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ABSTRACT: The time-dependent degradation of titanium bioactivity (i.e., the biological aging of titanium) has been reported in previous studies. This phenomenon is caused by the loss of hydrophilicity and the inevitable occurrence of progressive contamination of titanium surfaces by hydrocarbons. In this study, we tested the hypothesis that gamma ray treatment, owing to its high energy to decompose and remove organic contaminants, enhances the bioactivity and osteoconductivity of titanium. Titanium disks were acid-etched and stored for 4 weeks. Rat bone marrow-derived osteoblasts (BMOs) were cultured on titanium disks with or without gamma ray treatment (30 kGy) immediately before experiments. The cell density at day 2 increased by 50% on gamma-treated surfaces, which reflected the 25% higher rate of cell proliferation. Osteoblasts on gamma-treated surfaces showed 30% higher alkaline phosphatase activity at day 5 and 60% higher calcium deposition at day 20. The strength of in vivo bone-implant integration increased by 40% at the early healing stage of week 2 for gamma-treated implants. Gamma ray-treated surfaces regained hydrophilicity and showed a lower percentage of carbon (35%) as opposed to 48% on untreated aged surfaces. The data indicated that gamma ray pretreatment of titanium substantially enhances its bioactivity and osteoconductivity, in association with the significant reduction in surface carbon and the recovery of hydrophilicity. The results suggest that gamma ray treatment could be an effective surface enhancement technology to overcome biological aging of titanium and improve the biological properties of titanium implants. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012.
Journal of Biomedical Materials Research Part B Applied Biomaterials 09/2012; 100(8):2279-87. · 2.15 Impact Factor
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ABSTRACT: This study aimed to analyze growth changes in mandibular body morphology and quantitative bone mineral content (QBMC) with eruption of mandibular third molars (M(3)s) and the relationship between those variables and posterior mandibular body length. Linear and angular measurements were conducted using standard lateral radiographs of 37 dried mandibles in Hellman's dental developmental stages IVA (14 specimens) to VA (23 specimens). Cortical and trabecular basal bone mineral contents (CBMC and TBMC) in the mandible were expressed in millimeter titanium equivalent values using a titanium step wedge. The largest significant change in the mandibular body morphology was an increase in the horizontal dimension (M(2)DP'-Go': 7.59mm), followed by vertical dimension - total height of the mandibular body (THOMB: 4.96mm) and mandibular cortical width (MCW: 1.22mm). The gonial angle (GA) decreased significantly by 6.72° between stages IVA and VA. The mandibular cortical index (MCI) was classified only as C1 or C2 in each stage. Among 4 types of line profile, types 1 and 2 were most commonly observed in both stages. Mean values for CBMC and TBMC increased significantly between stages IVA and VA. Posterior mandibular body length (MeF'-Go') correlated positively with M(2)DP'-Go', THOM, MCW, and CBMC (r=0.816, 0.698, 0.595, and 0.507), respectively and negatively with GA (r=-0.582). These results demonstrated that the morphological changes in the posterior mandibular body and the QBMC increased significantly with M(3) eruption, while the GA became significantly smaller. The posterior mandibular body length had a linear correlation with these variables.
Annals of anatomy = Anatomischer Anzeiger: official organ of the Anatomische Gesellschaft 09/2012; · 0.88 Impact Factor
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ABSTRACT: The topographic and physicochemical features of implant surfaces influence the process of osseointegration. The biologic properties of implant surfaces have been considered to remain stable over time, ie, the capability of osseointegration of implant surfaces presumably does not change over time after manufacturing. However, recent reports have demonstrated that titanium surfaces undergo a progressive change in their biologic characteristics over time, resulting in a significant decrease in osseointegration capability. In comparison to newly prepared titanium surfaces, 4-week-old titanium surfaces (ie, stored for 4 weeks after processing) required more than twice as much healing time to achieve a similar strength of osseointegration. The boneimplant contact percentage for the 4-week-old surfaces was less than 60%, as opposed to more than 90% for the new surfaces. In vitro, the 4-week-old surfaces showed only 20% to 50% of the levels of recruitment, attachment, settlement, and proliferation of osteogenic cells versus new surfaces. On the other hand, a series of recent papers reported the generation of highly cell-attractive and osteoconductive titanium surfaces by ultraviolet (UV) light treatment. The phenomenon, defined as photofunctionalization, caused a fourfold acceleration in the process of osseointegration and resulted in nearly 100% bone-implant contact. Remarkably enhanced behavior and response of osteogenic cells around UV-treated surfaces exceeded the levels observed for the newly prepared surfaces. These studies indicated that UV treatment reverses the time-dependent biologic degradation of titanium and even enhances the surface beyond its innate potential. The present paper summarizes the findings about the aging-like time-dependent biologic degradation of titanium surfaces as well as about the discovery of UV photofunctionalization as a solution for this phenomenon. It also provides a novel understanding of osseointegration and calls for immediate attention to a new avenue of exploration in the science and therapeutics of implant dentistry.
