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1.
São Paulo; s.n; 20210523. 75 p.
Thesis in Portuguese | LILACS, BBO | ID: biblio-1369753

ABSTRACT

A cartilagem de Meckel é uma estrutura transitória embrionária presente durante os estágios iniciais da formação da mandíbula, localizada em toda sua extensão e dividida em três porções, anterior, intermediária e posterior. O enfoque deste trabalho foi direcionado à elucidação do destino final da porção intermediária por meio de um estudo temporal sequenciado. Por isso, foi investigado a presença de células de reabsorção e a presença de fibras colágenas, bem como da proteína óssea osteopontina (OPN) na cartilagem de Meckel na região do germe do 1º molar inferior e no seu entorno. Foram utilizados fetos de ratos Wistar em períodos gestacionais pré-estabelecidos, G18 a G21 (grupos de dias intrauterinos), bem como P0 e P1 (recém-nascidos) para remoção das cabeças. Em sequência, os espécimes foram fixados em solução de formaldeído 4% + glutaraldeído 0,1% com tampão fosfato 0,1M, descalcificados em EDTA 4,13%, desidratados em concentrações crescentes de etanol e incluídos em parafina. As amostras foram coradas em hematoxilina e eosina (HE) e tricrômico de Mallory para análise histológica. Adicionalmente, os grupos G19 a P0 foram submetidos à reação histoquímica de TRAP para determinação da presença de células clásticas. Além disso, os grupos G21 e P0 (dia do nascimento) passaram por reações de imunomarcação para análise da expressão de OPN. Foi observado a degeneração gradual da cartilagem com a observação de mudanças estruturais, a justaposição de células clásticas na superfície da cartilagem por reação histoquímica TRAP a partir do G21, o aparecimento de colágeno tipo I nas fases terminais da degeneração, assim como a marcação positiva para a osteopontina na superfície de G21 e em todo o remanescente da cartilagem de Meckel no grupo P0. O estudo apontou um processo de degeneração da cartilagem com evidências de formação de matriz mineralizada de natureza óssea, a qual foi reabsorvida por células clásticas, sugerindo a ossificação da porção intermediária da cartilagem de Meckel.


Subject(s)
Osteoclasts , Cartilage , Osteopontin , Mandible
2.
Braz. dent. sci ; 24(4, suppl 1): 1-8, 2021. tab, ilus
Article in English | LILACS, BBO | ID: biblio-1352634

ABSTRACT

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


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


Subject(s)
Humans , Periodontitis , Bone Resorption , Diabetes Mellitus, Type 2 , Osteopontin
3.
Chinese Medical Journal ; (24): 1093-1100, 2021.
Article in English | WPRIM | ID: wpr-878150

ABSTRACT

BACKGROUND@#Although osteopontin (OPN) is expressed in the liver and pigment gallstones of patients with hepatolithiasis, its role in pigment gallstone formation remains unclear. This study aimed to explore the function of OPN in pigment gallstone formation.@*METHODS@#Rats were fed a chow diet (CD) or lithogenic diet (LD) for 10 consecutive weeks; blocking tests were then performed using an OPN antibody (OPN-Ab). Incidence of gallstones and levels of several bile components, OPN, tumor necrosis factor alpha (TNF-α), and cholesterol 7 alpha-hydroxylase (CYP7A1) were analyzed. To determine TNF-α expression in hepatic macrophages and both CYP7A1 and bile acid (BA) expression in liver cells, recombinant rat OPN and recombinant rat TNF-α were used to treat rat hepatic macrophages and rat liver cells, respectively. Chi-square or Fisher exact tests were used to analyze qualitative data, Student t-test or one-way analysis of variance were used to analyze qualitative data.@*RESULTS@#Incidence of gallstones was higher in LD-fed rats than in CD-fed rats (80% vs. 10%, P < 0.05). BA content significantly decreased in bile (t = -36.08, P < 0.01) and liver tissue (t = -16.16, P < 0.01) of LD-fed rats. Both hepatic OPN protein expression (t = 9.78, P < 0.01) and TNF-α level (t = 8.83, P < 0.01) distinctly increased in the LD group; what's more, CYP7A1 mRNA and protein levels (t = -12.35, P < 0.01) were markedly down-regulated in the LD group. Following OPN-Ab pretreatment, gallstone formation decreased (85% vs. 25%, χ2 = 14.55, P < 0.01), liver TNF-α expression (F = 20.36, P < 0.01) was down-regulated in the LD group, and CYP7A1 expression (F = 17.51, P < 0.01) was up-regulated. Through CD44 and integrin receptors, OPN promoted TNF-α production in macrophage (F = 1041, P < 0.01), which suppressed CYP7A1 expression (F = 48.08, P < 0.01) and reduced liver BA synthesis (F = 119.4, P < 0.01).@*CONCLUSIONS@#We provide novel evidence of OPN involvement in pigmented gallstone pathogenesis in rats.


