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Cell Attachment (cell + attachment)
Selected AbstractsEffect of Silicate-Substitution on Attachment and Early Development of Human Osteoblast-Like Cells Seeded on Microporous Hydroxyapatite Discs,ADVANCED ENGINEERING MATERIALS, Issue 1-2 2010Katharina Guth Hydroxyapatite (HA) is a well-established graft material used in bone repair. Silicon-substituted hydroxyapatite (SA; 0.8,wt% Si) has shown greater bone ingrowth and bone coverage than phase pure HA. To assess the effect of microporosity on sensitivity of cell attachment to surface physiochemistry, microporous SA and HA discs, and control Thermanox (TMX) discs were incubated with osteoblast-like cells (5,×,104 HOS-TE85 cells) under differing tissue culture conditions. To investigate early cellular attachment, organization, and differentiation, cells were also stained for integrin,,5,1, actin, and runt-related transcription factor (RUNX-2), respectively, after incubation on HA, SA, and TMX discs for 3 days. No significant differences emerged between HA, SA, and TMX discs in mean numbers of cells attached in serum free medium (SFM) over 90,min incubation. In contrast, significantly more cells were attached to SA than HA after 180,min incubation in complete medium (C-MEM) containing fetal calf serum (p,<,0.05). Cell attachment to SA and HA discs pre-conditioned in SFM supplemented with fibronectin (FN) was lower than discs pre-conditioned in C-MEM, suggesting sensitivity of an active FN conformation to the presence of co-adsorbates. Confocal microscopy demonstrated significantly more co-localization of integrin ,5,1 and actin on SA than HA. Translocalization of RUNX-2 to the nucleus was stronger in cells incubated on SA. Microporosity did not diminish the effect of surface physiochemistry on cell adhesion, and enhanced cell attachment for SA appears to be mediated by differences in the quality of adsorbed protein rather than via direct effects of substrate chemistry. [source] Polypyrrole Thin Films Formed by Admicellar Polymerization Support the Osteogenic Differentiation of Mesenchymal Stem CellsMACROMOLECULAR BIOSCIENCE, Issue 8 2004Harold Castano Abstract Summary: The objective of this study was to evaluate the attachment, proliferation, and differentiation of rat mesenchymal stem cells (MSC) toward the osteoblastic phenotype seeded on polypyrrole (PPy) thin films made by admicellar polymerization. Three different concentrations of pyrrole (Py) monomer (20, 35, and 50,×,10,3M) were used with the PPy films deposited on tissue culture polystyrene dishes (TCP). Regular TCP dishes and PPy polymerized on TCP by chemical polymerization without surfactant using 5,×,10,3M Py, were used as controls. Rat MSC were seeded on these surfaces and cultured for up to 20 d in osteogenic media. Surface topography was characterized by atomic force microscopy, X-ray photoelectron spectroscopy, and static contact angle. Cell attachment, proliferation, alkaline phosphatase (ALP) activity, and calcium content were measured to evaluate the ability of MSC to adhere and differentiate on PPy-coated TCP. Increased monomer concentrations resulted in PPy films of increased thickness and surface roughness. PPy films generated by different monomer concentrations induced drastically different cellular events. A wide spectrum of cell attachment characteristics (from excellent cell attachment to the complete inability to adhere) were obtained by varying the monomer concentration from 20 m to 50,×,10,3M. In particular the 20,×,10,3M PPy thin films demonstrated superior induction of MSC osteogenicity, which was comparable to standard TCP dishes, unlike PPy films of similar thickness prepared by chemical polymerization without surfactant. Adhesion of mesenchymal stem cells on tissue culture plates (TCP) coated with polypyrrole thin films made by admicellar polymerization. [source] Accurate control of oxygen level in cells during culture on silicone rubber membranes with application to stem cell differentiationBIOTECHNOLOGY PROGRESS, Issue 3 2010Daryl E. Powers Abstract Oxygen level in mammalian cell culture is often controlled by placing culture vessels in humidified incubators with a defined gas phase partial pressure of oxygen (pO2gas). Because the cells are consuming oxygen supplied by diffusion, a difference between pO2gas and that experienced by the cells (pO2cell) arises, which is maximal when cells are cultured in vessels with little or no oxygen permeability. Here, we demonstrate theoretically that highly oxygen-permeable silicone rubber membranes can be used to control pO2cell during culture of cells in monolayers and aggregates much more accurately and can achieve more rapid transient response following a disturbance than on polystyrene and fluorinated ethylene-propylene copolymer membranes. Cell attachment on silicone rubber was achieved by physical adsorption of fibronectin or Matrigel. We use these membranes for the differentiation of mouse embryonic stem cells to cardiomyocytes and compare the results with culture on polystyrene or on silicone rubber on top of polystyrene. The fraction of cells that are cardiomyocyte-like increases with decreasing pO2 only when using oxygen-permeable silicone membrane-based dishs, which contract on silicone rubber but not polystyrene. The high permeability of silicone rubber results in pO2cell being equal to pO2gas at the tissue-membrane interface. This, together with geometric information from histological sections, facilitates development of a model from which the pO2 distribution within the resulting aggregates is computed. Silicone rubber membranes have significant advantages over polystyrene in controlling pO2cell, and these results suggest they are a valuable tool for investigating pO2 effects in many applications, such as stem cell differentiation. