RESUMO
Presentamos un paciente de 63 años con cáncer renal y aumento de fosfatasa alcalina sérica de tipo óseo de acuerdo con su reactividad con anticuerpos monoclonales específicos. Se descartaron las causas conocidas de aumento de la isoenzima, incluyendo metástasis óseas. Los niveles enzimáticos cayeron abruptamente con la remoción del tumor, por lo que consideramos a este último como su origen. Diversas isoenzimas de fosfatasa alcalina pueden ser producidas y secretadas por tumores como manifestación paraneoplásica. El conocimiento de esto puede, en ocasiones, orientarnos hacia la presencia de una neoplasia oculta. Además, los cambios en los niveles séricos de esas isoenzimas pueden ser indicadores de respuesta al tratamiento o de recidiva tumoral. (AU)
A 63-year old man was seen in the outpatient clinic because of renal cancer and elevation in bone alkaline phosphatase measured by monoclonal antibodies assay. Known causes of bone isoenzyme augmentation, including bone metastases, were ruled out. The tumoral origin of the isoenzyme was diagnosed because after removal of the tumor the enzymatic levels fell sharply. Several alkaline phosphatase isoenzymes can be produced and secreted by tumors as a paraneoplasic manifestation and their elevation could be a manifestation of an occult neoplasia. Furthermore the monitoring of their blood levels can be useful means of treatment response and a tool to monitoring recurrence if a sharp decrease after removal of the tumor is observed. (AU)
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Fosfatase Alcalina/biossíntese , Neoplasias Renais/metabolismo , Osteíte Deformante/diagnóstico por imagem , Atenolol/uso terapêutico , Biomarcadores , Eritropoetina/uso terapêutico , Sinvastatina/uso terapêutico , Fosfatase Alcalina/análise , Fosfatase Alcalina/efeitos da radiação , Fosfatase Alcalina/fisiologia , Everolimo/uso terapêutico , Sunitinibe/uso terapêutico , Ácido Zoledrônico/uso terapêutico , Hipercolesterolemia/tratamento farmacológico , Hipertensão/tratamento farmacológico , Ílio/diagnóstico por imagem , Anemia/tratamento farmacológico , Neoplasias Renais/patologia , Neoplasias Renais/tratamento farmacológico , Neoplasias Renais/diagnóstico por imagem , Anticorpos Monoclonais/efeitos da radiaçãoRESUMO
INTRODUCTION: The purpose of this study was to compare the cell viability of dental pulp cells treated with Biodentine (Septodont, Saint-Maur, France) and mineral trioxide aggregate (MTA) and the in vitro and in vivo expression of mineralization markers induced by the 2 materials. METHODS: Human dental pulp cells isolated from 6 permanent teeth were stimulated with Biodentine and MTA extracts. Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and quantitative reverse-transcriptase polymerase chain reaction was used to determine the expression of mineralization markers. Specimens of teeth from dogs treated with Biodentine and MTA after pulpotomy were used to determine the presence of osteopontin and alkaline phosphatase by immunohistochemistry and runt-related transcription factor 2 by immunofluorescence. RESULTS: No significant differences in cell viability were found between MTA and Biodentine extracts and controls after 24 and 48 hours (P > .05). After 48 hours, osteopontin (SPP1), alkaline phosphatase (ALP), and runt-related transcription factor 2 (RUNX2) expression was higher in MTA and Biodentine than in controls (P < .05). Osteopontin staining was more intense and spread over a greater number of areas in Biodentine than in MTA samples (P < .0001). Alkaline phosphatase staining of a mineralized tissue bridge was significantly different between materials (P < .0001), but no difference in alkaline phosphatase staining of pulp tissue was found between MTA and Biodentine (P = .2). Also, no significant difference in the number of cells labeled for runt-related transcription factor 2 by immunofluorescence was observed between materials (P > .05). CONCLUSIONS: Biodentine stimulated similar markers as MTA, but staining was more intense and spread over a larger area of the pulp tissue.
