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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

EFFECT OF VARIOUS MECHANICAL TREATMENTS ON TITANIUM PLASMA SPRAYED IMPLANT SURFACES (Titanium plasma sprayed implant에 관한 여러가지 기계적 표면처리방법이 implant표면조도에 미치는 영향)

  • Yu, Hyeon-Seok;Park, Jae-Wan;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.23 no.3
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    • pp.493-506
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    • 1993
  • For maintenance of an ailing or failed implant it is essential to treat the implant fixture surface so as to remove bacterial endotoxin and make a surface tolerated by surrounding soft and hard tissue. Thus in this study the method that makes the smoothest surface treated with a high speed diamond bur, a low speed diamond bur, a stone, a rubber point or Jetpolisher was studied. With the profilomenter a mean value of $R_{max}$ was measured. The hight speed diamond, the rubber point and the Jetpolisher showed a mean $R_{max}\;7.77{\mu}m$. The low speed diamond bur, the rubber, point and the Jetolisher made a mean value of $R_{max}\;8.44{\mu}m$. The stone, the rubber point and the Jetolisher showed the smoothest surfaces with a mean value of $R_{max}\;6.24{\mu}m$. TPS (titanium plasma sprayed) areas showed a mean vlaue of $R_{max}\;24.42{\mu}m$, and the smooth surfaced titanium disc manufactured by the company (IMZ, Germany) shows a mean value of $R_{max}\;3.00{\mu}m$. Under the SEM examination the disc treated with a high speed diamond bur, a rubber point and a Jetpolisher showed partially remaining TPS particles, but the height of these particles were reduced remarkably compared with those of the original TPS. The disc treated with a low speed diamond bur, a rubber point and a Jetpolisher showed a rough topography with remaining TPS on the entire surface. A stone, a rubber point and a Jetpolisher removed almost TPS and the bulk titanium metal was exposed and some scratches were made by the stone. All treated discs were revealed as rougher than the smooth surface disc manufactured by the company. An untreated TPS disc shows a very irregular surface and a $40{\mu}m$ height of the plasma sprayed areas.

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A comparative study on the fracture behavior of zironia, glass infiltrated alumina and PFM full crown system (지르코니아, 유리침투알루미나 및 PFM 전부관 시스템의 파절 경향에 관한 비교연구)

  • Lee, Sang-Hyeok;Ahn, Jin-Soo;Kim, Myung-Ho;Lim, Bum-Soon
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.4
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    • pp.235-242
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    • 2012
  • Purpose: The purpose of this study was to compare the fracture behavior of Zironia, glass infiltrated Alumina and PFM full crown system. Materials and methods: Fifteen crowns for each of 3 experimental groups (Zironia, glass infiltrated Alumina and PFM full crown) were made by the conventional method. The crowns mounted on the testing jig were inclined in 30 degrees to the long axis of the tooth and the universal testing machine was used to measure the fracture strength. Results: 1. The mean fracture strengths were $588.3{\pm}49.6MPa$ for zirconia system, $569.1{\pm}61.8MPa$ for PFM system and $551.0{\pm}76.5MPa$ for glass-infiltrated alumina system (P>.05). 2. The mean shear bond strengths were $25.5{\pm}5.6MPa$ for zirconia system, $38.9{\pm}5.0MPa$ for Ni-Cr alloy system and $39.4{\pm}5.1MPa$ for glass-infiltrated alumina system. 3. The chemical bonding was observed at interfaces between PFM or glass-infiltrated alumina and veneering porcelain, however, no chemical bonding was observed at interface between zirconia and veneering porcelain. Conclusion: With the study, the fracture strengths of PFM crown system had a higher fracture strength than conventional zirconia system crown and glass-infiltrated alumina crowns. and than the shear bond strengths glass-infiltrated alumina system had a higher shear bond strength than conventional PFM system and zirconia system.