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] Influence of modifying and veneering the surface of ceramic abutments on cellular attachment and proliferationCLINICAL ORAL IMPLANTS RESEARCH, Issue 11 2008Kamal Mustafa Abstract Objectives: This in vitro study was aimed to investigate the attachment, spreading and proliferation of human gingival fibroblasts to milled and polished non-veneered ceramic surfaces in alumina and zirconia and to ceramic surfaces veneered by two different types of porcelain baseliners. Materials and methods: Fibroblasts were cultured on discs of pressed alumina or zirconia, on discs which had been milled, on discs comprising alumina or zirconia which had been polished, on discs of alumina veneered with NobelRondo baseliner Al, on discs of zirconia veneered with Cercon-S baseliner, and on alumina or zirconia discs veneered with the above baseliners and then polished. The surfaces were analyzed using an optical interferometer and scanning electron microscopy (SEM). Cell profile areas were measured using SEM and an image analyzer. Cell attachment was determined after 3 and 24 h as a ratio of the cell profiles and the total micrograph area and was expressed as percent of attachment. MTT analyses were undertaken to determine cellular attachment after 3 h of incubation and cellular proliferation after 7 days. Results: The polished zirconia specimens had the smoothest surface in terms of average height deviation (Sa=0.03 ,m): the roughest were the zirconia specimens with milled surfaces (Sa=0.36 ,m). The application of the baseliners resulted in surfaces smoother than those of the non-veneered discs. The milled surfaces of both alumina and zirconia had significantly higher percentages of cell attachment and proliferation than the other surfaces whereas the milled surfaces in zirconia demonstrated better cellular attachment after 3 and 24 h of culture than the one in alumina. Fibroblasts attached and grew effectively on the surfaces veneered with NobelRondo throughout the experiments, whereas the zirconia surfaces veneered with Cercon-S had the lowest percentage of cell attachment and proliferation. Conclusions: Although the roughness of all surfaces investigated was <0.4 ,m, the study disclosed significant differences in cellular attachment and proliferation associated with the various surface modifications. [source] Vinculin is proteolyzed by calpain during platelet aggregation: 95 kDa cleavage fragment associates with the platelet cytoskeletonCYTOSKELETON, Issue 4 2004Katherine Serrano Abstract The focal adhesion protein vinculin contributes to cell attachment and spreading through strengthening of mechanical interactions between cell cytoskeletal proteins and surface membrane glycoproteins. To investigate whether vinculin proteolysis plays a role in the influence vinculin exerts on the cytoskeleton, we studied the fate of vinculin in activated and aggregating platelets by Western blot analysis of the platelet lysate and the cytoskeletal fractions of differentially activated platelets. Vinculin was proteolyzed into at least three fragments (the major one being ,95 kDa) within 5 min of platelet activation with thrombin or calcium ionophore. The 95 kDa vinculin fragment shifted cellular compartments from the membrane skeletal fraction to the cortical cytoskeletal fraction of lysed platelets in a platelet aggregation-dependent manner. Vinculin cleavage was inhibited by calpeptin and E64d, indicating that the enzyme responsible for vinculin proteolysis is calpain. These calpain inhibitors also inhibited the translocation of full-length vinculin to the cytoskeleton. We conclude that cleavage of vinculin and association of vinculin cleavage fragment(s) with the platelet cytoskeleton is an activation response that may be important in the cytoskeletal remodeling of aggregating platelets. Cell Motil. Cytoskeleton 58:242,252, 2004. © 2004 Wiley-Liss, Inc. [source] Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteinsCYTOSKELETON, Issue 3 2001Nathalie F. Worth Abstract Smooth muscle cells (SMC) exhibit a functional plasticity, modulating from the mature phenotype in which the primary function is contraction, to a less differentiated state with increased capacities for motility, protein synthesis, and proliferation. The present study determined, using Western analysis, double-label immunofluorescence and confocal microscopy, whether changes in phenotypic expression of rabbit aortic SMC in culture could be correlated with alterations in expression and distribution of structural proteins. "Contractile" state SMC (days 1 and 3 of primary culture) showed distinct sorting of proteins into subcellular domains, consistent with the theory that the SMC structural machinery is compartmentalised within the cell. Proteins specialised for contraction (,-SM actin, SM-MHC, and calponin) were highly expressed in these cells and concentrated in the upper central region of the cell. Vimentin was confined to the body of the cell, providing support for the contractile apparatus but not co-localising with it. In line with its role in cell attachment and motility, ,-NM actin was localised to the cell periphery and basal cortex. The dense body protein ,-actinin was concentrated at the cell periphery, possibly stabilising both contractile and motile apparatus. Vinculin-containing focal adhesions were well developed, indicating the cells' strong adhesion to substrate. In "synthetic" state SMC (passages 2,3 of culture), there was decreased expression of contractile and adhesion (vinculin) proteins with a concomitant increase in cytoskeletal proteins (,-non-muscle [NM] actin and vimentin). These quantitative changes in structural proteins were associated with dramatic changes in their distribution. The distinct compartmentalisation of structural proteins observed in "contractile" state SMC was no longer obvious, with proteins more evenly distributed throughout