Assuntos
Fosfatase Alcalina/biossíntese , Compostos de Alumínio/farmacologia , Compostos de Cálcio/farmacologia , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/metabolismo , Osteopontina/biossíntese , Óxidos/farmacologia , Silicatos/farmacologia , Animais , Biomarcadores/metabolismo , Calcificação Fisiológica/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/biossíntese , Polpa Dentária/patologia , Cavidade Pulpar/efeitos dos fármacos , Cavidade Pulpar/metabolismo , Cavidade Pulpar/patologia , Cães , Combinação de Medicamentos , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase , Ápice Dentário/efeitos dos fármacos , Ápice Dentário/metabolismo , Ápice Dentário/patologiaRESUMO
We aimed at evaluating the effect of titanium (Ti) with nanotopography (Nano) on the endogenous expression of BMP-2 and BMP-4 and the relevance of this process to the nanotopography-induced osteoblast differentiation. MC3T3-E1 cells were grown on Nano and machined (Machined) Ti surfaces and the endogenous BMP-2/4 expression and the effect of BMP receptor BMPR1A silencing in both osteoblast differentiation and expression of genes related to TGF-ß/BMP signaling were evaluated. Nano supported higher BMP-2 gene and protein expression and upregulated the osteoblast differentiation compared with Machined Ti surface. The BMPR1A silencing inhibited the osteogenic potential induced by Nano Ti surface as indicated by reduced alkaline phosphatase (ALP), osteocalcin and RUNX2 gene expression, RUNX2 protein expression and ALP activity. In addition, the expression of genes related to TGF-ß/BMP signaling was deeply affected by BMPR1A-silenced cells grown on Nano Ti surface. In conclusion, we have demonstrated for the first time that nanotopography induces osteoblast differentiation, at least in part, by upregulating the endogenous production of BMP-2 and modulating BMP signaling pathway. J. Cell. Biochem. 117: 1718-1726, 2016. © 2015 Wiley Periodicals, Inc.
Assuntos
Proteína Morfogenética Óssea 2/biossíntese , Proteína Morfogenética Óssea 4/biossíntese , Diferenciação Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Osteoblastos/metabolismo , Transdução de Sinais/efeitos dos fármacos , Titânio/farmacologia , Fosfatase Alcalina/biossíntese , Animais , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Linhagem Celular , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Camundongos , Propriedades de SuperfícieRESUMO
OBJECTIVE: Fresh-frozen bone allograft (FFBA) is an alternative to autogenous bone (AB) for reconstructing maxillary bone. Despite the promising clinical results, cell responses to FFBA and AB were not evaluated. Thus, our aim was to compare cells harvested from maxillary reconstructed sites with either AB or FFBA in terms of osteoblast differentiation and to evaluate the effect of culturing cells in contact with FFBA. METHODS: Cells harvested from three patients submitted to bilateral maxillary reconstruction with AB and FFBA were cultured to evaluate: proliferation, alkaline phosphatase activity, extracellular matrix mineralization and gene expression of osteoblastic markers. The effect of FFBA on osteoblast differentiation was studied by culturing cells harvested from AB in contact with FFBA and evaluating the same parameters. Data were compared using either two-way ANOVA followed by Tukey-b test or Student's t test (p≤0.05). RESULTS: Cell proliferation was higher in cultures from AB grafted sites and extracellular matrix mineralization was higher in cultures derived from FFBA grafted sites. The gene expression of alkaline phosphatase, RUNX2, bone sialoprotein and osteocalcin was higher in cells derived from FFBA compared with cells from AB grafted sites. However, the exposure of cells derived from AB to FFBA particles did not have any remarkable effect on osteoblast differentiation. CONCLUSIONS: These results indicate the higher osteogenic activity of cells derived from FFBA compared with AB reconstructed sites, offering an explanation at cellular level of why FFBA could be a suitable alternative to AB for reconstructing maxillary bone defects.