Subject(s)
Animals , Diet/adverse effects , Gallstones/etiology , Lithiasis , Liver , Liver Diseases , Osteopontin/genetics , Rats
4.
Int. j. morphol ; 38(5): 1398-1404, oct. 2020. tab, graf
Article in English | LILACS | ID: biblio-1134455

ABSTRACT

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


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


Subject(s)
Animals , Male , Rats , Tibia/surgery , Tibia/drug effects , Dexamethasone/administration & dosage , Bone Transplantation , Tibia/pathology , Bone Regeneration , Immunohistochemistry , Osteonectin/physiology , Bone Remodeling , Rats, Wistar , Disease Models, Animal , Osteopontin/physiology
5.
Pesqui. vet. bras ; 40(3): 210-219, Mar. 2020. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1135610

ABSTRACT

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


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


Subject(s)
Animals , Female , Dogs , Carcinoma , Biomarkers , Mammary Neoplasms, Animal , Dog Diseases , Osteopontin , Immunohistochemistry
6.
Braz. j. otorhinolaryngol. (Impr.) ; 85(2): 150-156, Mar.-Apr. 2019. tab, graf
Article in English | LILACS | ID: biblio-1001542

ABSTRACT

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


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


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Young Adult , Granuloma, Giant Cell/pathology , Jaw Diseases/pathology , Matrix Metalloproteinase 2/analysis , Osteopontin/analysis , Reference Values , Severity of Illness Index , Immunohistochemistry , Sex Factors , Retrospective Studies , Age Factors , Statistics, Nonparametric , Streptavidin
7.
Pesqui. bras. odontopediatria clín. integr ; 19(1): 4991, 01 Fevereiro 2019. tab, graf
Article in English | LILACS, BBO | ID: biblio-998272

ABSTRACT

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


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Periodontitis , RNA, Messenger , Root Planing/methods , Osteopontin , Case-Control Studies , Statistics, Nonparametric , Indonesia
8.
Frontiers of Medicine ; (4): 250-258, 2019.
Article in English | WPRIM | ID: wpr-771315

ABSTRACT

Biomarkers for hepatocellular carcinoma (HCC) following curative resection are not currently sufficient for prognostic indication of overall survival (OS) and disease-free survival (DFS). The aim of this study was to investigate the prognostic performance of osteopontin (OPN), matrix metalloproteinase 7 (MMP7), and pregnancy specific glycoprotein 9 (PSG9) in patients with HCC. A total of 179 prospective patients with HCC provided plasma before hepatectomy. Plasma OPN, MMP7, and PSG9 levels were determined by enzyme-linked immunosorbent assay. Correlations between plasma levels, clinical parameters, and outcomes (OS and DFS) were overall analyzed. High OPN ( ⩾ 149.97 ng/mL), MMP7 ( ⩾ 2.28 ng/mL), and PSG9 ( ⩾ 45.59 ng/mL) were prognostic indicators of reduced OS (P < 0.001, P < 0.001, and P = 0.007, respectively). Plasma PSG9 protein level was an independent factor in predicting OS (P = 0.008) and DFS (P = 0.038). Plasma OPN + MMP7 + PSG9 elevation in combination was a prognostic factor for OS (P < 0.001). OPN was demonstrated to be a risk factorassociated OS in stage I patients with HCC and patients with low α-fetoprotein levels ( < 20 ng/mL). These findings suggested that OPN, MMP7, PSG9 and their combined panels may be useful for aiding in tumor recurrence and mortality risk prediction of patients with HCC, particularly in the early stage of HCC carcinogenesis.


