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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
81

Atividade de novos derivados de tiazolidinodionas sobre a diferenciação de pré-osteoblastos e pré-adipócitos / Activity of new derivatives of thiazolidinediones on differentiation of preosteoblasts and preadipocytes

Cristiane Akemi Iamamoto Saito 06 February 2015 (has links)
As tiazolidinodionas (TZDs) são sensibilizadores de insulina utilizados no tratamento do diabetes mellitus tipo 2. Contudo, apesar dos efeitos benéficos sobre a glicemia, importantes efeitos adversos incluindo perda óssea e aumento de adiposidade são relatados com o uso clínico das TZDs. Assim, é necessário o desenvolvimento de novos derivados de TZDs com potenciais efeitos benéficos sobre a hiperglicemia e menos efeitos adversos. Neste estudo, investigamos os efeitos de 5 novos derivados de TZDs (LYSO-7, GQ-89, GQ-150, GQ-177 e SF-3) sobre a diferenciação celular de pré-osteoblastos murinos MC3T3-E1, pré-adipócitos murinos 3T3-L1 e pré-adipócitos SGBS de linhagem humana. Seus potenciais efeitos sobre a utilização de glicose, a produção de adipocinas e mediadores pró-inflamatórios também foram avaliados, utilizando linhagens murinas e humanas de adipócitos, e macrófagos THP-1 de linhagem humana. O principal achado de nosso estudo foi que os novos derivados de TZDs estimulam a utilização celular de glicose, porém não alteram o processo de diferenciação celular de pré-osteoblastos e pré-adipócitos, quando comparados com a TZD clássica Rosiglitazona. Conforme esperado, o tratamento com Rosiglitazona na concentração de 5 μM inibiu a osteogênese de pré-osteoblastos murinos MC3T3-E1. No entanto, o tratamento com 2 novos derivados de TZDs (GQ-89 e GQ-177) na mesma concentração não afetou a diferenciação celular, sendo possível observar níveis de mineralização de matriz extracelular similares aos do grupo controle. Além disso, enquanto a GQ-89 estimulou a atividade da fosfatase alcalina, a GQ-177 não modulou sua atividade enzimática e induziu a expressão gênica de osteocalcina. Contudo, ambos inibiram a expressão de Runx2 e colágeno. Por sua vez, quando os efeitos foram avaliados sobre a diferenciação de adipócitos, foi possível observar que ao contrário do efeito pró-adipogênico constatado com a Rosiglitazona na concentração de 1 μM, as TZDs GQ-150, GQ-177, LYSO-7 e SF-3 foram incapazes de induzir o acúmulo lipídico em pré-adipócitos murinos e humanos. Além disso, a GQ-150 inibiu a expressão gênica de C/EBPα, assim como a expressão gênica e os níveis protéicos de CD36, enquanto que a SF-3 estimulou a expressão gênica de C/EBPα e de FABP4 e diminuiu a expressão gênica e os níveis protéicos de CD36, os quais não foram modificados pela LYSO-7 em pré-adipócitos murinos 3T3-L1. No entanto, em pré-adipócitos SGBS de linhagem humana, nenhum efeito sobre os marcadores de fenótipo adipogênico C/EBPα e FABP4 foi observado com os novos derivados de TZDs. Ademais, os novos derivados de TZDs não interferiram na via de sinalização de Wnt, não apresentaram qualquer efeito sobre a expressão de adipocinas (adiponectina, resistina e leptina) e mediadores pró-inflamatórios (IL-6, CCL2/MCP-1, TNF-α e JNK), bem como não ativaram o fator de transcrição PPARγ no ensaio de gene repórter. Por sua vez, a LYSO-7, GQ-150 e SF-3 aumentaram o consumo de glicose em presença de insulina em adipócitos 3T3-L1 e modificaram a atividade de enzimas mitocondriais em adipócitos SGBS e macrófagos THP-1. Entretanto, o efeito sensibilizador de insulina foi confirmado somente com a GQ-177 pelo aumento da captação de glicose e somente a LYSO-7 e a SF-3 foram capazes de inibir o consumo de oxigênio e modificar a taxa de glicólise em macrófagos, sugerindo que também poderiam alterar os níveis de ATP/ADP. Considerando que baixos níveis de ATP estimulam a via de sinalização de AMPK, essa via também foi investigada em nosso estudo. Entretanto, os resultados sobre a ativação de AMPK foram inconclusivos. Desse modo, nossos resultados apontam que os novos derivados de TZDs não atuam como ligantes de PPARγ, apresentam atividade sensibilizadora de insulina in vitro, e que exercem