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Decreased Neutrophil Apoptosis in Patients with Sepsis is Related to the Activation of NF-κB (패혈증 환자에서 NF-κB 활성화에 의한 호중구 아포프토시스의 억제)

  • Kwon, Sung Youn;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.495-509
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    • 2003
  • Background : Neutrophil-mediated inflammation is usually self-limiting, because neutrophils have a remarkably short life span. Prolonged neutrophil survival, which is caused by decreased spontaneous apoptosis, leads to persistent inflammation in sepsis. Because many inflammatory cytokines, which generate signals that delay apoptosis, are regulated by nuclear factor-${\kappa}B$ transcription factor, we hypothesized that nuclear factor-${\kappa}B$ might be related to the reduced neutrophil apoptosis observed in sepsis. Methods : Neutrophils of healthy volunteers and sepsis patients were freshly isolated from venous blood. Neutrophil apoptosis was assayed with two approaches : by counting apoptotic cells under a microscope and by flow cytometry using Annexin V. The activity of nuclear factor-${\kappa}B$ was assessed by immunofluorescent staining or electrophoretic mobility shift assay. Expression of X-linked inhibitor of apoptosis was measured by western blot assay. Results : We confirmed reduced spontaneous neutrophil apoptosis in patients with sepsis. The number of apoptotic neutrophils in patients with sepsis increased to the level of that in healthy controls after cycloheximide treatment, suggesting that decreased spontaneous neutrophil apoptosis is dependent on de novo protein synthesis. In patients with sepsis, basal neutrophil nuclear factor-${\kappa}B$ was activated compared to the level in healthy controls. Moreover, a blockade of nuclear factor-${\kappa}B$ activity reversed the decreased spontaneous neutrophil apoptosis in sepsis patients. Meanwhile, X-linked inhibition of apoptosis expression, which is regulated by nuclear factor-${\kappa}B$, decreased 24 hours after incubation in healthy persons, but persisted for 24 hours in patients with sepsis. Conclusion : These observations suggest that the reduced spontaneous neutrophil apoptosis observed in patients with sepsis may be related to the induction of survival protein by nuclear factor-${\kappa}B$.

Comparison of flexural strength according to thickness between CAD/CAM denture base resins and conventional denture base resins (CAD/CAM 의치상 레진과 열중합 의치상 레진의 두께에 따른 굴곡 강도 비교)

  • Lee, Dong-Hyung;Lee, Joon-Seok
    • Journal of Dental Rehabilitation and Applied Science
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    • v.36 no.3
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    • pp.183-195
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    • 2020
  • Purpose: The purpose of this study is to compare the flexural strength of CAD/CAM denture base resins with conventional denture base resins based on their thicknesses. Materials and Methods: For the conventional denture base resins, Lucitone 199® (C-LC) was used. DIOnavi - Denture (P-DO) and DENTCA Denture Base II (P-DC) were taken for the 3D printing denture base resins. For the prepolymerized PMMA resins, Vipi Block Gum (M-VP) and M-IVoBase® CAD (M-IV) were used. The final dimensions of the specimens were 65.0 mm x 12.7 mm x 1.6 mm / 2.0 mm / 2.5 mm. The 3-point bend test was implemented to measure the flexural strength and flexural modulus. Microscopic evaluation of surface of fractured specimen was conducted by using a scanning electron microscope (SEM). After testing the normality of the data, one-way ANOVA was adopted to evaluate the differences among sample groups with a significance level of P = 0.05. The Tukey HSD test was performed for post hoc analysis. Results: Under the same thicknesses, there are significant differences in flexural strength between CAD/CAM denture base resins and conventional denture base resins except for P-DO and C-LC. M-VP showed higher flexural strength than conventional denture base resins, P-DC and M-IV displayed lower flexural strength than conventional denture base resins. Flexural modulus was highest in M-VP, followed by C-LC, P-DO, P-DC, M-IV, significant differences were found between all materials. In the comparison of flexural strength according to thickness, flexural strength of 2.5 mm was significantly higher than that of 1.6 mm in C-LC. Flexural strength of 2.5 mm and 2.0 mm was significantly higher than that of 1.6 mm in P-DC and M-VP. In M-IV, as the thickness increases, significant increase in flexural strength appeared. SEM analysis illustrates different fracture surfaces of the specimens. Conclusion: The flexural strength of different CAD/CAM denture base resins used in this study varied according to the composition and properties of each material. The flexural strength of CAD/CAM denture base resins was higher than the standard suggested by ISO 20795-1:2013 at a thickness of 1.6 mm or more though the thickness decreased. However, for clinical use of dentures with lower thickness, further researches should be done regarding other properties at lower thickness of denture base resins.