Assuntos
Transplante Ósseo/métodos , Criopreservação , Reconstrução Mandibular/métodos , Fosfatase Alcalina/biossíntese , Aloenxertos/transplante , Regeneração Óssea/genética , Transplante Ósseo/efeitos adversos , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Células Cultivadas , Feminino , Humanos , Pessoa de Meia-Idade , Osteoblastos/citologia , Osteoblastos/fisiologia , Osteocalcina/biossínteseRESUMO
In this work we characterized the infection of a primary culture of rat osteoblastic lineage cells (OBCs) with measles virus (MeV) and the effect of infection on cell differentiation and maturation. Infection of OBCs with MeV led to high titers of infectivity released early after infection. Also, analysis of mRNAs corresponding to osteogenic differentiation markers like alkaline phosphatase (ALP), bone sialo-protein (BSP) and bone morphogenetic proteins (BMPs) 1-4-5-7 in OBCs revealed higher values (2-75-fold of increment) for infected cells in comparison with uninfected controls. Differentiation of OBCs in osteogenic medium prior to infection influenced the level of stimulation induced by MeV. Furthermore, treatment of OBCs with Ly294002, a PI3K/AKT inhibitor, increased viral titers, whereas treatment with 10µM or 100µM ATPγS diminished MeV multiplication. In addition, increments of osteogenic differentiation markers induced by MeV infection were not modified either by treatment with Ly294002 or ATPγS. These data provide the first evidence demonstrating that MeV can infect osteoblasts in vitro leading to osteoblastic differentiation, a key feature in bone pathogenic processes like otosclerosis.
Assuntos
Vírus do Sarampo/fisiologia , Osteoblastos/virologia , Osteogênese/fisiologia , Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/farmacologia , Fosfatase Alcalina/biossíntese , Animais , Proteína Morfogenética Óssea 4/biossíntese , Osso e Ossos/metabolismo , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Cromonas/farmacologia , Morfolinas/farmacologia , Osteoblastos/fisiologia , Otosclerose/etiologia , Inibidores de Fosfoinositídeo-3 Quinase , Ratos , Replicação ViralRESUMO
Chitosan/poly(DL-lactide-co-glycolide) (Ch/DL PLG) composite scaffolds were fabricated by freeze-drying lyophilization, and were evaluated and compared for use as a bone regeneration scaffold through measurements of the compression mechanical properties of the porous scaffolds. Also, In vitro cell culture of Sprague-Dawley rat's osteoblasts were used to evaluate the phenotype expression of cells in the scaffolds, characterizing the cellular adhesion, proliferation and alkaline phosphatase activity. The gene expression of osteocalcin, sialoprotein, alkaline phosphatase, Type I collagen and TGFß1 were confirmed in the samples; moreover, it was confirmed, the mineralization by IR spectra and EDS analysis. Our results thus show that Ch/DL PLG scaffolds are suitable for biological applications.
Assuntos
Quitosana/química , Teste de Materiais/métodos , Poliglactina 910/química , Engenharia Tecidual/métodos , Fosfatase Alcalina/biossíntese , Fosfatase Alcalina/metabolismo , Animais , Adesão Celular , Proliferação de Células , Colágeno Tipo I/metabolismo , Liofilização , Masculino , Osteoblastos/metabolismo , Osteocalcina/biossíntese , Fenótipo , Ratos , Ratos Sprague-Dawley , Sialoglicoproteínas/biossíntese , Fator de Crescimento Transformador beta1/metabolismoRESUMO
OBJECTIVE: The aim of this study was to assess the effect of cigarette smoke inhalation (CSI) on gene expression in alveolar bone healing sites. STUDY DESIGN: Wistar rats were randomly assigned to the groups: control [animals not exposed to CSI (n = 20)] and test [animals exposed to CSI, starting 3 days before teeth extraction and maintained until killing them (n = 20)]. First mandibular molars were bilaterally extracted, and the expression of alkaline phosphatase, bone morphogenetic protein (BMP) 2 and 7, receptor activator of nuclear factor κB ligand, osteoprotegerin, and d2 isoform of vacuolar adenosine triphosphatase V(0) domain were assessed by quantitative polymerase chain reaction in the newly formed tissue in the sockets. RESULTS: Overall, data analysis demonstrated that CSI significantly affected the expression pattern of all of the studied genes except BMP-7. CONCLUSION: The expression of key genes for bone healing may be affected by CSI in tooth extraction sites.