A Study of Effect on Quality of Life of Cancer Patient's Caregiver : Focusing on the Mediating Effect of Feeling of Burden and Growth (사회적 지지와 암환자 가족의 삶의 질의 관계에서 돌봄부담감과 내적성장의 매개효과)

  • Rhee, Young-Sun
    • Korean Journal of Social Welfare
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    • v.61 no.2
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    • pp.325-348
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    • 2009
  • This study intends to investigate the main and mediating effects which caregiving appraisal and positive reappraisal exert on quality of life (QOL) of primary family caregivers of cancer patient considering the relationship with social support. The processes of this study areas follows. First, the variables which research model were chosen on the basis of stress-appraisal-coping theory through reviews of the previous studies. Second, a survey was conducted upon 295 primary caregiver of patient with cancer at National Cancer Center. Collected data were analyzed by SPSS 12.0 and SEM (Structural Equation Modeling) method using AMOS 5.0. The summary of the result is as follows. First, the entire model including measurement and structural model shows sufficient fit index of CFI(.951), TLI(.940) and RMSEA(.062). Second, the results of analysis of direct effects among variables are as follows. The 'Social support' has statistically significant direct effect on the 'feeling of burden' and 'growth'. The 'feeling of burden' has statistically significant direct effect on the 'growth' and 'QOL-mental and physical'. The 'growth' has statistically significant direct effect on the 'QOL-mental'. Third, the results of analysis of mediating effects of the 'social support and QOL' and 'feeling of burden and QOL' are as follows. The effects of 'social support' on 'QOL-mental' are significantly mediated by the 'feeling of burden' and 'growth'. The effects of 'social support' on 'QOL-physical' are significantly mediated by the 'feeling of burden'. The effects of 'feeling of burden' on 'QOL-mental' are significantly mediated by 'growth'. Through this research, these implications in social work study and practice are found: (1) this study extended the scope of study in the caregiver's health area from negative sides into positive ones by using growth variables as positive reappraisalof caregiving in research model, which has not been tried on the Korean family caregivers of the cancer patient. (2) The effects of positive reappraisal on QOL-mental can provide a foundational necessity for social workers to help family caregivers find positive meaning in their caregiving experience. This approach of social work practice will improve QOL of family caregivers. (3) This study present a framework including social support, negative appraisal, positive reappraisal, and QOL variables available to social work practice and explaining affective relationships among these variables in various aspects.

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EFFECT OF NERVE GROWTH FACTOR GENE INJECTION ON THE NERVE REGENERATION IN RAT LINGUAL NERVE CRUSH-INJURY MODEL (백서 설신경 압박손상모델에서 신경성장인자 유전자 주입이 신경재생에 미치는 영향)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Soung-Min;Kim, Yun-Hee;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.375-395
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    • 2006
  • Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

Antibacterial effects of Chitosanon-ascorbate Treated Kwamaegi Prepared on Microorganism Contamination (Chitosan-ascorbate 처리 과메기에 있어서 오염미생물에 대한 저해효과)

  • Kim, Young-Sook;oh, Seung-Hee;Kim, Soon-Dong
    • KSBB Journal
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    • v.24 no.2
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    • pp.156-162
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    • 2009
  • We examined saury, herring, gizzard shad kwamaegi to measure of microbic contamination rate of kwamegi that are sold in the market now. In the total bacteria, staphylococcus, peroxide value, and microorganisms is inhibited that from sample that we treated a substance with chitosan-ascorbate (CA) and other orders deep water (DW), ginseng steamed red and wine (GRW), NT (not treated). When we compared between SGRW and SNT, SCA show us more inhibition effect 0.22-0.49 log cycle in the total aerobacter. When we compared between HDW and HNT, HCA restraint 0.05-0.45log cycle, and when we compared between GDW and GNT, GCA inhibited 0.45 log cycle. In the coliform and E. coli, growths of microorganisms were inhibited followed order by treatment of CA, NT, and DW. GDW, HCA and HNT checked enough amount of water from the moisture measurement; but SGW, GCA, HEW and SCA showed 7-15% lack of moisture, and SNT and GNT have 10% more moisture. Peroxide value is changed to 41-51meq/kg when we did treat CA in there and a side that didn't add antimicrobial expressed the result numerically that 56-58meq/kg. In the sensory evaluation, customer gave preference to followed by Saury kwamaegi, herring, and gizzard shad kwamaegi. We have a point of view when kwamaeki manufactured if we add natural antibiotic and it uses to vacuum drying, we would inhibited of multiplication of microorganism, and of peroxides.