menores efeitos antiosteoblásticos e adipogênicos quando comparados com a Rosiglitazona. Mais estudos são necessários para elucidar os mecanismos responsáveis por esses efeitos, bem como para estabelecer se os novos derivados de TZDs são mais seguros in vivo, com relação ao risco de fraturas ósseas e ganho de massa adiposa. / Thiazolidinediones (TZDs) are insulin sensitizers used in the treatment of type 2 diabetes mellitus. However, despite the beneficial effects on blood glucose, significant adverse effects including bone loss and increased adiposity are reported with the clinical use of TZDs. Thus, it is necessary to develop new derivatives of TZDs with potential beneficial effects on hyperglycemia and fewer adverse effects. In this study, we investigated the effects of 5 new derivatives of TZDs (LYSO-7, GQ-89, GQ-150, GQ-177 e SF-3) on cellular differentiation in murine MC3T3-E1 preosteoblasts, murine 3T3-L1 preadipocytes, and SGBS preadipocytes from human lineage. Potential effects on glucose consumption, adipokines, and pro-inflammatory mediators were also assessed using murine and human strains of adipocytes, and macrophages from human THP-1 lineage. The main finding of this study was that new derivatives of TZDs stimulate glucose consumption, but do not change the cell differentiation process of preosteoblasts and preadipocytes compared to classical TZD Rosiglitazone. As expected, the treatmet with Rosiglitazone, at 5μM, inhibited the osteogenesis in murine MC3T3-E1 preosteoblasts. However, the treatment with 2 new derivatives of TZDs (GQ-89 and GQ-177) at the same concentration did not affect cell differentiation, and levels of mineralization of the extracellular matrix similar to the control group were observed. In addition, whereas the GQ-89 stimulated the activity of alkaline phosphatase, GQ-177 does not modulate its enzymatic activity and induced gene expression of osteocalcin. However, both of them inhibit the expression of Runx2 and collagen. In turn, when the effects were assessed on the adipocyte differentiation, unlike the proadipogenic effect observed with Rosiglitazone at a concentration of 1 μM, the new TZDs GQ-150, GQ-177, LYSO-7 and SF-3 were unable to induce lipid accumulation in human and murine preadipocytes. In addition, GQ-150 inhibited the gene expression of C/EBPα , as well as the gene expression and protein levels of CD36, whereas SF-3 stimulated the gene expression of C/EBPα and FABP4 and decreased gene expression and protein levels of CD36, which was not modified by LYSO-7 on murine 3T3- L1 preadipocytes. However, no effect on markers of adipogenic phenotype C/EBPα and FABP4 has been observed with the novel derivatives of TZDs in human SGBS preadipocytes. Furthermore, the new derivatives of TZDs do not interfere with the Wnt signaling pathway, showed no effect on the adipokines expression (adiponectin, resistin and leptin) and proinflammatory mediators (IL-6, CCL2 / MCP-1, TNF α and JNK) and did not activate the transcription factor PPARγ in the gene reporter assay. In turn, LYSO-7, GQ-150, and SF-3 increased glucose consumption in the presence of insulin in 3T3-L1 adipocytes and modified the activity of mitochondrial enzymes in SGBS adipocytes and THP-1 macrophages. However, the effect on insulin sensitization was confirmed only to GQ-177 that increased glucose uptake and just LYSO-7 and SF-3 were able to inhibit oxygen consumption and modify the rate of glycolysis in macrophages, suggesting that they could also alter the levels of ATP/ADP. Since low levels of ATP could stimulate AMPK pathway, this signaling pathway was also investigated in our study. However, the results on the AMPK activation were inconclusive. Thus, our results demonstrate that the new derivatives of TZDs do not act as PPARγ ligands, present insulin sensitizing activity in vitro, and display minor antiosteoblastic and adipogenic effects when compared to Rosiglitazone. More studies are needed to elucidate the exact mechanisms responsible for these effects, as well as to establish whether the safety of the new TZDs with respect to the risk of bone fractures and body mass gain using in vivo models.
82

