Academic literature on the topic 'Osteoblasts'
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Journal articles on the topic "Osteoblasts"
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Kim, Jung Ha, Kabsun Kim, Inyoung Kim, Semun Seong, Jeong-Tae Koh, and Nacksung Kim. "The ATF3–OPG Axis Contributes to Bone Formation by Regulating the Differentiation of Osteoclasts, Osteoblasts, and Adipocytes." International Journal of Molecular Sciences 23, no. 7 (March 23, 2022): 3500. http://dx.doi.org/10.3390/ijms23073500.
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Activating transcription factor 3 (ATF3) has been identified as a negative regulator of osteoblast differentiation in in vitro study. However, it was not associated with osteoblast differentiation in in vivo study. To provide an understanding of the discrepancy between the in vivo and in vitro findings regarding the function of ATF3 in osteoblasts, we investigated the unidentified roles of ATF3 in osteoblast biology. ATF3 enhanced osteoprotegerin (OPG) production, not only in osteoblast precursor cells, but also during osteoblast differentiation and osteoblastic adipocyte differentiation. In addition, ATF3 increased nodule formation in immature osteoblasts and decreased osteoblast-dependent osteoclast formation, as well as the transdifferentiation of osteoblasts to adipocytes. However, all these effects were reversed by the OPG neutralizing antibody. Taken together, these results suggest that ATF3 contributes to bone homeostasis by regulating the differentiation of various cell types in the bone microenvironment, including osteoblasts, osteoclasts, and adipocytes via inducing OPG production.
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Bauer, Omri, Amnon Sharir, Ayako Kimura, Shay Hantisteanu, Shu Takeda, and Yoram Groner. "Loss of Osteoblast Runx3 Produces Severe Congenital Osteopenia." Molecular and Cellular Biology 35, no. 7 (January 20, 2015): 1097–109. http://dx.doi.org/10.1128/mcb.01106-14.
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Congenital osteopenia is a bone demineralization condition that is associated with elevated fracture risk in human infants. Here we show thatRunx3, likeRunx2, is expressed in precommitted embryonic osteoblasts and that Runx3-deficient mice develop severe congenital osteopenia. Runx3-deficient osteoblast-specific (Runx3fl/fl/Col1α1-cre), but not chondrocyte-specific (Runx3fl/fl/Col1α2-cre), mice are osteopenic. This demonstrates that an osteoblastic cell-autonomous function of Runx3 is required for proper osteogenesis. Bone histomorphometry revealed that decreased osteoblast numbers and reduced mineral deposition capacity in Runx3-deficient mice cause this bone formation deficiency. Neonatal bone and cultured primary osteoblast analyses revealed a Runx3-deficiency-associated decrease in the number of active osteoblasts resulting from diminished proliferation and not from enhanced osteoblast apoptosis. These findings are supported by Runx3-null culture transcriptome analyses showing significant decreases in the levels of osteoblastic markers and increases in the levels of Notch signaling components. Thus, while Runx2 is mandatory for the osteoblastic lineage commitment, Runx3 is nonredundantly required for the proliferation of these precommitted cells, to generate adequate numbers of active osteoblasts. HumanRUNX3resides on chromosome 1p36, a region that is associated with osteoporosis. Therefore, RUNX3 might also be involved in human bone mineralization.
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Park, Yu-Seong, Hyun-Woo Kim, Jin-Hyeon Hwang, Jung-Young Eom, Dong-Ha Kim, Jinho Park, Hyun-Jin Tae, et al. "Plum-Derived Exosome-like Nanovesicles Induce Differentiation of Osteoblasts and Reduction of Osteoclast Activation." Nutrients 15, no. 9 (April 27, 2023): 2107. http://dx.doi.org/10.3390/nu15092107.
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Osteoblasts and osteoclasts play crucial roles in bone formation and bone resorption. We found that plum-derived exosome-like nanovesicles (PENVs) suppressed osteoclast activation and modulated osteoblast differentiation. PENVs increased the proliferation, differentiation, and mineralization of osteoblastic MC3T3-E1 cells and osteoblasts from mouse bone marrow cultures. Notably, PENVs elevated the expression of osteoblastic transcription factors and osteoblast differentiation marker proteins in MC3T3-E1 cells. Higher levels of phosphorylated BMP-2, p38, JNK, and smad1 proteins were detected in PENV-treated MC3T3-E1 cells. Additionally, the number of TRAP-positive cells was significantly decreased in PENV-treated osteoclasts isolated from osteoblasts from mouse bone marrow cultures. Importantly, osteoclastogenesis of marker proteins such as PPAR-gamma, NFATc1, and c-Fos were suppressed by treatment with PENVs (50 μg/mL). Taken together, these results demonstrate that PENVs can be used as therapeutic targets for treating bone-related diseases by improving osteoblast differentiation and inhibiting osteoclast activation for the first time.
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Kim, Jung Ha, Kabsun Kim, Inyoung Kim, Semun Seong, Hyun Kook, Kyung Keun Kim, Jeong-Tae Koh, and Nacksung Kim. "Bifunctional Role of CrkL during Bone Remodeling." International Journal of Molecular Sciences 22, no. 13 (June 29, 2021): 7007. http://dx.doi.org/10.3390/ijms22137007.
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Coupled signaling between bone-forming osteoblasts and bone-resorbing osteoclasts is crucial to the maintenance of bone homeostasis. We previously reported that v-crk avian sarcoma virus CT10 oncogene homolog-like (CrkL), which belongs to the Crk family of adaptors, inhibits bone morphogenetic protein 2 (BMP2)-mediated osteoblast differentiation, while enhancing receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation. In this study, we investigated whether CrkL can also regulate the coupling signals between osteoblasts and osteoclasts, facilitating bone homeostasis. Osteoblastic CrkL strongly decreased RANKL expression through its inhibition of runt-related transcription factor 2 (Runx2) transcription. Reduction in RANKL expression by CrkL in osteoblasts resulted in the inhibition of not only osteoblast-dependent osteoclast differentiation but also osteoclast-dependent osteoblast differentiation, suggesting that CrkL participates in the coupling signals between osteoblasts and osteoclasts via its regulation of RANKL expression. Therefore, CrkL bifunctionally regulates osteoclast differentiation through both a direct and indirect mechanism while it inhibits osteoblast differentiation through its blockade of both BMP2 and RANKL reverse signaling pathways. Collectively, these data suggest that CrkL is involved in bone homeostasis, where it helps to regulate the complex interactions of the osteoblasts, osteoclasts, and their coupling signals.
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Ducy, P., and G. Karsenty. "Two distinct osteoblast-specific cis-acting elements control expression of a mouse osteocalcin gene." Molecular and Cellular Biology 15, no. 4 (April 1995): 1858–69. http://dx.doi.org/10.1128/mcb.15.4.1858.
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Osteoblasts are cells of mesodermal origin that play a pivotal role during bone growth and mineralization. The mechanisms governing osteoblast-specific gene expression are still unknown. To understand these mechanisms, we analyzed the cis-acting elements of mouse osteocalcin gene 2 (mOG2), the best-characterized osteoblast-specific gene, by DNA transfection experiments in osteoblastic and nonosteoblastic cell lines and by DNA-binding assays. 5' deletion analysis of an mOG2 promoter-luciferase chimeric gene showed that a region located between -147 and -34 contained most if not all of the regulatory elements required for osteoblast-specific expression. Three different binding sites, called A, B, and C, for factors present in nuclear extracts of osteoblasts were identified in this short promoter by DNase I footprint assays. In gel retardation assays, the A element, located between bp -64 and -47, bound a factor present only in nuclear extracts of osteoblastic cell lines and nonmineralizing primary osteoblasts. The B element, located between bp -110 and -83, bound a ubiquitously expressed factor. The C element, located between bp -146 and -132, bound a factor present only in nuclear extracts of osteoblastic cell lines and nonmineralizing and mineralizing primary osteoblasts. When cloned upstream of a minimum osteocalcin promoter or a heterologous promoter, multimers of the A element strongly increased the activities of these promoters in osteoblastic cell lines at two different stages of differentiation but in no other cell line; we named this element osteocalcin-specific element 1 (OSE1). Multimers of the C element increased the activities of these promoters predominantly in a differentiated osteoblastic cell line; we named this element OSE2. This study demonstrates that two distinct cis-acting elements are responsible for osteoblast expression of mOG2 and provides for the first time a functional characterization of osteoblast-specific cis-acting elements. We speculate that these two elements may be important at several stages of osteoblast differentiation.
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Moriishi, Takeshi, Yosuke Kawai, Ryo Fukuyama, Yuki Matsuo, You-Wen He, Haruhiko Akiyama, Izumi Asahina, and Toshihisa Komori. "Bcl2l1 Deficiency in Osteoblasts Reduces the Trabecular Bone Due to Enhanced Osteoclastogenesis Likely through Osteoblast Apoptosis." International Journal of Molecular Sciences 24, no. 24 (December 10, 2023): 17319. http://dx.doi.org/10.3390/ijms242417319.
