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Cellular Aging Inhibitory Effect of Perilla Leaf Extract on D-Galactose Induced C2C12 Myoblasts (D-갈락토스 유도 C2C12 근원세포에 대한 자소엽 추출물의 세포 노화 억제 효과)

  • Song-Mi Park;Sung-Woo Cho;Yung-Hyun Choi
    • Journal of Korean Medicine Rehabilitation
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    • v.34 no.2
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    • pp.15-28
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    • 2024
  • Objectives We used the D-galactose (D-gal) induced C2C12 myoblast senescence model to investigate whether ethanol extract of Perilla. fructescens leaves (EEPF) could delay cellular senescence and regulate related mechanisms. Methods C2C12 myogenic cells were cultured in an incubator under 37 ℃ and 5% CO2 conditions. EEPF, dried perilla leaves were pulverized and extracted at 1:10 (v/v) at 50 ℃ for 4 hours. Cell counting kit-8 and western blot analysis was performed. Annexin V-FITC apoptosis detection kit and DAPI staining was applied. Catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and malondialdehyde analysis kits were used. To measure the level of reactive oxygen species generation, staining and flow cytometry was used. To analyze the mitochondrial activity, membrane potential changes were measured using JC-1. 𝛽-gal activity was analyzed using SA-𝛽-gal staining solution, and DNA damage was analyzed by using 𝛾-H2AX. Quantikine ELISA kit was used to analyze inflammatory cytokine production. Results According to the results of this study, EEPF significantly alleviated the decrease in cell viability in C2C12 cells treated with D-gal and suppressed the decrease in the expression of proliferating cell nuclear antigen. EEPF also markedly blocked D-gal-induced C2C12 cell apoptosis and restored reduced activity of CAT, GSH-Px, T-AOC, SOD. In addition, EEPF suppressed the decrease in 𝛽-galactosidase activity, the induction of DNA damage and the increase in expression of senescence-associated secretory phenotype proteins such as p16, p53 and p21 in D-gal-treated C2C12 cells. Furthermore, EEPF significantly attenuated D-gal-induced production and expression of inflammatory cytokines such as interleukin (IL)-6 and IL-18. Conclusions The results of this study indicate that EEPF can be used as a potential candidate for the prevention and treatment of muscle aging.

20(S)-ginsenoside Rh2 ameliorates ATRA resistance in APL by modulating lactylation-driven METTL3

  • Siyu Cheng;Langqun Chen;Jiahui Ying;Ying Wang;Wenjuan Jiang;Qi Zhang;Hong Zhang;Jiahe Wang;Chen Wang;Huimin Wu;Jing Ye;Liang Zhang
    • Journal of Ginseng Research
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    • v.48 no.3
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    • pp.298-309
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    • 2024
  • Background: 20(S)-ginsenoside Rh2(GRh2), an effective natural histone deacetylase inhibitor, can inhibit acute myeloid leukemia (AML) cell proliferation. Lactate regulated histone lactylation, which has different temporal dynamics from acetylation. However, whether the high level of lactylation modification that we first detected in acute promyelocytic leukemia (APL) is associated with all-trans retinoic acid (ATRA) resistance has not been reported. Furthermore, Whether GRh2 can regulate lactylation modification in ATRA-resistant APL remains unknown. Methods: Lactylation and METTL3 expression levels in ATRA-sensitive and ATRA-resistant APL cells were detected by Western blot analysis, qRT-PCR and CO-IP. Flow cytometry (FCM) and APL xenograft mouse models were used to determine the effect of METTL3 and GRh2 on ATRA-resistance. Results: Histone lactylation and METTL3 expression levels were considerably upregulated in ATRA-resistant APL cells. METTL3 was regulated by histone lactylation and direct lactylation modification. Overexpression of METTL3 promoted ATRA-resistance. GRh2 ameliorated ATRA-resistance by downregulated lactylation level and directly inhibiting METTL3. Conclusions: This study suggests that lactylation-modified METTL3 could provide a promising strategy for ameliorating ATRA-resistance in APL, and GRh2 could act as a potential lactylation-modified METTL3 inhibitor to ameliorate ATRA-resistance in APL.