Gewinnung und Charakterisierung von humanen Zementoblasten / Sourcing and characterisation of human cementoblasts

Bernhardt, Katharina 14 May 2018 (has links)
No description available.
83

Attachment, polarity and communication characteristics of bone cells

Ilvesaro, J. (Joanna) 26 March 2001 (has links)
Abstract Bone resorbing osteoclasts require tight attachment of their plasma membrane to the bone surface in order to retain the specific microenvironment and thus to be able to dissolve the bone matrix underneath. Cadherins are transmembrane glycoproteins usually mediating homophilic calcium-dependent cell-cell adhesion. In the present work we have studied the effects of the cadherin CAR sequence HAV-containing hexapeptide AHAVSE on osteoclasts. The primary attachment of osteoclasts to bone surface is not affected by the peptide, suggesting that it is not mediated by cadherins. Treatment of osteoclast cultures with AHAVSE decreased the number of resorption pits and the total resorbed area. Furthermore, we show rapid inactivation of osteoclasts with AHAVSE, which is seen as a decrease in the percentage of osteoclasts with actin rings. Pan-cadherin antibodies localized cadherin-like molecule in the sealing zone area of osteoclasts. These results suggest that cadherin-like molecules may mediate the tight attachment of osteoclasts in the sealing zone area and that the decrease of resorption in AHAVSE-treated osteoclast cultures is due to prevention of sealing zone formation. We studied the polarity of mesenchymal osteoblasts using osteosarcoma cell line UMR-108 and endosteal osteoblasts in situ in bone tissue cultures. Immunofluorescence confocal microscopy revealed that the vesicular stomatitis virus glycoprotein (VSV G) was targeted to the culture medium-facing surface. In endosteal osteoblasts, VSV G protein was found in the surface facing the bone marrow and circulation. On the contrary, Influenza virus hemagglutinin (HA) was localized to the bone substrate-facing surface of the UMR-108 cells. Electron microscopy showed that VSV particles were budding from the culture medium-facing surface, whereas Influenza viruses budded from the bone substrate-facing plasma membrane. These findings suggest the bone attaching plasma membrane of osteoblasts is apical, and the circulation or bone marrow facing plasma membrane is basolateral in nature. Gap junctions often mediate communication between different cells and cell types. In the present work, we demonstrate that rat osteoclasts show connexin-43 staining localizing in the plasma membrane of the cells in cell-cell contacts and over the basolateral membrane of osteoclasts. The effects of heptanol and Gap 27, known gap- junctional inhibitors, were studied using the well-characterized pit formation assay. The inhibitors decreased the number and activity of osteoclasts, suggesting a defect in the fusion of mononuclear osteoclast precursors to multinucleated mature osteoclasts. Furthermore, the total resorbed area and the number of resorption pits also decreased in the cultures. These results suggest that gap-junctional connexin-43 plays a functional role in osteoclasts, and that the blocking of gap junctions decreases both the number and the activity of osteoclasts.
84

Biocompatibility of orthopaedic implants on bone forming cells

Kapanen, A. (Anita) 22 February 2002 (has links)
Abstract Reindeer antler was studied for its possible use as a bone implant material. A molecular biological study showed that antler contains a growth factor promoting bone formation. Ectopic bone formation assay showed that antler is not an equally effective inducer as allogenic material. Ectopic bone formation assay was optimised for biocompatibility studies of orthopaedic NiTi implants. Ti-6Al-4V and stainless steel were used as reference materials. The assay showed differences in bone mineral densities, with superior qualities in NiTi. The rate of endochondral ossification varied between the implants, NiTi ossicles had larger cartilage and bone areas than ossicles of the two other materials. The cytocompatibility of NiTi was studied with three different methods. Cell viability, cell adhesion and TGF-β1 concentration were assessed in ROS-17/2.8 cell cultures. Cells grown on NiTi had better viability than cells grown on pure nickel or stainless steel. Cell attachment on the materials was studied with paxillin staining of focal contacts. The number of focal contacts was clearly higher in cells grown on NiTi than in cells grown on pure titanium, pure nickel or stainless steel. TGF-β1 concentration was measured with ELISA. The results showed that there was only some minor variation between NiTi, pure titanium and stainless steel. Nickel showed a lower TGF-β1 concentration. Taken together, these results suggest that NiTi is well tolerated by ROS-17/2.8 cells. The cytocompatibility of stainless steel is not so good as that of NiTi. The same tests were used to study the effects of the surface roughness of the implant on cytocompatibility. Three different surface roughness grades were compared in cell cultures on NiTi and titanium alloy discs. Titanium alloy was subjected to two different heat treatments, to compare the effects of the treatments on cytocompatibility. The studies showed that NiTi had a lesser impact on cell viability and attachment than titanium alloy. Further, rough NiTi was found to be a better tolerated surface than the others. In this study, heat treatment of titanium alloy at +850° C did not interfere with cell viability or attachment, as did the +1050° C treatment of the alloy. On the contrary, TGF-β1 concentrations decreased on the +850° C treated alloy and were approximately same on the +1050° C treated alloy and on NiTi.
85

