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Analysis of p53 and Retinoblasoma(Rb) Gene Polymorphisms in Relation to Lung Cancer in Koreans (한국인 폐암 환자에 대한 p53 및 Rb유전자의 다형성 분석)

  • Lee, Kyung-Sang;Sohn, Jang-Won;Yang, Suck-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo;Lee, Jung-Hee;Lee, Chun-Geun;Cho, Youl-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.534-546
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    • 1997
  • Background : The p53 and retinoblastoma(Rb) tumor suppressor genes are associated with the pathogenesis of several types of human cancer. Substantial proportion of the primary lung cancers or cell lines have been reported to have the p53 and/or the Rb gene mutations. But, so far there is no report on the analysis of the Rb gene polymorphism as one of the genetic susceptibility marker. This study was undertaken to establish the gene frequencies of the polymorphic genotypes of the p53 and Rb genes in Koreans to evaluate the possible involvement of these genotypes as a risk factor of lung cancer. Methods : In this study 145 controls without previous and present tumor history and 128 lung cancer patients were subjected to analysis. The two intragenic polymorphisms of the p53 gene(exon 4/ AccII, intron 6/MspI) and one intron 17/XbaI polymorphism of the Rb gene were analysed by the method of polymersae chain reaction- restriction fragment length polymorphisms(PCR-RFLPs). The genotype of the intron 3/16 bp repeat polymorphism of p53 was determined by PCR and direct gel electrophoresis. Results : There were no significant differences in the genotype distributions of the p53 gene between lung cancer patients and controls. But heterozygotes(Arg/Pro) of the exon 4/AccII polymorphisms were slightly over-represented than controls, especially in the Kreyberg type I cancer, which was known to be associated with smoking. The intron 3/16 bp duplication and the intron 6/MspI polymorphisms were in complete linkage disequilibrium. About 95% of the individuals were homozygotes of the common alleles both in the 16 duplication and MspI polymorphisms, and no differences were deteced in the genotype distributions between lung cancer patients and controls. Overall genotype distributions of the Rb gene polymorphisms between lung cancer patients and controls were not significantly different However, the genotype distributions in the Kreyberg type I cancer were significantly different from those of controls(p = 0.0297) or adenocarcinomas(p = 0.0008). It was noticeable that 73.4% of the patients with adenocarcinomas were heterozygotes(r1/r2) whereas 39.2% of the Kreyberg type I cancer were heterozygous at this polymorphisms. In the lung cancer patients, significant differences were also noted between the high dose smokers and low dose smokers including non-smokers(p = 0.0258). The relative risk to Kreyberg type I cancer was significantly reduced in the individuals with the genotype of r1/r2(odds ratio = 0.46, 95% C.I. = 0.25-0.86, p = 0.0124). The combined genotype distribution of the exon 4 AccII of the p53 and the intron 17 Rb gene polymorphisms in Kreyberg type I cancers were significantly different from dose of controls or adenocarcinomas. The highest odds ratio were observed in the individuals with the genotypes of Arg/Pro and r2/r2(odds ratio = 1.97,95% C.I. = 0.84-4.59) and lowest one was in the patients with Arg/Arg, r1/r2 genotype(odds ratio = 0.54, 95% C.I. = 0.25-1.14). Conclusion : The p53 and the Rb gene polymorphisms modulate the risk of smoking induced lung cancer development in Koeans. However, the exact mechanism of risk modulation by these polymorphism remains to be determined. For more discrete clarification of associations between specific genotypes and lung cancer risk, the evaluations of these polymorphisms in other ethnics and more number of patients will be needed.

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An Analysis on the Value Chain and the Value System of the Korean Wind Power Industry (한국 풍력산업의 가치사슬 및 가치시스템 분석)

