[Show abstract][Hide abstract] ABSTRACT: Titanium surfaces processed by wire-type electric discharge machining (EDM) are microfabricated surfaces with an irregular morphology, and they exhibited excellent in vitro bone biocompatibility. In this study, the efficiency of in vivo osteogenesis on EDM surfaces was investigated by surgically placing screw-shaped EDM-processed and machined-surface implants into the femurs of four Japanese white rabbits. The volume and process of new bone formation were evaluated by an X-ray micro-CT scanner, coupled with histopathological observations at 1, 2, and 4 weeks post-implantation. Before surgical implantation, the surface topography and contact angle of each implant surface were examined. Bone formation increased over time on both implant surfaces, with both implant types yielding statistically equivalent bone volume at 4 weeks post-implementation. However, at 1 week post-implantation, amount of new bone at EDM-processed implant was markedly greater than that at machined-surface implant. Moreover, new bone appeared to initiate directly from the EDM surfaces, while new bone appeared to generate from pre-existing host bone to the machined surfaces. Thus, EDM seemed to be a promising method for surface modification of titanium implants to support enhanced osteogenesis.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study is to investigate the surface characteristics and biocompatibility of titanium (Ti) surfaces modified by wire electrical discharge machining (EDM). EDM surface characteristics were evaluated by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), thin-film X-ray diffractometry (XRD) and contact angle measurements. MC3T3-E1 cell morphology, attachment and proliferation, as well as analysis of osteoblastic gene expressions, on machined surfaces and EDM surfaces were also evaluated. EDM surfaces exhibited high super hydrophilicity, due to high surface energy. XPS and XRD revealed that a passive oxide layer with certain developing thickness onto. EDM surfaces promoted cell attachment, but restrained proliferation. Counted cell numbers increased significantly on the machined surfaces as compared to the EDM surfaces. Real-time PCR analyses showed significantly higher relative mRNA expression levels of osteoblastic genes (ALP, osteocalcin, Runx2, Osterix) in cells cultured on the EDM surfaces as compared to cells cultured on the machined surfaces.
[Show abstract][Hide abstract] ABSTRACT: This study aimed to develop a contamination-free porous titanium scaffold by a plasma-activated sintering within an originally developed TiN-coated graphite mold. The surface of porous titanium sheet with or without a coated graphite mold was characterized. The cell adhesion property of porous titanium sheet was also evaluated in this study. The peak of TiC was detected on the titanium sheet processed with the graphite mold without a TiN coating. Since the titanium fiber elements were directly in contact with the carbon graphite mold during processing, surface contamination was unavoidable event in this condition. The TiC peak was not detectable on the titanium sheet processed within the TiN-coated carbon graphite mold. This modified plasma-activated sintering with the TiN-coated graphite mold would be useful to fabricate a contamination-free titanium sheet. The number of adherent cells on the modified titanium sheet was greater than that of the bare titanium plate. Stress fiber formation and the extension of the cells were observed on the titanium sheets. This modified titanium sheet is expected to be a new tissue engineering material in orthopedic bone repair.
Journal of tissue engineering. 01/2011; 2010:425402.
[Show abstract][Hide abstract] ABSTRACT: Wire-type electric discharge machining has been applied to the manufacture of endosseous titanium implants as this computer associated technique allows extremely accurate complex sample shaping with an optimal micro textured surface during the processing. Since the titanium oxide layer is sensitively altered by each processing, the authors hypothesized that this technique also up-regulates biological responses through the synergistic effects of the superficial chemistry and micro topography. To evaluate the respective in vitro cellular responses on the superficial chemistry and micro topography of titanium surface processed by wire-type electric discharge, we used titanium-coated epoxy resin replica of the surface. An oxide layer on the titanium surface processed by wire-type electric discharge activated the initial responses of osteoblastic cells through an integrin-mediated mechanism. Since the mRNA expression of ALP on those replicas was up-regulated compared to smooth titanium samples, the micro topography of a titanium surface processed by wire-type electric discharge promotes the osteogenic potential of cells. The synergistic response of the superficial chemistry and micro topography of titanium processed by wire-type electric discharge was demonstrated in this study.
Bio-medical materials and engineering 01/2011; 21(2):113-21. · 1.09 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Photo-functionalized radical reactions on TiO(2) have been correlated with adsorption of organic impurities and decreasing hydrophilicity of titanium-based biomaterials. Such reactive oxygen species (ROS) spontaneously generated on oxidized titanium surfaces may also have important roles against time-dependent degradation of biological ability and adherent micro-organisms. This study examined in vitro biological ability as a function of time and antimicrobial activity on oxidized titanium surfaces without photo-functionalization. Mechanically polished titanium and thermally oxidized titanium surfaces that had been stored for 4 wks showed adsorbed organic impurities with decreased surface hydrophilicity. Even after the storage period, anodically oxidized titanium surfaces enabled super-hydrophilicity without adsorption of organic impurities, because of the ROS and the hydrophilic functional groups generated on the surfaces. The osteogenic gene expressions of osteoblasts cultured on anodically oxidized titanium surfaces with or without storage were significantly higher than those on thermally oxidized titanium and polished titanium surfaces. Titanium surfaces anodically oxidized in a solution with chloride achieved antimicrobial activity against an oral microorganism due to the amount of ROS generated on the surface. Thus, titanium anodically oxidized in solution with chloride may have potential use for titanium-based internal fixation devices.
