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Estudo das características físico-químicas e biológicas pela adesão de osteoblastos em superfícies de titânio modificadas pela nitretação em plasma / Study of physical-chemical and osteoblast adhesion characteristics of titanium surfaces modified by plasma nitridingSilva, José Sandro Pereira da 27 January 2009 (has links)
INTRODUÇÃO: Superfícies de titânio modificadas por diferentes métodos foram estudadas com base nos parâmetros físicos e químicos de caracterização superficial e sua influência no comportamento de células pré-osteoblásticas (MC3T3) in vitro. MÉTODOS: Discos de titânio comercialmente puro grau II foram submetidos a três métodos de modificação de superfície (polimento, nitretados em plasma em configuração planar e gaiola catódica). As diferentes superfícies foram caracterizadas para observar o efeito do processamento na estrutura da camada superficial, na rugosidade e molhabilidade. Ensaios de adesão e proliferação celular usando linhagens de células pré-osteoblásticas MC3T3 foram realizados para avaliar o efeito das novas superfícies no comportamento celular in vitro. RESULTADOS: Os resultados demonstraram que a nitretação em plasma na configuração de gaiola catódica produz superfícies mais rugosas (p<0,02) e com menores ângulos de contato com a água. CONCLUSÕES: A adesão celular é maior nas superfícies mais rugosas do que nas superfícies polidas (p<0,05) e reagem de modo diferente a composição química do substrato e à topografia da superfície. / PURPOSE: The aim of this study was to evaluated the physico-chemical properties of different titanium surfaces modified by means of low temperature plasma nitridind on rat osteoblast cell adhesion and proliferation. METHODS: Pure Titanium discs grade II was submitted to three different surface preparations (polishing, glow discharge plasma nitriding in planar and cathodic cage configurations). Surface parameters as roughness, wettability and chemichal composition was determined to compare influency of gas mixture on the modified surface material properties. Cellular morphology was observed by scanning electron microscopy. To evaluate the effect of the surface on cellular response, osteoblast cells (MC3T3) adhesion and proliferation was quantified and data analised by Kruskal-Wallis and Friedman statistical tests. RESULTS: plasma nitriding discs shows rougher surfaces( p<0,02) in cathodic cage configuration and lower contact angle values. MC3T3 cells attached on rough surfaces produced by cathodic cage configuration was statistically significant p<0,05 compared to polished discs. CONCLUSIONS: Glow discharge plasma nitriding improve titanium surface roughness and wettability. MC3T3 cell adhesion behavior is related to substrate chemical composition and topography.
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Osteotropic cytokines : expression in human gingival fibroblasts and effects on bonePalmqvist, Py January 2006 (has links)
Bone metabolism is regulated by endocrine and paracrine signalling molecules influencing bone cells in the continuously remodelling bone tissue. These molecules include a variety of osteotropic stimulatory and inhibitory cytokines. Degradation of alveolar bone in periodontal disease is believed to be a result of local release of such osteotropic cytokines, although the relative importance of particular cytokines and their cellular origin is currently unknown. The aim of the present project was to study if, and how, pro-inflammatory cytokines in the interleukin-6 (IL-6) family of cytokines, and anti-inflammatory IL-4 and IL-13 type of cytokines, can affect osteoclast differentiation and bone resorption. Additionally, the objective was to study if gingival fibroblasts may influence alveolar bone resorption through secretion of IL-6 type cytokine release and if the secretion is regulated by pro-inflammatory as well as anti-inflammatory mediators such as IL-4 and IL-13. IL-6 in combination with its soluble receptor (sIL-6R) was found to stimulate mouse calvarial bone resorption. Similarly, two other IL-6 family members, leukemia inhibitory factor (LIF) and oncostatin M (OSM) were found to stimulate bone resorption. The stimulatory effect on bone resorption induced by the three cytokines was associated with increased expression of receptor activator of NF- κB ligand (RANKL), a cytokine which is essential in osteoclast formation and activation through binding to receptor activator of NF- κB (RANK) on osteoclastic cells. The interaction between RANKL and RANK can be inhibited by binding of the decoy receptor osteoprotegerin (OPG) to RANKL, and the expression of OPG was also regulated by