the cytoplasm to accommodate altered cell function. Thus, SMC phenotypic modulation involves not only quantitative changes in contractile and cytoskeletal proteins, but also reorganisation of these proteins. Since the cytoskeleton acts as a spatial regulator of intracellular signalling, reorganisation of the cytoskeleton may lead to realignment of signalling molecules, which, in turn, may mediate the changes in function associated with SMC phenotypic modulation. Cell Motil. Cytoskeleton 49:130,145, 2001. © 2001 Wiley-Liss, Inc. [source] Homocysteine enhances cardiac neural crest cell attachment in vitro by increasing intracellular calcium levelsDEVELOPMENTAL DYNAMICS, Issue 8 2008David J. Heidenreich Abstract Elevated homocysteine (Hcys) increases the risk of neurocristopathies. Previous studies show Hcys inhibits neural crest (NC) cell migration in vivo. However, the mechanisms responsible for this effect are unknown. Here, we evaluated the effect of Hcys on NC cell attachment in vitro and determined if any of the effects were due to altered Ca2+ signaling. We found Hcys enhanced NC cell attachment in a dose and substrate-dependent manner. Ionomycin mimicked the effect of Hcys while BAPTA-AM and 2-APB blocked the effect of Hcys on NC attachment. In contrast, inhibitors of plasma membrane Ca2+ channels had no effect on NC attachment. Hcys also increased the emission of the intracellular Ca2+ -sensitive probe, Fluo-4. These results show Hcys alters NC attachment by triggering an increase in intracellular Ca2+ possibly by generating inositol triphosphate. Hence, the teratogenic effect ascribed to Hcys may be due to perturbation of intracellular Ca2+ signaling. Developmental Dynamics 237:2117,2128, 2008. © 2008 Wiley-Liss, Inc. [source] Axon extension in the fast and slow lanes: Substratum-dependent engagement of myosin II functionsDEVELOPMENTAL NEUROBIOLOGY, Issue 10 2007Andrea R. Ketschek Abstract Axon extension involves the coordinated regulation of the neuronal cytoskeleton. Actin filaments drive protrusion of filopodia and lamellipodia while microtubules invade the growth cone, thereby providing structural support for the nascent axon. Furthermore, in order for axons to extend the growth cone must attach to the substratum. Previous work indicates that myosin II activity inhibits the advance of microtubules into the periphery of growth cones, and myosin II has also been implicated in mediating integrin-dependent cell attachment. However, it is not clear how the functions of myosin II in regulating substratum attachment and microtubule advance are integrated during axon extension. We report that inhibition of myosin II function decreases the rate of axon extension on laminin, but surprisingly promotes extension rate on polylysine. The differential effects of myosin II inhibition on axon extension rate are attributable to myosin II having the primary function of mediating substratum attachment on laminin, but not on polylysine. Conversely, on polylysine the primary function of myosin II is to inhibit microtubule advance into growth cones. Thus, the substratum determines the role of myosin II in axon extension by controlling the functions of myosin II that contribute to extension. © 2007 Wiley Periodicals, Inc. Develop Neurobiol, 2007. [source] Comparison of biodegradation kinetic parameters for naphthalene in batch and sand column systems by pseudomonas putidaENVIRONMENTAL PROGRESS & SUSTAINABLE ENERGY, Issue 2 2001Jeong-Hun Park Kinetic parameters for the degradation of naphthalene by Pseudomonas putida ( ATCC 17484) were estimated in both batch and column assays, in order to evaluate the role of flow and cell attachment on biodegradation rates. Suspended cells and cells attached to Ottawa sand were used under a variety of biomass levels, column flow-rates, and substrate concentrations. In batch systems, degradation followed zero order kinetics across the entire concentration range, while the columns exhibited decreased rates at concentrations less than 100 (,g/L), describable by Michaelis-Menten kinetics. This is reflected in elevated values of the half-saturation constant, Ks, in columns. We offer the explanation that this may have resulted from reactive heterogeneity within the porous media, imposing a distribution of length-scales for transfer of substrate to the cell surfaces. Well-mixed batch systems are expected to have both shorter and more uniform transfer distances. When kinetic parameters obtained in batch system are used for prediction of degradation in columns, at least two factors,exposed reduction of exposed cell surface are a and heterogeneity of cell distribution,will likely reduce overall column degradation rates. [source] The Influence of Tetracycline Loading on the Surface Morphology and Biocompatibility of Films Made from P(3HB) Microspheres,ADVANCED ENGINEERING MATERIALS, Issue 7 2010Lydia Francis Tetracycline, an antibiotic used against a broad range of Gram positive and Gram negative bacteria was encapsulated in microspheres made of poly(3-hydroxybutyric acid) P(3HB), a microbial biodegradable polymer isolated from Bacillus cereus SPV. The drug loaded microspheres were prepared using an oil emulsion technique and compressed uniaxially to produce films. Although the same fabrication conditions were used for preparing the drug loaded and unloaded microspheres, the presence of the drug changed the surface morphology and roughness of the films. The surface morphology of the drug loaded films appeared uneven and coarser and the roughness, with an average root mean square value of 5.89,µm, was significantly higher than that of the unloaded film. The in vitro biocompatibility of the films was investigated using a human keratinocyte cell line (HaCaT) by comparing cell viability on the films to that on conventional tissue culture plastics. Both films appear to support cell growth but cell