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Bcl2l1 (Bcl-XL) belongs to the Bcl-2 family, Bcl2 and Bcl2-XL are major anti-apoptotic proteins, and the apoptosis of osteoblasts is a key event for bone homeostasis. As the functions of Bcl2l1 in osteoblasts and bone homeostasis remain unclear, we generated osteoblast-specific Bcl2l1-deficient (Bcl2l1fl/flCre) mice using 2.3-kb Col1a1 Cre. Trabecular bone volume and the trabecular number were lower in Bcl2l1fl/flCre mice of both sexes than in Bcl2l1fl/fl mice. In bone histomorphometric analysis, osteoclast parameters were increased in Bcl2l1fl/flCre mice, whereas osteoblast parameters and the bone formation rate were similar to those in Bcl2l1fl/fl mice. TUNEL-positive osteoblastic cells and serum TRAP5b levels were increased in Bcl2l1fl/flCre mice. The deletion of Bcl2l1 in osteoblasts induced Tnfsf11 expression, whereas the overexpression of Bcl-XL had no effect. In a co-culture of Bcl2l1-deficient primary osteoblasts and wild-type bone-marrow-derived monocyte/macrophage lineage cells, the numbers of multinucleated TRAP-positive cells and resorption pits increased. Furthermore, serum deprivation or the deletion of Bcl2l1 in primary osteoblasts increased apoptosis and ATP levels in the medium. Therefore, the reduction in trabecular bone in Bcl2l1fl/flCre mice may be due to enhanced bone resorption through osteoblast apoptosis and the release of ATP from apoptotic osteoblasts, and Bcl2l1 may inhibit bone resorption by preventing osteoblast apoptosis.
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Sutton, Amelia L. M., Xiaoxue Zhang, Diane R. Dowd, Yogendra P. Kharode, Barry S. Komm, and Paul N. MacDonald. "Semaphorin 3B Is a 1,25-Dihydroxyvitamin D3-Induced Gene in Osteoblasts that Promotes Osteoclastogenesis and Induces Osteopenia in Mice." Molecular Endocrinology 22, no. 6 (June 1, 2008): 1370–81. http://dx.doi.org/10.1210/me.2007-0363.
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Abstract The vitamin D endocrine system is important for skeletal homeostasis. 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] impacts bone indirectly by promoting intestinal absorption of calcium and phosphate and directly by acting on osteoblasts and osteoclasts. Despite the direct actions of 1,25(OH)2D3 in bone, relatively little is known of the mechanisms or target genes that are regulated by 1,25(OH)2D3 in skeletal cells. Here, we identify semaphorin 3B (SEMA3B) as a 1,25(OH)2D3-stimulated gene in osteoblastic cells. Northern analysis revealed strong induction of SEMA3B mRNA by 1,25(OH)2D3 in MG-63, ST-2, MC3T3, and primary osteoblastic cells. Moreover, differentiation of these osteogenic cells enhanced SEMA3B gene expression. Biological effects of SEMA3B in the skeletal system have not been reported. Here, we show that osteoblast-derived SEMA3B alters global skeletal homeostasis in intact animals and osteoblast function in cell culture. Osteoblast-targeted expression of SEMA3B in mice resulted in reduced bone mineral density and aberrant trabecular structure compared with nontransgenic littermates. Histomorphometry studies indicated that this was likely due to increased osteoclast numbers and activity. Indeed, primary osteoblasts obtained from SEMA3B transgenic mice stimulated osteoclastogenesis to a greater extent than nontransgenic osteoblasts. This study establishes that SEMA3B is a 1,25(OH)2D3-induced gene in osteoblasts and that osteoblast-derived SEMA3B impacts skeletal biology in vitro and in vivo. Collectively, these studies support a putative role for SEMA3B as an osteoblast protein that regulates bone mass and skeletal homeostasis.
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Giuliani, Nicola, Francesca Morandi, Sara Tagliaferri, Mirca Lazzaretti, Sabrina Bonomini, Monica Crugnola, Cristina Mancini, et al. "The proteasome inhibitor bortezomib affects osteoblast differentiation in vitro and in vivo in multiple myeloma patients." Blood 110, no. 1 (July 1, 2007): 334–38. http://dx.doi.org/10.1182/blood-2006-11-059188.
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The proteasome inhibitor bortezomib may increase osteoblast-related markers in multiple myeloma (MM) patients; however, its potential osteoblastic stimulatory effect is not known. In this study, we show that bortezomib significantly induced a stimulatory effect on osteoblast markers in human mesenchymal cells without affecting the number of osteoblast progenitors in bone marrow cultures or the viability of mature osteoblasts. Consistently we found that bortezomib significantly increased the transcription factor Runx2/Cbfa1 activity in human osteoblast progenitors and osteoblasts without affecting nuclear and cytoplasmatic active β-catenin levels. Consequently a stimulatory effect of bortezomib on bone nodule formation was also demonstrated in osteoblast progenitors. These in vitro observations were confirmed in vivo by the finding of a significant increase in the number of osteoblastic cells × mm2 of bone tissue and in the number of Runx2/Cbfa1-positive osteoblastic cells that was observed in MM patients who responded to bortezomib. Our in vitro and in vivo observations support the hypothesis that a direct stimulatory effect on bone formation process could occur during bortezomib treatment.
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Ogata, Naoshi, Hiroshi Kawaguchi, Ung-il Chung, Sanford I. Roth, and Gino V. Segre. "Continuous Activation of Gαq in Osteoblasts Results in Osteopenia through Impaired Osteoblast Differentiation." Journal of Biological Chemistry 282, no. 49 (September 5, 2007): 35757–64. http://dx.doi.org/10.1074/jbc.m611902200.
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We explored the role of Gαq-mediated signaling on skeletal homeostasis by selectively expressing a constitutively active Gαq (mutation of Q209L) in osteoblasts. Continuous signaling via Gαq in mouse osteoblastic MC3T3-E1 cells impaired differentiation. Mice that expressed the constitutively active Gαq transgene in cells of the osteoblast lineage exhibited severe osteopenia in cortical and trabecular bones. Osteoblast number, bone volume, and trabecular thickness were reduced in transgenic mice, but the osteoclasts were unaffected. Osteoblasts from transgenic mice showed impaired differentiation and matrix formation. In the presence of a protein kinase C inhibitor GF109203X, this impairment was not seen, indicating mediation by the protein kinase C pathway. We propose that continuous activation of the Gαq signal in osteoblasts plays a crucial, previously unrecognized role in bone formation.
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Taichman, Russell S. "Blood and bone: two tissues whose fates are intertwined to create the hematopoietic stem-cell niche." Blood 105, no. 7 (April 1, 2005): 2631–39. http://dx.doi.org/10.1182/blood-2004-06-2480.
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AbstractThe mechanisms of bone and blood formation have traditionally been viewed as distinct, unrelated processes, but compelling evidence suggests that they are intertwined. Based on observations that hematopoietic precursors reside close to endosteal surfaces, it was hypothesized that osteoblasts play a central role in hematopoiesis, and it has been shown that osteoblasts produce many factors essential for the survival, renewal, and maturation of hematopoietic stem cells (HSCs). Preceding these observations are studies demonstrating that the disruption or perturbation of normal osteoblastic function has a profound and central role in defining the operational structure of the HSC niche. These observations provide a glimpse of the dimensions and ramifications of HSC-osteoblast interactions. Although more research is required to secure a broader grasp of the molecular mechanisms that govern blood and bone biology, the central role for osteoblasts in hematopoietic stem cell regulation is reviewed herein from the perspectives of (1) historical context; (2) the role of the osteoblast in supporting stem cell survival, proliferation, and maintenance; (3) the participation, if any, of osteoblasts in the creation of a stem cell niche; (4) the molecules that mediate HSC-osteoblast interactions; (5) the role of osteoblasts in stem cell transplantation; and (6) possible future directions for investigation.
Dissertations / Theses on the topic "Osteoblasts"
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Teixeira, Lucas Novaes 1981. "Estudo da expressão de osteopontina em sistemas de coculturas de células osteoblásticas e epiteliais neoplásicas humanas e seus efeitos sobre o fenótipo neoplásico e a ativação osteoclástica." [s.n.], 2015. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288466.