Subject(s)
Adult , Aged , Biomarkers, Tumor , Blood , Carcinoma, Hepatocellular , Blood , Mortality , Enzyme-Linked Immunosorbent Assay , Female , Hepatectomy , Humans , Liver Neoplasms , Blood , Mortality , Male , Matrix Metalloproteinase 7 , Blood , Middle Aged , Osteopontin , Blood , Pregnancy-Specific beta 1-Glycoproteins , Prognosis , Prospective Studies , Risk Assessment , Survival Analysis
9.
Acta cir. bras ; 34(7): e201900704, 2019. tab, graf
Article in English | LILACS | ID: biblio-1038112

ABSTRACT

Abstract Purpose: The effects of resveratrol administration on calvarial bone defects with alloplastic graft material was investigated for osteoinductive reaction and bone development in rats. Methods: Healthy male rats were randomly divided into 3 groups consisting of 10 rats. Groups were as follows: control (defect) group, defect + graft group, and defect + graft + resveratrol group. A calvarial bone defect was created in all groups, alloplastic bone grafts were applied to the defect in the 2nd and 3rd group, resveratrol (5 mg/kg/day) was added to the drinking water of the animals following graft application for 28 days in the 3rd group. Results: Increase in osteoclasts and necrotic changes were observed histopathologically in the control group. In the 2nd group, reduction of inflammation, congestion of blood vessels, increased osteblastic activity, osteoinductive effect, progression of osteocyte development and increased collagen fibers in connective tissue were observed. In the 3rd group, osteoblasts seemed to secrete bone matrix and accelerate osteoinductive effect with increased osteopregenitor activity and positive osteopontin and osteonectin expressions. Conclusion: Resveratrol treatment was thought to be an alternative and supportive drug for implant application by inducing new bone formation in the calvaral defect region as a result of short-term treatment.


Subject(s)
Animals , Male , Rats , Skull/surgery , Bone Regeneration/drug effects , Bone Transplantation/methods , Bone Substitutes/administration & dosage , Resveratrol/administration & dosage , Osteoblasts/drug effects , Osteogenesis/drug effects , Skull/drug effects , Drug Administration Schedule , Osteonectin/administration & dosage , Osseointegration/drug effects , Bone Substitutes/therapeutic use , Disease Models, Animal , Osteopontin/administration & dosage
10.
J. appl. oral sci ; 27: e20180103, 2019. graf
Article in English | LILACS, BBO | ID: biblio-1002400

ABSTRACT

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


Subject(s)
Humans , Animals , Male , Rats , Bone Regeneration/drug effects , Bone Regeneration/physiology , Plant Extracts/pharmacology , Dental Pulp/cytology , Mesenchymal Stem Cell Transplantation/methods , Aloe/chemistry , Osteogenesis/drug effects , Osteogenesis/physiology , Tibia/drug effects , Tibia/physiology , Tibia/pathology , Time Factors , Immunohistochemistry , Hemostatics/pharmacology , Cell Survival/drug effects , Cell Survival/physiology , Cells, Cultured , Reproducibility of Results , Collagen/pharmacology , Treatment Outcome , Osteopontin/analysis , Flow Cytometry , Microscopy, Fluorescence
11.
J. appl. oral sci ; 27: e20180014, 2019. graf
Article in English | LILACS, BBO | ID: biblio-975888