Hyperoxia-Induced ΔR1: MRI Biomarker of Histological Infarction in Acute Cerebral Stroke

  • Kye Jin Park;Ji-Yeon Suh;Changhoe Heo;Miyeon Kim;Jin Hee Baek;Jeong Kon Kim
    • Korean Journal of Radiology
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    • v.23 no.4
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    • pp.446-454
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    • 2022
  • Objective: To evaluate whether hyperoxia-induced ΔR1 (hyperO2ΔR1) can accurately identify histological infarction in an acute cerebral stroke model. Materials and Methods: In 18 rats, MRI parameters, including hyperO2ΔR1, apparent diffusion coefficient (ADC), cerebral blood flow and volume, and 18F-fluorodeoxyglucose uptake on PET were measured 2.5, 4.5, and 6.5 hours after a 60-minutes occlusion of the right middle cerebral artery. Histological examination of the brain was performed immediately following the imaging studies. MRI and PET images were co-registered with digitized histological images. The ipsilateral hemisphere was divided into histological infarct (histological cell death), non-infarct ischemic (no cell death but ADC decrease), and nonischemic (no cell death or ADC decrease) areas for comparisons of imaging parameters. The levels of hyperO2ΔR1 and ADC were measured voxel-wise from the infarct core to the non-ischemic region. The correlation between areas of hyperO2ΔR1-derived infarction and histological cell death was evaluated. Results: HyperO2ΔR1 increased only in the infarct area (p ≤ 0.046) compared to the other areas. ADC decreased stepwise from non-ischemic to infarct areas (p = 0.002 at all time points). The other parameters did not show consistent differences among the three areas across the three time points. HyperO2ΔR1 sharply declined from the core to the border of the infarct areas, whereas there was no change within the non-infarct areas. A hyperO2ΔR1 value of 0.04 s-1 was considered the criterion to identify histological infarction. ADC increased gradually from the infarct core to the periphery, without a pronounced difference at the border between the infarct and non-infarct areas. Areas of hyperO2ΔR1 higher than 0.04 s-1 on MRI were strongly positively correlated with histological cell death (r = 0.862; p < 0.001). Conclusion: HyperO2ΔR1 may be used as an accurate and early (2.5 hours after onset) indicator of histological infarction in acute stroke.

A Study on the Effects and Mechanisms of the Combination Extract of Ephedrae Herba and Coicis Semen on Lipid Accumulation and Glucose Absorption in Non-Alcoholic Fatty Liver Disease (마황과 의이인 혼합추출물이 비알콜성 지방간 모델에서 지질 축적 및 포도당 흡수에 미치는 효과 및 기전 연구)

  • Ga-Ram Yu;Hye-Lin Jin;Dong-Woo Lim;Won-Hwan Park
    • Journal of Korean Medicine for Obesity Research
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    • v.24 no.1
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    • pp.1-12
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    • 2024
  • Objectives: Ephedrae herba (EH) and Coicis semen (CS) has been frequently prescribed for the treatment of obesity. However, effects of combinational extracts of these two herbs on non-alcoholic fatty liver disease are unknown. The aim of the present study was to investigate the effects of EH and CS on lipid accumulation and glucose absorption in free fatty acids (FFAs) or palmitic acid (PA)-treated HepG2 cells. Methods: Five samples of EH and CS were extracted by combination ratios (S1=0:100, S2=25:75, S3=50:50, S4=75:25, S5=100:0). Oil Red O staining was used to measure lipid accumulation in FFAs-induced steatosis cells. Intracellular triglycerides and total cholesterol levels were measured in FFAs-induced steatotic HepG2 cells. In PA-treated cells, intracellular 2-NBDG was detected using a fluorescence microplate reader and flow cytometry. Phosphorylation of key metabolism-related factors of AMP-activated protein kinase and acetyl-CoA carboxylase, expression of key lipid synthesis-related factors carnitine palmitoyltransferase 1 alpha (CPT1α), sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT enhancer binding protein alpha (C/EBPα) were confirmed by western blot. Results: Treatment of EH-CS combination in the FFAs-induced steatotic HepG2 cells significantly reduced lipid accumulation. As the relative ratio of Ephedrae herba increased, the lipid-lowering effects of the combination were increased. However, S1 and S5 of Ephedrae herba and Coicis semen did not significantly reduce triglycerides and total cholesterol induced by FFAs. However, the combination of Ephedrae herba and Coicis semen restored glucose absorption in PA-induced HepG2 cells. Major makers of SREBP1, PPARγ, C/EBPα, and CPT1α expression tended to decrease with EH ratio. Conclusions: The EH-CS combination has advantages over sole EH and CS extracts in improving lipid and glucose metabolism in liver steatosis models.