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Orientador: Paulo Tambasco de OliveiraTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: O carcinoma espinocelular (CEC) oral representa a neoplasia maligna mais prevalente das estruturas bucais, podendo invadir o tecido ósseo e promover sua reabsorção em até 56% dos casos. A expressão da proteína matricelular osteopontina (OPN) tem sido relacionada a uma maior agressividade de neoplasias malignas, incluindo o CEC oral. No tecido ósseo, a OPN representa a proteína mais abundante da matriz não colágena, concentrada nas interfaces ósseas, i.e. lâminas limitantes, linhas de cimentação e reversas, sendo essencial para adesão e funções de osteoblastos e osteoclastos. Apesar de no microambiente tumoral a OPN estar associada a um fenótipo neoplásico mais agressivo, ainda não está estabelecido o papel da OPN secretada por osteoblastos sobre células neoplásicas derivadas de CEC oral e o impacto sobre osteoclastos. O presente estudo teve como objetivos avaliar a expressão de OPN em sistemas de coculturas de células osteoblásticas e epiteliais neoplásicas malignas humanas e os efeitos da expressão de OPN secretada por osteoblastos sobre o fenótipo neoplásico in vitro. Adicionalmente, avaliou-se o efeito das coculturas sobre a atividade osteoclástica. Células epiteliais neoplásicas malignas derivadas de CEC oral (SCC9) foram plaqueadas sobre membranas de Transwell®, recobertas ou não por uma camada fina e uniforme de Matrigel, e cocultivadas com células osteoblásticas (SAOS-2) durante seu pico de expressão de OPN (10o dia de cultura). Células SCC9 expostas a culturas SAOS-2 silenciadas para OPN por RNA de interferência (RNAi) e células SCC9 cultivadas isoladamente foram usadas como controles. Após 24 h de cocultivo, células SCC9 foram avaliadas, quantitativamente, para adesão, proliferação, migração e invasão de Matrigel. A atividade de osteoclastos derivados de células monocíticas U-937 foi avaliada, quantitativamente, por meio dos ensaios de reabsorção de fosfato cálcio e de dosagem de citocinas em eluentes obtidos de células SCC9 e SAOS-2 após o cocultivo durante o pico de OPN ou com o seu silenciamento. A análise estatística foi realizada pelo teste não-paramétrico de Kruskal-Wallis (p < 0,05). Os resultados indicaram indução recíproca na expressão de OPN em SAOS-2 e SCC9 em cocultura. A OPN secretada por células SAOS-2 afetou o fenótipo de culturas SCC9, promovendo a adesão e a proliferação celulares e a invasão de Matrigel, a qual também estava aumentada, mas em menor intensidade, com o silenciamento para OPN. A migração celular não foi afetada. O cocultivo com SAOS-2, principalmente durante o pico de OPN, resultou na sobre-expressão das citocinas IL 6 e IL 8 pelas células SCC9, aumentando a capacidade de células osteoclásticas em reabsorver fosfato de cálcio. Conjuntamente, esses resultados sugerem que a OPN derivada de osteoblastos afeta as interações entre células epiteliais neoplásicas malignas, osteoblastos e osteoclastos, possivelmente contribuindo para a progressão de lesões ósseas do CEC oral
Abstract: The oral squamous cell carcinoma (OSCC) is the most prevalent malignant neoplasm of the oral structures. It may invade bone in up to 56% of the cases and promote osteoclast-mediated bone extracellular matrix (ECM) resorption. Expression of the matricellular protein osteopontin (OPN) in malignant neoplasms, including OSCC, has been positively correlated with aggressive tumor behavior. OPN is the most abundant non collagenous ECM protein in bone, where it preferentially accumulates at interfaces, including cement lines, laminae limitantes and reversal lines, being essential for the adhesion and function of osteoblasts and osteoclasts. Despite the importance attributed to OPN in the tumor microenvironment, indicative of more aggressive neoplastic phenotypes, the effects of osteoblast-derived OPN on OSCC cells and on OSCC-induced osteoclast activity are still not fully understood. The present in vitro study aimed to evaluate temporal expression of OPN in cocultures of human osteoblastic cells and malignant neoplastic epithelial cells and the effects of osteoblast-derived OPN on the neoplastic cell phenotype. Additionally, the effects of cocultures on osteoclastic activity were evaluated. Human OSCC-derived epithelial cells (SCC9 cell line) were plated on Transwell® membranes coated or not by a thin uniform layer of Matrigel and cocultured with human osteoblastic cells (SAOS-2 cell line) during its peak of OPN expression (day 10 of SAOS-2 culture). SCC9 cells exposed to OPN-silenced SAOS-2 cultures by means of interference RNA and SCC9 cells cultured alone were used as controls. At 24 h of coculture, SCC9 cells were quantitatively evaluated for cell adhesion, proliferation, migration and invasion of Matrigel. The impact of coculturing SCC9 and SAOS-2 cells either during the OPN peak expression or under the silencing of OPN was quantitatively evaluated in terms of calcium phosphate resorption by U-937-derived osteoclastic cells and expression of cytokines in the culture medium by ELISA assay. The statistical analyses were carried out using the non-parametric Kruskal-Wallis test (p < 0.05). The results showed a reciprocal induction of SAOS-2 and SCC9 cells in terms of OPN expression over the coculture interval. SAOS-2-secreted OPN altered the SCC9 cell phenotype, leading to enhanced cell adhesion and proliferation and higher Matrigel invasion, which was also enhanced, but to a lesser degree, by SAOS-2 cultures silenced for OPN. Cell migration was not affected. Cocultures with SAOS-2, mainly during the peak expression of OPN, resulted in overexpression of IL 6 and IL 8 by SCC9 cells, which corresponded with an enhanced resorptive capacity of osteoclastic cells. Taken together, the results suggest that osteoblast-derived OPN affects the interactions among malignant neoplastic epithelial cells, osteoblasts and osteoclasts, likely contributing to the progression of bone lesions in OSCC
Doutorado
Patologia
Doutor em Estomatopatologia
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Prele, Cecilia Marie Antoinette. "Vesicular trafficking in osteoblasts." Thesis, University College London (University of London), 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.271058.
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Bond, Alistair. "Purinergic signalling in osteoblasts." Thesis, University of Liverpool, 2012. http://livrepository.liverpool.ac.uk/11293/.
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ATP is now well established as an extracellular signalling molecule and has been shown to play a role in many different tissues including bone. ATP acting on P2 receptors has been reported to cause an increase in osteoblast proliferation, apoptosis, IL-6 production, AA release and a decrease in bone mineralization. It has also been shown in vivo, by using KO mice, that ATP can cause both an anabolic and catabolic response depending on the P2 receptor involved, with P2Y2 KO mice showing an increased bone formation whilst P2X7 mice have a decrease in the periosteal bone formation. ATP is released from osteoblasts in vitro basally, following trauma to the cell and in response to mechanical strain. This mechanical strain comes in the form of medium displacements, where the medium is taken up in a pipette and then immediately washed over the cells. Work carried out in this thesis looked to refine the experimental procedure of these tests in an attempt to reduce the variation of ATP supernatant measurements following medium displacement experiments. A new experimental procedure has now been implicated that ensures that the luciferase enzyme, used to measure ATP, is kept under constant conditions, the measurements from the wells are taken from the same area and basal levels are measured in each well as it was determined that ATP measurements are not consistent throughout a single well. Further to this a sensitive measure of cell death was developed in order to ensure that the observed ATP release was not due to cell trauma. This used qPCR to measure mtDNA present in the supernatant. Tests were also carried out to investigate the synergy between ATP and PTH as ATP synergises with PTH to increase the expression of c-fos in osteoblasts. c-fos is a master regulator gene that causes an increase in osteoblast proliferation and differentiation. It also causes the secretion of RANKL and decreases secretion of OPG, the RANKL decoy receptor, thus osteoclast fusion is also upregulated with an increase in c-fos production. Given that ATP is a local hormone that is released by mechanical stress and acts in an autocrine/paracrine manner and that PTH is an indiscriminate system wide hormone this synergy could be one mechanism whereby Wolff’s Law is achieved. That is, BMD is increased in areas of higher loading. The mechanism of this synergy is still to be elucidated and is likely to be different between cell types. Experiments on UMR-106 cells has shown that ATP and PTH can cause a potentiation of intracellular calcium release that ultimately leads to increased c-fos transcription. In SaOS-2 cells two separate signalling pathways converge on different areas of the c-fos promoter to cause an increase in c-fos. Work carried out for this project now shows for the first time that ATP sensitises osteoblasts to the action of PTH. Given the short half life of both molecules this increases the likelihood that they do play a significant role in bone remodelling in vivo. The mechanism behind this sensitisation appears to be independent of the known mechanisms described above, as it was shown that the sensitisation was conferred in the medium and was not due to ATP break down products ADP, AMP and adenosine. Thus it is likely another, longer lasting, paracrine factor is involved. BL-1249 is a putative K+ channel opener and inhibits ATP+PTH induced c-fos expression. The inhibition was shown not to be due to BL-1249s effect on K+ channels. It appears more likely that it acts on the cAMP/PKA pathway to inhibit c-fos transcription, as BL-1249 did not decrease FCS induced c-fos expression but did inhibit sp-cAMP induced c-fos expression. Further to this BL-1249 at concentrations of 100 μM caused a massive increase in cell death to the osteosarcoma cell lines SaOS-2, MG-63 and TE-85, however it had no effect on primary human osteoblasts. The apoptotic effect of BL-1249 is very likely due to the activation of K+ channels as it has been reported that opening K+ channels will cause apoptosis through K+ efflux and subsequent cell shrinkage. This was further confirmed by use of TEA, a K+ channel blocker, which acted to reverse the effect of BL-1249. Thus BL-1249 can act to decrease osteosarcoma cell number via inhibition of c-fos and, via K+ xiii channels, cause apoptosis. It is still unclear how apoptosis is targeted to osteosarcoma lines however it is likely through BL-1249s effect on c-fos expression. Occasionally the true effect of a gene or protein can only be elucidated when studying it in the context of a pathology. It has been reported that P2X7 can cause astrocytes to be toxic to motor neurones by secreting factors that contribute towards their death. In situations of increased oxidative stress, such as those found in SOD1 -/- mice, this effect can be increased and cause basal levels of ATP to activate the neurotoxic phenotype of the astrocytes. SOD1 -/- mice have also been examined for the bone and muscle phenotypes. They were shown to have a massive reduction in muscle mass with the peak difference being less than 50% their wild-type counterparts, their bones also had a decrease in BMD, a decreased length of their femur and had a decreased stiffness. Given this link between SOD1, P2 signalling and an altered bone phenotype SOD1-/- mice were examined in more detail. μCT scans were used to analyse the bone structure of these mice, however in this study no differences were observed. P2X4 was increased in SOD1-/- calvarial cells whilst P2Y2 was decreased in these cells suggesting that oxidative stress does cause an altered P2 receptor expression and this may change the BMD of bone.