attachment and percentage cell viability were greater on the drug loaded films (32% of control) compared to the unloaded film (10% of control), possibly as a result of the non-uniform surface morphology and increased roughness of the drug loaded film. Thus, the above results illustrate that the drug loaded films, in addition to being a suitable matrix for drug delivery, represent an improved substrate for keratinocyte cell attachment. [source] Surface Physiochemistry Affects Protein Adsorption to Stoichiometric and Silicate-Substituted Microporous Hydroxyapatites,ADVANCED ENGINEERING MATERIALS, Issue 4 2010Katharina Guth An important factor in the bioactivity and success of a bone-graft substitute is the nature of the adsorbed protein layer, which plays a vital role in orchestrating cell attachment and development through the presence of adhesion proteins such as fibronectin (Fn) and vitronectin (Vn). In this study, microporous hydroxyapatite (HA) and silicate-substituted hydroxyapatite (SA) discs with matched porosity and surface morphology are developed to mimic the topography found in commercial bone-graft substitutes in order to identify whether the introduction of microporosity and associated surface roughness eliminates the beneficial effect that silicate substitution has on protein adsorption. The introduction of microporosity does not abolish the relative enrichment of the protein layer that is adsorbed to the microporous SA discs, as opposed to HA, but appears to accelerate it. Fibronectin and Vn adsorption in a range of competitive environments at physiological temperatures confirm that the microporous SA discs have a greater affinity for Fn and Vn compared with HA, suggesting differences in the mechanisms behind the surface affinity to SA. Thus, development of a surface protein layer on SA and HA is likely to be dependent on the nature of the local protein environment and a combination of factors that are associated with the addition of silicate: the surface charge, the nature of the ionic species at the interface and the resultant hydrophilicity of the surface. Total protein adsorption is not found to be a good indicator of potential implant performance, particularly at early time points. [source] Effect of Silicate-Substitution on Attachment and Early Development of Human Osteoblast-Like Cells Seeded on Microporous Hydroxyapatite Discs,ADVANCED ENGINEERING MATERIALS, Issue 1-2 2010Katharina Guth Hydroxyapatite (HA) is a well-established graft material used in bone repair. Silicon-substituted hydroxyapatite (SA; 0.8,wt% Si) has shown greater bone ingrowth and bone coverage than phase pure HA. To assess the effect of microporosity on sensitivity of cell attachment to surface physiochemistry, microporous SA and HA discs, and control Thermanox (TMX) discs were incubated with osteoblast-like cells (5,×,104 HOS-TE85 cells) under differing tissue culture conditions. To investigate early cellular attachment, organization, and differentiation, cells were also stained for integrin,,5,1, actin, and runt-related transcription factor (RUNX-2), respectively, after incubation on HA, SA, and TMX discs for 3 days. No significant differences emerged between HA, SA, and TMX discs in mean numbers of cells attached in serum free medium (SFM) over 90,min incubation. In contrast, significantly more cells were attached to SA than HA after 180,min incubation in complete medium (C-MEM) containing fetal calf serum (p,<,0.05). Cell attachment to SA and HA discs pre-conditioned in SFM supplemented with fibronectin (FN) was lower than discs pre-conditioned in C-MEM, suggesting sensitivity of an active FN conformation to the presence of co-adsorbates. Confocal microscopy demonstrated significantly more co-localization of integrin ,5,1 and actin on SA than HA. Translocalization of RUNX-2 to the nucleus was stronger in cells incubated on SA. Microporosity did not diminish the effect of surface physiochemistry on cell adhesion, and enhanced cell attachment for SA appears to be mediated by differences in the quality of adsorbed protein rather than via direct effects of substrate chemistry. [source] Hydroxyapatite/SiO2 Composites via Freeze Casting for Bone Tissue Engineering,ADVANCED ENGINEERING MATERIALS, Issue 11 2009Silke Blindow Freeze casting is a fabrication method that allows producing near-net-shaped ceramics with variable porosity. Hydroxyapatite (HA) was modified by the addition of different amounts of SiO2 nanoparticles during freeze cast preparation. The addition of SiO2 introduced a partial phase transformation of HA to , -tricalcium phosphate and improved the form stability due to less shrinkage after sintering. The impact of surface roughness of pure HA ceramics and the influence of SiO2 introduction during freeze casting on adhesion, proliferation, and differentiation of human osteoblast-like cells (MG-63) was investigated. While both cell attachment and proliferation of smooth pressed HA was significantly enhanced compared to rough freeze cast HA, the addition of SiO2 improved the cell numbers of the latter. The expression of cell differentiation markers osteocalcin and collagen I was found to be supported by rough surfaces (Ra,=,5,6,µm) in particular on ceramics containing SiO2 [source] Proteolysis of the tumour suppressor hDlg in response to osmotic stress is mediated by caspases and independent of phosphorylationFEBS JOURNAL, Issue 2 2009Francisco A. Ińesta-Vaquera Human disc-large (hDlg) is a scaffold protein critical for the maintenance of cell polarity and adhesion. hDlg is a component of the p38, MAP kinase pathway, which is important for the adaptation of mammalian cells to changes in environmental osmolarity. Here we report a strong decrease in the levels of hDlg protein in the human epithelial cell line HeLa when exposed to osmotic shock. This is independent of the phosphorylation state of hDlg, is prevented