PREPARATION OF AMORPHOUS CARBON NITRIDE FILMS AND DLC FILMS BY SHIELDED ARC ION PLATING AND THEIR TRIBOLOGICAL PROPERTIES

  • Takai, Osamu
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2000.11a
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    • pp.3-4
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    • 2000
  • Many researchers are interested in the synthesis and characterization of carbon nitride and diamond-like carbon (DLq because they show excellent mechanical properties such as low friction and high wear resistance and excellent electrical properties such as controllable electical resistivity and good field electron emission. We have deposited amorphous carbon nitride (a-C:N) thin films and DLC thin films by shielded arc ion plating (SAIP) and evaluated the structural and tribological properties. The application of appropriate negative bias on substrates is effective to increase the film hardness and wear resistance. This paper reports on the deposition and tribological OLC films in relation to the substrate bias voltage (Vs). films are compared with those of the OLC films. A high purity sintered graphite target was mounted on a cathode as a carbon source. Nitrogen or argon was introduced into a deposition chamber through each mass flow controller. After the initiation of an arc plasma at 60 A and 1 Pa, the target surface was heated and evaporated by the plasma. Carbon atoms and clusters evaporated from the target were ionized partially and reacted with activated nitrogen species, and a carbon nitride film was deposited onto a Si (100) substrate when we used nitrogen as a reactant gas. The surface of the growing film also reacted with activated nitrogen species. Carbon macropartic1es (0.1 -100 maicro-m) evaporated from the target at the same time were not ionized and did not react fully with nitrogen species. These macroparticles interfered with the formation of the carbon nitride film. Therefore we set a shielding plate made of stainless steel between the target and the substrate to trap the macropartic1es. This shielding method is very effective to prepare smooth a-CN films. We, therefore, call this method "shielded arc ion plating (SAIP)". For the deposition of DLC films we used argon instead of nitrogen. Films of about 150 nm in thickness were deposited onto Si substrates. Their structures, chemical compositions and chemical bonding states were analyzed by using X-ray diffraction, Raman spectroscopy, X-ray photoelectron spectroscopy and infrared spectroscopy. Hardness of the films was measured with a nanointender interfaced with an atomic force microscope (AFM). A Berkovich-type diamond tip whose radius was less than 100 nm was used for the measurement. A force-displacement curve of each film was measured at a peak load force of 250 maicro-N. Load, hold and unload times for each indentation were 2.5, 0 and 2.5 s, respectively. Hardness of each film was determined from five force-displacement curves. Wear resistance of the films was analyzed as follows. First, each film surface was scanned with the diamond tip at a constant load force of 20 maicro-N. The tip scanning was repeated 30 times in a 1 urn-square region with 512 lines at a scanning rate of 2 um/ s. After this tip-scanning, the film surface was observed in the AFM mode at a constant force of 5 maicro-N with the same Berkovich-type tip. The hardness of a-CN films was less dependent on Vs. The hardness of the film deposited at Vs=O V in a nitrogen plasma was about 10 GPa and almost similar to that of Si. It slightly increased to 12 - 15 GPa when a bias voltage of -100 - -500 V was applied to the substrate with showing its maximum at Vs=-300 V. The film deposited at Vs=O V was least wear resistant which was consistent with its lowest hardness. The biased films became more wear resistant. Particularly the film deposited at Vs=-300 V showed remarkable wear resistance. Its wear depth was too shallow to be measured with AFM. On the other hand, the DLC film, deposited at Vs=-l00 V in an argon plasma, whose hardness was 35 GPa was obviously worn under the same wear test conditions. The a-C:N films show higher wear resistance than DLC films and are useful for wear resistant coatings on various mechanical and electronic parts.nic parts.