IL-6, LIF and OSM (Paper I). The two related cytokines IL-4 and IL-13 were found to inhibit osteoclastogenesis and mouse calvarial bone resorption by mechanisms involving a decreased RANKL/OPG ratio in osteoblasts and decreased RANK expression in osteoclastic cells. The results further demonstrated that IL-4 and IL-13 exert their effects on both osteoblasts and osteoclasts by a mechanism involving the transcription factor signal transducer and activator of transcription 6 (STAT6) (Paper II). Constitutional expression of IL-6, LIF and another member of the IL-6 family of cytokines, IL-11, was demonstrated in human gingival fibroblasts. IL-6 type cytokine expression levels were found to be enhanced by IL-1β and tumour necrosis factor-α (TNF-α) (Paper III), whereas IL-4 and IL-13 inhibited IL 11 and LIF release from gingival fibroblasts (Paper IV). In conclusion, IL 6 type cytokines were found to be stimulators and IL-4 and IL-13 inhibitors of bone resorption in vitro via mechanisms involving RANK/RANKL/OPG interactions. Additionally, gingival fibroblasts were able to secrete several cytokines in the IL-6 family. Secretion was further enhanced by pro-inflammatory mediators and inhibited by IL-4 and IL- 13. These findings support the view that resident cells may influence the pathogenesis of periodontal disease through osteotropic cytokine production.
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The role of surface chemistry and wettability of microtextured titanium surfaces in osteoblast differentiationPark, Jung Hwa 11 May 2012 (has links)
Biomaterial surface energy, chemical composition, charge, wettability and roughness all play an important role in determining the degree of the direct bone-to-implant interface, termed osseointegration. Surface chemistry, which is influenced by surface energy, wettability, and composition, is another factor that determines osteoblast phenotype and regulates osteoblast maturation. Increased surface energy is desirable for bone implants due to enhanced interaction between the implant surface and the biological environment. The extent of bone formation in vivo is also increased with increasing water wettability of implants.
The physiological role of implant surface chemistry is important in determining the success of implant osseointegration because of molecular rearrangements, surface reactions, contamination, and release of toxic or biologically active ions that are determined by the starting chemistry. However, the role of surface chemistry on osteoblast response is not fully studied.
Therefore, the overall goal of this dissertation is to understand how the surface chemistry, including wettability, chemical composition, and charge density, of titanium biomaterials impacts osteoblast maturation (in vitro). This study focuses on the general hypothesis that modifications of surface chemistry of titanium surfaces with sterilization or polyelectrolyte coating on titanium surfaces regulate osteoblast response.
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Osteoblast Behaviour on Injectable Biomaterials Intended for Augmentation of Vertebral Compression FracturesRamstedt, Sandra January 2007 (has links)
Biomaterials used for stabilization of compressed vertebraes due to osteoporosis, have mainly been based on resin materials, like PMMA (polymethyl methacrylate), but have recently expanded to consist of injectable ceramics, such as calcium-aluminate. In this in vitro study human osteoblast-like cells, MG-63, were cultured on three different injectable biomaterials based on: Ca-aluminate, Bis-GMA (bisphenol A-glycidylmethacrylate) and PMMA, to investigate the cellular response elicited by these materials. Cell proliferation was measured by the NucleoCounter® system, cell viability was investigated by LDH (lactate dehydrogenase) analysis, cell differentiation and mineralization was evaluated by mRNA gene expression of the osteoblastic markers: ALP (alkaline phosphatase), OC (osteocalcin) and COLL-I (collagen type I) by qPCR (quantitative polymerase chain reaction) analysis. Two control materials were used: TCP (tissue culture polystyrene, negative control) and PVC (polyvinyl chloride, positive control). The results showed that all the bone cement materials were non-toxic and biocompatible, i.e. they provided good cell viability and proliferation of the MG-63 cells. They are specific for bone cells, since they expressed high values of the osteoblast-specific differentiation markers, and are thus promising as injectable bone cement materials. Among the bone cements, Xeraspine appears to be the most biocompatible material for bone cells. It is followed by Cortoss and then Vertebroplastic.