by preincubating the cell with the caspase inhibitor z-VAD and is part of the apoptotic process triggered by cellular stress. Although, both caspase 3 and caspase 6 are strongly activated by osmotic shock, the time course of caspase 6 activation parallels hDlg degradation, suggesting that this caspase may be responsible for the proteolysis. Mutating hDlg Asp747 to Ala abolishes caspase-induced cleavage, but does not affect the early stage of apoptosis or cell attachment. Our findings show that osmotic stress triggers hDlg degradation through a mechanism different from the one mediated by proteasomes, and we identify hDlg as a caspase substrate during the apoptotic process, although its proteolysis may not be implicated in the progression of early apoptosis. [source] Characterization of a Novel Fiber Composite Material for Mechanotransduction Research of Fibrous Connective TissuesADVANCED FUNCTIONAL MATERIALS, Issue 5 2010Hazel R. C. Screen Abstract Mechanotransduction is the fundamental process by which cells detect and respond to their mechanical environment, and is critical for tissue homeostasis. Understanding mechanotransduction mechanisms will provide insights into disease processes and injuries, and may support novel tissue engineering research. Although there has been extensive research in mechanotransduction, many pathways remain unclear, due to the complexity of the signaling mechanisms and loading environments involved. This study describes the development of a novel hydrogel-based fiber composite material for investigating mechanotransduction in fibrous tissues. By encapsulating poly(2-hydroxyethyl methacrylate) rods in a bulk poly(ethylene glycol) matrix, it aims to create a micromechanical environment more representative of that seen in vivo. Results demonstrated that collagen-coated rods enable localized cell attachment, and cells are successfully cultured for one week within the composite. Mechanical analysis of the composite indicates that gross mechanical properties and local strain environments could be manipulated by altering the fabrication process. Allowing diffusion between the rods and surrounding matrix creates an interpenetrating network whereby the relationships between shear and tension are altered. Increasing diffusion enhances the shear bond strength between rods and matrix and the levels of local tension along the rods. Preliminary investigation into fibroblast mechanotransduction illustrates that the fiber composite upregulates collagen I expression, the main protein in fibrous tissues, in response to cyclic tensile strains when compared to less complex 2D and 3D environments. In summary, the ability to create and manipulate a strain environment surrounding the fibers, where combined tensile and shear forces uniquely impact cell functions, is demonstrated. [source] Carbon Nanotube Coatings on Bioglass-Based Tissue Engineering ScaffoldsADVANCED FUNCTIONAL MATERIALS, Issue 15 2007R. Boccaccini Abstract The coating of highly porous Bioglass® based 3D scaffolds with multi-walled carbon nanotubes (CNT) was investigated. Foam like Bioglass® scaffolds were fabricated by the replica technique and electrophoretic deposition was used to deposit homogeneous layers of CNT throughout the scaffold pore structure. The optimal experimental conditions were determined to be: applied voltage 15,V and deposition time 20 minutes, utilizing a concentrated aqueous suspension of CNT with addition of a surfactant and iodine. The scaffold pore structure remained invariant after the CNT coating, as assessed by SEM. The incorporation of CNTs induced a nanostructured internal surface of the pores which is thought to be beneficial for osteoblast cell attachment and proliferation. Bioactivity of the scaffolds was assessed by immersion studies in simulated body fluid (SBF) for periods of up to 2 weeks and the subsequent determination of hydroxyapatite (HA) formation. The presence of CNTs can enhance the bioactive behaviour of the scaffolds since CNTs can serve as template for the ordered formation of a nanostructured HA layers, which does not occur on uncoated Bioglass® surfaces. [source] Dynamic Hydrogels: Switching of 3D Microenvironments Using Two-Component Naturally Derived Extracellular Matrices (Adv. Mater.ADVANCED MATERIALS, Issue 6 20106/2010) The front cover image depicts a two-component extracellular matrix (ECM) in which one component acts as a stable structural element (which supports cell attachment and migration) and another component gels or dissolves reversibly (a modulatory component). Samuel K. Sia and co-workers show on p. 686 that by dynamically adding or removing crosslinks in the modulatory component, properties of the composite ECM, such as the ability of cells to migrate and the rate of diffusive transport, can be altered. [source] Assessment of bioactive and bio-adhesive therapies to enhance stem cell attachment to root surface dentineINTERNATIONAL ENDODONTIC JOURNAL, Issue 7 2009M. A. Elseed Abstract Aim, To compare bioactive and bio-adhesive therapies to enhance stem cell attachment to the root dentine of human teeth. Methodology, Dentine slabs (n = 72) were cut from the lower 3 mm of the roots of extracted human permanent teeth. The root dentine slabs were untreated, or coated with bio-adhesive, or human recombinant transforming growth factor-beta1 (hrTGF-B1), or human recombinant bone morphogenic protein-2 (hrBMP-2). The dentine slabs were placed with the root surface in contact with confluent periodontal stem cell (PSC) cultures using aseptic techniques. The cells and dentine slabs were submerged in culture media for 4, 24 and 72 h. The specimens were fixed in formalin, dehydrated and processed for scanning electron microscopy (SEM). Results, SEM micrographs at ×2000 magnification revealed PSC extensive adherence to root dentine for all of the bio-adhesive and bioactive treatments. The