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  • 1
    ISSN: 0021-9304
    Keywords: implant ; titanium ; osteoblasts ; surface roughness ; 1α,25- (OH)2D3 ; differentiation ; local factor ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Surface roughness has been shown to affect differentiation and local factor production of MG63 osteoblast-like cells. This study examined whether surface roughness alters cellular response to circulating hormones such as 1α,25-(OH)2D3. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then were machined and acid-etched (MA). Ti disks also were sandblasted (SB), sandblasted and acid etched (CA), or plasma sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were: PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on standard tissue culture polystyrene (plastic) or the Ti surfaces and then treated for 24 h with either 10-8M or 10-7M 1α,25-(OH)2D3 or vehicle (control). Cellular response was measured by assaying cell number, cell layer alkaline phosphatase specific-activity, and the production of osteocalcin, latent (L) TGFβ, and PGE2. Alkaline phosphatase activity was affected by surface roughness; as the surface became rougher, the cells showed a significant increase in alkaline phosphatase activity. Addition of 1α,25-(OH)2D3 to the cultures caused a dose-dependent stimulation of alkaline phosphatase activity that was synergistic with the effect caused by surface roughness alone. 1α,25-(OH)2D3 also caused a synergistic increase in osteocalcin production as well as local factor (LTGFβ and PGE2) production on the rougher CA, SB, and PS surfaces, but it had no effect on the production on smooth surfaces. The inhibitory effect of surface roughness on cell number was not affected by 1α,25-(OH)2D3 except on the SB surface. 1α,25-(OH)2D3 decreased cell number, increased alkaline phosphatase activity and osteocalcin production, and had no effect on LTGFβ or PGE2 production by MG63 cells grown on tissue culture polystyrene. These data suggest that bone cell response to systemic hormones is modified by surface roughness and that surface roughness increases the responsiveness of MG63 cells to 1α,25-(OH)2D3. They also suggest that the endocrine system is actively involved in normal bone healing around implants. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 39, 77-85, 1998.
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  • 2
    ISSN: 0021-9304
    Keywords: implant ; titanium ; osteoblasts ; prostaglandin ; indomethacin ; surface roughness ; 1α,25-(OH)2D3 ; differentiation ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Surface roughness affects proliferation, differentiation (alkaline phosphatase and osteocalcin), local factor production [transforming growth factor (TGFβ) and prostaglandin E2 (PGE2)], and response to 1,25-(OH)2D3 (1,25) of MG63 osteoblast-like cells. In this study, we examined whether the effect of surface roughness on MG63 cells is mediated by prostaglandins produced by the cells. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then machined and acid-etched (MA). Disks were also coarse grit-sandblasted (SB), coarse grit-sandblasted and acid-etched (CA), or plasma-sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on the Ti disks in the presence or absence of 10-7M indomethacin (Indo), a specific inhibitor of cyclooxygenase activity, resulting in decreased prostaglandin production. When the cells reached confluence, cell number, cell layer alkaline phosphatase specific activity (ALPase), and osteocalcin (OC) and latent TGFβ (LTGFβ) production were determined. In addition, confluent cultures which had been grown in the absence of Indo were exposed to 10-7M 1,25, 10-7M Indo, or a combination of the two for 24 h. On the rougher surfaces, cell number was decreased and ALPase, OC, and LTGFβ were increased. When indomethacin was present throughout the culture period, the effect of surface roughness on cell number, OC, and LTGFβ was abolished. ALPase was reduced, but surface roughness-dependent effects were still observed. Addition of indomethacin to confluent cultures for 24 h had no effect on any of the parameters examined, with one exception: Cells cultured on MA surfaces exhibited a more differentiated phenotype. 1,25 increased all parameters examined on SB, CA, and PS surfaces. When indomethacin was added with 1,25, the 1,25-dependent effects on cell number and OC and LTGFβ production were abolished; however, ALPase was unaffected. This indicates that bone cell response to systemic hormones may be modified by implant surface roughness. This effect may be mediated, at least in part, by prostaglandins produced by the same cells. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 41, 489-496, 1998.