Preparation and Reactivity of Cu-Zn-Al Based Hybrid Catalysts for Direct Synthesis of Dimethyl Ether by Physical Mixing and Precipitation Methods (물리혼합 및 침전법에 의한 DME 직접 합성용 Cu-Zn-Al계 혼성촉매의 제조 및 반응특성)

  • Bang, Byoung Man;Park, No-Kuk;Han, Gi Bo;Yoon, Suk Hoon;Lee, Tae Jin
    • Korean Chemical Engineering Research
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    • v.45 no.6
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    • pp.566-572
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    • 2007
  • Two hybrid catalysts for the direct synthesis of DME were prepared and the catalytic activity of these catalysts were investigated. The hybrid catalyst for the direct synthesis of DME was composed as the catalytic active components of methanol synthesis and dehydration. The methanol synthesis catalyst was formed from the precursor contained Cu and Zn, the methanol dehydration catalyst was used ${\gamma}-Al_2O_3$. As PM-CZ+D and CP-CZA/D, Two hybrid catalysts were prepared by physical mixing method (PM-CZ+D) and precipitation method (CP-CZA/D), respectively. PM-CZ+D was prepared by physically mixing methanol synthesis catalyst and methanol dehydration catalyst, CP-CZA/D was prepared by depositing Cu-Zn or Cu-Zn-Al components on ${\gamma}-Al_2O_3$. The crystallinity and the surface morphology of synthesized catalyst were analyzed by X-ray diffraction (XRD) and scanning electron microscope (SEM) to investigate the physical property of prepared catalyst. And BET surface area by $N_2$ adsorption and the surface area of Cu by $N_2O$ chemisorption were investigated about the hybrid catalysts. In addition, catalytic activity of these hybrid catalysts was examined with varying reaction conditions. At that time, the reaction temperature of $250{\sim}290^{\circ}C$, the reaction pressure of 50~70 atm, the $[H_2]/[CO]$ mole ratio of 0.5~2.0 and the space velocity of $1,500{\sim}6,000h^{-1}$ were investigated the catalytic activity. From these results, it was confirmed that the reactivity of CP-CZA/D was higher than that of PM-CZ+D. When the conditions of reaction temperature, pressure, $[H_2]/[CO]$ ratio and space velocity were $260^{\circ}C$, 50 atm and 1.0, $3,000h^{-1}$ respectively, CO conversion using CP-CZA/D hybrid catalyst was 72% and the CO conversion of CP-CZA/D was more than 20% compared with the CO conversion of PM-CZ+D. It was known that Cu surface area of CP-CZA/D hybrid catalyst was higher than that of hybrid PM-CZ+D catalyst using $N_2O$ chemisorption. It was assumed that the catalytic activity was improved because Cu particle of hybrid catalyst prepared by precipitation method was well dispersed.

The Physical and Thermal Properties Analysis of the VOC Free Composites Comprised of Epoxy Resin, and Dicyandiamide (VOC Free Epoxy Resin/Dicyandiamide 경화물의 배합비 변화에 따른 물리적 특성 및 열적특성 분석)

  • Kim, Daeyeon;Kim, Soonchoen;Park, Young IL;Kim, Young Chul;Lim, Choong-Sun
    • Clean Technology
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    • v.21 no.1
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    • pp.76-82
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    • 2015
  • Volatile organic compounds (VOC) free adhesives have been interested by many scientists and engineers due to environmental regulations and the safety of industrial workers. In this work, a series of composites composed with bisphenol A epoxy resin used as solvent, dicyandiamide, and promoter were prepared to investigate the most appropriate molar ratio for steel-steel adhesion. The cured test specimen of each composite were measured with universal testing machine (UTM) to figure out mechanical properties such as tensile strength, Young’s modulus, and elongation. Furthermore, the lap shear strength of the specimen was tested with UTM while impact resistance was measured with Izod impact tester. The composite whose molar ratio of epoxy resin to curing agent is 1 : 0.9 (sample 3), showed better tensile strength, coefficient of elastic modulus, elongation, and impact strength than other composites did. The highest tanδ from dynamic mechanical analysis (DMA) was observed from sample 2 (epoxy resin: dicy = 1 : 0.7) while sample 3 showed slightly lower tanδ than that of 2. The morphology of the fracture surface of the cured composites from SEM showed that the number of subtle lines on the surface caused by impact increase as the contents of amine curing agent accrete. Furthermore, the viscosity change of sample 5 (epoxy resin: dicy = 1 : 1.3) was observed to confirm its storage stability.