Assuntos
Regeneração Óssea/genética , Fumar/efeitos adversos , Fosfatase Alcalina/biossíntese , Fosfatase Alcalina/genética , Animais , Proteína Morfogenética Óssea 2/biossíntese , Proteína Morfogenética Óssea 2/genética , Regulação da Expressão Gênica , Isoenzimas/biossíntese , Isoenzimas/genética , Masculino , Osteoprotegerina/biossíntese , Osteoprotegerina/genética , Ligante RANK/biossíntese , Ligante RANK/genética , Distribuição Aleatória , Ratos , Ratos Wistar , Extração Dentária , ATPases Vacuolares Próton-Translocadoras/biossíntese , ATPases Vacuolares Próton-Translocadoras/genéticaRESUMO
BACKGROUND: The aim of this study is to investigate the potential use of periosteum-derived cells (PCs) for tissue engineering in peri-implant defects. METHODS: Bone marrow cells (BMCs) and PCs were harvested from seven adult beagle dogs, cultured in vitro, and phenotypically characterized with regard to their osteogenic properties. The animals were then subjected to teeth extraction, and 3 months later, two implant sites were drilled, bone dehiscences created, and dental implants placed. Dehiscences were randomly assigned to one of two groups: PCs (PCs + carrier) and BMCs (BMCs + carrier). After 3 months, the animals were sacrificed and the implants with adjacent hard tissues were processed for undecalcified sections. Bone-to-implant contact, bone fill within the limits of implant threads, and new bone area in a zone lateral to the implant were histometrically obtained. RESULTS: In vitro, phenotypic characterization demonstrated that both cell populations presented osteogenic potential, as identified by the mineral nodule formation and the expression of bone markers. Histometrically, an intergroup analysis showed that both cell-treated defects had similar bone fill within the limits of implant threads and bone-to-implant contact (P >0.05), and although a trend toward higher new bone area values was found for the PC group, there was no significant difference between the experimental groups (P >0.05). CONCLUSIONS: Periosteal and bone marrow cells presented a similar potential for bone reconstruction. As such, periosteum may be considered as an alternative source of osteogenic cells in implant dentistry.
Assuntos
Transplante de Células , Osseointegração , Osteogênese , Periósteo/citologia , Engenharia Tecidual/métodos , Fosfatase Alcalina/biossíntese , Animais , Transplante de Medula Óssea , Adesão Celular , Proliferação de Células , Células Cultivadas , Colágeno Tipo I/biossíntese , Implantação Dentária Endóssea , Cães , Sialoproteína de Ligação à Integrina , Periósteo/metabolismo , Distribuição Aleatória , Sialoglicoproteínas/biossíntese , Deiscência da Ferida Operatória/terapia , Alicerces TeciduaisRESUMO
The aim of this study was to investigate the effects of low-level laser therapy (LLLT) by using gallium aluminum arsenide (GaAlAs) diode laser on human osteoblastic cells grown on titanium (Ti). Osteoblastic cells were obtained by enzymatic digestion of human alveolar bone and cultured on Ti discs for up to 17 days. Cells were exposed to LLLT at 3 J/cm2 (wavelength of 780 nm) at days 3 and 7 and non-irradiated cultures were used as control. LLLT treatment did not influence culture growth, ALP activity, and mineralized matrix formation. Analysis of cultures by epifluorescence microscopy revealed an area without cells in LLLT treated cultures, which was repopulated latter with proliferative and less differentiated cells. Gene expression of ALP, OC, BSP, and BMP-7 was higher in LLLT treated cultures, while Runx2, OPN, and OPG were lower. These results indicate that LLLT modulates cell responses in a complex way stimulating osteoblastic differentiation, which suggests possible benefits on implant osseointegration despite a transient deleterious effect immediately after laser irradiation.