[Show abstract][Hide abstract] ABSTRACT: A new strategy with bone tissue engineering by mesenchymal stem cell transplantation on titanium implant has been dawn attention. The surface scaffold properties of titanium surface play an important role in bone regenerative potential of cells. The surface topography and chemistry are postulated to be two major factors increasing the scaffold properties of titanium implants. This study aimed to evaluate the osteogenic gene expression of mesenchymal stem cells on titanium processed by wire-type electric discharge machining. Some amount of roughness and distinctive irregular features were observed on titanium processed by wire-type electric discharge machining. The thickness of suboxide layer was concomitantly grown during the processing. Since the thickness of oxide film and micro-topography allowed an improvement of mRNA expression of cells, titanium processed by wire-type electric discharge machining is a promising candidate for mesenchymal stem cell based functional restoration of implants. Comment: 6 pages, 4 figures
[Show abstract][Hide abstract] ABSTRACT: The biomechanical stability of osseointegrated implants is of particular importance, especially the stability which is achieved from structural manipulation at the interface between the implant surface and the bone tissues. Nanoscale beta-tricalcium phosphate-immobilized titanium was prepared by discharge into a physiological buffered saline solution. Compared with hydroxyapatite, it has been shown to be effective in generating a bone-like chemical structure on the surface by cooperative interaction between osteoblastic cells and the beta-tricalcium phosphate. The present study, after cell cultivation, investigates the nanostructures and biomechanical property differences of a mineralized layer formed on two samples of nano-calcium phosphate-immobilized titanium. A scanning probe microscope study revealed that the mineralized tissue formed on the beta-tricalcium phosphate samples after 1 week of cell culture showed significantly higher roughness, compared with hydroxyapatite samples. Nanoindentation micromechanical evaluation of the in vitro generated multilayered structures exhibited thicker bone-like mineralized layers on the beta-tricalcium phosphate samples. A successful modification of titanium implants through the cooperative interaction between osteoblastic cells and nano beta-tricalcium phosphate is anticipated.
Journal of Biomechanics 12/2008; 41(16):3414-8. · 2.72 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Laboratory-designed biocomposites structured by organic matrices impregnated with oriented biominerals have been significantly progressed by mimicking biological processes, although several problems associated with their formulation or antigenicity remain to be solved. Here, we describe a new strategy for the formulation of bioinspired nanostructures that involves spontaneous mediation by cooperative interactions between inorganic nanocrystals and host cells without the complex procedures required for laboratory-designed biocomposites. In the present study, osteoblastic cells were cultured on hydroxyapatite and beta-tricalcium phosphate nanocrystals prepared by discharging in electrolytes. Specifically, a high level of assembly of collagenous proteins associated with cell proliferation was achieved on nanoscale beta-tricalcium phosphate crystals by catalysis of polyphosphate chains produced during cell culture. Furthermore, a spatial structure that was practically composed of natural biocomposites found in bone and teeth was obtained on the nanocrystals due to increased cross-linking between inorganic molecules and biomolecules. Suggestions for the spontaneous formulation of bioinspired nanostructures in a living body mediated by inorganic biomaterials are also discussed.
Journal of Biomedical Materials Research Part A 04/2008; 84(4):869-74. · 2.83 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Reconstitution of carious dentin has been recognized as difficult, because it progresses by loss of collagen polymerization and by demineralization under acidic conditions. Recently, colloidal alkaline nano-calcium phosphate, prepared by electrical discharge in a buffered physiological saline solution, has been shown to be effective in the formulation of a bone-like biocomposite by simply being mixed with acidic collagen solution. It was hypothesized that colloidal calcium phosphate was suitable for the reconstitution of carious dentin. Natural caries lesions in dentin from permanent teeth were exposed to colloidal hydroxyapatite and beta-tricalcium phosphate for 10 days. The micromechanical properties of these tissues were evaluated by nano-indentation. The elastic modulus of human carious dentin improved after samples were immersed in colloidal beta-tricalcium phosphate. The mineral density of carious dentin exposed to beta-tricalcium phosphate increased more than that immersed in hydroxyapatite. However, since it was not directly proportional to micromechanical recovery, mineral density alone was not a sufficient indicator of mechanical behavior.
Journal of Dental Research 04/2008; 87(3):233-7. · 3.83 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The influence of sintering temperature on the in vitro proliferation of osteoblast-like cells to sintered tricalcium phosphate (TCP) sheets prepared by tape-casting was investigated. Green sheets of tape-cast beta-TCP were sintered for 2h in a furnace at atmospheric pressure at five different sintering temperatures: 900, 1000, 1100, 1150 and 1200 degrees C. The number of osteoblast-like (MC3T3-E1) cells deposited onto TCP sheets was counted after cell cultivation for 1week and was found to have increased with increasing sintering temperature. The TCP sheets sintered at 900 degrees C exhibited a significantly lower cell number than TCP sheets sintered at 1000, 1100, 1150 and 1200 degrees C. In the attenuated total reflection infrared spectra, the peaks around 900-1150cm(-1), corresponding to the P-O vibration mode of the phosphate group, gradually decreased and shifted to lower wavenumbers with increasing sintering temperature. Meanwhile, the zeta potential of TCP sintered at 900 degrees C showed a highly negative charge when compared with the other groups. This would suggest that the higher solubility of the TCP sheets sintered at 900 degrees C exerted the higher negative charge obtained from zeta potential measurement. Within the limitations of this study, it was indicated that osteoblast-like cell proliferation increased with increasing sintering temperature. The biological stability of the sintered TCP sheet surface was considered to have affected cell proliferation.