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Energy Restriction Effects on Estrogen Status and the Skeletal Response to LoadingSwift, Sibyl Nichole 2010 August 1900 (has links)
Moderate energy restriction in young, exercising women attenuates the positive
effects of exercise on bone density. Studies have shown that in the absence of adequate
levels of circulating estrogen, there may not be enough functional estrogen receptor-a
(ER-a) to respond adequately to loading. The experiment described in this document is
significant because this model has not been explored under conditions of energy
restriction (EnR) which are known to reduce circulating estrogen levels; it has been
tested only in ovariectomized animals. The central hypothesis of this research is that
reductions in estrogen due to EnR limit the ability of bone to respond to mechanical
loading (LOAD) through a down-regulation of ER-a.
Study one determined which nutrient’s (calcium or energy) restriction (-40 percent)
had the greatest negative effects on the skeletal integrity of exercising female rats and
whether exercise (EX) could mitigate these deleterious changes. EnR caused
detrimental effects in many of the structural properties of bone; however EX attenuated
losses in cancellous bone.
Study two ascertained whether EX maintained cancellous bone mass in female
rats subjected to graded EnR (-20 or -40 percent) and whether changes in endocrine factors
were related. EX preserved cancellous bone volume and osteoblast activity under both
levels of EnR, in addition to total body lean mass and bone mineral content. A similar
maintenance of serum insulin-like growth factor and estradiol occurred in the
EX EnR(40 percent) group suggesting that these changes may be related to the protective
effects of EX.
Study three determined the effects of 40 percent EnR on bone formation rate to LOAD
in young adult female rats and tracked alterations in ovarian function (estradiol).
Although higher than non-loaded animals, the response of bone to LOAD in EnR
animals was dampened in comparison to energy-replete animals.
The experiments described in this document are significant because these are the
first experiments to explore the relationship between EnR and estrogen levels on
cancellous bone response to LOAD. This is particularly important for physically active,
energy restricted women because cancellous bone in these women will not experience
the same effects of loading which can increase their risk for developing osteoporosis.
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Adverse effects of bone morphogenic protein-2 during osseointegrationHyzy, Sharon Leigh 21 May 2012 (has links)
Modifications of biomaterial surface properties are employed to increase osteoblast differentiation and bone formation. Microtextured metallic surfaces promote osteoblast differentiation and high surface energy- achieved by controlling surface hydrocarbon contamination- increases osteoblast differentiation and peri-implant bone formation. Recombinant human bone morphogenic protein 2 (BMP2) is approved to induce bone formation in a number of applications. It is used clinically in combination with biomaterials to improve peri-implant bone formation and osseointegration. The amount of BMP2 that is required is large and inflammatory (swelling/seroma) and bone-related (ectopic bone/bone resorption) complications have been reported after BMP2 treatment. The aim of this study was to examine potential deleterious effects of BMP2 on the inflammatory environment and apoptosis of osteoblasts.
Surface roughness and energy decreased pro-inflammatory interleukins and increased anti-inflammatory interleukins. In contrast, BMP2 abolished the surface effect, increasing pro-inflammatory interleukin (IL) 6, IL8, and IL17 in a surface roughness-dependent fashion and decreasing anti-inflammatory IL10 on rough surfaces. 5Z-7-Oxozeaenol and Dorsomorphin, but not H-8, blocked the effect of BMP2 on IL1A expression. There was an increase in expression of IL6 when treated with BMP2 for the control and H-8 groups, but both 5Z-7-Oxozeaenol and Dorsomorphin blocked the effect. Both 5Z-7-Oxozeaenol and H-8 blocked the effect of BMP2 on IL10 expression.
BMP2 treatment had little effect on apoptosis in human mesenchymal stem cells (MSCs). Exogenous BMP2 had no effect on TUNEL. Caspase-3 activity was increased only at 200ng/ml BMP2. BAX/BCL2 decreased in MSCs treated with 50 and 100ng/ml BMP2. In contrast, BMP2 increased caspase-3 activity and TUNEL at all doses in normal human osteoblasts (NHOst). BAX/BCL2 increased in NHOst treated with BMP2 in a dose-dependent manner. Cells treated with 200 ng/ml BMP2 had an 8-fold increase in BAX/BCL2 expression in comparison with untreated cells. Similarly, BMP2 increased DNA fragmentation in NHOst cells. The BMP2-induced increase in DNA fragmentation was eliminated by 5-Z7-Oxozeaenol and Dorsomorphin.