addition of bioactive molecules did not improve PSC attachment. Few cells attached to the negative control treatments. Conclusions, Bio-adhesive and bioactive growth factors were not needed to promote PSC attachment to the root dentine of human teeth, because it already appears to have good natural properties to promote PSC attachment. This suggests PSC can be used for the clinical replantation of avulsed teeth without the need for bio-adhesive and bioactive treatments. [source] Effects of lithium carbonate on rat seminiferous tubules: an ultrastructural studyINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 6 2006O. Zarnescu Summary Lithium salts are commonly used for treatment of bipolar disorder but prolonged treatment with therapeutic doses induces substantial toxic effects. In the present study we examined the effects of lithium carbonate on the ultrastructure of rat seminiferous tubules. Rats were exposed to lithium carbonate at doses of 35 mg/kg/day for 21 days. After lithium treatment, the tunica propria widened and folded together with convolutions of the basement membrane, myoid cells and lymphatic endothelium. In the seminiferous epithelium loss of germ cell attachment and appearance of expanded intercellular spaces between spermatogenic cells were observed. Early stages of spermatogenic cells showed nuclear protrusions or swellings because of an extensive enlargement of the outer nuclear membrane. Round spermatids exhibited abnormally shaped acrosomes and dilation of the subacrosomal space. Many abnormal, degenerated late spermatids with random orientation were seen towards the basal and adluminal compartments of the seminiferous epithelium. In addition, spermatids exhibited alteration in F-actin bundle ectoplasmic specialization and contained many mitochondria-associated granular bodies. [source] Bioceramic Bone Graft Substitutes: Influence of Porosity and ChemistryINTERNATIONAL JOURNAL OF APPLIED CERAMIC TECHNOLOGY, Issue 3 2005Karin A. Hing Bioceramics have been considered for use as synthetic bone graft substitutes (BGSs) for over 30 years, throughout which there have been two primary areas of research: (i) optimization of the physical pore structure and (ii) formulation of an appropriate bioceramic chemistry. While it is well recognized that both the rate of integration and the final volume of regenerated bone are primarily dependent on the macroporosity, there still seems to be some dispute regarding the optimum "type" of porosity. The rate and quality of bone integration have, in turn, been related to a dependence on pore size, porosity volume fraction, and interconnection size and interconnection density, both as a function of structural permeability and mechano-transduction. Moreover, the role of strut microstructure and pore geometry have been considered with respect to their influence on entrapment and recruitment of growth factors (GFs) in addition to its influence on scaffold mechanics. Deconvoluting the relative affects of these parameters is complicated by the use of both resorbable and nonresorbable bioactive bioceramics, which are believed to mediate bioactivity in the osseous environment through two principal mechanisms: (i) directly through dissolution and release of ionic products in vivo, elevating local concentrations of soluble species that interact directly with local cells or influence cell behavior by their effect on local pH, (ii) indirectly through the influence that surface chemistry will have on protein adsorption, GF entrapment, and subsequent cell attachment and function. This article aims to review some of the recent developments in bioceramic BGSs, with a view to understanding how the various physiochemical parameters may be optimized to promote bone healing. [source] Biodegradable Xylitol-Based Polymers,ADVANCED MATERIALS, Issue 10 2008Joost P. Bruggeman Synthetic polymers composed of metabolites endogenous to the mammalian organism are designed. The design is based on the monomer xylitol, which possesses a wide range of physical properties that are biologically relevant. Xylitol-based hydrogels and elastomers are biocompatible in vitro and in vivo, compared to the prevalent synthetic polymer poly(L-lactic- co - glycolic acid) (PLGA). It furthermore provides a platform to tune mechanical properties, degradation profiles, and cell attachment. [source] The effect of the Vector® scaler system on human teeth: a systematic reviewINTERNATIONAL JOURNAL OF DENTAL HYGIENE, Issue 3 2008DE Slot Abstract:, Aim:, To review the available literature, considering the effect of instrumentation with the Vector® ultrasonic scaler on human teeth in vitro and in vivo compared to conventional ultrasonic instruments and/or hand instrumentation. The assessed effects are calculus removal, time of instrumentation, root surface aspects, cell attachment, patients' perception, bleeding upon probing, pocket depth, clinical attachment loss and microbiological effects. Materials and methods:, MEDLINE,PubMed and the Cochrane Central register of controlled trials (CENTRAL) were searched up through January 2008 to identify appropriate studies. Results:, Independent screening of the titles and abstracts of 270 MEDLINE,PubMed and 15 Cochrane papers resulted in 15 suitable publications. The studies differed in design and outcome, so this review summarizes the outcomes in a descriptive manner. Comparisons are presented against conventional ultrasonic system and scaling and root planing. Conclusion:, The Vector® ultrasonic scaler provided comparable clinical and microbiological periodontal healing results as scaling and root planing and conventional ultrasonic system in moderately deep pockets. The Vector® ultrasonic scaler may be used as a gentle root debridement device for supportive periodontal therapy, as an alternative to other conventional ultrasonic system. The operator should however