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  • 3
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 40 (1998), S. 324-335 
    ISSN: 0021-9304
    Keywords: titanium ; bioactive coating ; immobilization ; silanization ; covalent attachment ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: The surface of implantable biomaterials is in direct contact with the host tissue and plays a critical role in determining biocompatibility. In order to improve the integration of implants, it is desirable to control interfacial reactions such that nonspecific adsorption of proteins is minimized and tissue-healing phenomena can be controlled. In this regard, our goal has been to develop a method to functionalize oxidized titanium surfaces by the covalent immobilization of bioactive organic molecules. Titanium first was chemically treated with a mixture of sulfuric acid and hydrogen peroxide to eliminate surface contaminants and to produce a consistent and reproducible titanium oxide surface layer. An intermediary aminoalkylsilane spacer molecule was then covalently linked to the oxide layer, followed by the covalent binding of either alkaline phosphatase or albumin to the free terminal NH2 groups using glutaraldehyde as a coupling agent. Surface analyses following coating procedures consisted of X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and atomic force microscopy (AFM). Enzymatic activity of coupled alkaline phosphatase was assayed colorimetrically, and surface coverage by bound albumin was evaluated by SEM visualization of colloidal gold immunolabeling. Our results indicate that the linkage of the aminoalkylsilane to the oxidized surface is stable and that bound proteins such alkaline phosphatase and albumin retain their enzymatic activity and antigenicity, respectively. The density of immunolabeling for albumin suggests that the binding and surface coverage obtained is in excess of what would be expected for inducing biological activity. In conclusion, this method offers the possibility of covalently linking selected molecules with known biological activity to oxidized titanium surfaces in order to guide and promote the tissue healing that occurs during implant integration in bone and soft tissues. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 324-335, 1998
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  • 4
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 43 (1998), S. 184-191 
    ISSN: 0021-9304
    Keywords: amalgams ; titanium ; galvanic corrosion ; cell culture ; atomic absorption ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: The potential exists for titanium and amalgams to become galvanically coupled in the oral cavity. While low galvanic corrosion rates have been measured in vivo for titanium-amalgam or mercury-free alloy couples, concerns exist over released corrosion products and adverse tissue responses. It was hypothesized in this study that coupling titanium to amalgams or gallium alloys increased the release of metallic corrosion products and decreased cellular activity and function. The effects of titanium coupled and uncoupled to a conventional amalgam, palladium-enriched spherical high copper amalgam, a dispersed type high copper amalgam, and a mercury-free gallium alloy were evaluated in 24-h cell culture tests. Viability, proliferation, and collagen synthesis were evaluated by the uptake of neutral red, 3H-thymidine, and immunoassay of procollagen, respectively, and compared to cells not exposed to any test material. The gallium alloy-titanium couple resulted in significant decreases in cellular viability, proliferation, and collagen synthesis as compared to the other coupled and uncoupled samples. Few differences in the cellular responses of the other coupled and uncoupled samples were observed. Atomic absorption analyses indicated increased release of metal ions from the amalgam and gallium alloy samples coupled to titanium as compared to their uncoupled condition, although the differences were not always significant. Galvanic corrosion of amalgam-titanium couples in the long term may become significant, and further research is needed. Coupling the gallium alloy to titanium may result in increased galvanic corrosion and cytotoxic responses. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res (Appl Biomater) 43: 184-191, 1998
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  • 5
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 40 (1998), S. 244-256 
    ISSN: 0021-9304
    Keywords: titanium ; oxide film ; implant ; H2O2 ; osteoblast ; cell culture ; XPS and EIS ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Variations of titanium oxide films induced by osteoblast-like cells in a rat calvaria culture system and the influence of an H2O2 pretreatment have been investigated by using X-ray photoelectron spectroscopy and electrochemical impedance spectroscopy. For abraded titanium, the results revealed that phosphate and calcium ions may incorporate into the surface oxide film during the cell culture, forming a precipitate with a Ca/P ratio near that of hydroxyapatite. Oxidized carbon also was found in the surface layer, most likely precipitated hydroxylcarbonated apatite (HCA). The H2O2 pretreatment of titanium in a phosphate-buffered saline solution results in a 10-fold thickened porous oxide film and large amounts of surface hydroxyl groups as well as a certain amount of phosphate ions inside the oxide film. During the cell culture, the H2O2-treated titanium surface favors the ion incorporation and precipitation of the HCA-like compound, which probably is inlaid into the oxide film. Osteoblast-like cells on the H2O2-treated titanium showed a more active morphology during the initial stage compared with cells on abraded titanium. Moreover, bone-like nodule formation and mineralization appear to be related to the precipitation of the HCA-like compound on the surface. The results are discussed with respect to corrosion resistance, ion incorporation and precipitation of the HCA-like compound on the surface, osseointegration, and bioactivity of titanium Implants. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 244-256, 1998.