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Vasconcelos, Daniel Fernando Pereira. "Efeito da administração intermitente do fragmento 1-34 do hormonio paratireoideo em defeito fenestrado na mandibula de ratos : analise histomorfometrica." [s.n.], 2008. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288498.
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Orientadores: Pedro Duarte Novaes, Marcelo Rocha MarquesTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: A administração intermitente de hormônio paratireóideo (PTH) é capaz de promover anabolismo ósseo, favorecendo a neoformação óssea em condições como osteoporose e reparo de fraturas. Sabendo-se que tanto os tecidos ósseos mandibulares e alveolares como os tecidos periodontais podem responder à administração de PTH de forma intermitente, o objetivo deste estudo foi avaliar histomorfometricamente o efeito da administração de PTH (1-34) em um modelo de defeito periodontal, do tipo fenestrado, na mandíbula de ratos. Foram utilizados neste estudo 32 ratos Wistar machos, com o objetivo de expor a região vestibular da raiz distal do primeiro molar inferior direito. A criação do defeito possibilitou a remoção parcial de ligamento periodontal e do cemento dentário. Após a cirurgia os animais foram separados aleatoriamente em 4 grupos (n=8): Grupo C14: administração intermitente (3 vezes por semana) do veículo de diluição do PTH por 14 dias; Grupo P14: administração intermitente de PTH (1-34) por 14 dias; Grupo C21: injeções intermitentes do veículo de diluição do PTH por 21 dias; Grupo P21: injeções intermitentes de PTH por 21 dias. Os animais foram mortos e preparados para avaliação histomorfométrica dos seguintes parâmetros: I - extensão inicial do defeito, II - extensão do defeito remanescente, III - área do defeito ósseo remanescente, IV - densidade do osso neoformado, V - área do calo formado e marcação para TRAP, VI - área do cemento neoformado, VII - análise do retardo óptico (birrefringência) do ligamento periodontal reinserido sobre a raiz operada. A análise intergrupo demonstrou que os defeitos tinham tamanhos similares inicialmente e que a administração de PTH reduziu significativamente (p<0,05) a extensão e a área do defeito remanescente. Além disso, o PTH aumentou significativamente (p<0,05) a densidade do osso neoformado, a área do calo, o número de osteoclastos presentes no calo formado, a área de cemento neoformado e a birrefringência do ligamento periodontal reinserido. Conclui-se que a administração intermitente de PTH (1-34), em modelo experimental de reparo em de ratos, pode influenciar positivamente o processo de reparação óssea e periodontal.
Abstract: The intermittent PTH administration is known to contribute to bone formation in cases of osteoporosis and bone fractures. Also, it has been reported to reduce periodontitis-related bone loss. The aim of this study was to evaluate the effect of anabolic PTH on periodontal repair and mandibular bone defect in rats. Fenestration defects were created unilaterally at the buccal surface of distal roots of lower first molars in Wistar rats (n=32), and both periodontal ligament and cementum were removed. Animals were then assigned to 4 groups (n=8): 1) C14 - placebo administration for 14 days; 2) P14 - PTH administration for 14 days; 3) C21 - placebo administration for 21 days; and 4) P21 - PTH administration for 21 days. All groups were treated intermittently (3 times a week). All animals were killed and prepared to histomorphometric analysis considering the following parameters: I - extension of the initial defect; II - extension of the remaining defect; III - area of the remaining defect; IV - density of neoformed bone; V - total callus area and staining for tartrate-resistant acidic phosphatase (TRAP); VI - area of the neoformed cementum; VII - polarized light microscopic analysis of root periodontal ligament reattachment. Intergroup analysis showed that the bone defects were initially similar in size (p>0.05). The intermittent PTH administration decreased (p<0.05) both extension and area of the remaining defect, and increased (p<0.05) the neoformed bone density and the total callus area. An increase in TRAP-positive cells was observed in the PTH treated groups (p<0.05). The area of the neoformed cementum and reattachment of the periodontal ligament were also observed to increase concerning PTH-treated groups (p<0.05). Data analysis suggests that the intermittent PTH administration might contribute to bone and periodontal repair in rats.
Doutorado
Histologia e Embriologia
Doutor em Biologia Buco-Dental
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Lage, Thais Claudino. "Análise in vitro da citotoxicidade em osteoblastos de dispositivos poliméricos incorporados com antimicrobianos para uso local." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/23/23147/tde-27112017-154525/.
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Os osteoblastos são células de origem mesenquimal envolvidas na formação óssea. Essas células podem sofrer alterações decorrentes de traumas, intervenções, e infecções. As infecções podem ser minimizadas a partir do uso de antimicrobianos. O poli(L-lactídeo) ou PLLA, é um polímero sintético que se destaca por sua biocompatibilidade e absorção, o qual pode ser utilizado como um liberador farmacológico local, como alternativa à terapêutica antimicrobiana sistêmica. Esse polímero também é empregado como matriz de suporte celular na engenharia de tecidos ósseos, por auxiliar na reparação e regeneração. A incorporação de partículas nesse polímero pode gerar reações adversas, portanto, devemos nos certificar que o dispositivo polimérico incorporado com antimicrobianos não seja citotóxico. Proposição: Analisar a estrutura e a citotoxidade em osteoblastos de dispositivos poliméricos de PLLA incorporados com antimicrobianos, sendo eles: Amoxicilina ou Azitromicina ou Clindamicina ou Metronidazol para uso local. Metodologia: Foram confeccionados 270 dispositivos poliméricos com 6mm de diâmetro composto de PLLA com a incorporação de antimicrobianos a 20%, Amoxicilina (AM) ou Azitromicina (AZ) ou Clindamicina (CL) ou Metronidazol (ME) sendo confeccionados através de dois métodos: eletrofiação (malhas) ou deposição (filmes). Posteriormente, foi realizado o teste de citotoxicidade direta MTT nesses dispositivos com a cultura de osteoblastos em 24, 48 e 72h de experimento. Para análise da estrutura do dispositivo, foram feitas análises macroscópicas através de fotografias digitais e microscópicas com Microscópio Eletrônico de Varredura (MEV). Resultados: A reação de citotoxidade mostrou que malhas e filmes incorporados à antimicrobianos são compatíveis com a cultura de osteoblastos, não apresentando citotoxicidade em nenhum momento do estudo (p<0.05). Na fotografia pudemos observar que os dispositivos apresentam coloração semelhante em relação às malhas e coloração diferenciada para filmes dependendo do tipo de antimicrobiano incorporado. No MEV, através da análise dos dispositivos pudemos notar que houve diferença no aspecto das superfícies dos filmes, sendo que os filmes de AM apresentaram aspecto irregular e poroso, enquanto AZ aparece liso com alguns grânulos, os de CL e ME possui superfícies ásperas e os de PLLA apresentaram superfície lisa. Quanto às malhas, notamos que todas as amostras apresentaram microfibras e poros que imitam a matriz extracelular, diferenciando-se apenas na espessura das fibras. Houve a presença de osteoblastos em todos os filmes confeccionados, mas os filmes de AM não induziram a proliferação, aparecendo apenas células isoladas. Enquanto nas malhas só foram observados osteoblastos em malhas de AM, ME e PLLA. Conclusão: Os dispositivos poliméricos confeccionados com PLLA incorporados com antimicrobianos podem ser usados na reparação e regeneração óssea uma vez que não apresentaram citotoxicidade em osteoblastos.Osteoblasts are mesenquima originated cells, which are involved in the bone formation. These cells may suffer alterations due to traumas, interventions and infeccions. The infections can be minimized by the handling of antimicrobials. Poly (L-lactide) or PLLA is a synthetic polymer known for its biocompatibility and absorption, which can be used as a local pharmacological releaser, as an alternative to the systemic antimicrobial therapy. This polymer also can be frequently used as a supporting structure to cellular matrix in the bone tissue engineering as it can be used for support in repair and regeneration. The particle incorporation in this polymer can create side effects, therefore, we need to certificate that the polymeric device incorporated with antimicrobials are not cytotoxic. Proposition: Analyse the structure and cytotoxicity in osteoblasts of PLLA polymeric devices associated with antimicrobials, being them: Amoxicillin, Azithromycin, Clindamycin and Metronidazole. Methods: For this study 270 polymerical devices were manufactured with 6mm diameter of PLLA with a 20% antimicrobials incorporation of Amoxicillin (AM), Azithromycin (AZ), Clindamycin (CL) and Metronidazole (ME) that have been produced through two methods: eletrospinning (mesh) or casting (film). Afterwards a MTT cytotoxicity test was made over the periods of 24, 48 and 72 hours of experiment. To make a structural analysis of the device a macroscopic analysis was performed through photographs and microscopic imaging with scanning electron microscope (SEM). Results: The cytotoxicity reaction exhibited that meshes and films incorporated with antimicrobials are comparable with the osteoblasts culture, indicating that there was no cytotoxicity in any moment (p < 0.05). In the phothograph we could observe that the devices showed a similar coloration among the meshes and different coloration for the films depending on the incorporated antimicrobial. The SEM analysis displayed a difference in the surface appearance of the films. The AM films displayed an irregular and porous appearance, meanwhile, AZ looked smooth with few grains, the CL and ME have rough surfaces and PLLA presents smooth surfaces. As for the meshes, we noticed that all the samples had microfibers and pores that mimic the extracellular matrix, differing only in the thickness of the fibers. Osteoblasts were present in all films but AM did not induce proliferation, with only isolated cells emerging. In meshes osteoblasts were only found in AM, ME and PLLA. Conclusion: Polymeric devices made with PLLA incorporated with antimicrobials can be used in bone repair and regeneration given that they did not offer cytotoxicity for osteoblasts.
