• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.299-308
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• 2008

Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

• The Journal of Korean Academy of Prosthodontics
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• v.45 no.6
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• pp.795-804
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• 2007

Statement of problem. Nano-scale calcium-phosphate coating on the anodizing titanium surface using ion beam-assisted deposition (IBAD) has been recently introduced to improve the early osseointegration. However, not much is known about their surface characteristics that have influence on tissue-implant interaction. Purpose. This study was aimed to investigate microtopography, surface roughness, surface composition, and wettability of the titanium surface modified by the anodic oxidation and calcium phosphate coating using IBAD. Material and methods. Commercially pure titanium disks were used as substrates. The experiment was composed of four groups. Group MA surfaces represented machined surface. Group AN was anodized surface. Group CaP/AN was anodic oxidized and calcium phosphate coated surfaces. Group SLA surfaces were sandblasted and acid etched surfaces. The prepared titanium discs were examined as follows. The surface morphology of the discs was examined using SEM. The surface roughness was measured by a confocal laser scanning microscope. Phase components were analyzed using thin-film x-ray diffraction. Wettability analyses were performed by contact angle measurement with distilled water, formamide, bromonaphtalene and surface free energy calculation. Results. (1) The four groups showed specific microtopography respectively. Anodized and calcium phosphate coated specimens showed multiple micropores and tiny homogeneously distributed crystalline particles. (2) The order of surface roughness values were, from the lowest to the highest, machined group, anodized group, anodized and calcium phosphate deposited group, and sandblasted and acid etched group. (3) Anodized and calcium phosphate deposited group was found to have titanium and titanium anatase oxides and exhibited calcium phosphorous crystalline structures. (4) Surface wettability was increased in the order of calcium phosphate deposited group, machined group, anodized group, sandblasted and acid etched group. Conclusion. After ion beam-assisted deposition on anodized titanium, the microporous structure remained on the surface and many small calcium phosphorous crystals were formed on the porous surface. Nanoscale calcium phosphorous deposition induced roughness on the microporous surface but hydrophobicity was increased.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.3
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• pp.307-319
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• 2008

STATEMENT OF PROBLEM: It is known that an anodic oxidation technique, one of the methods for the implant surface treatment, remarkably increased surface area, enhanced wettability and accelerated the initial bone healing. Purpose: This study was performed to evaluate the wettability of anodized titanium surface which has a nanotubular structure, to assess osseointegration after the placement of implant with nano-size tubes on tibia of rats and to analyze quantitatively transferable rhBMP-2 on each surface. MATERIAL AND METHOD: Four different kinds of surface-treated titanium discs (polished (machined surface) group, micro (blasting surface) group, nano (anodizedmachined surface) group, and nano-micro (anodized-blasting surface) group) were fabricated (n=10). Three different media were chosen to measure the surface contact angles; distilled water, plasma and rhBMP-2 solution. After a single drop (0.025 $m{\ell}$) of solution, the picture was taken with the image camera, and contact angle was measured by using image analysis system. For the test of osseointegration, 2 kinds of anodized surface (anodized-machined surface, anodized-blasting surface) implants having 2.0 mm in diameter and 5.0 mm in length inserted into the tibia of Wistar rats. After 3 weeks, tibia were harvested and the specimens were stained with hematoxylin and eosin for histological analysis. To test the possibility of drug delivery, after soaking sample groups in the concentration of 250 ng/$m{\ell}$l of rhBMP-2 for 48 hours, the excess solution of rhBMP-2 were removed. After that, they were lyophilized for 24 hours, and then the rhBMP-2 on the surface of titanium was resolved for 72 hours in PBS. All the extracted solution was analyzed by ELISA. One-way analysis of variance (ANOVA) was performed on the data. RESULTS: The wettability is improved by anodic oxidation. The best wettability was shown on the nano-micro group, and it was followed by nano group, micro group, and polished group. In the histological findings, all implants showed good healing and the new bone formation were observed along the implant surface. After 3 days, nano-micro group delivered the most amount of rhBMP-2, followed by nano group, micro group, and polished group. CONCLUSION: It indicated that anodic oxidation on blasting surface produce functionally graded nano-micro porous structure and enhance hydrophilicity of the surface and osseointegration. The findings suggest that the nano-micro porous structure could be a useful carrier of osteogenic molecules like rhBMP-2.