The International journal of oral & maxillofacial implants 07/2012; 27(4):753-61. · 1.78 Impact Factor
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ABSTRACT: OBJECTIVE: The role of nanoscale/submicron morphological features in the process of osseointegration is largely unknown. This study reports the creation of a unique submicrofeatured titanium surface by a combination of anodic oxidation and sandblasting and determines how the addition of this submicrofeature to a microroughened surface affects the early-stage process of osseointegration. MATERIALS AND METHODS: Nonmicroroughened implants were prepared by machining Ti-6Al-4V alloy in a cylindrical form (1 mm diameter and 2 mm long). Microroughened implants were prepared by sandblasting machined implants, while submicrofeatured implants were created by anodic oxidation of the sandblasted implants. Implants were placed into rat femurs and subjected to biomechanical, interfacial, and histological analyses at 1 and 2 weeks post-implantation (n = 6). RESULTS: The submicrotopography was characterized by 50-300 nm nodules and pits in addition to other submicron-level irregularities formed entirely within the sandblast-created microstructures. The biomechanical strength of osseointegration increased continuously from week 1 to 2 for the submicrofeatured implants but not for the microroughened implants. A significant increase in bone-implant contact and bone volume, as well as a reduction in soft tissue intervention, were commonly found for the microroughened surface and the submicrofeatured surface compared with the nonmicroroughened surface. However, there were no differences in these parameters between the microroughened surface and the submicrofeatured surface. An extensive area of bone tissue at the submicrofeatured implant interface was retained intact after biomechanical shear testing, while the microroughened implant-tissue interface showed a gap along the entire axis of the implant, leading to clear separation of the tissue during the shear procedure. CONCLUSIONS: This study demonstrates that a submicrofeatured titanium surface created by a combination of sandblasting and anodic oxidation enhances the strength of early-stage osseointegration, primarily because of the increased resistance of peri-implant bone tissue against external force rather than modulation of bone morphogenesis.
Clinical Oral Implants Research 06/2012; · 2.51 Impact Factor
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ABSTRACT: The survival of cemented endosseous implants can be improved by enhancing the bond between the implant and the cement. We
hypothesized that the light-inducible generation of super-hydrophilicity of titanium positively affects its bone cement-philicity
and bone cement–titanium bonding. Commercially pure titanium disks with machined surface and acid-etched micro roughened surfaces
were prepared. Ultra-violet (UV) light treatment (0.1mW/cm2 UVA and 0.03mW/cm2 UVB for 48h) created a super-hydrophilic surface for both surface types. The area of poly-methyl methacrylate (PMMA)-based
bone cement spread increased by 30% and 20% on the light-treated machined titanium and acid-etched titanium surfaces, respectively,
compared to the matched untreated ones. The contact angle of the bone cement decreased significantly after the light treatment,
confirming the enhanced wettability of bone cement by the light treatment. Interfacial tensile stress between the bone cement
material and titanium was increased 100% for the machined surface and 50% for the acid-etched surface by light treatment.
Interfacial shear stress measured by a push-out test of titanium rods also revealed a 40% increase for the machined surface
and 25% increase for the acid-etched surface. In conclusion, the pre-UV light treatment of titanium enhances the wettability
and bonding strength of poly-methyl-methacrylate-based bone cement.