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Valdivia, Maria Alejandra Medina. "Cultura e caracterização de células da granulação óssea in vitro: efeitos proliferativos estimulados por diferentes biomateriais." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/25/25146/tde-03092013-162521/.
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O objetivo deste estudo foi estabelecer cultura primária de células derivadas da granulação óssea (GO) de seres humanos para determinar seu padrão de crescimento in vitro e determinar os efeitos biológicos de três membranas reabsorvíveis feitas de colágeno (BioGide®, GenDerm®, CollaTape®) em culturas de fibroblastos gengivais humanos (FGH) e células da granulação óssea (GO). Foram coletadas amostras de tecido ósseo presente no alvéolo de cicatrização de dois pacientes adultos saudáveis sistemicamente com indicação de cirurgia periodontal regenerativa pela técnica do enxerto ósseo em neoformação. Imediatamente após a coleta, as amostras foram transportadas ao laboratório de cultura de células para estabelecimento da cultura primária. As células foram cultivadas em atmosfera úmida, contendo 5% CO2 a 37oC. A curva de crescimento das células foi determinada por meio de contagem de células viáveis. Após a caracterização da curva de crescimento, foram realizadas a caracterização da amostra por meio de determinação da atividade de fosfatase alcalina e de mineralização. Posteriormente, os efeitos de três diferentes tipos de membranas colágenas sobre a proliferação de células GO e FGH foram investigados por meio do teste MTT. As amostras foram divididas em oito grupos: (1) células FGH em meio DMEM (C-FGH); (2) células FGH em meio DMEM condicionado com membrana GenDerm (GD-FGH); (3) células FGH em meio DMEM condicionado com membrana BioGide (BG-FGH); (4) células FGH em meio DMEM condicionado com membrana ColaTape (CT-FGH); (5) células GO em meio DMEM (C-GO); (6) células GO em meio DMEM condicionado com membrana GenDerm (GD-GO); (7) células GO em meio DMEM condicionado com membrana BioGide (BG-GO); (8) células GO em meio DMEM condicionado com membrana CollaTape (CT-GO). O teste de proliferação celular mostrou que houve aumento significativo (p< 0.05; ANOVA para medidas repetidas) do número de células vitais presentes na cultura nos dias 3 (90,8%), 5 (132,50%), 7 (137,50%) e 10 (227,50%) em relação ao controle (dia 0). Foram observadas atividade de fosfatase alcalina e de mineralização in vitro. Houve aumento do número de células FGH e GO viáveis em todos os grupos (p< 0.05; ANOVA para medidas repetidas). Houve maior efeito proliferativo nas células FGH e GO nos grupos GD e CT, com diferenças estatisticamente significantes entre os grupos (p< 0.05; Mann Whitney) apenas no período de 96 horas. Esses achados sugerem que as células GO apresentam alta atividade de proliferação e síntese, sendo compatíveis com células de linhagem osteoblástica. Duas membranas colágenas testadas exerceram maior ação proliferativa tardia sobre osteoblastos, indicando sua eficácia na regeneração dos tecidos periodontais.The aim of this study was to establish primary culture of cells derived from human bone granulation tissue (GO) in order to determine its growth pattern in vitro and the biological effects of three absorbable collagen membranes (BioGide®, GenDerm®, CollaTape®) in human gingival fibroblasts (FGH) and human bone granulation (GO) cell cultures. Samples of bone tissue present at healing sockets of two systemically healthy adults with indication of periodontal regenerative therapy by the newly forming bone were collected. Immediately after, samples were transported to the laboratory of cell culture to the establishment of primary cultures. Cells were cultivated in humid atmosphere with 95% CO2 at 37oC. Cells growth pattern were determined by counting of viable cells. After characterization of growth pattern, samples were characterized according to alkaline phosphatase activity and mineralization detected by alizarin red. Afterwards, the effects of three different types of collagen membranes on GO and FGH cells were investigated by MTT test. Samples were divided into eight groups: (1) FGH cells in DMEM (C-FGH); (2) FGH in DMEM conditioned by GenDerm® membrane (GD-FGH); (3) FGH in DMEM conditioned with BioGide® (BG-FGH); (4) FGH in DMEM conditioned by CollaTape® (CT-FGH); (5) GO cells in DMEM (C-GO); (6) GO cells in DMEM conditioned by GenDerm® (GD-GO); (7) GO cells in DMEM conditioned by BioGide® (BG-GO); (8) GO cells in DMEM conditioned by CollaTape® (CT-GO). Cell proliferation test showed a significant (p< 0.05; ANOVA for repeated measures) increase in the number of vital cells present in the culture at days 3 (90.8%), 5 (132.50%), 7 (137.50%) and 10 (227.50%) compared to control (dia 0). It was observed alkaline phosphatase activity and mineralization in vitro. There was an increase in the number of FGH and GO viable cells at all groups (p< 0.05; ANOVA for repeated measures). Greater proliferative effect at FGH and GO cells at GD and CT groups, with significant differences between groups (p< 0.05; Mann Whitney) only at 96 hs. Two of the collagen membranes tested exerted greater late proliferative effects on osteoblasts, suggesting its efficacy in the regeneration of periodontal tissues.
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Mello, Daphne de Camargo Reis. "Interações biológicas e microbiológicas da liga Ti-35Nb-7Zr oxidadas e não oxidadas : estudo in vitro /." São José dos Campos, 2017. http://hdl.handle.net/11449/150556.
Full textAbstract:
Orientador: Luana Marotta Reis de VasconcellosBanca: Luciane Dias de Oliveira
Banca: Sandra Giacomin Schneider
Resumo: O objetivo neste estudo foi avaliar in vitro a influência da liga Ti-35Nb-7Zr e de seus elementos básicos, submetidos ou não ao processo de oxidação, na atividade de osteoblastos e na formação de biofilmes monotípicos. As amostras foram confeccionadas com diferentes materiais: a) titânio puro (Ti - controle); b) liga Ti-35Nb-7Zr (L); c) Nb (nióbio); d) Zr (zircônio); e) Ti oxidado (TiO); f) liga Ti-35Nb-7Zr oxidada (LO); g) Nb oxidado (NbO); h) Zr oxidado(ZrO). Previamente ao estudo in vitro, todas as amostras foram caracterizadas por microscopia eletrônica de varredura (MEV) e espectroscopia de dispersão de energia (EDS) para observar a topografia de superfície e a presença dos elementos básicos. Células mesenquimais obtidas de fêmures de rato, após diferenciação em osteoblastos foram cultivadas sobre as amostras. Após períodos pré-determinados, foram realizados os testes de citotoxicidade, atividade de fosfatase alcalina, produção de proteína total, formação e quantificação dos nódulos de mineralização adesão e proliferação celular. Para análise da formação dos biofilmes monotípicos, suspensões padronizadas (106 céls/mL) com micro-organismos de S. aureus, S. mutans, P. aeruginosa e C. albicans foram cultivados por 24 h sobre as amostras e submetidos ao teste de MTT. Todas as amostras permitiram o espraiamento celular. O Nb e Zr exibiram uma adesão celular mais madura com prolongamentos celulares evidenciados. O Ti e a L apresentaram maior viabilidade celular sendo estatísti... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The objective of this study was to evaluate in vitro the influence of the Ti-35Nb- 7Zr alloy and its basic elements, whether or not subjected to the oxidation process, on the osteoblast activity and on the formation of monotypic biofilms. The samples were made with different materials: a) pure titanium (Ti - control); B) Ti-35Nb-7Zr (A) alloy; C) Nb (niobium); D) Zr (zirconium); E) Oxidized Ti (TiO); F) oxidized Ti-35Nb-7Zr (AO); G) Nb oxidized (NbO); H) Oxidized Zr (ZrO). Prior to the in vitro study, all samples were characterized by scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) to observe the surface topography and the presence of the basic elements. Mesenchymal cells obtained from mouse femurs after differentiation into osteoblasts were cultured in the samples. After pre-determined periods, cytotoxicity tests, alkaline phosphatase activity, total protein production, formation and quantification of the mineralization nodules, adhesion and cell proliferation were performed. To analyze the formation of monotypic biofilms, standardized suspensions (106 cells / ml) with S. aureus, S. mutans, P. aeruginosa and C. albicans microorganisms were cultured for 24 h on the samples and submitted to the MTT test. All samples allowed for cell spreading. Nb and Zr exhibited more mature cell adhesion with evidenced cell extensions. The Ti and A presented greater cell viability being statistically different from the others (p <0.05). ZrO presented lower viability exhibiting statistical difference with Ti and A. The NbO expressed the highest amount of total protein with a statistical difference of L, Zr, and AO (p <0.05) while Zr showed the lowest result with a statistical difference of Ti, Nb, TiO, NbO, and ZrO. In the quantification of the alkaline phosphatase activity, the Zr presented the best result and was statistically different from all the others ..... (Resumo completo, clicar acesso eletrônico abaixo)
Mestre
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Bomfim, Fernando Russo Costa do [UNIFESP]. "Laser de baixa intensidade na expressão de genes ligados a mineralização óssea e ao cálcio em cultura de osteoblastos adultos." Universidade Federal de São Paulo (UNIFESP), 2014. http://repositorio.unifesp.br/handle/11600/23253.