consider the extra time needed for instrumentation. [source] Organic-soluble chitosan/polyhydroxybutyrate ultrafine fibers as skin regeneration prepared by electrospinningJOURNAL OF APPLIED POLYMER SCIENCE, Issue 6 2010Guiping Ma Abstract In the present contribution, the ultrafine fiber membranes of polyhydroxybutyrate (PHB) and organic-soluble chitosan(O-CS) was prepared by electrospinning. The structure and thermal stability were studied by infrared (FTIR) and thermogravimetric analysis (TG). The surface properties of ultrafine fibers were estimated by contact angle measurements using water. The morphology was observed by scanning electron microscopy (SEM). The cytotoxicity assessment with mouse fibroblast cells (L929) was also investigated. Cell culture results showed that it benefits promoting the cell attachment and proliferation. The results showed it could be as tissue engineering for skin regeneration. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010 [source] Physicochemical and biological evaluation of plasma-induced graft polymerization of acrylamide onto polydimethylsiloxaneJOURNAL OF APPLIED POLYMER SCIENCE, Issue 4 2008A. Parvin Abstract Polydimethylsiloxane (PDMS) rubbers exhibit good mechanical properties for biomedical and industrial applications, but their inherently high hydrophobicity limits biomedical applications of this material despite its favorable mechanical properties. In this work, surface modification of PDMS by radio-frequency glow discharge and subsequently graft polymerization of acrylamide was studied. PAAm-grafted, oxygen plasma-treated, and control (untreated) PDMS rubbers were characterized using attenuated total reflectance Fourier transform infrared, scanning electron microscopy, dynamic mechanical thermal analyses, zeta potential, and contact angle techniques. Fibroblast (L929) cell attachment and growth onto these surfaces were examined by optical microscopy. The data from in vitro assays showed that cell attachment onto control surface was very negligible while significant cell attachment and growth was observed onto oxygen plasma-treated and PAAm-grafted PDMS surfaces. The method developed in this work offers a convenient way of surface modifications of biomaterials to improve attachment of cells onto substrates. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2008 [source] Effects on titanium implant surfaces of chemical agents used for the treatment of peri-implantitisJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2010Krisztina Ungvári Abstract The treatment of peri-implantitis, which causes tissue deterioration surrounding osseointegrated implants, involves surface decontamination and cleaning. However, chemical cleaning agents may alter the structure of implant surfaces. We investigated three such cleaning solutions. Commercially pure (grade 4) machined titanium discs (CAMLOG Biotechnologies AG, Switzerland) were treated with 3% H2O2 (5 min), saturated citric acid (pH = 1) (1 min) or chlorhexidine gel (5 min), and their surface properties were examined by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). Human epithelial cell attachment (24-h observation) and proliferation (72-h observation) were investigated via dimethylthiazolyl-diphenyltetrazolium bromide (MTT) and bicinchoninic acid (BCA) protein content assays. AFM revealed no significant difference in roughness of the three treated surfaces. XPS confirmed the constant presence of typical surface elements and an intact TiO2 layer on each surface. The XPS peaks after chlorhexidine gel treatment demonstrated CO and/or CO bond formation, due to chlorhexidine digluconate infiltrating the surface. MTT and BCA assays indicated similar epithelial cell attachments in the three groups; epithelial cell proliferation being significantly higher after H2O2 than after chlorhexidine gel treatment (not shown by BCA assays). These agents do not harm the Ti surface. Cleaning with H2O2 slightly enhances human epithelial cell growth, in contrast to chlorhexidine gel. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010. [source] Physically crosslinked composite hydrogels of PVA with natural macromolecules: Structure, mechanical properties, and endothelial cell compatibilityJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2009Y. Liu Abstract Polyvinyl alcohol (PVA) hydrogels have been considered potentially suitable for applications as engineered blood vessels because of their structure and mechanical properties. However, PVA's hydrophilicity hinders its capacity to act as a substrate for cell attachment. As a remedy, PVA was blended with chitosan, gelatin, or starch, and hydrogels were formed by subjecting the solutions to freeze-thaw cycles followed by coagulation bath immersion. The structure-property relationships for these hydrogels were examined by measurement of their swelling, rehydration, degradation, and mechanical properties. For the case of pure PVA hydrogels, the equilibrium swelling ratio was used to predict the effect of freeze thaw cycles and coagulation bath on average molecular weights between crosslinks and on mesh size. For all hydrogels, trends for the reswelling ratio, which is indicative of the crosslinked polymer fraction, were consistent with relative tensile properties. The coagulation bath treatment increased the degradation resistance of the hydrogels significantly. The suitability of each hydrogel for cell attachment and proliferation was examined by protein adsorption and bovine vascular endothelial cell culture experiments. Protein adsorption and cell proliferation was highest on the PVA/gelatin hydrogels. This study demonstrates that the potential of PVA hydrogels for artificial blood vessel applications can be improved by the addition of natural polymers, and that freeze-thawing and coagulation bath treatment can be utilized for fine adjustment of the physical characteristics. