and the underlying mechanism needs to be further explored.

The influence of Aloe vera with mesenchymal stem cells from dental pulp on bone regeneration: characterization and treatment of non-critical defects of the tibia in rats

Abstract Objective This study aimed to evaluate the inflammatory effect and bone formation in sterile surgical failures after implantation of a collagen sponge with mesenchymal stem cells from human dental pulp (hDPSCs) and Aloe vera. Material and Methods Rattus norvegicus (n=75) were divided into five experimental groups according to treatment: G1) control (blood clot); G2) Hemospon®; G3) Hemospon® in a culture medium enriched with 8% Aloe vera; G4) Hemospon® in a culture medium containing hDPSCs and G5) Hemospon® in a culture medium enriched with 8% Aloe vera and hDPSCs. On days 7, 15 and 30, the animals were euthanized, and the tibia was dissected for histological, immunohistochemistry and immunofluorescence analyses. The results were analyzed using nonparametric Kruskal-Wallis test and Dunn's post-test. Results On days 7 and 15, the groups with Aloe vera had less average acute inflammatory infiltrate compared to the control group and the group with Hemospon® (p<0.05). No statistically significant difference was found between the groups regarding bone formation at the three experimental points in time. Osteopontin expression corroborated the intensity of bone formation. Fluorescence microscopy revealed positive labeling with Q-Tracker® in hDPSCs before transplantation and tissue repair. Conclusion The results suggest that the combination of Hemospon®, Aloe vera and hDPSCs is a form of clinical treatment for the repair of non-critical bone defects that reduces the inflammatory cascade's effects.