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Previous issue date: 2014Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
A reducao ou minimizacao das consequencias das fraturas osseas e a regeneracao tecidual e objetivo de recursos clinicos e terapeuticos. O tecido osseo tem funcao mecanica, de deposito mineral e hematopoietica sendo composto por tres tipos celulares, osteoblastos, osteocitos e osteoclastos. Diversas substancias sao secretadas pelos osteoblastos, entre elas o calcio, que tem papel fundamental na homeostase celular e na producao do tecido osseo mineralizado e em cujo fluxo estao envolvidos os genes S100A6, PMCA1b e a osteocalcina ligados ao processo transporte e de mineralizacao. O objetivo deste trabalho foi avaliar os efeitos do laser de baixa intensidade, no que concerne a sinalizacao de calcio e mineralizacao ossea, na expressao dos genes S100A6, PMCA1b e osteocalcina em celulas osteoblasticas adultas. Trinta fragmentos mediais de femures com aproximadamente 5mm foram coletados cirurgicamente de ratos Wistar e distribuidos em dois grupos A (n=15, laser) e B (n=15, sem laser). Os fragmentos foram digeridos mecanica e enzimaticamente com colageno tipo II. Apos sete dias de cultura o grupo A foi irradiado com laser de baixa intensidade de Arseneto de Galio e Aluminio λ=808nm, 250mW de potencia nominal, densidade de potencia de 200mW/cm2, densidade de energia de 2000mJ/cm2, dose de energia de 2J/cm2, diametro de feixe de 0,02mm, tempo de 5s em 1 ponto de aplicacao durante 6 dias. Em seguida, o RNA foi extraido, quantificado, submetido a sintese de cDNA e realizada quantificacao de Ct comparativo pela Reacao em Cadeia da Polimerase em Tempo Real. Os valores foram submetidos a analise estatistica de teste Mann-Whitney com p<0,05. A analise em relacao a quantidade de RNA dos grupos A (mediana 3,80) e B (mediana 3,80) e os valores das medianas de ΔCt dos tres genes, S100A6 (A=2,22 e B=2,89), osteocalcina (A=4,06 e B=3,45) e PMCA1b (A=1,85 e B=4,16) nao mostraram diferencas significativas. O laser de baixa intensidade em osteoblastos adultos nao alterou a expressao dos genes S100A6, PMCA1b e osteocalcina descartando a relacao, no periodo estudado, destes genes com a mineralizacao e regeneracao ossea
The aim of clinical and therapeutic features is to reduce or minimize the consequences of fractures and bone regeneration. The bone has mechanical, mineral deposit and hematopoietic functions due to three types of cells, osteoblasts, osteocytes and osteoclasts. Several substances are secreted by osteoblasts, including calcium, which plays a fundamental role in cellular homeostasis and production of mineralized bone tissue and in whose flow are involved the S100A6, PMCA1b and osteocalcin genes related to the transport and mineralization process. This study was designed to evaluate the effects of low-level laser, when it comes to calcium signaling and bone mineralization, in S100A6, PMCA1b and osteocalcin genes expression in adult osteoblast cells. Thirty medial femoral fragments with approximately 5mm were surgically collected from Wistar rats and distributed into two groups A (n=15, laser) and B (n=15 without laser). The fragments were digested mechanical and enzymatically with type II collagen. After seven days of culture group A was irradiated with low intensity Gallium-Aluminum-Arsenide laser λ=808nm, 250mW nominal power, power density 200mW/cm2, energy density 2000mJ/cm2 dose of energy 2J/cm2 beam diameter 0,02 mm, length of 5s in one application point for 6 days. Then, RNA was extracted, quantified, underwent to cDNA synthesis and quantification performed by the comparative Ct by Real Time Polymerase Chain Reaction. The values were underwent to statistical analysis by Mann-Whitney test with p<0,05. The analysis for the amount of RNA of group A (median 3,80) and B (median 3,80) and ΔCt median values of the three genes, S100A6 (A=2.22 and B=2.89), osteocalcin (A=4.06 and B=3.45) and PMCA1b (A=1.85 and B=4.16) showed no significant differences. Low-level laser irradiation in adult osteoblasts did not modify the expression of S100A6, PMCA1b and osteocalcin genes discarding the relationship of these genes with the mineralization and bone regeneration in the studied period.
BV UNIFESP: Teses e dissertações
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McManus, Lindsay L. "A study of human mesenchymal stem cells, human primary osteoblasts and osteoblast-like cells using Raman spectroscopy." Thesis, University of Ulster, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.551188.
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Raman spectroscopy is a vibrational spectroscopy technique that provides a global biochemical signature and has been shown to have utility in the analysis of biological cells for bone tissue engineering applications. Traditionally, sample analysis in this field employs destructive biological methods that require the use of biomarkers, however, Raman has since become an essential tool in various areas of bio-industry and by incorporating the technique into biological laboratories these perturbing methodologies are no longer the only means of analysis. Therefore the focus of this study was to investigate the capability of Raman spectroscopy as a tool for the in vitro characterisation of the sub-cellular composition and osteogenic potential of human mesenchymal stem cells. As with most biological samples a high degree of heterogeneity is often found, therefore in order to extract the desired information from the biological studies multivariate analysis tools were utilised. The reliability and consistency of the vibrational analysis was confirmed by means of comparison with current gold-standard techniques such as, alizarin red staining, real-time polymerase chain reaction and immunocytochemistry. A further novelty was introduced with the use of Raman spectroscopy to determine the suitability of the U20S osteoblast-like cell line for use as a model for human primary osteoblasts with emphasis on the ability of these cell types to replicate their tissue of origin. Investigation of the U20S osteoblast-like cell line provided evidence of dense multilayered mineralised regions that corresponded more closely to native bone, which has not been previously reported on. This finding contradicts previous reports on U20S osteoblast-like cells which have consistently been described as non-osteoinductive. When Raman spectroscopy is coupled with biological and multivariate analyses techniques, it shows further novelty when employed to monitor mineralisation of human mesenchymal stem cells, human primary osteoblasts and osteoblast-like cells. This body of work culminates the success of a truly multidisciplinary approach and provides the potential for further work on bone cell analysis and the applications of spectroscopy for biological studies and bone tissue engineering applications.
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Hempel, Ute, Carolin Preissler, Sarah Vogel, Stephanie Möller, Vera Hintze, Jana Becher, Matthias Schnabelrauch, Martina Rauner, Lorenz C. Hofbauer, and Peter Dieter. "Artificial Extracellular Matrices with Oversulfated Glycosaminoglycan Derivatives Promote the Differentiation of Osteoblast-Precursor Cells and Premature Osteoblasts." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-165309.
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Sulfated glycosaminoglycans (GAG) are components of the bone marrow stem cell niche and to a minor extent of mature bone tissue with important functions in regulating stem cell lineage commitment and differentiation. We anticipated that artificial extracellular matrices (aECM) composed of collagen I and synthetically oversulfated GAG derivatives affect preferentially the differentiation of osteoblast-precursor cells and early osteoblasts. A set of gradually sulfated chondroitin sulfate and hyaluronan derivatives was used for the preparation of aECM. All these matrices were analysed with human bone marrow stromal cells to identify the most potent aECM and to determine the influence of the degree and position of sulfate groups and the kind of disaccharide units on the osteogenic differentiation. Oversulfated GAG derivatives with a sulfate group at the C-6 position of the N-acetylglycosamine revealed the most pronounced proosteogenic effect as determined by tissue nonspecific alkaline phosphatase activity and calcium deposition. A subset of the aECM was further analysed with different primary osteoblasts and cell lines reflecting different maturation stages to test whether the effect of sulfated GAG derivatives depends on the maturation status of the cells. It was shown that the proosteogenic effect of aECMwasmost prominent in early osteoblasts. [ABSTRACT FROM AUTHOR]
Books on the topic "Osteoblasts"
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Bland, Rosemary. Studies of thyroid hormone action in osteoblasts. Birmingham: University of Birmingham, 1996.
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Makinistoglu, Munevver. HDAC4 Integrate PTH and Sympathetic Signaling In Osteoblasts. [New York, N.Y.?]: [publisher not identified], 2014.
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Kung, Vanessa. Estrogen related receptor [alpha], ERR[alpha], regulation of target genes in osteoblasts. Ottawa: National Library of Canada, 2003.