Aspects of bone sugar biology:pectin nanocoatings of hard tissue implants

Kokkonen, H. (Hanna) 24 November 2009 (has links)
Abstract The improvement of implant biocompatibility is constantly under investigation. Titanium is a standard biomaterial that performs well in dental and orthopedic implantations. However, detrimental adverse effects resulting from e.g. biomaterial properties, inflammatory responses and surgical procedures occasionally occur. Coating the biomaterials aims at increasing the proportion of successful operations. Pectins, large plant cell wall polysaccharides, are innovative, modifiable, and potentially anti-inflammatory candidates for biomaterial nanocoatings. In this thesis, covalently-grafted pectin fragments (modified hairy regions, MHRs) modified either in vitro (from apple) or in vivo (from potato) were tested. Cell culture vessels and titanium substratum coated with the apple-MHRs, MHR-A and a further-tailored fragment type, MHR-B, were compared with controls for their ability to support proliferation and differentiation of osteoclasts and osteoblasts. Cells grew and differentiated on MHR-B and on the control surfaces; MHR-A did not perform well in these assays. Genetically-engineered potato MHRs did not support bone cell growth to the same extent as apple MHR-B, but nonetheless the possibility to manipulate cellular proliferation with specific in vivo – modifications of pectins was introduced. When implanted into rat soft tissues, neither of the apple MHRs provoked severe acute inflammatory reactions, which indicates good in vivo - tolerance of these botanical macromolecules. These studies illustrate the biocompatibility of MHRs, and the directions towards which they could be further tailored. In terms of clinical use, their tolerability in vivo is especially significant.
86

Biomimetické modifikace titanu v tkáňovém inženýrství kostí. / Biomimetic modifications of titanium in bone tissue engineering.

Krýslová, Markéta January 2015 (has links)
When the big joints like a knee or hip joint are damaged, the solution of this problem is an artificial substitute. The replacement of damaged joints with endoprotesis helps to reduce the pain and to move normally. In the design of the implant is necessary to fulfil all requirements on the properties of the material. The surface of implant is important, because it is directly connected to bone tissue. After implantation, the negative effect include infection, inflammation or release of the implant due to limited osseointegration, may appear. The osseointegration can be improved by modifying the material surface. This thesis is focused on development and evaluation of advanced materials imitating the bone structure, especially nanoroughness and the presence of biomimetic component, such as hydroxyapatite. In this study is evaluated adhesion, proliferation, viability, differentiation, and synthesis of specific proteins of human osteoblasts like Saos-2 on titanium modified with nanotubes and plasma sprayed hydroxyapatite compared with smooth surfaces. Key words: titanium, nanotubes, osteoblasts, hydroxyapatite, nanoroughness
87

Osteoblastic PLEKHO1 contributes to joint inflammation in rheumatoid arthritis

Dang, Lei 14 June 2019 (has links)
Background: Osteoblasts participating in the inflammation regulation gradually obtain concerns. However, its role in joint inflammation of rheumatoid arthritis (RA) is largely unknown. Here, we investigated the role of osteoblastic pleckstrin homology domain-containing family O member 1 (PLEKHO1), a negative regulator of osteogenic lineage activity, in regulating joint inflammation in RA. Methods: The level of osteoblastic PLEKHO1 in RA patients and collagen-induced arthritis (CIA) mice was examined. The role of osteoblastic PLEKHO1 in joint inflammation was evaluated by a CIA mice model which was induced in osteoblast-specific Plekho1 conditional knockout mice and mice expressing high Plekho1 exclusively in osteoblasts, respectively. The effect of osteoblastic PLEKHO1 inhibition was explored in a CIA mice model. The mechanism of osteoblastic PLEKHO1 in regulating joint inflammation was performed by a series of in vitro studies. Results: PLEKHO1 was highly expressed in osteoblasts from RA patients and CIA mice. Osteoblastic Plekho1 deletion ameliorated joint inflammation, whereas overexpressing Plekho1 only within osteoblasts exacerbated local inflammation in CIA mice. PLEKHO1 was required for TNF receptor-associated factor 2 (TRAF2)-mediated the ubiquitination of receptor-interacting serine/threonine-protein kinase 1 (RIP1) to activate nuclear factor kappa-light-chain-enhancer of activated B (NF-kB) pathway for inducing inflammatory cytokines production in osteoblasts. Moreover, osteoblastic PLEKHO1 inhibition improved joint inflammation and attenuated bone formation reduction in CIA mice. Conclusions: These data strongly suggest that highly expressed PLEKHO1 in osteoblasts mediates joint inflammation in RA. Targeting osteoblastic PLEKHO1 may exert dual therapeutic action of alleviating joint inflammation and promoting bone repair in RA.
88