ABSTRACT

Abstract Stanozolol (ST) is a synthetic androgen with high anabolic potential. Although it is known that androgens play a positive role in bone metabolism, ST action on bone cells has not been sufficiently tested to support its clinical use for bone augmentation procedures. Objective: This study aimed to assess the effects of ST on osteogenic activity and gene expression in SaOS-2 cells. Material and Methods: SaOS-2 deposition of mineralizing matrix in response to increasing doses of ST (0-1000 nM) was evaluated through Alizarin Red S and Calcein Green staining techniques at 6, 12 and 24 days. Gene expression of runt-related transcription factor 2 (RUNX2), vitamin D receptor (VDR), osteopontin (SPP1) and osteonectin (ON) was analyzed by RT-PCR. Results: ST significantly influenced SaOS-2 osteogenic activity: stainings showed the presence of rounded calcified nodules, which increased both in number and in size over time and depending on ST dose. RT-PCR highlighted ST modulation of genes related to osteogenic differentiation. Conclusions: This study provided encouraging results, showing ST promoted the osteogenic commitment of SaOS-2 cells. Further studies are required to validate these data in primary osteoblasts and to investigate ST molecular pathway of action.


Subject(s)
Humans , Osteogenesis/drug effects , Stanozolol/pharmacology , Gene Expression/drug effects , Anabolic Agents/pharmacology , Osteoblasts/drug effects , Time Factors , Calcification, Physiologic/drug effects , Linear Models , Osteonectin/analysis , Osteonectin/drug effects , Reproducibility of Results , Analysis of Variance , Receptors, Calcitriol/analysis , Receptors, Calcitriol/drug effects , Cell Line, Tumor/drug effects , Core Binding Factor Alpha 1 Subunit/analysis , Core Binding Factor Alpha 1 Subunit/drug effects , Osteopontin/analysis , Osteopontin/drug effects , Real-Time Polymerase Chain Reaction
12.
Int. j. morphol ; 36(1): 206-211, Mar. 2018. tab, graf
Article in English | LILACS | ID: biblio-893212

ABSTRACT

SUMMARY: The purpose of this study was to investigate effects of diabetes mellitus (DM) on the alveolar bone with histopathological and immunohistochemical methods. Wistar rats were divided into two groups, control and diabetes group. Control group was fed standard rat chow and drinking water for 8 weeks. Single dose (Streptozotocin) STZ (55 mg/kg), was dissolved in sodium citrate buffer and introduced intraperitoneal injection. Diabetes group and control group were compared in terms of glucose values. The blood glucose concentration in diabetic rats was significantly high (p <0.05). In diabetes group; periodontal membrane and the dilation of blood vessels, hemorrhage has also been a significant increase in inflammatory cells. In the diabetes group, osteonectin showed positive expression in periodontal membrane and showed negative expression in osteocytes of alveolar bone. Osteopontin expression in fibroblast cells and periodontal membrane collagen fibrils was positive, alveolar cells, osteocytes and bone matrix bone was found positive. Diabetes results showed that there formed periodontitis; due to the increase in inflammation inhibiting bone formation delaying the development of early bone cells.


RESUMEN: El objetivo de este estudio fue investigar los efectos de la diabetes mellitus (DM) sobre el hueso alveolar con métodos histopatológicos e inmunohistoquímicos. Las ratas Wistar se dividieron en dos grupos, grupo control y grupo de diabetes. El grupo control fue alimentado con comida estándar y agua potable durante 8 semanas. La dosis única Streptozotocina (STZ) (55 mg/ kg), se disolvió en tampón de citrato de sodio y se introdujo mediante inyección intraperitoneal. El grupo diabetes y el grupo control se compararon en términos de valores de glucosa. La concentración de glucosa en sangre en ratas diabéticas fue significativamente alta (p <0,05). En el grupo diabetes hubo un aumento significativo de la membrana periodontal y dilatación de los vasos sanguíneos y hemorragia, con un aumento significativo de células inflamatorias. En el grupo diabetes, la osteonectina mostró una expresión positiva en la membrana periodontal además se observó expresión negativa en los osteocitos del hueso alveolar. La expresión de osteopontina en fibroblastos y fibrillas de colágeno en membrana periodontal fue positiva, las células alveolares, osteocitos y hueso de la matriz ósea dio positivo. Los resultados de la diabetes mostraron que existía periodontitis, debido al aumento de la inflamación que inhibió la formación ósea retardando el desarrollo de células óseas tempranas.