  • Ryu, Jae-Ho;Choi, Ta-Gwan;Park, Jung-Gu
    • Journal of Energy Engineering
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    • v.23 no.1
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    • pp.46-57
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    • 2014
  • This study analyzes whether the value-added structure of Korean wind power industry exhibits a virtuous cycle through the value chain(VC) within wind power firms and the value system(VS) among the wind power industries, using a regression analysis based on a survey about Korean wind power companies. According to the VC, the government's R&D support is analyzed to have contributed to an increase in the R&D investments of the wind power companies. An increase in corporates' R&D investments has led to an increase in corporates' R&D outputs, and in turn, induced a remarkable increase in the amounts of production. But an increase in production has not led to a decrease in the costs of production, not resulting in an increase in profit rates per sales amount. In addition, while an increase in profit rates is analyzed to have contributed to an increase in production, this did not induce further investments in corporate's R&D. The virtuous cycle of the value chain in Korean wind power firms is, therefore, analyzed to be weak. Next, the VS is analyzed by dividing the whole chain into the system group including rotor blades, gear boxes, and power generators, and the structure group, such as towers. Two groups are analyzed to have mutually positive effects in the processes of the government's support for corporates' R&D, corporates' investment in R&D, R&D outputs, and profit rates per sales amount. Such mutual positive effects are, however, not found in the processes of the amounts of production and the costs of production. These results demonstrates that the value system of Korean wind power industry is not completed. This study has a policy implication to need further efforts to create the virtuous cycle in the VC and VS of Korean wind power industry.

The relation between angiotensin converting enzyme (ACE) gene polymorphism and neonatal hyperbilirubinemia in Korea (한국인 신생아 황달과 안지오텐신 전환효소 유전자의 다형성)

  • Kim, Mi Yeoun;Lee, Jae Myoung;Kim, Ji Sook;Kim, Eun Ryoung;Lee, Hee Jae;Yoon, Seo Hyun;Chung, Joo Ho
    • Clinical and Experimental Pediatrics
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    • v.50 no.1
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    • pp.28-32
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    • 2007
  • Purpose : Human angiotensin converting enzyme (ACE) gene shows an insertion/deletion polymorphism in 16 intron, and three genotypes are determined by whether a 287 bp fragment of the DNA is present or not; II, ID and DD genotype. DD genotype has been suggested as a risk factor of chronic nephrotic disease such as IgA nephropathy and diabetic nephropathy, various cardiovascular diseases and several other diseases. ACE activity increases in acute hepatitis, chronic persistent hepatitis, chronic active hepatitis and cirrhosis. On the other hand, patients with fatty livers have normal ACE activity. This study was designed to find out the relation between polymorphsims of the ACE genes and neonatal hyperbilirubinemia in Koreans. Methods : The genomic DNA was isolated from 110 full-term Korean neonates who had hyperbilirubinemia with no obvious causes (serum bilirubin$${\geq_-}12mg/dL$$) and 164 neonates of a control population (serum bilirubin <12 mg/dL). We performed polymerase chain reaction (PCR) to see the allele of the ACE gene. Electrophoresis was done in the PCR products in 1.5 percent agarose gel, and then DNA patterns were directly visualized under ethidium bromide staining. Results : ACE genotypes in the hyperbilirubinemia group are as follows; 26.36 percent for II, 53.64 percent for ID, 20.00 percent for DD, 0.532 for I allele and 0.468 for D allele. These distributions were not significantly different from those in the control group; 24.39 percent for II, 51.83 percent for DI, 23.78 percent for DD, 0.503 for I allele and 0.497 for D allele. Conclusion : In this study, ACE gene polymorphism was detected in the neonatal hyperbilirubinemia and control group. The most frequent genotype was ID. Our results indicate that the ACE gene polymorphism is not associated with the prevalence of neonatal hyperbilirubinemia in Koreans.

Quantitative Assessment and Ligament Traceability of Volume Isotropic Turbo Spin Echo Acquisition (VISTA) Ankle Magnetic Resonance Imaging: Fat Suppression versus without Fat Suppression (발목관절 VISTA 자기공명영상에서 정량평가와 인대의 Traceability: 지방억제 대비 지방억제기법)

  • Cho, Kyung Eun;Yoon, Choon-Sik;Song, Ho-Taek;Lee, Young Han;Lim, Daekeon;Suh, Jin-Suck;Kim, Sungjun
    • Investigative Magnetic Resonance Imaging
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    • v.17 no.2
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    • pp.110-122
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    • 2013
  • Purpose : To compare the image quality and ligament traceability in ankle images obtained using Volume Isotropic Turbo Spin Echo Acquisition (VISTA) MRI with and without fat suppression. Materials and Methods: The signal-to-noise ratios (SNRs) in images from a phantom and from the ankle of a volunteer were compared. Ten ankles from 10 non-symptomatic volunteers were imaged for comparisons of contrast ratio (CR) and ligament traceability. All examinations were performed using VISTA sequences with and without fat suppression on a 3T MRI scanner. The SNRs were obtained from images with subjects and without subjects (noise-only). Contrast ratios from images of the 10 ankles were acquired between fluid and tendon (F-T), F-cartilage (C), F-ligament (L), fat (f)-T, f-C and f-L. Two musculoskeletal radiologists independently scored the traceability of 7 ligaments, in sagittal, axial and coronal images respectively, based on a 4-point scale (1 as not traceable through 4 as clearly traceable). The Wilcoxon signed-rank test was used to compare the CR. Fisher's exact test and Pearson's chi-squared test were used to compare the ligament traceability. Results: The SNRs did not differ significantly between the two sequences except in bone marrow. VISTA SPAIR showed the higher CR only in F-T (p = 0.04), whereas VISTA showed higher CR in f-T (p = 0.005), f-C (p = 0.005) and f-L (p = 0.005). The calcaneofibular ligament traceability with VISTA was superior to that obtained with VISTA SPAIR (p < 0.05) in all planes. Conclusion: VISTA showed significant superiority to VISTA SPAIR in tracing CFL due to the superior CR between fat and ligament.