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Compston, Juliet. Osteoporosis and bone biology: The state of the art. London: International Medical Press, 2000.
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Shelton, Richard Michael. The effect of substratum charge on the interfacial behaviour of osteoblasts in vitro. Birmingham: University of Birmingham, 1989.
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Mayeenuddin, Linnia H. Modulation of PTH-mediated signal transduction in osteoblasts by factors regulating cellular growth. Ottawa: National Library of Canada = Bibliothèque nationale du Canada, 1999.
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National Institute on Aging/National Institute of Dental Research Workshop on Human Models of Skeletal Aging (1994 Washington, D.C.). National Institute on Aging/National Institute of Dental Research Workshop on Human Models of Skeletal Aging: Washington, DC, March 1-2, 1994. Edited by Robey Pamela Gehron 1952-, Sherman Sherry, National Institute of Dental Research (U.S.), and National Institute on Aging. New York, NY: Springer International, 1995.
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Walker, Lesley Margaret. The effect of mechanical and hormonal stimuli on femur derived osteoblasts; intracellular calcium fluxes and calcium channels. Birmingham: University of Birmingham, 1998.
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Crothers, Kristina A. Effects of Tamoxifen on Prostaglandin E2 induced insulin-like growth factor I expression in fetal rat osteoblasts. [New Haven, Conn: s.n.], 1997.
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David, Evered, Harnett Sara, and Ciba Foundation, eds. Cell and molecular biology of vertebrate hard tissues. Chichester, UK: Wiley, 1988.
Find full textBook chapters on the topic "Osteoblasts"
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Rhodes, Steven D. "Osteoblasts." In Encyclopedia of Systems Biology, 1616–17. New York, NY: Springer New York, 2013. http://dx.doi.org/10.1007/978-1-4419-9863-7_881.
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Baak, Marleen A., Bernard Gutin, Kim A. Krawczewski Carhuatanta, Stephen C. Woods, Heinz W. Harbach, Megan M. Wenner, Nina S. Stachenfeld, et al. "Osteoblasts." In Encyclopedia of Exercise Medicine in Health and Disease, 671. Berlin, Heidelberg: Springer Berlin Heidelberg, 2012. http://dx.doi.org/10.1007/978-3-540-29807-6_2793.
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Pavelka, Margit, and Jürgen Roth. "Osteoblasts and Osteocytes." In Functional Ultrastructure, 296–97. Vienna: Springer Vienna, 2010. http://dx.doi.org/10.1007/978-3-211-99390-3_152.
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Martin, T. J., D. M. Findlay, J. K. Heath, and K. W. Ng. "Osteoblasts: Differentiation and Function." In Physiology and Pharmacology of Bone, 149–83. Berlin, Heidelberg: Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-77991-6_4.
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Franceschi, Renny T., Chunxi Ge, and Christopher G. Wilson. "Osteoblasts of Craniofacial Bone." In Mineralized Tissues in Oral and Craniofacial Science, 43–57. West Sussex, UK: John Wiley & Sons, Inc.,, 2013. http://dx.doi.org/10.1002/9781118704868.ch6.
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Perpétuo, Inês P., Lucie E. Bourne, and Isabel R. Orriss. "Isolation and Generation of Osteoblasts." In Methods in Molecular Biology, 21–38. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-8997-3_2.
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Yoshida, Takashi, Yuki Miyajima, and Minoru Wakamori. "Mechanosensitive TRP channels in osteoblasts." In Interface Oral Health Science 2009, 205–6. Tokyo: Springer Japan, 2010. http://dx.doi.org/10.1007/978-4-431-99644-6_49.
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Krauze, Michal T., and G. David Roodman. "Bortezomib and Osteoclasts and Osteoblasts." In Bortezomib in the Treatment of Multiple Myeloma, 43–52. Basel: Springer Basel, 2010. http://dx.doi.org/10.1007/978-3-7643-8948-2_3.
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Hughes-Fulford, Millie. "Growth and Repair of Mineralized Osteoblasts." In Spinal Instability, 3–19. New York, NY: Springer New York, 1993. http://dx.doi.org/10.1007/978-1-4613-9326-9_1.
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Gartland, Alison, Robin M. H. Rumney, Jane P. Dillon, and James A. Gallagher. "Isolation and Culture of Human Osteoblasts." In Methods in Molecular Biology, 337–55. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-367-7_22.
Full textConference papers on the topic "Osteoblasts"
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Li, K., Y. Xie, M. You, L. Huang, and X. Zheng. "Preparation and In Vitro Evaluation of Plasma Sprayed Cerium Oxide Coatings." In ITSC 2016, edited by A. Agarwal, G. Bolelli, A. Concustell, Y. C. Lau, A. McDonald, F. L. Toma, E. Turunen, and C. A. Widener. DVS Media GmbH, 2016. http://dx.doi.org/10.31399/asm.cp.itsc2016p0730.
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Abstract This work investigates the effects of cerium oxide (CeO2) coatings on the response of osteoblasts to H2O2-induced oxidative stress. The results show that the coatings have a protective effect, promoting both osteoblast growth and differentiation. This indicates that the CeO2 coating reduces the production of reactive oxygen species in H2O2-treated osteoblasts. These coatings, with their antioxidant properties, appear quite promising for bone regeneration.
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Tuan, Rocky S. "Functional Analysis of Bone-Biomaterial Interface." In ASME 2000 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2000. http://dx.doi.org/10.1115/imece2000-2675.
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Abstract Proper function and long-term stability of orthopaedic implants depend on the intimate association between bone cells and the implant biomaterial, a process known as osseointegration. Understanding the processes responsible for the establishment and maintenance of a functional bone-biomaterial interface and how these processes may be enhanced is crucial to the rational design and optimization of prosthetic devices. We have utilized cellular, molecular, and high-resolution imaging approaches to analyze the mechanistic basis of bone-biomaterial interactions. Specifically, we have characterized the initial adhesion of osteoblasts in terms of kinetics and relationship to the surface topography and chemistry of the biomaterials, particularly the cobalt-chrome and titanium alloys commonly used to fabricate orthopaedic prostheses. Results from these studies indicate that the long-term performance of osteoblasts adherent to biomaterials is crucially dependent on the characteristics of the initial adhesion step. Furthermore, osteoactive factors such as members of the transforming growth factor-β superfamily, including TGF-β1 and BMP-2, significantly enhance osteoblast cell adhesion. The molecular components responsible for the adhesion process include extracellular matrix proteins (e.g. fibronectin and collagen type I) and their cognate membrane receptors, the integrins. Our recent studies reveal that specific downstream, intracellular signaling events are also activated as a result of osteoblast adhesion, and that these signaling events are coupled to signal transduction mechanisms mediating growth factor activity. These events in combination regulate the continued expression and maintenance of the osteoblastic phenotype of the adherent cells, resulting in matrix maturation and mineralization, hallmarks of the bony tissue. Our current efforts focus on defining the target molecular pathways responsible for bone cell functioning on biomaterials, and the identification of critical biological and material parameters to optimize long-term osteoblast function and interaction with orthopaedically relevant biomaterials. The information gathered from these studies should provide a rational basis for the design of optimal implant biomaterials. (Supported in part by the NIH and the Annenberg Foundation)
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Cox, Lieke G. E., Corrinus C. van Donkelaar, Bert van Rietbergen, Rik Huiskes, and Keita Ito. "Osteocytes in Bone Can Regulate the Turnover of Adjacent Mineralized Growth Plate Cartilage." In ASME 2009 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2009. http://dx.doi.org/10.1115/sbc2009-206251.
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It is generally believed that bone remodeling is controlled by osteocytes, which act as mechanosensors and regulate the activity of osteoblast and osteoclast cells [1,2]. Osteocytes seem suitable for this function, since they are the most abundant cell type of bone and they form an extensive network of cellular processes by gap junction connections to each other, lining cells, and osteoblasts [1,3].
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Coughlin, Thomas R., Matthew Haugh, Muriel Voisin, Evelyn Birmingham, Laoise M. McNamara, and Glen L. Niebur. "Primary Cilia Knockdown Reduces the Number of Stromal Cells in Three Dimensional Ex Vivo Culture." In ASME 2013 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/sbc2013-14723.
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Mesenchymal stem cells (MSCs) are multipotent stromal cells that reside in the bone marrow and differentiate into connective cell lines, such as adipocytes and osteoblasts [1]. An appropriate balance of MSC differentiation toward adipocytes and osteoblasts is vital to bone homeostasis [6]. In vitro work demonstrates that differentiation of MSCs is influenced by mechanical stimuli [2, 3]. In a mouse model, the ratio of adipocytes to MSCs in the marrow was 19% lower compared to controls following treatment by low magnitude mechanical signals (LMMS) [4]. In mice, LMMS increased MSC number by 46% and the differentiation capacity of MSCs was biased towards osteoblastic compared to adipogenic differentiation [5]. Thus, mechanobiological stimuli may play an important role in maintaining balanced MSC differentiation.
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Liu, X., and C. Ding. "Cytocompatibility of Plasma Sprayed Bioceramic Coatings." In ITSC2005, edited by E. Lugscheider. Verlag für Schweißen und verwandte Verfahren DVS-Verlag GmbH, 2005. http://dx.doi.org/10.31399/asm.cp.itsc2005p0600.