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  • 6
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 41 (1998), S. 120-130 
    ISSN: 0021-9304
    Keywords: implant surface ; titanium ; fibronectin ; atomic force microscopy ; contact angle ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Protein binding on metallic implant surfaces, such as titanium, is governed by the physico-chemical nature of the metallic surface. Human plasma fibronectin (HPF) is an important matrix glycoprotein that mediates cell and protein attachment to each other or to the extracellular matrix present during wound healing. The objective of this study was to investigate the adsorption of HPF onto polished commercially pure titanium (cpTi) by using atomic force microscopy (AFM) and electron spectroscopy for chemical analysis (ESCA) and to measure the resultant surface contact angle before and after HPF binding. Two types of cpTi disks, one highly polished in our laboratory (HSS) and one commercially prepared (31), were reacted with HPF solutions of varying concentrations (1 μg/mL - 10 ng/mL). ESCA survey spectra of samples coated with 1 μg/mL of fibronectin showed an increase in organic nitrogen and carbon compared with uncoated controls. Contact angle measurements of HSS and 3I cpTi disks showed no significant difference in average contact angle (36.3° ± 3.5 and 39.1° ± 3.1) despite differences in local root mean square (RMS) surface roughness (4.45 ± 0.46 nm and 22.37 ± 4.17 nm) as measured by AFM. Images obtained by AFM showed that 3I specimens were more irregular, with large parallel polishing grooves. Adsorbed HPF appeared in a globular form with an average length of 16.5 ± 1.0 nm, a height of 2.5 ± 0.5 nm, and a width of 9.6 ± 1.2 nm. Fibronectin coating on both HSS and 3I cpTi specimens resulted in a significant increase in hydrophobicity compared to uncoated specimens. These results indicate the significance of HPF on cpTi and may explain how cpTi implants function in situ. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 41, 120-130, 1998.
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  • 7
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 39 (1998), S. 437-445 
    ISSN: 0021-9304
    Keywords: titanium ; surface topography ; MMP-2 ; molecular biocompatability ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: The chemical and topographic effects of commercially pure titanium on cell morphology and the regulation of matrix metalloproteinase-2 (MMP-2) gene expression, synthesis, and activity were investigated in early passage human gingival fibroblasts. Scanning electron microscopy showed that on smooth titanium (Ti), fibroblasts remained well spread and randomly oriented throughout the culture period. In contrast, cells on V-shaped grooved titanium (VTi) were oriented along the grooves by 16 h and proliferated in this organization throughout the culture period. The effects of substratum surface chemistry on MMP-2 expression were found to be distinct from those of topography. Northern hybridization analysis of fibroblasts cultured on Ti revealed an MMP-2 mRNA time-course expression pattern parallel to that observed on the tissue culture plastic (TCP) dishes, but at significantly lower levels at each time-point. The Ti mRNA levels were decreased by 34% at 16 h, 55% at 40 h, and 45% at 90 h relative to TCP. In contrast, MMP-2 mRNA expression on VTi showed both an altered time-course expression pattern and altered levels compared to Ti and TCP. Relative to TCP, VTi MMP-2 mRNA levels were ∼80% less at 16 h and ∼50% less at 40 h, but not significantly different at 90 h. Relative to Ti, VTi MMP-2 mRNA levels were ∼75% less at 16 h, but ∼40% greater at 40 h and ∼70% greater at 90 h. These differences may be explained in part by the observed changes in MMP-2 mRNA half-life which decreased by ∼40% on Ti but increased by over fourfold on VTi relative to TCP. The smooth Ti also showed an approximate twofold increase of MMP-2 secretion in the late cultures over TCP controls. These results indicate that substratum surface chemistry and topography-induced changes in cell shape can alter MMP-2 expression in normal fibroblasts. The molecular approach to investigating the major molecules involved in tissue degradation may provide sensitive indicators of tissue remodeling at the tissue-biomaterial interface. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 39, 437-445, 1998.
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  • 8
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 40 (1998), S. 646-659 
    ISSN: 0021-9304
    Keywords: titanium ; surface treatment ; surface roughness ; surface composition ; surface energy ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Surgical implant finishing and sterilization procedures were investigated to determine surface characteristics of unalloyed titanium (Ti). All specimens initially were cleaned with phosphoric acid and divided into five groups for comparisons of different surface treatments (C = cleaned as above, no further treatment; CP = C and passivated in nitric acid; CPS = CP and dry-heat sterilized; CPSS = CPS and resterilized; CS = C and dry-heat sterilized). Auger (AES), X-ray photoelectron (XPS), and Raman spectroscopic methods were used to examine surface compositions. The surface oxides formed by all treatments primarily were TiO2, with some Ti2O3 and possibly TiO. Significant concentrations of carbonaceous substances also were observed. The cleaning procedure alone resulted in residual phosphorus, primarily as phosphate groups along with some hydrogen phosphates. A higher percentage of physisorbed water appeared to be associated with the phosphorus. Passivation (with HNO3) alone removed phosphorus from the surface; specimens sterilized without prior passivation showed the thickest oxide and phosphorus profiles, suggesting that passivation alters the oxide characteristics either directly by altering the oxide structure or indirectly by removing moieties that alter the oxide. Raman spectroscopy showed no crystalline order in the oxide. Carbon, oxygen, phosphorus, and nitrogen presence were found to correlate with previously determined surface energy. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 646-659, 1998.
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