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CONFOCAL LASER SCANNING MICROSCOPIC MORPHOLOGY OF DENTIN-RESIN INTERFACE AND ITS RELATIONSHIP WITH SHEAR BOND STRENGTH (상아질-레진 계면의 공초점 현미경적 형태 및 전단결합강도와의 관계)

  • Choi, Nak-Won;Cho, Byeong-Hoon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.24 no.2
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    • pp.310-321
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    • 1999
  • In this in vitro study, confocal laser scanning microscopic morphology of dentin-resin interface and its relationship to shear bond strength were investigated after the exposed dentin surfaces were treated with 3 different kinds of dentin adhesive systems[three-step; Scotchbond Multi-Purpose Plus(SMPP), self-priming bonding resin; Single Bond(SB), self-etching primer; Clearfil Liner Bond 2(LB2)]. 52 extracted human molar teeth without caries and/or restorations. The experimental teeth were randomly divided into three groups of seventeen teeth each. In five teeth of each group, class V cavities(depth: 1.5mm) with 900 cavosurface angles were prepared at the cementoenamel junction on buccal and lingual surfaces. Bonding resins of each dentin adhesive system were mixed with rhodamine B. Primer of SMPP was mixed with fluorescein. In group 1. the exposed dentin was conditioned with etchant, applied with above primer and bonding resin of SMPP. In group 2, with etchant and self-priming bonding agent of SB. In group 3, with self-etching primer and bonding agent of LB2. After treatment with dentin adhesive systems, composite resin were applied and photocured. The experimental teeth were cut longitudinally through the center line of restoration and grounded so that about $90{\mu}m$-thick wafers of buccolingually orientated dentin were obtained. And, $70{\sim}80{\mu}m$-thick wafers sectioned horizontally, thus presenting a dentinal tubules at 900 to the cut surface of a remaining tooth, were obtained. Primer of SMPP mixed with rhodamine B was applied to these wafers. Confocal laser scanning microscopic investigations of these wafers were done within of 24 hours after treatment. To measure shear bond strength, the remaining twelve teeth of each group were grounded horizontally below the dentinoenamel junction, so that no enamel remained. After applying dentin adhesive systems on the dentin surface, composite was applied in the shape of cylinder. The cylinder was 5mm in diameter, and 2mm in thickness. Shear bond strength was measured using Instron with a cross-head speed of 0.5mm/min. It was concluded as follows ; 1. Hybrid layer of SMPP(mean: $4.56{\mu}m$) was thicker than that of any other groups. This value was not statistically significant thicker than that of SB(mean: $3.41{\mu}m$, p>0.05), and significant thicker than that of LB2(mean: $1.56{\mu}m$, p<0.05). There was a statistical difference between SB and LB2(p<0.05). 2. Although there were variations in the length of resin tag even in a sample, and in a group, most samples in SMPP and SB showed resin tags extending above $20{\mu}m$. But samples in LB2 showed resin tags of $10{\mu}m$ at best. 3. Besides primer's infiltration into demineralized peritubular dentin and dentinal tubules, fluorophore of primer was detected in the lateral branches of dentinal tubules. 4. All groups demonstrated statistically significant differences from one another(p<0.05), with shear bond strengths given in descending order as follows: SMPP(18.3MPa), SB(16.0MPa) and LB2(12.4MPa). 5. LB2 having thinnest hybrid layer($1.56{\mu}m$) showed the lowest shear bond strength(12.4MPa).