Journal of Materials Science 04/2012; 43(5):1552-1558. · 2.02 Impact Factor
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ABSTRACT: This study aimed to determine the differences in 5 anatomical parameters of the mandibular body on dental panoramic radiographs (DPRs) of 234 dentulous Japanese subjects (21-78years; mean, 25.76; median, 27 teeth) according to gender and gonial angle (GA) size, and to analyse the correlations between GA size and each variable by gender. The subjects were divided into low (LGA≤120°) and high gonial angle (HGA≥125°) groups. Linear, angular, and morphological measurements were obtained using the DPRs. Statically significant differences were determined using the Mann-Whitney U and χ(2) tests and simple regression analysis. Mandibular cortical width - MCW, antegonial notch depth - AD and angle - AA, and panoramic mandibular index - PMI in the LGA group were significantly larger than those of the HGA group. In men, a significant difference was observed in the distribution of the 3 mandibular cortical index categories between both GAs. Sexual dimorphism for MCW and AD was significantly greater in men than in women. There were significantly negative low correlations between the GA and variables of the MCW, AD, and PMI. These results suggest that the MCW and AD can show significantly different morphology according to gender and GA size.
Annals of anatomy = Anatomischer Anzeiger: official organ of the Anatomische Gesellschaft 02/2012; 194(5):446-51. · 0.88 Impact Factor
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ABSTRACT: The mechanism by which hydroxyapatite (HA)-coated titanium promotes bone-implant integration is largely unknown. Furthermore, refining the fabrication of nano-structured HA to the level applicable to the mass production process for titanium implants is challenging. This study reports successful creation of nanopolymorphic crystalline HA on microroughened titanium surfaces using a combination of flame spray and low-temperature calcination and tests its biological capability to enhance bone-implant integration. Sandblasted microroughened titanium implants and sandblasted + HA-coated titanium implants were subjected to biomechanical and histomorphometric analyses in a rat model. The HA was 55% crystallized and consisted of nanoscale needle-like architectures developed in various diameters, lengths, and orientations, which resulted in a 70% increase in surface area compared to noncoated microroughened surfaces. The HA was free from impurity contaminants, with a calcium/phosphorus ratio of 1.66 being equivalent to that of stoichiometric HA. As compared to microroughened implants, HA-coated implants increased the strength of bone-implant integration consistently at both early and late stages of healing. HA-coated implants showed an increased percentage of bone-implant contact and bone volume within 50 μm proximity of the implant surface, as well as a remarkably reduced percentage of soft tissue intervention between bone and the implant surface. In contrast, bone volume outside the 50 μm border was lower around HA-coated implants. Thus, this study demonstrated that the addition of pure nanopolymorphic crystalline HA to microroughened titanium not only accelerates but also enhances the level of bone-implant integration and identified the specific tissue morphogenesis parameters modulated by HA coating. In particular, the nanocrystalline HA was proven to be drastic in increasing osteoconductivity and inhibiting soft tissue infiltration, but the effect was limited to the immediate microenvironment surrounding the implant.
International Journal of Nanomedicine 01/2012; 7:859-73. · 3.13 Impact Factor
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ABSTRACT: This study introduces nanopolymorphic features of alkali- and heat-treated titanium surfaces, comprising of tuft-like, plate-like, and nodular structures that are smaller than 100 nm and determines whether and how the addition of these nanofeatures to a microroughened titanium surface affects bone-implant integration. A comprehensive assessment of biomechanical, interfacial, and histological analyses in a rat model was performed for machined surfaces without microroughness, sandblasted-microroughened surfaces, and micro-nano hybrid surfaces created by sandblasting and alkali and heat treatment. The microroughened surface accelerated the establishment of implant biomechanical fixation at the early healing stage compared with the non-microroughened surface but did not increase the implant fixation at the late healing stage. The addition of the nanopolymorphic features to the microroughened surface further increased implant fixation throughout the healing time. The area of the new bone within 50 μm proximity of the implant surfaces, which was increased 2-3-fold using microroughened surfaces, was further increased 2-fold using nanopolymorphic surfaces. In contrast, the bone area in a 50-200 μm zone was not influenced by either microroughened or nanopolymorphic surfaces. The percentage of bone-implant contact, which was increased 4-5-fold, using microroughened surfaces, was further increased substantially by over 2-fold throughout the healing period. The percentage of soft tissue intervention between bone and implant surfaces, which was reduced to half by microroughened surfaces, was additionally reduced by the nanopolymorphic surfaces to between one-third and one-fourth, resulting in only 5-7% soft tissue intervention compared with 60-75% for the non-microroughened surface. Thus, using an exemplary alkali- and heat-treated nanopolymorphic surface, this study identified critical parameters necessary to describe the process and consequences of bone-implant integration, for which nanofeatures have specific and substantial roles beyond those of microfeatures. Nanofeature-enhanced osteoconductivity, which resulted in both the acceleration and elevation of bone-implant integration, has clearly been demonstrated.