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009 [source] Novel 3D collagen scaffolds fabricated by indirect printing technique for tissue engineeringJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2008C. Z. Liu Abstract This article reports the mechanical properties and in vitro evaluation of a collagen scaffold fabricated using an indirect 3D printing technique. Collagen scaffolds, featuring predefined internal channels and capillary networks, were manufactured using phase change printing. It was observed that the collagen scaffolds featured internal channels and a hierarchical structure that varied over length scales of 10,400 ,m. In vitro evaluation using hMSCs demonstrated that the resultant collagen based scaffolds have the ability to support hMSC cell attachment and proliferation; cells can migrate and survive deep within the structure of the scaffold. The cell numbers increased 2.4 times over 28 days in culture for the lysine treated scaffolds. The cells were spread along the collagen fibers to form a 3D structure and extracellular matrix was detected on the surface of the scaffolds after 4 weeks in culture. The crosslinking treatment enhanced the biostability and dynamic properties of the collagen scaffolds significantly. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008 [source] Fibroblast cell attachment and growth on nanoengineered sculptured thin filmsJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2007M. C. Demirel Abstract Nanoengineered parylene-C sculptured thin films (STFs) are deposited on glass and silicon substrates using a direct one-step growth technique. The deposited STFs support fibroblast cell attachment and proliferation in vitro, which is an early indication of biocompatibility and bioactivity of this emerging class of biomaterials. Surface modification of endoprostheses of the small joints of the hand, which heal with fibrous stabilization, may be greatly enhanced by such nanoengineered biomaterials. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006 [source] Characterization of Tissue Transglutaminase in Human Osteoblast-like CellsJOURNAL OF BONE AND MINERAL RESEARCH, Issue 8 2001Deborah J. Heath Abstract Tissue transglutaminase (tTG) is a calcium-dependent and guanosine 5,-triphosphate (GTP) binding enzyme, which catalyzes the post-translational modification of proteins by forming intermolecular ,(,-glutamyl)lysine cross-links. In this study, human osteoblasts (HOBs) isolated from femoral head trabecular bone and two osteosarcoma cell lines (HOS and MG-63) were studied for their expression and localization of tTG. Quantitative evaluation of transglutaminase (TG) activity determined using the [1,414C]-putrescine incorporation assay showed that the enzyme was active in all cell types. However, there was a significantly higher activity in the cell homogenates of MG-63 cells as compared with HOB and HOS cells (p < 0.001). There was no significant difference between the activity of the enzyme in HOB and HOS cells. All three cell types also have a small amount of active TG on their surface as determined by the incorporation of biotinylated cadaverine into fibronectin. Cell surface-related tTG was further shown by preincubation of cells with tTG antibody, which led to inhibition of cell attachment. Western blot analysis clearly indicated that the active TG was tTG and immunocytochemistry showed it be situated in the cytosol of the cells. In situ extracellular enzyme activity also was shown by the cell-mediated incorporation of fluorescein cadaverine into extracellular matrix (ECM) proteins. These results clearly showed that MG-63 cells have high extracellular activity, which colocalized with the ECM protein fibronectin and could be inhibited by the competitive primary amine substrate putrescine. The contribution of tTG to cell surface/matrix interactions and to the stabilization of the ECM of osteoblast cells therefore could by an important factor in the cascade of events leading to bone differentiation and mineralization. [source] Valproic acid blocks adhesion of renal cell carcinoma cells to endothelium and extracellular matrixJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 8b 2009Jon Jones Abstract Treatment strategies for metastatic renal cell carcinoma (RCC) have been limited due to chemotherapy and radiotherapy resistance. The development of targeted drugs has now opened novel therapeutic options. In the present study, anti-tumoral properties of the histone deacetylase inhibitor valproic acid (VPA) were tested in vitro and in vivo on pre-clinical RCC models. RCC cell lines Caki-1, KTC-26 or A498 were treated with various concentrations of VPA to evaluate tumour cell adhesion to vascular endothelial cells or to immobilized extracellular matrix proteins. In vivo tumour growth was conducted in subcutaneous xenograft mouse models. VPA was also combined with low dosed interferon-, (IFN-,) and the efficacy of the combination therapy, as opposed to VPA monotherapy, was compared. VPA significantly and dose-dependently prevented tumour cell attachment to endothelium or matrix proteins, accompanied by elevated histones H3 and H4 acetylation. VPA altered integrin-, and -, subtype expression, in particular ,3, ,5 and ,3, and blocked integrin-dependent signalling. In vivo, VPA significantly inhibited the growth of Caki-1 in subcutaneous xenografts with the 200 mg/kg being superior to the 400 mg/kg dosing schedule. VPA-IFN-, combination markedly enhanced the effects of VPA on RCC adhesion, and in vivo tumour growth was further reduced by the 400 mg/kg but not by the 200 mg/kg VPA dosing schedule. VPA profoundly blocked the interaction of RCC cells with endothelium and extracellular matrix and reduced tumour growth in vivo. Therefore, VPA should be considered an attractive candidate for clinical trials. [source] | |||||||||||||||||||||||||||||||