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The Localization of the Excretory, Purified and Infected Antigenic Protein in the Tissue of Trichinella spiralis Larval Worm (선모충(Trichinella spiralis) 유충의 조직 내 배설, 분리 및 감염항원 단백의 분포)

  • Kim, Soo-Jin;Joo, Kyoung-Hwan;Chung, Myung-Sook;Rho, Young-Bok
    • Applied Microscopy
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    • v.37 no.1
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    • pp.43-52
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    • 2007
  • In order to observe the localization of excretory, purified and infected antigenic protein in the tissue of Trichinella spiralis larvae, immunogoldlabeling methodology using IgG and protein A-gold complex was implemented. T. spiralis larvae obtained from rat muscle were initially cultured in medium, and secreted excretory antigen was collected for 1 or 3 days. Purified antigenic protein was obtained from homogenized T. spiralis larvae. Rabbits were then immunized with 1 or 3 days secreted excretory protein and purified 45 kDa protein, and IgG was purified from collected serum. Serum, against infected antigen, collected from rat on 1 and 4 weeks after infection with T. spiralis larvae, and IgG was purified from collected serum. T. spiralis larvae were embedded in Lowicryl HM20 medium. Then they were finally treated with immunized IgG and protein A-gold complex (particle size; 15 nm) and observed under electron microscope. In T. spiralis larvae tissue, the tissue antigen reacted with rabbit IgC antigen Day 1 secreted excretory protein, infected antigenic protein and purified 45 kDa protein. But different distribution pattern of labeled gold particles were observed. When Day 1 secreted excretoy protein was used, gold particle labeling was observed specifically on the cuticle, basal layer, esophagus interstitial matrix (EIM) and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. In a separate group of tissue, the antigen reacted with rabbit IgG against Day 3 secreted excretory protein. Labeled gold particles were specifically distributed on the surface layer of cuticle, EIM and ${\alpha}_0$ granules of stichocyte of the worm. In case of using infected antigenic protein, gold particle labeling was specifically distributed on the cuticle and EIM of the worm. When purifed 45 kDa protein was used gold particle labeling was specifically distributed on the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. Therefore, excretory antigens appeared to originate from the cuticle and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte for the first day but the cuticle layer associated with globular proteins and ${\alpha}_0$ granules of stichocyte after 3 days and infected antigens appeared to originate from the cuticle for 1 and 4 weeks after infection. These results suggest that excretory and infection specific antigens are secreted into the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte and 45 kDa protein may be contained these specific antigens.

Fine Structural Changes of the Renal Corpuscle of the Mice following the Administration of 5-Fluorouracil or Mitomycin C (5-Fluorouracil 및 Mitomycin C 투여후 생쥐 콩팥소체의 미세구조 변화)

  • Ko, Jeong-Sik;Oh, Won-Young;Kim, Jin-Gook;Park, Kyung-Ho;Ahn, E-Tay
    • Applied Microscopy
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    • v.29 no.1
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    • pp.25-41
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    • 1999
  • The experiment was performed to study the morphological responses of the renal glomeruli of the mice after administration of 5-fluorouracil or mitomycin C. 5-fluorouracil (60 mg/kg) or mitomycin-C $(400{\mu}g/kg)$ were injected subcutaneously to the animals every other day, and animals were sacrificed at 4 days or 7 days following the first injections. Pieces of tissues were observed with a JEM 100CX-II electron microscope. The observed results were as follows: 1. In the fourth day following the first injection of 5-fluorouracil or mitomycin C, components of the renal glomeruli of the mice are looked compact since they were filled with the widened the mesangium, and showed narrowing lumen of glomerular capillaries and of urinary spaces. The changes were more significant in the mitomycin C treated mice. 2. In the 5-fluorouracil treated mice, morphological changes of glomeruli were generally recovered in the seventh day, whereas the glomeruli of the mitomycin C treated mice have not shown general recovery. 3. In the fourth and seventh days following the first injection of mitomycin C, in the renal glomeruli of the mice, swollen endothelial cells, and protruded mesangeal cells into the capillary lumen are frequently observed. 4. In the fourth day following the first injection of mitomycin C, in the glomerular basal lamina of the mice, the electron densities of the lamina rara interna and the lamina rara externa were similar to the density of the lamina densa and the expanded lamina rara interna were often seen. From the above results, it is suggested that the cytotoxic effects of the mitomycin C on renal glomeruli are more severe as compared with those of 5-fluorouracil.