Biomaterials 10/2011; 32(30):7297-308. · 7.40 Impact Factor
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ABSTRACT: Abstract The aim of this study was to examine the bactericidal effects and bactericidal time of an acidified sodium chlorite compound gel (ASC-Gel) on bacteria isolated from the peri-implant sulci of 10 patients who received implants 3-27 years previously, and the depth of each peri-implant sulcus was 5 mm or less. Porphyromonas gingivalis (ATCC33277) (P. gingivalis) was used as the control bacterium. Five ASC-Gel preparations were created by adding 3.3%, 5.0%, 7.0%, 9.0%, and 11.0% citric acid (CA) (condition a, b, c, d, and e, respectively) into an oral moisturizing gel containing sodium chlorite. The concentrations of ClO2 generated in ASC-Gel under conditions (ae) were 12.1, 14.1, 17.2, 21.2, and 39.3 ppm, respectively. We examined the bactericidal effects of the 5 ASC-Gel preparations at volumes of 0.5, 1.0, and 2.0 ml, and measured the bactericidal time when 2.0 ml of ASC-Gel was used under condition (e). The bactericidal effects of ASC-Gel became significantly greater with increased concentrations of CA and ClO2 and with increased usage (0.52.0 ml) of the gel. All bacteria were killed by using 2.0 ml of ASC-Gel under condition (e). ASC-Gel also needed between 45 and 90 min to kill all microbes under condition (e). Within the limits of the present investigation, these results suggest that ASC-Gel is useful as a chemical disinfectant against bacteria in the peri-implant sulcus. Further studies are also required to protect teeth, the surface of hydroxyapatite-coated implants, and the surrounding soft tissues from effects of chemical dissolution such as acid erosion due to the low pH of ASC-Gel.
Journal of Oral Implantology 09/2011; · 1.53 Impact Factor
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ABSTRACT: The role of nanofeatured titanium surfaces in a number of aspects of in vivo bone-implant integration, and, in particular, their potential advantages over microfeatured titanium surfaces, as well as their specific contribution to osteoconductivity, is largely unknown. This study reports the creation of a unique nanobimorphic titanium surface comprised of nanotrabecular and nanotuft-like structures and determines how the addition of this nanofeature to a microroughened surface affects bone-implant integration. Machined surfaces without microroughness, sandblasted microroughened surfaces, and micro-nano hybrid surfaces created by sandblasting and alkali and heat treatment of Ti-15Mo-5Zr-3Al alloy were subjected to biomechanical, interfacial and histological analyses in a rat model. The presence of microroughness enabled accelerated establishment of biomechanical implant fixation in the early stages of healing compared to the non-microroughened surfaces; however, it did not increase the implant fixation at the late stages of healing. The addition of nanobimorphic features to the microroughened surfaces further increased the implant fixation by as much as 60-100% over the healing time. Bone area within 50 μm of the implant surface, but not beyond this distance, was significantly increased by the presence of nanobimorphic features. Although the percentage of bone-implant contact was also significantly increased by the addition of nanobimorphic features, the greatest improvement was found in the soft tissue intervention between the bone and the implant, which was reduced from >30% to <5%. Mineralized tissue densely deposited with calcium-binding globular proteins was observed in an extensive area of nanobimorphic surfaces after biomechanical testing. This study clearly demonstrates the nanofeature-enhanced osteoconductivity of titanium by an alkali- and heat-treated nanobimorphic surface compared to that by microfeatured surfaces, which results not only in an acceleration but also an improvement of bone-implant integration. The identified biological parameters that successfully detect the advantages of nanofeatures over microfeatures will be useful in evaluating new implant surfaces in future studies.