Assuntos
Matriz Óssea/crescimento & desenvolvimento , Expressão Gênica/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Osseointegração/efeitos da radiação , Osteoblastos/efeitos da radiação , Fosfatase Alcalina/biossíntese , Fosfatase Alcalina/genética , Análise de Variância , Proteína Morfogenética Óssea 7/biossíntese , Proteína Morfogenética Óssea 7/genética , Células Cultivadas/efeitos da radiação , Colágeno Tipo I/biossíntese , Colágeno Tipo I/genética , Subunidade alfa 1 de Fator de Ligação ao Core/biossíntese , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Humanos , Sialoproteína de Ligação à Integrina/biossíntese , Sialoproteína de Ligação à Integrina/genética , Molécula 1 de Adesão Intercelular/biossíntese , Molécula 1 de Adesão Intercelular/genética , Lasers Semicondutores/uso terapêutico , Osteoblastos/metabolismo , Osteocalcina/biossíntese , Osteocalcina/genética , Osteopontina/biossíntese , Osteopontina/genética , Osteoprotegerina/biossíntese , Osteoprotegerina/genética , Ligante RANK/biossíntese , Ligante RANK/genética , Estatísticas não Paramétricas , TitânioRESUMO
The aim of this study was to investigate the effects of low-level laser therapy (LLLT) by using gallium aluminum arsenide (GaAlAs) diode laser on human osteoblastic cells grown on titanium (Ti). Osteoblastic cells were obtained by enzymatic digestion of human alveolar bone and cultured on Ti discs for up to 17 days. Cells were exposed to LLLT at 3 J/cm2 (wavelength of 780 nm) at days 3 and 7 and non-irradiated cultures were used as control. LLLT treatment did not influence culture growth, ALP activity, and mineralized matrix formation. Analysis of cultures by epifluorescence microscopy revealed an area without cells in LLLT treated cultures, which was repopulated latter with proliferative and less differentiated cells. Gene expression of ALP, OC, BSP, and BMP-7 was higher in LLLT treated cultures, while Runx2, OPN, and OPG were lower. These results indicate that LLLT modulates cell responses in a complex way stimulating osteoblastic differentiation, which suggests possible benefits on implant osseointegration despite a transient deleterious effect immediately after laser irradiation.
Este estudo teve como objetivo investigar o efeito do laser diodo de gálio-alumÃnio-arsênio (GaAlAs) em células osteoblásticas humanas cultivadas sobre discos de Ti. Para tanto, células osteoblásticas foram obtidas por digestão enzimática de osso alveolar humano e cultivadas sobre discos de Ti por 17 dias. As células foram submetidas à irradiação no 3º e 7º dias na dose de 3 J/cm2 e comprimento de onda de 780 nm e células não irradiadas foram usadas como controle. A irradiação não alterou a proliferação celular, atividade de ALP e formação de matriz mineralizada. Microscopia por epifluorescência indicou que após 24 h da aplicação do laser, as culturas irradiadas apresentaram áreas sem células, que mais tarde foram repovoadas por células em fase de proliferação e menos diferenciadas. O laser aumentou a expressão gênica relativa da ALP, OC, BSP e BMP-7 e reduziu a de RUNX2, OPN e OPG. Os resultados indicam que a terapia com laser modula de forma complexa as respostas celulares, estimulando a diferenciação osteoblástica. Assim, é possÃvel sugerir possÃveis benefÃcios do laser na osseointegração de implantes de Ti apesar do efeito deletério à s células imediatamente após a irradiação.