• The Journal of Advanced Prosthodontics
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• v.1 no.2
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• pp.75-84
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• 2009

STATEMENT OF PROBLEM. A number of studies about the nano-treated surfaces of implants have been conducting along with micro-treated surfaces of implants. PURPOSE. The purpose of this study was to get information for the clinical use of nano-treated surfaces compared with micro-treated surfaces by measuring removal torque and analyzing histological characteristics after the placement of various surface-treated implants on femurs of dogs. MATERIAL AND METHODS. Machined surface implants were used as a control group. 4 nano-treated surface implants and 3 micro-treated surface implants [resorbable blast media surface (RBM), sandblast and acid-etched surface (SAE), anodized RBM surface] were used as experimental groups. Removal torque values of implants were measured respectively and the histological analyses were conducted on both 4weeks and 8weeks after implant surgery. The surfaces of removed implants after measuring removal torque values were observed by scanning electron microscopy (SEM) at 8 weeks. RESULTS. 1. Removal torque values of the nano-treated groups were lower than those of micro-treated groups. 2. Removal torque values were similar in the anodized RBM surface groups. 3. On the histological views, there was much of bone formation at 8 weeks, but there was no difference between 4 and 8 weeks, and between the types of implant surfaces as well. CONCLUSION. it is suggested that implant topography is more effective in removal torque test than surface chemistry. To get better clinical result, further studies should be fulfilled on the combined effect of surface topography and chemistry for the implant surface treatments.

• Journal of Periodontal and Implant Science
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• v.43 no.4
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• pp.198-205
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• 2013

Purpose: The aim of this study was to evaluate the surface properties and biological response of an anodized titanium surface by cell proliferation and alkaline phosphatase activity analysis. Methods: Commercial pure titanium (Ti) disks were prepared. The samples were divided into an untreated machined Ti group and anodized Ti group. The anodization of cp-Ti was formed using a constant voltage of 270 V for 60 seconds. The surface properties were evaluated using scanning electron microscopy, X-ray photoelectron spectroscopy, and an image analyzing microscope. The surface roughness was evaluated by atomic force microscopy and a profilometer. The contact angle and surface energy were analyzed. Cell adhesion, cell proliferation, and alkaline phosphatase activity were evaluated using mouse $MC_3T_3-E_1$ cells. Results: The anodized Ti group had a more porous and thicker layer on its surface. The surface roughness of the two groups measured by the profilometer showed no significant difference (P>0.001). The anodized Ti dioxide ($TiO_2$) surface exhibited better corrosion resistance and showed a significantly lower contact angle than the machined Ti surface (P>0.001). Although there was no significant difference in the cell viability between the two groups (P>0.001), the anodized $TiO_2$ surface showed significantly enhanced alkaline phosphatase activity (P<0.001). Conclusions: These results suggest that the surface modification of Ti by anodic oxidation improved the osteogenic response of the osteoblast cells.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.499-518
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• 2003

The surface characteristics of titanium have been shown to have an important role in contact ossseointegration around the implant. Anodizing at high voltage produces microporous structure and increases thickness of surface titanium dioxide layer. The aim of present study was to analyse the response of rat calvarial osteoblast cell to commercially pure titanium and Ti-6A1-4V anodized in 0.06 mol/l ${\beta}$-glycerophosphate and 0.03 mol/l sodium acetate. In this study, rat calvarial osteoblasts were used to assay for cell viability and cell proliferation on the implant surface at 1,2,4,7 days. 1. Surface roughness was 1.256${\mu}m$ at 200V, and 1.745${\mu}m$ at 300V. 2. The thickness of titanium oxide layer was increased 1 ${\mu}m$ with the increase of 50V. 3. The proliferation rate of osteoblastic cells was increased with the increase of the surface roughness and the thickness of titanium oxide layer. 4. There was no difference in cell viability and cell proliferation between commercially pure titanium and Ti-6A1-4V anodized at the same condition. In conclusion, the titanium surface modified by anodizing was biocompatible, produced enhanced osteoblastic response. The reasons of enhanced osteoblast response might be due to reduced metal ion release by thickened and stabilized titanium dioxide layer and microporous rough structures.