Subject(s)
Animals , Rats , Alveolar Process/metabolism , Alveolar Process/pathology , Diabetes Mellitus, Experimental/pathology , Blood Glucose , Blotting, Western , Diabetes Mellitus, Experimental/metabolism , Immunohistochemistry , Osteonectin/metabolism , Osteopontin/metabolism , Rats, Wistar
13.
Article in English | WPRIM | ID: wpr-739495

ABSTRACT

Osteopontin (OPN) is a phosphorylated glycoprotein secreted into body fluids by various cell types. OPN contains arginine-glycine-aspartate (RGD) and serine-leucine-alanine-tyrosine (SLAY) motifs that bind to several integrins and mediate a wide range of cellular processes. In the present study, the proangiogenic effects of a 20-amino-acid OPN peptide (OPNpt20) containing RGD and SLAY motifs were examined in human umbilical vein endothelial cells (HUVECs) and in a rat focal cerebral ischemia model. OPNpt20 exerted robust proangiogenic effects in HUVECs by promoting proliferation, migration and tube formation. These effects were significantly reduced in OPNpt20-RAA (RGD->RAA)-treated cells, but only slightly reduced in OPNpt20-SLAA (SLAY->SLAA)-treated cells. Interestingly, a mutant peptide without both motifs failed to induce these proangiogenic processes, indicating that the RGD motif is crucial and that SLAY also has a role. In OPNpt20-treated HUVEC cultures, AKT and ERK signaling pathways were activated, but activation of these pathways and tube formation were suppressed by anti-αvβ3 antibody, indicating that OPNpt20 stimulates angiogenesis via the αvβ3-integrin/AKT and ERK pathways. The proangiogenic function of OPNpt20 was further confirmed in a rat middle cerebral artery occlusion model. Total vessel length and vessel densities were markedly greater in OPNpt20-treated ischemic brains, accompanied by induction of proangiogenic markers. Together, these results demonstrate that the 20-amino-acid OPN peptide containing RGD and SLAY motifs exerts proangiogenic effects, wherein both motifs have important roles, and these effects appear to contribute to the neuroprotective effects of this peptide in the postischemic brain.


Subject(s)
Animals , Body Fluids , Brain Ischemia , Brain , Glycoproteins , Human Umbilical Vein Endothelial Cells , Infarction, Middle Cerebral Artery , Integrins , MAP Kinase Signaling System , Neuroprotective Agents , Osteopontin , Rats
14.
Article in Korean | WPRIM | ID: wpr-713738