Trends identification of species distribution modeling study in Korea using text-mining technique (텍스트마이닝을 활용한 종분포모형의 국내 연구 동향 파악)

  • Dong-Joo Kim;Yong Sung Kwon;Na-Yeon Han;Do-Hun Lee
    • Korean Journal of Environmental Biology
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    • v.41 no.4
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    • pp.413-426
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    • 2023
  • Species distribution model (SDM) is used to preserve biodiversity and climate change impact. To evaluate biodiversity, various studies are being conducted to utilize and apply SDM. However, there is insufficient research to provide useful information by identifying the current status and recent trends of SDM research and discussing implications for future research. This study analyzed the trends and flow of academic papers, in the use of SDM, published in academic journals in South Korea and provides basic information that can be used for related research in the future. The current state and trends of SDM research were presented using philological methods and text-mining. The papers on SDM have been published 148 times between 1998 and 2023 with 115 (77.7%) papers published since 2015. MaxEnt model was the most widely used, and plant was the main target species. Most of the publications were related to species distribution and evaluation, and climate change. In text mining, the term 'Climate change' emerged as the most frequent keyword and most studies seem to consider biodiversity changes caused by climate change as a topic. In the future, the use of SDM requires several considerations such as selecting the models that are most suitable for various conditions, ensemble models, development of quantitative input variables, and improving the collection system of field survey data. Promoting these methods could help SDM serve as valuable scientific tools for addressing national policy issues like biodiversity conservation and climate change.

Pilot-scale Applications of a Well-type Reactive Barrier using Autotrophic Sulfur-oxidizers for Nitrate Removal (독립영양 황탈질 미생물을 이용한 관정형 반응벽체의 현장적용성 연구)

  • Lee, Byung-Sun;Um, Jae-Yeon;Lee, Kyu-Yeon;Moon, Hee-Sun;Kim, Yang-Bin;Woo, Nam-C.;Lee, Jong-Min;Nam, Kyoung-Phile
    • Journal of Soil and Groundwater Environment
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    • v.14 no.3
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    • pp.40-46
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    • 2009
  • The applicability of a well-type autotrophic sulfur-oxidizing reactive barrier (L $\times$ W $\times$ D = $3m\;{\times}\;4\;m\;{\times}\;2\;m$) as a long-term treatment option for nitrate removal in groundwater was evaluated. Pilot-scale (L $\times$ W $\times$ D = $8m\;{\times}\;4\;m\;{\times}\;2\;m$) flow-tank experiments were conducted to examine remedial efficacy of the well-type reactive barrier. A total of 80 kg sulfur granules as an electron donor and Thiobacillus denitrificans as an active bacterial species were prepared. Thiobacillus denitrificans was successfully colonized on the surface of the sulfur granules and the microflora transformed nitrate with removal efficiency of ~12% (0.07 mM) for 11 days, ~24% (1.3 mM) for 18 days, ~45% (2.4 mM) for 32 days, and ~52% (2.8 mM) for 60 days. Sulfur granules attached to Thiobacillus denitrificans were used to construct the well-type reactive barrier comprising three discrete barriers installed at 1-m interval downstream. Average initial nitrate concentrations were 181 mg/L for the first 28 days and 281 mg/L for the next 14 days. For the 181 mg/L (2.9 mM) plume, nitrate concentrations decreased by ~2% (0.06 mM), ~9% (0.27 mM), and ~15% (0.44 mM) after $1^{st}$, $2^{nd}$, and $3^{rd}$ barriers, respectively. For the 281 mg/L (4.5 mM) plume, nitrate concentrations decreased by ~1% (0.02 mM), ~6% (0.27 mM), and ~8% (0.37 mM) after $1^{st}$, $2^{nd}$, and $3^{rd}$ barriers, respectively. Nitrate plume was flowed through the flow-tank for 49 days by supplying $1.24\;m^3/d$ of nitrate solution. During nitrate treatment, flow velocity (0.44 m/d), pH (6.7 to 8.3), and DO (0.9~2.8 mg/L) showed little variations. Incomplete destruction of nitrate plume was attributed to the lack of retention time, rarely transverse dispersion, and inhibiting the activity of denitrification enzymes caused by relatively high DO concentrations. For field applications, it should be considered increments of retention time, modification of well placements, and intrinsic DO concentration.