• The Journal of Korean Academy of Prosthodontics
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• v.43 no.6
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• pp.751-763
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• 2005

Statement of problem. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. The surface quality of the implant depends on the chemical, physical, mechanical and topographical properties of the surface. The different properties will interact with each other and a change in thickness of the oxide layer may also result in a change in surface energy, the surface topography and surface, chemical composition. However, there is limited the comprehensive study with regard to changed surface and biologic behavior of osteoblast by anodization. Purpose of study. The aim of this study was to analyze the characteristics of an oxide layer formed and to evaluate the cellular biologic behaviors on titanium by anodic oxidation (anodization) by cellular proliferation, differentiation, ECM formation and gene expression. And the phospholipase activity was measured on the anodized surface as preliminary study to understand how surface properties of Ti implant are transduced into downstream cellular events. Methods and Materials. The surface of a commercially pure titanium(Grade 2) was modified by anodic oxidation. The group 1 samples had a machined surface and other three experimental specimens were anodized under a constant voltage of 270 V(Group 2), 350 V(Group 3), and 450 V(Group 4). The specimen characteristics were inspected using the following five categories; the surface morphology, the surface roughness, the thickness of oxide layer, the crystallinity, and the chemical composition of the oxide layer. Cell numbers were taken as a marker for cell proliferation. While the expression of alkaline phosphatase and Runx2 (Cbfa1) was used as early differentiation marker for osteoblast. The type I collagen production was determined, which constitutes the main structural protein of the extracellular matrix. Phospholipase $A_2$ and D activity were detected. Results. (1) The anodized titanium had a porous oxide layer, and there was increase in both the size and number of pores with increasing anodizing voltage. (2) With increasing voltage, the surface roughness and thickness of the oxide film increased significantly (p<0.01), the $TiO_2$phase changed from anatase to rutile. During the anodic oxidization, Ca and P ions were more incorporated into the oxide layer. (3) The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity, type I collagen production, and Cbfa 1 gene expression increased significantly (p<0.01), while the cell proliferation decreased. (4) In preliminary study on the relation of surface property and phospholipase, PLD activity was increased but $PLA_2$ activity did not changed according to applied voltage. Conclusion. The anodized titanium shows improved surface characteristics than the machined titanium. The surface properties acquired by anodization appear to give rise more mature osteoblast characteristics and might result in increased bone growth, and contribute to the achievement of a tight fixation. The precise mechanism of surface property signaling is not known, may be related to phospholipase D.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.363-372
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• 2008

Purpose: This study compared the effects of coating implants with hydroxyapatite (HA) using an ion beam-assisted deposition (IBAD) method prepared with machined, anodized, sandblasted and large-grit acid etched (SLA) surfaces in minipigs, and verified the excellency of coating method with HA using IBAD. Material and Methods: 4 male Minipigs(Prestige World Genetics, Korea), 18 to 24 months old and weighing approximately 35 to 40 kg, were chosen. All premolars and first molars of the maxilla were carefully extracted on each side. The implants were placed on the right side after an 8 week healing period. The implant stability was assessed by resonance frequency analysis (RFA) at the time of placement. 40 implants were divided into 5 groups; machined, anodized, anodized plus IBAD, SLA, and SLA plus IBAD surface implants. 4 weeks after implantation on the right side, the same surface implants were placed on the left side. After 4 weeks of healing, the minipigs were sacrificed and the implants were analyzed by RFA, histology and histometric. Results: RFA showed a mean implant stability quotient (ISQ) of $75.625{\pm}5.021$, $76.125{\pm}3.739$ ISQ and $77.941{\pm}2.947$ at placement, after 4 weeks healing and after 8 weeks, respectively. Histological analysis of the implants demonstrated newly formed, compact, mature cortical bone with a nearby marrow spaces. HA coating was not separated from the HA coated implant surfaces using IBAD. In particular, the SLA implants coated with HA using IBAD showed better contact osteogenesis. Statistical and histometric analysis showed no significant differences in the bone to implant contact and bone density among 5 tested surfaces. Conclusion: We can conclude that rough surface implants coated with HA by IBAD are more biocompatible, and clinical, histological, and histometric analysis showed no differences when compared with the other established implant surfaces in normal bone.