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Abstract In this paper, the Al2O3, ZrO2, TiO2, wollastonite, dicalcium silicate and their composite coatings were deposited onto Ti-6Al-4V substrates by an atmosphere plasma spray system. The rat osteoblasts were seeded onto the surface of the coatings to evaluate the growth behavior of osteoblasts on different implant materials. Scanning electron microscopy was used to observe the morphologies of the osteoblasts seeded on the surface of the coating for different time. The SEM observation indicated the osteoblasts are difficult to survive and proliferate on the Al2O3 coating surface. Osteoblasts can survive and proliferate slowly on the TiO2 and ZrO2 coating surface. Osteoblasts grow and proliferate very well on the surface of the wollastonite and dicalcium silicate coating, which present the two coatings possess excellent cytocompatibility. The addition of wollastonite and dicalcium silicate into ZrO2 and TiO2 can improve their cytocompatibility.
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Lu, X. Lucas, Bo Huo, Andrew D. Baik, and X. Edward Guo. "Calcium Signaling in Bone Cell Networks Induced by Fluid Flow." In ASME 2009 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2009. http://dx.doi.org/10.1115/sbc2009-206043.
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Mechanical stimuli such as fluid flow can induce robust multiple intracellular calcium ([Ca2+]i) peaks in connected bone cell networks [1]. This fluid flow induced oscillation of [Ca2+]i can come from two sources: intracellular Ca2+ stores (e.g., endoplasmic reticulum, ER) and the extracellular environment. Moreover, [Ca2+]i signaling is mediated by various molecular pathways, such as IP3, ATP, PGE2, and NO. Osteocytes are believed to comprise a sensory network in bone tissue that monitors in vivo mechanical loading and triggers appropriate adaptive responses from osteoblasts and osteoclasts [2]. It is also well recognized that osteoblasts, the cells responsible for bone formation, can directly sense and respond to mechanical stimulation (e.g., fluid flow). In the present study, two types of cell networks were constructed in vitro with osteocyte-like and osteoblast-like cells, respectively, by using microcontact printing and self assembled monolayer (SAM) technologies. The calcium responses of the two types of cell networks to fluid flow were recorded, quantitatively analyzed, and compared. Then we examined how the [Ca2+]i response in the osteocyte cell network was influenced by gap junctions, intra/extracellular calcium sources, and other various molecular pathways.
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Horiguchi, Atsushi, and Toshihiko Shiraishi. "Study on a Cell Mechanosensing System by Measuring Structural Deformation and Biochemical Response." In ASME 2015 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2015. http://dx.doi.org/10.1115/imece2015-51456.
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Mechanical stimulation induces new bone formation in vivo and promotes the metabolic activity and the gene expression of osteoblasts in vitro. It was reported that biochemical signals of osteoblasts to sense mechanical stimulation are activated according to their actin cytoskeletal deformation. However, there have been not so many researches on the relationship between cytoskeletal deformation and biochemical response. Here we show an original method to investigate a cell mechanosensing system and the quantitative relationship between the deformation of cytoskeletal structure and the change of intracellular calcium ion concentration as biochemical response in a living cell stimulated by a micropipette. Gene transfection of green fluorescent protein to osteoblastic cells enabled visualization of actin in cells. When local deformation was applied to a single osteoblastic cell by a micropipette, the displacement distribution of cytoskeletal structure in the whole cell was automatically obtained from the two images of the cell before and after deformation by using Kanade-Lucas-Tomasi (KLT) method. Intracellular calcium ion response to mechanical stimulation was measured as the spatial and temporal changes of intensity of Fura Red loaded to a cell. As a result, we obtained the quantitative relationship between structural deformation and biochemical response of a cell and found that the change of calcium ion concentration increases with increasing the displacement of actin cytoskeleton. It indicates that the deformation of actin cytoskeleton is highly related to the cell mechanosensing system.
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Shiraishi, Toshihiko, Akinori Ishii, and Shin Morishita. "Effects of Mechanical Vibration on Multilayering of Cultured Osteoblasts." In ASME 2014 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2014. http://dx.doi.org/10.1115/imece2014-37731.
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This paper describes the mechanism of cell proliferation promotion by mechanical vibration focusing on multilayering of cultured osteoblasts. After osteoblasts were cultured under the mechanical vibration of 0.5 G and 12.5 Hz, the saturated cell density reached approximately twice as high as control. In the vibration group, multilayer formation of osteoblasts was observed by fluorescent microscopy in contrast with almost single layer in the control group. Fluorescent staining demonstrated that the expression of N-cadherin, which plays an important role of cell-cell adhesion, was lower under mechanical vibration than control. Therefore, the application of mechanical vibration to osteoblasts can downregulate the expression of N-cadherin, resulting in weakening of cell-cell adhesion and multilayer formation followed by promotion of cell proliferation.
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Song, Wang, Yang Yueqin, and Chen Guoqing. "Effect of mechanical stress on formation of osteoblast in vitro — A new way to culture osteoblasts." In 2009 ISECS International Colloquium on Computing, Communication, Control, and Management (CCCM). IEEE, 2009. http://dx.doi.org/10.1109/cccm.2009.5267860.
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Uemura, Toshimasa, Atsuko Nemoto, Jing An, Yin-kun Liu, Takafumi Yoshikawa, Hajime Ohgushi, Yoshinori Kuboki, Takashi Ushida, and Tetsuya Tateishi. "OSTEOPONTIN INVOLVEMENT IN OSTEOBLAST DIFFERENTIATION AND ITS EFFECT ON IN VIVO OSTEOGENIC POTENTIAL OF BONE MARROW DERIVED OSTEOBLASTS." In Proceedings of the 12th International Symposium on Ceramics in Medicine. WORLD SCIENTIFIC, 1999. http://dx.doi.org/10.1142/9789814291064_0064.
Full textReports on the topic "Osteoblasts"
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Gerstenfeld, Louis C. Mechanisms of Mechano-Transduction Within Osteoblasts. Fort Belvoir, VA: Defense Technical Information Center, September 2001. http://dx.doi.org/10.21236/ada415975.
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Gerstenfeld, Louis C. Mechanisms of Mechano-Transduction within Osteoblasts. Fort Belvoir, VA: Defense Technical Information Center, September 2000. http://dx.doi.org/10.21236/ada391000.
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Gerstenfeld, Louis. Mechanisms of Mechano-Transduction within Osteoblasts. Fort Belvoir, VA: Defense Technical Information Center, September 1999. http://dx.doi.org/10.21236/ada376489.
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Vélez, Rómulo Andrés, Alejandro Fereño Caceres, Wilson Daniel Bravo Torres, Daniela Astudillo Rubio, and Jacinto José Alvarado Cordero. Primary stability with the osseodensification drilling technique for dental implants in low density bone in humans: a systematic review. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, September 2022. http://dx.doi.org/10.37766/inplasy2022.9.0066.
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Review question / Objective: - Does the osseodensification drilling technique increase primary stability in low-density bone? - The aim of the present investigation was to evaluate primary stability in dental implants in people with low density bone using the osseodensification technique. Condition being studied: The replacement of missing teeth through dental implants is currently the most practiced in dental clinics. The main criterion for determining the success of an implant is osseointegration, which is a direct structural and functional connection between vital bone and the prosthetic load-bearing surface of an implant. In the same way, primary stability must be obtained for a good lasting clinical result of the implant and to achieve this purpose, the bone density must be evaluated where the dental implant is to be placed. Salah Huwais in 2013 introduced a new osteotomy procedure (Oseodensification) for site preparation without removal and bone preservation. The Osseodensification process produces an autograft layer around the implant with the osteotomy surface, the autologous bone comes into contact through an endosteal device that accelerates osseointegration due to the nucleation of osteoblasts in the instrumented bone adjacent to the implant and has a greater primary stability due to contact between the device and the bone.
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Mercer, Robyn R. Breast Cancer Metastasis to Bone Affects Osteoblast Differentiation. Fort Belvoir, VA: Defense Technical Information Center, May 2004. http://dx.doi.org/10.21236/ada426273.
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Mercer, Robyn R., and Andrea M. Mastro. Breast Cancer Metastasis to Bone Affects Osteoblast Differentiation. Fort Belvoir, VA: Defense Technical Information Center, May 2003. http://dx.doi.org/10.21236/ada416623.
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Karaplis, Andrew. Osteoblast-Derived PTHRP and Breast Cancer Bone Metastasis. Fort Belvoir, VA: Defense Technical Information Center, November 2004. http://dx.doi.org/10.21236/ada625302.
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Navone, Nora M. Osteoblast-Prostate Cancer Cell Interaction in Prostate Cancer Bone. Fort Belvoir, VA: Defense Technical Information Center, February 2000. http://dx.doi.org/10.21236/ada391088.
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Paranavitana, Chrysanthi. In Vitro Osteoblast Model for Bone Wound Infections and Antimicrobial Therapy. Fort Belvoir, VA: Defense Technical Information Center, January 2013. http://dx.doi.org/10.21236/ada608594.
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Sanders, Jennifer L. Actions of Tamoxifen and Estrogen on Osteoblast Protein Kinase C Expression. Fort Belvoir, VA: Defense Technical Information Center, July 1995. http://dx.doi.org/10.21236/ada306529.
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