The results suggest that while surface features modulate an initial controlled inflammatory response, the addition of BMP2 induces a pro-inflammatory response. The effect of BMP2 on apoptosis depends on cell maturation state, inducing apoptosis in committed osteoblasts. BMP2 together with microtextured orthopaedic and dental implants may increase inflammation and possibly delay bone formation. Dose, location, and delivery strategies are important considerations in BMP2 as a therapeutic and must be optimized to minimize complications.
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Osteoblast Behaviour on Injectable Biomaterials Intended for Augmentation of Vertebral Compression FracturesRamstedt, Sandra January 2007 (has links)
<p>Biomaterials used for stabilization of compressed vertebraes due to osteoporosis, have mainly been based on resin materials, like PMMA (polymethyl methacrylate), but have recently expanded to consist of injectable ceramics, such as calcium-aluminate. In this in vitro study human osteoblast-like cells, MG-63, were cultured on three different injectable biomaterials based on: Ca-aluminate, Bis-GMA (bisphenol A-glycidylmethacrylate) and PMMA, to investigate the cellular response elicited by these materials. Cell proliferation was measured by the NucleoCounter® system, cell viability was investigated by LDH (lactate dehydrogenase) analysis, cell differentiation and mineralization was evaluated by mRNA gene expression of the osteoblastic markers: ALP (alkaline phosphatase), OC (osteocalcin) and COLL-I (collagen type I) by qPCR (quantitative polymerase chain reaction) analysis. Two control materials were used: TCP (tissue culture polystyrene, negative control) and PVC (polyvinyl chloride, positive control). The results showed that all the bone cement materials were non-toxic and biocompatible, i.e. they provided good cell viability and proliferation of the MG-63 cells. They are specific for bone cells, since they expressed high values of the osteoblast-specific differentiation markers, and are thus promising as injectable bone cement materials. Among the bone cements, Xeraspine appears to be the most biocompatible material for bone cells. It is followed by Cortoss and then Vertebroplastic.</p>
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Mechanisms of Hematopoietic-Mesenchymal Cell ActivationLemieux, Justin Michael 03 November 2009 (has links)
As the prevalence of osteoporosis is expected to increase over the next few decades, the development of novel therapeutic strategies to combat this disorder becomes clinically imperative. These efforts draw extensively from an expanding body of knowledge pertaining to the physiologic mechanisms of skeletal homeostasis. To this body of knowledge, we contribute that cells of hematopoietic lineage may play a crucial role in balancing osteoblastic bone formation against osteoclastic resorption. Specifically, our laboratory has previously demonstrated that megakaryocytes can induce osteoblast proliferation in vitro, but do so only when direct cell-to-cell contact is permitted. To further investigate the nature of this interaction, we have effectively neutralized several adhesion molecules known to function in the analogous interaction of megakaryocytes with another cell-type of mesenchymal origin - the fibroblast. Our findings implicate the involvement of fibronectin/RGD-binding integrins including á3â1 (VLA-3) and á5â1 (VLA-5) as well as glycoprotein IIb (CD41), all of which are known to be expressed on megakaryocyte membranes. Furthermore, we demonstrate that IL-3 can enhance megakaryocyte-induced osteoblast activation in vitro, as demonstrated in the megakaryocyte-fibroblast model system. Taken together, these results suggest that although their physiologic and clinical implications are very different, these two models of hematopoietic-mesenchymal cell activation are mechanistically analogous.
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Growth hormone signaling and action in osteoblastsDigirolamo, Douglas J. January 2008 (has links) (PDF)
Thesis (Ph.D.)--University of Alabama at Birmingham, 2008. / Title from PDF title page (viewed on Feb. 4, 2010). Includes bibliographical references.
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Design of synthetic peptides that display cell binding and signaling sequences on calcium phosphate surfaces /Gilbert, Michele. January 2001 (has links)
Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 180-209).
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