Glycerol Steam Reforming for Hydrogen Production on Metal-ceramic Core-shell CoAl2O4@Al Composite Structures (금속-세라믹 Core-Shell CoAl2O4@Al 구조체를 적용한 불균일계 촉매의 글리세롤 수소전환 반응특성)

  • Kim, Jieun;Lee, Doohwan
    • Clean Technology
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    • v.21 no.1
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    • pp.68-75
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    • 2015
  • In this study, we investigated the structure and properties of a highly heat conductive metal-ceramic core-shell CoAl2O4@Al micro-composite for heterogeneous catalysts support. The CoAl2O4@Al was prepared by hydrothermal surface oxidation of Al metal powder, which resulted in the structure with a high heat conductive Al metal core encapsulated by a high surface area CoAl2O4 shell. For comparison, CoAl2O4 was also prepared by co-precipitation method and also utilized for a catalyst support. Rh catalysts supported on CoAl2O4@Al and CoAl2O4 were prepared by incipient wetness impregnation and characterized by N2 adsorption, X-ray diffraction (XRD), scanning electron microscopy (SEM), CO chemisorption, and temperature-programmed reduction (TPR). The properties of catalysts were investigated for glycerol steam reforming reaction for hydrogen production at 550 ℃. Rh/CoAl2O4@Al exhibited about 2.8 times higher glycerol conversion turnover frequency (TOF) than Rh/CoAl2O4 due to facilitated heat transport through the core-shell structure. The CoAl2O4@Al and CoAl2O4 also showed some catalytic activities due to a partial reduction of Co on the support, and a higher catalytic activity was also found on the CoAl2O4@Al core-shell than CoAl2O4. These catalysts, however, displayed deactivation on the reaction stream due to carbon deposition on the catalysts surface.

The Effect of Acid Beverages Purchased on the Retail Market upon the Surface of Tooth Enamel (수종시판 청량음료가 법랑질 표면에 미치는 영향)

  • Ahn, Yong-Soon;Kim, Eun-Ha;Min, Ji-Hye;Min, Su-Ji;Park, Hyun-Sook;Seo, Si-Nae;Lee, Sun-Min;Lee, Ji-Eun;Park, Mi-Ae;Han, Keum-A;Hong, Ju-Hee;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.10 no.6
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    • pp.451-457
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    • 2010
  • It is well known that most of the beverages on sale contain acid. Among these beverages, one of the most typically purchased products is Coca-Cola ($Coke^{(R)}$), which affects the demineralization of the tooth enamel surface. However other beverages also affect the erosion of the tooth enamel surface. Therefore, the aim of this study was to observe the influence that acid containing beverages have upon the tooth enamel surface. In this study, 30 enamel specimens were obtained from extracted maxillary third molars without cracks. The study measured the pH concentrations and the Ca and P concentration levels of the acidic beverages. The control without exposure was kept in distilled water, and each enamel specimen was stored for 30-min, 60-min, and 120-min. The erosion effect of the enamel surface was observed by SEM. The results obtained from this study can be summarized as follows: 1. The pH concentration of acidic beverages, without milk, was 3.105 on average. The beverages eroded the enamel surface and the erosion degree was determined by the pH concentration of beverages. 2. The erosion degree significantly increased by as the exposure time increased. The difference in the erosion degree between the 30-min group and the 60-min and the 120-min group was significant, but the erosion degree of the 120-min group was significantly less than that of the initial group. 3. When compared with the beverages containing similar pH concentrations, the demineralization degree of the beverages containing higher concentrations of Ca and P was lower.