Origin of limestone conglomerates in the Choson Supergroup(Cambro-Ordovician), mid-east Korea

  • Kwon Y.K.;Chough S.K.;Choi D.K.;Lee D.J.
    • 한국석유지질학회:학술대회논문집
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    • autumn
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    • pp.63-65
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    • 2001
  • The Chosen Supergroup (Cambro-Ordovician), mid-east Korea consists mainly of shallow marine carbonates and contains a variety of limestone conglomerates. These conglomerates largely comprise oligomictic, rounded lime-mudstone clasts of various size and shape (equant, oval, discoidal, tabular, and irregular) and dolomitic shale matrices. Most clasts are characterized by jigsaw-fit (mosaic), disorganized, or edgewise fabric and autoclastic lithology. Each conglomerate layer is commonly interbedded with limestone-dolomitic shale couplets and occasionally underlain by fractured limestone layer, capped by calcareous shale. According to composition, characteristic sedimentary structures, and fabric, limestone conglomerates in the Hwajol, Tumugol, Makkol, and Mungok formations of Chosen Supergroup can be classified into 4 types: (1) disorganized polymictic conglomerate (Cd), (2) horizontally stratified polymictic conglomerate (Cs), (3) mosaic conglomerate (Cm), and (4) disorganized/edgewise oligomictic conglomerate (Cd/e). These conglomerates are either depositional (Cd and Cs) or diagenetic (Cm and Cd/e) in origin. Depositional conglomerates are interpreted as storm deposits, tidal channel fills, or transgressive lag deposits. On the other hand, diagenetic conglomerates are not deposited by normal sedimentary processes, but formed by post-depositional diagenetic processes. Diagenetic conglomerates in the Chosen Supergroup are characterized by autoclastic and oligomictic lithology of lime-mudstone clasts, jigsaw-fit (mosaic) fabric, edgewise fabric, and a gradual transition from the underlying bed (Table 1). Autoclastic and oligomictic lithologies may be indicative of subsurface brecciation (fragmentation). Consolidation of lime-mudstone clasts pre-requisite for brecciation may result from dissolution and reprecipitation of CaCO3 by degradation of organic matter during burial. Jigsaw-fit fabric has been considered as evidence for in situ fragmentation. The edgewise fabric is most likely formed by expulsion of pore fluid during compaction. The lower boundary of intraformational conglomerates of depositional origin is commonly sharp and erosional. In contrast, diagenetic conglomerate layers mostly show a gradual transition from the underlying unit, which is indicative of progressive fragmentation upward (Fig. 1). The underlying fractured limestone layer also shows evidence for in situ fragmentation such as jigsaw-fit fabric and the same lithology as the overlying conglomerate layer (Fig, 1). Evidence from the conglomerate beds in the Chosen Supergroup suggests that diagenetic conglomerates are formed by in situ subsurface fragmentation of limestone layers and rounding of the fragments. In situ subsurface fragmentation may be primarily due to compaction, dewatering (upward-moving pore fluids), and dissolution, accompanying volume reduction. This process commonly occurs under the conditions of (1) alternating layers of carbonate-rich and carbonate-poor sediments and (B) early differential cementation of carbonate-rich layers. Differential cementation commonly takes place between alternating beds of carbonate-rich and clay-rich layers, because high carbonate content promotes cementation, whereas clay inhibits cementation. After deposition of alternating beds and differential cementation, with progressive burial, upward-moving pore fluid may raise pore-pressure in the upper part of limestone layers, due to commonly overlying impermeable shale layers (or beds). The high pore-pressure may reinforce propagation of fragmentation and cause upward-expulsion of pore fluid which probably produces edgewise fabric of tabular clasts. The fluidized flow then extends laterally, causing reorientation and further rounding of clasts. This process is analogous to that of autobrecciation, which can be analogously termed autoconglomeration. This is a fragmentation and rounding process whereby earlier semiconsolidated portions of limestone are incorporated into still fluid portions. The rounding may be due mainly to immiscibility and surface tension of lime-mud. The progressive rounding of the fragmented clasts probably results from grain attrition by fluidized flow. A synthetic study of limestone conglomerate beds in the Chosen Supergroup suggests that very small percent of the conglomerate layers are of depositional origin, whereas the rest, more than $80\%$, are of diagenetic origin. The common occurrence of diagenetic conglomerates warrants further study on limestone conglomerates elsewhere in the world.