• The Journal of Korean Academy of Prosthodontics
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• v.42 no.3
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• pp.307-326
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• 2004

Statement of problem. The long-term success of implants is the development of a stable direct connection between bone and implant surface, which must be structural and functional. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. Among them, altering the surface properties can modify cellular responses such as cell adhesion, cell motility and bone deposition. Purpose. This study was to evaluate the cellular behaviors on the surface-modified titanium by morphological observation, cellular proliferation and differentiation. Material and methods. Specimens were divided into five groups, depending on their surface treatment: electropolishing(EP) anoclizing(AN), machining(MA), blasting with hydroxyapatite particle(RBM) and electrical discharge machining(EDM). Physicochemical properties and microstructures of the specimens were examined and the responses of osteoblast-like cells were investigated. The microtopography of specimens was observed by scanning electron microscopy(SEM). Surface roughness was measured by a three-dimensional roughness measuring system. The microstructure was analyzed by X-ray diffractometer(XRD) and scanning auger electron microscopy(AES). To evaluate cellular responses to modified titanium surfaces, osteoblasts isolated from neonatal rat were cultured. The cellular morphology and total protein amounts of osteoblast-like cell were taken as the marker for cellular proliferation, while the expression of alkaline phosphatase was used as the early differentiation marker for osteoblast. In addition, the type I collagen production was determined to be a reliable indicator of bone matrix synthesis. Results. 1. Each prepared specimen showed specific microtopography at SEM examination. The RBM group had a rough and irregular pattern with reticulated appearance. The EDM-treated surface had evident cracks and was heterogeneous consisting of broad sheet or plate with smooth edges and clusters of small grains, deep pores or craters. 2. Surface roughness values were, from the lowest to the highest, electropolished group, anodized group, machined group, RBM group and EDM group. 3. All groups showed amorphous structures. Especially anodized group was found to have increased surface oxide thickness and EDM group had titaniumcarbide(TiC) structure. 4. Cells on electropolished, anodized and machined surfaces developed flattened cell shape and cells on RBM appeared spherical and EDM showed both. After 14 days, the cells cultured from all groups were formed to be confluent and exhibited multilayer proliferation, often overlapped or stratified. 5. Total protein amounts were formed to be quite similar among all the group at 48 hours. At 14 days, the electropolished group and the anodized group induced more total protein amount than the RBM group(P<.05). 6. There was no significant difference among five groups for alkaline phosphatase(ALP) activity at 48 hours. The AN group showed significantly higher ALP activity than any other groups at 14 days(P<.05). 7. All the groups showed similar collagen synthesis except the EDM group. The amount of collagen on the electropolished and anodized surfaces were higher than that on the EDM surface(P<.05).

• The Journal of Korean Academy of Prosthodontics
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• v.43 no.6
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• pp.745-750
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• 2005

Statement of problem. Resonance frequency analysis has been increasingly served as a non-invasive and objective method for clinical monitoring of implant stability. Many clinical studies must be required for standardized data using RFA. Purpose. This study was performed to evaluate RFA value changes in two anodized implant groups. Material and method. Among a total of 24 implants, twelve screw shaped implants as a test group (H2-R8.5) were manufactured, which had a pitch-height of 0.4 mm, an outer diameter of 4.3 mm, a length of 8.5 mm, and external hexa-headed, were turned from 5 mm rods of commercially pure titanium (ASTM Grade IV, Warantec Co., Seoul, Korea), and another twelve implants as a control group were $Br{\aa}nemark$ Ti-Unite MK4 (diameter 4.0 mm, length 8.5 mm). Each group was installed in tibia of rabbit. Two implants were placed in each tibia (four implants per rabbit). Test two implants were inserted in right side and control two in left side. ISQ values were measured using $Osstell^{TM}$ (Integration Diagnostics Ltd. Sweden) during fixture installation, and 12 weeks later and evaluated the RFA changes. Results. Mean and SD of baseline ISQ values of test group were $75.0{\pm}3.4$ and $68.7{\pm}8.1$ for control group. Mean and SO of ISQ values 12 weeks after implant insertion were $73.2{\pm}4.7$ for test group and $72.6{\pm}3.9$ for control group. There were no statistically significant differences between groups in ISQ values after 3 months (P>0.05). From the data, RFA gains after 3 months were calculated, and there was statistically significant difference between groups (P<0.05). Conclusion. Although there were RFA changes between groups, implant stability after experimental period shows alike tendency and good bone responses.