Microbial profile of asymptomatic and symptomatic teeth with primary endodontic infections by pyrosequencing (원발성 치근단 치주염을 갖는 감염근관에서 증상유무에 따른 세균분포의 pyrosequencing 분석)

  • Lim, Sang-Min;Lee, Tae-Kwon;Kim, Eun-Jeong;Park, Jun-Hong;Lee, Yoon;Bae, Kwang-Shik;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.36 no.6
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    • pp.498-505
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    • 2011
  • Objectives: The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing. Materials and Methods: Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing. Results: On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing identified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Spirochetes, and Synergistetes. In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth (p < 0.05). Conclusions: GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diversity in primary endodontic infections.

The Genotypes of Helicobacter pylori, Gastric Epithelial Cell Proliferation and Apoptosis in Children (소아 Helicobacter pylori 감염에서 균의 유전형, 위 상피세포의 증식과 세포사)

  • Jung, Ji-Ah;Lee, Mi-Ae;Seo, Jeong-Wan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.6 no.1
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    • pp.1-9
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    • 2003
  • Purpose: To investigate the relation of the gastric epithelial cell proliferation, apoptosis and genotypes of H. pylori in children. Methods: Histologic grading by updated Sydney system, PCNA immunostaining, TUNEL method and the genotypes (cagA, picB and iceA) by PCR were performed in H. pylori positive (N=20) and negative (N=20) gastric biopsy specimens. Results: PCNA index was significantly different between H. pylori positive children ($77.4{\pm}13.12$) and H. pylori negative children ($52.3{\pm}12.20$) (p=0.000). There were positive correlations between PCNA index and H. pylori density (r=0.624, p=0.000), polymorphonuclear neutrophil activity (r=0.460, p=0.005) and chronic inflammation (r=0.433, p=0.009). Apoptosis index of H. pylori positive children ($0.70{\pm}0.411$) was significantly higher than of H. pylori negative children ($0.14{\pm}0.201$) (p=0.000). Positive correlations between apoptosis index and H. pylori density (r=0.691, p=0.000), polymorphonuclear neutrophil activity (r=0.585, p=0.000) and chronic inflammation (r=0.535, p=0.001) were noted. As PCNA index increased, apoptosis index significantly increased (r=0.527, p=0.001). The positive rates of genotypes were cagA 90%, picB 75%, iceA1 60% and iceA2 15%, respectively. There were no significant correlations between the status of the genotypes and PCNA index, apoptosis index, the endoscopic findings and the histologic findings. Conclusion: PCNA index and apoptosis index in H. pylori positive children were higher than in H. pylori negative children but were not related to H. pylori genotypes. This study suggested that correlatively increased gastric epithelial cell proliferation and apoptosis are important to pathogenesis of H. pylori infection in children.

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Anti-inflammatory Activity of Antimicrobial Peptide Zophobacin 1 Derived from the Zophobas atratus (아메리카왕거저리 유래 항균 펩타이드 조포바신 1의 항염증활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.9
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    • pp.804-812
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    • 2020
  • The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.