Acta biomaterialia 08/2011; 7(12):4267-77. · 3.98 Impact Factor
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ABSTRACT: Prevention of local infection from wound pathogens such as Staphylococci and Streptococci is crucial for tissue regeneration. N-acetyl cysteine (NAC), an anti-oxidant amino acid derivative, has anti-microbial potential against various species. This in vitro study evaluated whether NAC prevented bacterial infection of gingival fibroblasts and osteoblasts on a scaffold. N-acetyl cysteine delayed growth of Staphylococcus aureus and Streptococcus pyogenes cultured in brain heart infusion (BHI) broth for 12 h in an almost dose-dependent manner (2.5, 5.0 or 10.0 mm). The number of rat gingival fibroblasts on collagen scaffolds with bacterial co-incubation was less than 30% of that in cultures without bacterial co-incubation at day 7. However, pre-addition of NAC to the scaffold yielded a number comparable with that in culture without bacteria. Fibroblasts on the scaffold with bacterial co-incubation were small, rounded and filled with bacteria and reactive oxygen species. Pre-addition of NAC, however, resulted in fibroblasts similar to those observed in culture without bacterial co-incubation. N-acetyl cysteine completely prevented devastating suppression of alkaline-phosphatase activity and extracellular matrix mineralization in osteoblastic culture on scaffolds with bacterial co-incubation. These results indicate that NAC can functionalize a scaffold with anti-infective capabilities, thus assisting healing of soft and hard tissues.
Biomaterials 08/2011; 32(33):8474-85. · 7.40 Impact Factor
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ABSTRACT: The independent, genuine role of surface chemistry in the biological properties of titanium is unknown. Although microtopography has been established as a standard surface feature in osseous titanium implants, unfavorable behavior and reactions of osteogenic cells are still observed on the surfaces. To further enhance the biological properties of microfeatured titanium surfaces, this study tested the hypotheses that (1) the surface chemistry of microroughened titanium surfaces can be controllably varied by coating with a very thin layer of TiO(2), without altering the existing topographical and roughness features; and (2) the change in the surface chemistry affects the biological properties of the titanium substrates. Using a slow-rate sputter deposition of molten TiO(2) nanoparticles, acid-etched microroughened titanium surfaces were coated with a TiO(2) layer of 300-pm to 6.3-nm thickness that increased the surface oxygen levels without altering the existing microtopography. The attachment, spreading behavior, and proliferation of osteoblasts, which are considered to be significantly impaired on microroughened surfaces compared with relatively smooth surfaces, were considerably increased on TiO(2)-coated microroughened surfaces. The rate of osteoblastic differentiation was represented by the increased levels of alkaline phosphatase activity and mineral deposition as well as by the upregulated expression of bone-related genes. These biological effects were exponentially correlated with the thickness of TiO(2) and surface oxygen percentage, implying that even a picometer-thin TiO(2) coating is effective in rapidly increasing the biological property of titanium followed by an additional mild increase or plateau induced by a nanometer-thick coating. These data suggest that a super-thin TiO(2) coating of pico-to-nanometer thickness enhances the biological properties of the proven microroughened titanium surfaces by controllably and exclusively modulating their surface chemistry while preserving the existing surface morphology. The improvements in proliferation and differentiation of osteoblasts attained by this chemical modification is of great significance, providing a new insight into how to develop new implant surfaces for better osseointegration, based on the established microtopographic surfaces.