ABSTRACT

PURPOSE: The purpose of this paper was to determine the ability of a mixture consisting of mesenchymal stem cells, beta-tricalcium phosphate β-TCP), and hydrogel, to support cells and form new tissue. MATERIALS AND METHODS: A composite was produced by adding β-TCP to hydrogel, and mesenchymal stem cells were cultivated in the composite. Then, reverse transcription polymerase chain reaction (RT-PCR) was conducted to measure the level of gene expression for the new bone formation in the cells. Moreover, a composite in which the mesenchymal stem cells were added was injected into the subcutaneous fat of sprague-dawley rats. After four weeks, H&E, Masson trichrome, silver nitrate staining, and osterix immunohistochemical staining were conducted by taking the tissue to evaluate whether the composite supported mesenchymal stem cells and formed new tissue. RESULTS: By using RT-PCR, we found that the level of gene expression became significantly higher in 3-dimensional gel culture with RUNX2 by 1.26 times, with osteopontin by 1.23 times, transforming growth factor-β by 2.12 times, osterix by 1.07 times, type I collagen by 1.3 times, and fibronectin by 1.3 times. In the animal experiment in which a composite was transplanted into the subcutaneous fat, newly formed tissue was observed. Also, it was found that the composite prevented mesenchymal stem cells from leaving and formed new tissue. Osteogenic differentiation cells in the tissue was observed through osterix immunostaining. CONCLUSION: It was identified that the composite prevented mesenchymal stem cells dispersal and contributed to the formation of neogenic tissue. Therefore we conclude that the composite plays a role of a scaffold to support the implanted cells and form neogenic tissue more effectively.


Subject(s)
Animal Experimentation , Collagen Type I , Fibronectins , Gene Expression , Hydrogels , Mesenchymal Stem Cells , Osteogenesis , Osteopontin , Polymerase Chain Reaction , Rats, Sprague-Dawley , Reverse Transcription , Silver Staining , Subcutaneous Fat
15.
Article in English | WPRIM | ID: wpr-772300

ABSTRACT

Amelogenin (AMG) is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG (rhAMG) on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts (OCCM-30) were treated with different concentrations (0.1, 1, 10, 100, 1000, 10,000, 100,000 ng · mL) of recombinant human AMG (rhAMG) and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen type I (COL I), osteopontin (OPN), runt-related transcription factor 2 (Runx2), cementum attachment protein (CAP), and alkaline phosphatase (ALP) genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer. Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomal-associated membrane protein-1 (LAMP-1), which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100,000 ng · mL increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG (100,000 ng · mL) upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100,000 ng · mL remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10,000 and 100,000 ng · mL rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue.


Subject(s)
Alkaline Phosphatase , Metabolism , Amelogenin , Physiology , Animals , Biomarkers , Metabolism , Calcification, Physiologic , Cell Adhesion Molecules , Metabolism , Cell Proliferation , Cementogenesis , Physiology , Collagen Type I , Metabolism , Core Binding Factor Alpha 1 Subunit , Metabolism , Gene Expression Regulation , In Vitro Techniques , Integrin-Binding Sialoprotein , Metabolism , Mice , Microscopy, Confocal , Osteocalcin , Metabolism , Osteopontin , Metabolism , Real-Time Polymerase Chain Reaction
16.
Braz. oral res. (Online) ; 32: e61, 2018. tab, graf
Article in English | LILACS | ID: biblio-974452