Expression Properties and Skin Permeability of Human Basic Fibroblast Growth Factor with or without PTD Fused to N- or C-terminus in Escherichia coli (대장균 발현시스템에서 단백질 전달 도메인 PTD가 인간 섬유아세포 성장인자(FGF2)의 N- 또는 C-말단에 결합 되었을 때 미치는 재조합 단백질 복합체의 발현 특성과 피부 투과능력)

  • Park, In-Sun;Choe, Chung-Hyeon;Kwon, Bo-Ra;Choi, Young-Ji;Kwon, Tae-Ho;Yu, Kang-Yeol;Lee, Juhyung;Choo, Young-Moo
    • Journal of Life Science
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    • v.28 no.3
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    • pp.275-283
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    • 2018
  • Human fibroblast growth factor (FGF) has the potential to be a commercially important therapeutic or cosmeceutical agent due to its ability to generate tissue and heal wounds. Granting permeability into skin tissues increases the therapeutic effects of FGF. Thus, several researchers have attempted the fusion of FGF conjugates with protein transduction domains (PTDs) to investigate the transduction ability and therapeutic effects of FGF. Less is known, however, about whether the location of PTD fused to the N- or C-terminus of FGF proteins has a significant impact on the folding and stability in Escherichia coli, and eventually, on transduction. Here, we report cloning of human basic fibroblast growth factor (FGF2) as a control and FGF2 with PTD fused to the N- or C-terminal ends of FGF proteins by an overlap extension PCR. We performed expression, verified expression properties of recombinant FGF2 without or with PTD fused to the N-terminus and the C-terminus, and investigated transduction ability into tissue by treating the dorsal skin of mice subjects. As a result, FGF2 and FGF2-PTD (fused to C-terminus) fusion protein were expressed as soluble forms suitable for straight-forward purification, unlike insoluble PTD-FGF2 (fused to N-terminus), but only FGF2-PTD fusion protein could transduce into the dorsal skin tissue of the mice subjects. Our results suggest that FGF2 with PTD fused to the C-terminus is more efficient than other options in terms of expression, purification, and delivery into skin tissue, as it does not require labor-intensive, costly, and time-consuming methods.

Epidemiological Study on Acute Diarrheal Disease of Children and Adolescents in the Jeju Region Using a Multiplex-PCR (Multiplex-PCR을 이용한 제주지역 소아청소년의 급성설사질환 역학조사)

  • Lee, Kyu-taeg;Kim, Sunmi;Chong, Moo Sang
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.2
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    • pp.141-149
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    • 2017
  • To examine the cause of acute diarrheal disease in children and adolescents, 521 fecal samples underwent multiple-PCR for a period of two years, between March 2015 and February 20178, in the Jeju region of Korea. Based on the analysis of 179 positive samples, 102 samples were male (56.98%) and 77 were female (43.02%), and highest positive rates were shown in the age group of 3~4 years (51.96%) and 5~6 years (12.29%). When 209 cases (40.12%), including double infection were analyzed, there were 88 cases (16.89%) of norovirus-GII infection, 26 cases (4.99%) of Campylobacter spp. infection, 18 cases (3.45%) of rotavirus infection, 18 cases (3.45%) of Clostridium difficile Toxin B infection, 17 cases (3.26%) of adenovirus infection, 16 cases (3.07%) of Clostridium perfringens infection, 11 cases (2.11%) of astrovirus infection, 5 cases (0.96%) of Salmonella spp. infection, 3 cases (0.58%) of norovirus-GI, Yersinia spp. and Aeromonas spp. infections, and 1 case (0.19%) of verocytotoxin-producing E. coli infection. Based on a seasonal separation of early childhood, norovirus-GI and norovirus-GII mainly prevailed during the winter, when the temperature is low. Typical enteritis with an increased prevalence of rotavirus during the spring. Astrovirus prevailed between the months of April and October, when norovirus-GI, norovirus-GII, and rotavirus did not prevail. With increasing age, acute diarrheal disease was not only induced by a virus, but also by bacteria. Although a test for virus is an effective method when trying to identify the cause during early childhood by multiplex-PCR, it would be desirable to undergo tests for both virus and bacteria concurrently as age increases.

The effect of blasting and anodizing-combined treatment of implant surface on response of osteoblast-like cell (분사처리 후 양극산화 처리한 임플란트 표면이 골모 유사 세포의 반응에 미치는 영향)

  • Seo, Bo-Yong;Kim, Young-Min;Choi, Jae-Won;Yun, Mi-Jung;Jeon, Young-Chan;Jeong, Chang-Mo;Kim, Gyu-Cheon;Huh, Jung-Bo
    • The Journal of Korean Academy of Prosthodontics
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    • v.53 no.1
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    • pp.9-18
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    • 2015
  • Purpose: The purpose of this study is to examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell). Materials and methods: Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05. Results: In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained $0.08{\pm}0.04{\mu}m$ in the control group, $0.52{\pm}0.14{\mu}m$ in ASD group and $1.45{\pm}0.25{\mu}m$ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase. Conclusion: In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.