Biomaterials 08/2011; 32(33):8374-84. · 7.40 Impact Factor
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ABSTRACT: This study addresses the control of the biological capabilities of titanium through specific nanosurface features and its potential modulation by UV photofunctionalization. Rat bone marrow derived osteoblasts were cultured on titanium disks with micropits alone, micropits with 100 nm nodules, micropits with 300 nm nodules, or micropits with 500 nm nodules, with or without UV treatment. After a 24 h incubation protein adsorption, as well as the attachment, retention, and spread of osteoblasts were examined in correlation with the topographical parameters of the titanium substrates. Each of the biological events was governed by a different set of multiple surface topographical factors with a distinctive pattern of regulation. For instance, without UV treatment the protein adsorption and cell attachment capability of titanium substrates increased linearly with increasing average roughness (Ra) and surface area of titanium disks, but increased polynomially with increasing nanonodule diameter. The cell retention capability increased polynomially with increasing nanonodular diameter and Ra, but increased linearly with increasing surface area. Consequently, the micropits with 300 nm nodules created the most favorable environment for this initial osteoblast behavior and response. UV treatment of the nanonodular titanium surfaces resulted in considerable enhancement of all biological events. However, the pattern of UV-mediated enhancement was disproportionate; exponential and overriding effects were observed depending upon the biological event and topographical parameter. As an example of overriding enhancement, the cell retention capability, which fluctuated with changes in various topographical parameters, became invariably high after UV treatment. The present data provide a basis for understanding how to optimize nanostructures to create titanium surfaces with increased biological capabilities and uncover a novel advantage of UV photofunctionalization of titanium substrates that synergistically increases its nanotopography enhanced biological capabilities whereby most of the initial biological events of osteoblasts were overwhelmingly enhanced beyond a simple proportional increase.
Acta biomaterialia 06/2011; 7(10):3679-91. · 3.98 Impact Factor
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ABSTRACT: The purpose of this study was to evaluate the cytotoxicity of mineral trioxide aggregate (MTA) and its potential detoxification by an antioxidant amino acid, N-acetylcysteine (NAC).
Rat dental pulp cells extracted from rat maxillary incisors were directly cultured on MTA with or without NAC in culture medium. The number of cells and their spreading behavior were both assessed 24 hours after seeding. The intracellular levels of reactive oxygen species (ROS) and glutathione (GSH) were also assessed after 24 hours of culture.
The number of cells attached to MTA was 60% greater when NAC was added to the culture medium. In addition, the area and perimeter of the cells were found to be 2-fold greater in the culture containing NAC. Cells cultured on MTA alone showed large ROS concentrations, which disappeared when the medium was supplemented with NAC. The intracellular GSH level, however, increased 3.5-fold with NAC addition.
This study demonstrated that the presence of NAC in environments can substantially improve attachment and spreading behaviors of dental pulp cells on MTA. This biological effect was associated with an improvement in the cellular redox system by NAC and warrants further exploration of NAC for determining its therapeutic value in improving the biocompatibility of MTA.
Journal of endodontics 05/2011; 37(5):637-41. · 2.95 Impact Factor
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ABSTRACT: Titanium surfaces with micro-nano hybrid topography (nanoscale nodules in microscale pits) have been recently demonstrated to show higher biological capability than those with microtopography alone. On the other hand, UV treatment of titanium surfaces, which is called UV photofunctionalization, has recently been introduced to substantially increase the biological capability and osteoconductivity of titanium surfaces. However, synergistic effects of these two advanced surface modification technologies and regulatory factors to potentially modulate the mutual effects have never been addressed. In this study, utilization of a recently discovered controllable self-assembly of TiO(2) nanonodules has enabled the exploration of the relative contribution of different sizes of nanostructures to determine the biological capability of titanium surfaces and their relative responsiveness to UV photofunctionalization. Rat bone marrow-derived osteoblasts were cultured on titanium disks with either micropits alone, micropits with 100-nm nodules, micropits with 300-nm nodules, or micropits with 500-nm nodules, with or without UV treatment. Although UV treatment increased the attachment, spread, proliferation, and mineralization of these cells on all titanium surfaces, these effects were more accentuated (3-5 times) on nanonodular surfaces than on surfaces with micropits alone and were disproportionate depending on nanonodule sizes. For instance, on UV-treated micro-nano hybrid surfaces, cell attachment correlated with nanonodule sizes in a quadratic approximation with its peak for 300-nm nodules followed by a decline for 500-nm nodules, while cell attachment exponentially correlated with surface roughness with its plateau achieved for 300-nm nodules without a subsequent decline. Moreover, cell attachment increased in a linear correlation with the surface area, while no significant effect of the inter-irregularities space or degree of hydrophilicity was observed on cell attachment. These results suggest that the effect of UV photofunctionalization can be multiplied on micro-nano hybrid titanium surfaces compared with the surfaces with micropits alone. This multiplication is disproportionately regulated by a selected set of topographical parameters of the titanium surfaces. Among the nanonodules tested in this study, 300-nm nodules seemed to create the most effective morphological environment for responding to UV photofunctionalization. The data provide a systematic platform to effectively optimize nanostructures on titanium surfaces in order to enhance their biological capability as well as their susceptibility to UV photofunctionalization.