ABSTRACT

Abstract To evaluate the impact of the GaAlAs diode laser with energy densities of 160 J/cm2, 320 J/cm2, and 640 J/cm2 on the periodontal tissues under continuous orthodontic force application and on the rate of orthodontic tooth movement in rats with type-2 diabetes mellitus. The intensity of primary alveolar bone formation was also investigated through the immune-positive osteocytes for OPN antibody. Forty adult male Wistar rats were divided into eight groups of 5 rats: normoglycemic (N), 160 J-laser-normoglycemic (160 J-LN), 320 J-laser-normoglycemic (320 J-LN), 640 J-laser-normoglycemic (640 J-LN), diabetic (D), 160 J-laser-diabetic (160 J-LD), 320 J-laser-diabetic (320 J-LD), and 640 J-laser-diabetic (640 J-LD) rats. Diabetes mellitus was induced by a single intravenous injection of 40 mg/kg monohydrated-alloxan. An orthodontic force magnitude of 20cN was applied. The laser parameters were continuous emission of 780-nm wavelength, output power of 20mW, and fiber probe with a spot size of 0.04 cm in diameter. Radiographic, histomorphological, and immunohistochemical analysis were performed after a period of 21 days. The photobiomodulation using the energy density of 640 J/cm2 strongly stimulated the alveolar bone formation and contributed the reorganization of the soft periodontal tissues, followed by the 320 J/cm2. Extensive alveolar bone loss, intense infiltration of inflammatory cells, and degradation of the PDJ tissue were mainly found in the D and 160 J-LD groups. The rate of orthodontic tooth movement was represented by the interdental distance between the cementoenamel junctions of the right mandibular first and second molars . This distance was larger in the diabetic groups (D: 39.98±1.97, 160 J-LD: 34.84±6.01, 320 J-LD: 29.82±1.73, and 640 J-LD: 35.47±4.56) than in the normoglycemic groups (N: 21.13±1.19; 160 J-LN: 22.69±0.72, 320 J-LN: 22.28±0.78, and 640 J-LN: 24.56±2.11). The number of osteopontin-positive osteocytes was significantly greater in the 640 J-LD (14.72 ± 0.82; p < 0.01) and 640 J-LN (13.62 ± 1.33; p < 0.05) groups than with D (9.82 ± 1.17) and 160 J-LD (9.77 ± 1.10) groups. Therefore, the energy density of 640 J/cm2 provided the best maintenance and integrity of the periodontal tissue microarchitecture under continuous orthodontic force when compared with the other dosages, mainly in the uncontrolled diabetic rats. The interdental distance was greater in the D and 160 J-LD groups due to presence of severe periodontitis caused by diabetes plus the mechanical stress generated by continuous orthodontic forces, implying, thus, an insufficient biostimulatory effect for the dosage of 160 J/cm2.


Subject(s)
Animals , Male , Tooth Movement Techniques/methods , Periodontium/radiation effects , Low-Level Light Therapy/methods , Diabetes Mellitus, Type 2/physiopathology , Orthodontic Appliances , Osteoclasts/radiation effects , Osteocytes/radiation effects , Osteogenesis/radiation effects , Radiation Dosage , Reference Values , Periodontium/pathology , Periodontium/diagnostic imaging , Immunohistochemistry , Radiography , Random Allocation , Reproducibility of Results , Alveolar Bone Loss/pathology , Rats, Wistar , Diabetes Mellitus, Experimental , Osteopontin/analysis , Lasers, Semiconductor/therapeutic use
17.
Braz. oral res. (Online) ; 32: e85, 2018. tab, graf
Article in English | LILACS | ID: biblio-952161

ABSTRACT

Abstract This study aimed to investigate the effects of different doses of systemic melatonin application on new bone formation during mandibular distraction osteogenesis (DO) in rats. Mandibular DO was performed on 30 adult female Sprague-Dawley rats, which were randomly divided into three groups: control group (CNT), melatonin dose 1 (MLT-D1), and melatonin dose 2 (MLT-D2). A five-day latent waiting period and a ten-day distraction phase followed the surgery. After the surgery, rats from the MLT-D1 and MLT-D2 groups received 25 and 50 mg/kg melatonin, respectively, at 7, 14, 21, 28, and 35 days. The animals were euthanised 28 days after distraction, i.e. at 43 days after surgery. Histological and histomorphometric analyses revealed that the distracted bone area was completely filled with new bone formation in all three groups. The MLT-D2 group exhibited the most new bone formation, followed by MLT-D1 and CNT. The melatonin groups had more osteoclasts than the CNT (p < 0.05). The number of osteoblasts was higher in the melatonin groups than in the CNT group, and the MLT-D2 had more osteoclasts than the MLT-D1 group (p < 0.05). Finally, the osteopontin (OPN) and vascular endothelial growth factor (VEGF) levels were higher in the melatonin groups than in the CNT group, and the MLT-D2 had higher OPN and VEGF levels than the MLT-D1 (p < 0.05). This study suggests that systemic melatonin application could increase new bone formation in DO.