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Study on the Vitrification of Porcine GV and M II Oocytes after Removal of Cytoplasmic Lipid Droplets (세포질 내 지방구 제거가 돼지 난포란의 유리화 동결에 미치는 영향)

  • 최인경;이승진;송해범
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.1-14
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    • 2003
  • This study was conducted to investigate that the immature and mature oocytes of porcine can be cryopreserved by vitrification. Oocytes were centrifuged to polarize the cytoplasmic lipid droplets. The lipids were removed from cytoplasm by micromanipulation. Delipated oocytes were centrifuged after being preincubated with cytochalasin B(CB) fer 10 min, and lipid droplets were removed. Centrifuged oocytes were treated with CB and centrifuged to polorize lipid droplets but not delipated and control oocytes is not-treatment. Oocytes of three types were vitrified in electron microscope(EM) grids. The results of survival, maturation and cleavage rates were as follows. 1 The survival rates of immature oocytes were 15.1%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated wassignificantly higher than Centrifuged and Control(P<.01). 2. The survival rates of mature oocytes were 12.21%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated was significantly higher than Centrifuged and Control(P<.01). 3 The maturation rates of immature oocytes were 37.5% and 68.9% for metaphase II in the Delipated after vitrification and Non-vitrification, respectively, and its rate of Non-vitrification was significantly higher than Delipated after vitrification(P<.01). 4. The cleavage rates of immature oocytes were 12.5%, 0%, 0% and 56.1% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05). 5 The cleavage rates of mature oocytes were 25.0%, 0%, 0% and 67.9% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05).

Characterization of Biopesticides (Bacillus thuringiensis) Produced in Korea (국내에서 생산된 Bacillus thuringiensis 살충제의 특성)

  • Kil, Mi-Ra;Kim, Da-A;Choi, Su-Yeon;Paek, Seung-Kyoung;Kim, Jin-Su;Jin, Da-Yong;Hwang, In-Chon;Yu, Yong-Man
    • The Korean Journal of Pesticide Science
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    • v.11 no.3
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    • pp.201-209
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    • 2007
  • Characteristics of the 5 biopesticides that included Bacillus thuringiensis and on the domestic markets were investigated. These products were contained different strains of B. thuringiensis, for examples; product A and E was B. thuringiensis subsp aizawai; product B was B. thuringiensis; product C was B. thuringiensis Berline var. kurstaki; product D was B. thuringiensis var. kurstaki. Number of active spores were counted because they could influence the bio-activity against target pests. Only product C are contained the fixed quantity as its label, however, product D and E were a tenth part, and product A and B were a hundredth part of their descriptions. The pHs of product A and B were measured 3.67 and 3.73, and C, D and E were 5, respectively. Typical bypyramidal crystals produced from B. thuringiensis was found in only product C under a phase contrast microscope. For the uniform formulation of products that conformed whether B. thuringiensis were equally spreaded on the crops, B. thuringiensis in the C, D and E were equally grown on the nutrient agar medium As a results, product A were more different from product C than any other products. When product A and C were bioassayed against different larval stages of diamondback moth, their mortalities with spraying application were showed 100% after 48 hours.