Factor inhibiting ATF4-mediated transcription is a novel leucine zipper transcriptional repressor that regulates bone mass

Yu, Vionnie Wing Chi. January 2007 (has links)
No description available.
89

In Vitro Growth of Osteoblasts on Poly Lactic-Co-Glycolic Acid Scaffolds Created via Gas Foaming

Thomas, Matthew James 01 September 2018 (has links) (PDF)
This study analyzed the feasibility of using gas foaming to create Poly Lactic-co-Glycolic Acid (PLGA) scaffolds for use as a substrate in bone tissue engineering and set out to determine whether the presence of osteoblasts on these scaffolds enhanced their material stiffness. The process of bone formation involves osteoblasts depositing extracellular matrix and calcifying this matrix with calcium phosphate crystals (Hasegawa et al., 2017) and pits between 30-40μm in diameter on tissue engineering scaffold surfaces have been shown to best promote osteogenic activity in the presence of bone-forming cells (Halai et al., 2014).The scaffolds were determined to contain pits within this 30-40μm range and the ability of osteoblasts to lay down and calcify extracellular matrix on gas foamed PLGA scaffolds was confirmed by the image analysis of inverted optical microscope images of Alizarin Red S-stained scaffold cryosectionsThe presence of osteogenic activity combined with the desired scaffold porosity led us to conclude that gas foaming PLGA scaffolds are a feasible method of scaffold fabrication for bone tissue engineering and allowed us to optimize the gas foaming apparatus as an instrument to be used in further bone tissue engineering experiments at California Polytechnic State University, San Luis Obispo.However, this study failed to determine whether the presence of osteoblasts improved the material stiffness of the PLGA scaffolds due to a lack of statistical significance in compression testing results.
90

Effect of radiofrequency glow discharge on proliferation and osteogenic behavior of normal human osteoblasts

Elbadawi, Lena 28 September 2016 (has links)
BACKGROUND: Implants have been widely used in the medical field. It was adopted in dentistry, offering patients replacement of missing teeth. Researchers have been investigating techniques to improve implants’ survival. Among these techniques is plasma glow discharge. Radio-frequency Glow discharge (RGD) is a surface treatment and sterilization technique with the aim to improve the titanium oxide layer for better osseointegration. Previous studies have evaluated its effect on non-human cell lines with promising results. Up to date, there is no report on how RGD surface treatment of titanium affects normal human osteoblasts. MATERIAL AND METHODS: Human bone fragments were obtained from dental extraction sites and were processed to culture normal human osteoblasts. Cells were seeded on three different surfaces at a concentration of 1x105 cells per plate; Titanium discs with and without Argon RGD (ARGD), and tissue culture plates (TCP). Dishes were allocated to 3 timelines: 16 hours, 7 days and 14 days. The outcome measures were cell attachment, cell number, alkaline phosphatase and osteocalcin levels. RESULTS: Data was analyzed using a one-way ANOVA test. Mean cell proliferation percentage for the ARGD group at 7 days was the highest (167.966%). The difference in means among the three groups at 7 days was statistically significant (p=0.0022). At 14 days, the highest mean of cell proliferation percentage was highest for the ARGD group. When testing all pairs, at 7 days the differences in means were statistically significant between (ARGD vs. no ARGD, and ARGD vs. TCP) (p=0.0018, and p=0.0286), respectively. At 14 days, the differences in means were statistically significant between (ARGD vs. TCP, p= 0.0003) and (no ARGD vs. TCP, p=0.0007). There was a significant difference in means for alkaline phosphatase and osteocalcin at 7 and 14 days between TCP and ARGD, and TCP and no ARGD groups (p < 0.05). CONCLUSIONS: The results of this study on normal human osteoblasts indicated that ARGD significantly enhanced cell proliferation. There was no significant difference in osteogenic behavior between with and without ARGD treatment on titanium surfaces within the time studied. A prolonged phase of cell proliferation was observed in ARGD treated groups.

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