Subject(s)
Animals , Female , Osteogenesis/drug effects , Bone Regeneration/drug effects , Osteogenesis, Distraction/methods , Melatonin/administration & dosage , Antioxidants/administration & dosage , Osteoblasts/physiology , Osteoclasts/physiology , Osteogenesis/physiology , Bone Regeneration/physiology , Immunohistochemistry , Random Allocation , Reproducibility of Results , Treatment Outcome , Rats, Sprague-Dawley , Vascular Endothelial Growth Factor A/analysis , Osteopontin/analysis , Mandible/surgery , Mandible/drug effects , Mandible/physiology , Mandible/pathology
18.
Acta odontol. latinoam ; 31(2): 110-116, 2018. ilus, graf
Article in English | LILACS | ID: biblio-970843

ABSTRACT

The in vivo response of osteocytes to different force magnitudes soon after they are applied remains to be elucidated. The aim of this study was to examine the early effects of applying a very light (LF: 0,16 N) and a very strong (SF: 2,26 N) orthodontic force during one hour on apoptosis and osteopontin (OPN) expression on alveolar bone osteocytes, in rats. Results: LF: compared to the control group, they showed a significant increase in OPN expression, and a significant decrease in the number of TUNELpositive osteocytes. SF: compared to the control group, they showed a significant increase in OPN expression and a significant decrease in the number of TUNELpositive osteocytes. Our results show that osteocytes respond very early to the application of tension and pressure forces of different magnitudes, and application of forces decreases the number of apoptotic osteocytes and increases OPN expression. These results allow concluding that osteocytes activate rapidly when subjected to locally applied forces, whether these forces be pressure or tension, light or strong forces. Grants: UBACyT 200201301002270 BA and School of Dentistry, University of Buenos Aires (AU)


Hasta el momento no se ha dilucidado la respuesta temprana in vivo de los osteocitos a la aplicación de fuerzas de diferentes magnitudes sobre el hueso. El objetivo de este estudio fue examinar la respuesta temprana de la aplicación de una fuerza ortodóncica muy liviana (FL: 0,16 N) y muy fuerte (FF: 2,26 N) durante una hora sobre la expresión de apoptosis y osteopontina (OPN) en los osteocitos del hueso alveolar, en ratas. Resultados: FL: en comparación con el grupo control, mostraron un aumento significativo en la expresión de OPN y una disminución significativa en el número de osteocitos TUNELpositivos. FF: en comparación con el grupo control, mostraron un aumento significativo en la expresión de OPN y una disminución signi ficativa en el número de osteocitos TUNELpositivos. Nuestros resultados muestran que los osteocitos responden muy temprano a la aplicación de fuerzas de tensión y presión de diferentes magnitudes, y la aplicación de fuerzas disminuye el número de osteocitos apoptóticos y aumenta la expresión de OPN. Estos resultados permiten concluir que los osteocitos se activan rápidamente cuando se los somete a fuerzas aplicadas localmente, ya sean estas fuerzas de presión o tensión, livianas o fuertes (AU)


Subject(s)
Animals , Rats , Osteocytes , Stress, Mechanical , Tooth Movement Techniques , Apoptosis , Osteopontin , Immunohistochemistry , Data Interpretation, Statistical , In Situ Nick-End Labeling , Mechanotransduction, Cellular , Alveolar Process
20.
J. appl. oral sci ; 26: e20170326, 2018. graf
Article in English | LILACS, BBO | ID: biblio-954523

ABSTRACT

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.


Subject(s)
Humans , Male , Wound Healing/physiology , Bone Remodeling/physiology , Tooth Socket/physiology , Tooth Socket/diagnostic imaging , Reference Values , Time Factors , Tooth Extraction , Transcription Factors/analysis , Immunohistochemistry , Gene Expression , Osteocalcin/analysis , Reproducibility of Results , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Core Binding Factor Alpha 1 Subunit/analysis , Osteopontin/analysis , RANK Ligand/analysis , Osteoprotegerin/analysis , X-Ray Microtomography , Tartrate-Resistant Acid Phosphatase/analysis
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