Biomaterials 03/2011; 32(19):4358-68. · 7.40 Impact Factor
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ABSTRACT: The independent role of the surface chemistry of titanium in determining its biological properties is yet to be determined. Although titanium implants are often in contact with muscle tissue, the interaction of muscle cells with titanium is largely unknown. This study tested the hypotheses that the surface chemistry of clinically established microroughened titanium surfaces could be controllably varied by coating with a minimally thin layer of TiO(2) (ideally pico-to-nanometer in thickness) without altering the existing topographical and roughness features, and that the change in superficial chemistry of titanium is effective in improving the biological properties of titanium.
Acid-etched microroughened titanium surfaces were coated with TiO(2) using slow-rate sputter deposition of molten TiO(2) nanoparticles. A TiO(2) coating of 300 pm to 6.3 nm increased the surface oxygen on the titanium substrates in a controllable manner, but did not alter the existing microscale architecture and roughness of the substrates. Cells derived from rat skeletal muscles showed increased attachment, spread, adhesion strength, proliferation, gene expression, and collagen production at the initial and early stage of culture on 6.3 nm thick TiO(2)-coated microroughened titanium surfaces compared with uncoated titanium surfaces.
Using an exemplary slow-rate sputter deposition technique of molten TiO(2) nanoparticles, this study demonstrated that titanium substrates, even with microscale roughness, can be sufficiently chemically modified to enhance their biological properties without altering the existing microscale morphology. The controllable and exclusive chemical modification technique presented in this study may open a new avenue for surface modifications of titanium-based biomaterials for better cell and tissue affinity and reaction.
International Journal of Nanomedicine 01/2011; 6:2191-203. · 3.13 Impact Factor
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ABSTRACT: Bioactivity and osteoconductivity of titanium degrade over time after surface processing. This time-dependent degradation is substantial and defined as the biological aging of titanium. UV treatment has shown to reactivate the aged surfaces, a process known as photofunctionalization. This study determined whether there is a difference in the behavior of biological aging for titanium with micro-nano-hybrid topography and titanium with microtopography alone, following functionalization. Titanium disks were acid etched to create micropits on the surface. Micro-nano-hybrid surfaces were created by depositioning 300-nm diameter TiO(2) nodules onto the micropits using a previously established self-assembly protocol. These disks were stored for 8 weeks in the dark to allow sufficient aging, then treated with UV light for 48 hours. Rat bone marrow-derived osteoblasts were cultured on fresh disks (immediately after UV treatment), 3-day-old disks (disks stored for 3 days after UV treatment), and 7-day- old disks. The rates of cell attachment, spread, proliferation, and levels of alkaline phosphatase activity, and calcium deposition were reduced by 30%-50% on micropit surfaces, depending on the age of the titanium. In contrast, 7-day-old hybrid surfaces maintained equivalent levels of bioactivity compared with the fresh surfaces. Both micropit and micro-nano-hybrid surfaces were superhydrophilic immediately after UV treatment. However, after 7 days, the micro-nano- hybrid surfaces became hydrorepellent, while the micropit surfaces remained hydrophilic. The sustained bioactivity levels of the micro-nano-hybrid surfaces were nullified by treating these surfaces with Cl(-)anions. A thin TiO(2) coating on the micropit surface without the formation of nanonodules did not result in the prevention or alleviation of the time-dependent decrease in biological activity. In conclusion, the micro-nano-hybrid titanium surfaces may slow the rate of time-dependent degradation of titanium bioactivity after UV photofunctionalization compared with titanium surfaces with microtopography alone. This antibiological aging effect was largely regulated by its sustained electropositivity uniquely conferred in TiO(2) nanonodules, and was independent of the degree of hydrophilicity. These results demonstrate the potential usefulness of these hybrid surfaces to effectively utilize the benefits of UV photofunctionalization and provide a model to explore the mechanisms underlying antibiological aging properties.