EFFECT OF TEMPORARY CEMENT ON TENSILE BOND STRENGTH OF DENTIN BONDING AGENT (Temporary Cement가 상아질 접착제의 접착성능에 미치는 영향)

  • Chang, Heon-Soo;Cho, Kyeu-Zeung
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.685-698
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    • 1995
  • This study was conducted to the effect of temporary cement on the adhesiveness of dentin bonding agent to dentin surface. One hundred freshly extracted bovine mandibular incisors were grinded to expose flat labial dentin surface. The dentin surfaces were temporarized with either eugenol-containing temporary cement(TemBond and Zinc Oxide Eugenol cement) or non-eugenol temporary cement(Nogenol and TempBond NE) for 7days, and then the temporarization was removed with surgical currette and the exposed dentin surfaces were water-rinsed. Bonding specimens were made by use of All-Bond 2 and Super-Bond C&B dentin bonding agent, and stored in $37^{\circ}C$ distilled water for 24hours. The tensile bond strenth and the cohesive failure rate were measured, and then the pretreated dentin surfaces which the temporary cement had been applied to and removed from and the fractured dentin surfaces after bonding test were examined under scanning electron microscope. The results were as follows : In case of bonding with All-Bond 2, tensile bond strength of each experimental group was lower than that of the control group(p<0.05), but there was no significant difference between the bond strengths of the control group and each experimental group in case of bonding with Super-Bond C&B(p>0.05). No significant difference between tensile bond strength of experimental group, whether temporary cement contains eugenol or not, was seen(p>0.05). In case of bonding with All-Bond 2, the control group showed cohesive-adhesive mixed failure mode and the experimental groups mainly showed adhesive failure mode, but in case of bonding with Super-Bond C&B, almost of the control and the experimental groups mainly showed cohesive failure mode. On SEM examination, all of the dentin specimens pretreated with either 10 % phosphoric acid or 10% citric acid after application of the temporary cements demonstrated remnants of temporary cement attached to dentin surface.

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The effect of cavity wall property on the shear bond strength test using iris method (Iris 법을 이용한 전단접착강도 측정에서 와동벽의 영향)

  • Kim, Dong-Hwan;Bae, Ji-Hyun;Cho, Byeong-Hoon;Lee, In-Bog;Baek, Seung-Ho;Ryu, Hyun-Mi;Son, Ho-Hyun;Um, Chung-Moon;Kwon, Hyuck-Choon
    • Restorative Dentistry and Endodontics
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    • v.29 no.2
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    • pp.170-176
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    • 2004
  • Objectives : In the unique metal iris method. the developing interfacial gap at the cavity floor resulting from the cavity wall property during polymerizing composite resin might affect the nominal shear bond strength values. The aim of this study is to evaluate that the iris method reduces the cohesive failure in the substrates and the cavity wall property effects on the shear bond strength tests using iris method. Materials and Methods : The occlusal dentin of 64 extracted human molars were randomly divided into 4 groups to simulate two different levels of cavity wall property (metal and dentin iris) and two different materials ($ONE-STEP^{\circledR}$ and $ALL-BOND^{\circledR}$ 2) for each wall property. After positioning the iris on the dentin surface. composite resin was packed and light-cured. After 24 hours the shear bond strength was measured at a crosshead speed of 0.5 mm/min. Fracture analysis was performed using a microscope and SEM. The data was analyzed statistically by a two-way ANOV A and t-test. Results : The shear bond strength with metal iris was significant higher than those with dentin iris (p=0.034). Using $ONE-STEP^{\circledR}$, the shear bond strength with metal iris was significant higher than those with dentin iris (p=0.005), but not in $ALL-BOND^{\circledR}$ 2 (p=0.774). The incidence of cohesive failure was very lower than other shear bond strength tests that did not use iris method. Conclusions:The iris method may significantly reduce the cohesive failures in the substrates. According to the bonding agent systems. the shear bond strength was affected by the cavity wall property.