International Journal of Nanomedicine 01/2011; 6:1327-41. · 3.13 Impact Factor
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ABSTRACT: Current dental restorative materials are only used to fill the defect of hard tissues, such as dentin and enamel, because of their cytotoxicity. Therefore, exposed dental pulp tissues in deep cavities must be first covered by a pulp capping material like calcium hydroxide to form a layer of mineralized tissue. However, this tissue mineralization is based on pathological reaction and triggers long-lasting inflammation, often causing clinical problems. This study tested the ability of N-acetyl cysteine (NAC), amino acid derivative, to reduce cytotoxicity and induce mineralized tissue conductivity in resin-modified glass ionomer (RMGI), a widely used dental restorative material having dual cure mechanism. Rat dental pulp cells were cultured on untreated or NAC-supplemented RMGI. NAC supplementation substantially increased the percentage of viable cells from 46.7 to 73.3% after 24-h incubation. Cell attachment, spreading, proliferative activity, and odontoblast-related gene and protein expressions increased significantly on NAC-supplemented RMGI. The mineralization capability of cells, which was nearly suppressed on untreated RMGI, was induced on NAC-supplemented RMGI. These improved behaviors and functions of dental pulp cells on NAC-supplemented RMGI were associated with a considerable reduction in the production of intracellular reactive oxygen species and with the increased level of intracellular glutathione reserves. These results demonstrated that NAC could detoxify and functionalize RMGIs via two different mechanisms involving in situ material detoxification and antioxidant cell protection. We believe that this study provides a new approach for developing dental restorative materials that enables mineralized tissue regeneration.
Biomaterials 10/2010; 31(28):7213-25. · 7.40 Impact Factor
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ABSTRACT: The objectives of this in vitro study were (1) to determine whether a commercially available collagen membrane (CM) or human demineralized freeze-dried bone (DFDB) particles adversely affected viability or function in cultured osteoblasts through oxidative stress, and, if so, (2) to determine whether N-acetyl cysteine (NAC) successfully prevented loss of viability and dysfunction in osteoblasts.
Rat calvaria-derived osteoblasts were seeded onto polystyrene and commercially available CM (Cytoplast ®) or DFDB (DynaGraft ™) with or without pretreatment with NAC solution. The osteoblastic response was evaluated using a flow cytometric cell viability assay, measurement of attached viable cell number, quantification of reactive oxygen species (ROS) and alkaline phosphatase (ALP) staining.
The percentage of viable cells on CM was <50% at 24 h after seeding. However, this increased to 70% by pretreatment with NAC. The numbers of attached osteoblasts on DFDB remained at 60% the level of that on polystyrene at 24 h after seeding, but increased to up to 90% the level of that on polystyrene with NAC pretreatment. Although collagen materials increased intracellular ROS generation 1.5-5 times that with polystyrene, this was significantly reduced by NAC pretreatment. The percentage of the ALP-positive area was consistently 7% or less on CM and DFDB at days 7 and 14, which was restored by NAC pretreatment up to 60% or more.
Commercially available CM and DFDB impaired osteoblastic viability and function and markedly increased intracellular ROS, indicating an oxidative stress-mediated negative impact on osteoblasts. Pretreatment with NAC substantially alleviated these cytotoxic effects.
Clinical Oral Implants Research 10/2010; 22(2):165-72. · 2.51 Impact Factor
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ABSTRACT: We monitored the quantity of airborne microorganisms at 11 points (points A to K) in a dental office on a routine day of use, and tested the bactericidal efficacy of chlorine dioxide (ClO₂) gas in the dental operatory after consulting hours. Fallen airborne microorganisms were collected under air-conditioning (AC) in the dental office, and under four conditions in the operatory. Specimens of the microbes were cultivated on nutrient and Sabouraud agar media (NAM and SAM). Many colonies were observed at the entrance hall and on the cabinet in a disinfection room in the NAM and SAM tests, respectively, while no colony was observed at the foot position of the operating table and treatment bed, and above the head position of the operating room in the NAM and SAM tests, respectively. In the bactericidal efficacy test using ClO₂ gas, the dental operatory could be kept clean by the use of 4 mg/L-ClO₂ gas in addition to the use of an AC with a plasma filter and the HEPA filter.
Biocontrol science 09/2010; 15(3):103-9. · 0.78 Impact Factor