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The Development of a Cryotherapy System (한냉물리치료기의 개발)

  • Kim, Yeong-Ho;Yang, Gil-Tae;Jang, Yun-Hui;Park, Si-Bok;Ryu, Jin-Sang
    • Journal of Biomedical Engineering Research
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    • v.19 no.6
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    • pp.617-622
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    • 1998
  • A cryotherapy system using cold air was developed. The developed system had superior low-temperature characteristics with various flow rates and nozzle sizes, and used R-404A, as a coolant, which has no destructive effects of Ozone layers. Flow rates and the treatment time can be easily altered during the operation. In addition, and alarm system was designed for the overload, overheat, and over-charge of the machine. For clinical applications, skin temperatures, intra-articular temperatures of the knee joint and intra-muscluar temperatures of the gluteal muscles were measured during and after the cryotherapy. After a 5-minute therapy, skin and intra-articular temperatures decreased by $23.3{\pm}4.7 and 4.1 {\pm}1.0^{circ}C$, respectively. A 5-minute cryotherapy was good enough to maintain low intra-articular temperatures for 2-3 hours. Resting intra-muscular temperatures in 2, 4, and 6cm deep in the gluteal muscle were $36.5{\pm}1.2, 36.9{\pm}0.2, 37.1{\pm}0.2^{circ}C$, respectively (p<0.05). Lowest temperatures in 2, 4, and 6cm depth were $35.1{\pm}0.7, 36.2{\pm}0.4, 36.9{\pm}0.3^{circ}C$, respectively (p<0.05). Temperatures after a 2-hour cold air application on the skin and in the muscle in dept도 of 2, 4, and 6cm were $32.2{\pm}1.1, 36.2{\pm}0.5, 36.6{\pm}0.3, 36.9{\pm}0.3^{circ}C$respectively (p<0.05). Temperatures on the skin and in the muscle significantly decreased after 2 hours, compared with before cold air application (p<0.05). The intra-muscular temperature was changed more slowly than the skin temperature, and the deeper the muscle, the lesser temperature changes. The effect of a 5-minute cold air application lasts up to 2 hours, and it seems that the rebound-rise of the temperature dut to the reactive vasodilatation does not occur in the gluteal muscle.

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Experimental Fetal Cardiopulmonary Bypass in the Fetal Lamb Model (태아양 모델을 이용한 실험적 태아 심폐우회술)

  • 이정렬;임홍국;김원곤;김종성;최정연;김용진
    • Journal of Chest Surgery
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    • v.32 no.6
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    • pp.495-503
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    • 1999
  • Background: We tested the technical feasibility of fetal cardiac bypass and collected baseline data on the fetal hemodynamics and placental functions related to the cardiopulmonary bypass in the fetal lamb model. Material and Method: Eleven fetuses at 120 to 150 days of gestation were subjected to bypass via trans-sternal approach with a 12 G pulmonary arterial cannula and 14 to 18 F venous cannula for 30 minutes. All ewes received general anesthesia with ketamine. In all the fetuses, no anesthetic agents were used except muscle relaxant. Eight served as a group in which placenta was excluded from the extracorporeal circulation by clamping the umbilical cord during the bypass(the oxygenator group) and in the remaining three, the placenta worked as the only source of oxygen supply(the placenta group). Observations were made every 10 minute during a 30-minute bypass and 30-minute post bypass period. No prostaglandin inhibitors were used both in ewes and in fetuses. Result: Weights of the fetuses ranged from 1.9 to 5.2 kg. In the oxygenator group, means of arterial pressure, PaO2, atrial pressure, heart rate, and bypass flow rate ranged 69.8 to 82.6 mmHg, 201.7 to 220.9 mmHg, 4.1 to 4.3 mmHg, 169 to 182/min, and 140.3 to 164.0 ml/kg/min, respectively during bypass, but rapid deterioration of the fetal cardiac functions and the placental gas exchange was observed after the cessation of bypass. In the placenta group, means of arterial pressure decreased from 44.7 to 14.4 mmHg and means of PaCO2 increased from 61.9 to 129.6 mmHg during bypass. Flow rate was suboptimal(74.3 to 97.0 ml/kg/min) during bypass. All hearts fibrillated immediately after the discontinuation of bypass. Conclusion: In this study, the technical feasibility of fetal cardiopulmonary bypass was confirmed in the fetal lamb model. However, further studies with modifications of the bypass including an addition of prostaglandin inhibitor, an application of the total spinal anesthesia on the fetus, a creation of more concise bypass circuit, and a use of active pump are mandatory to improve the outcome.

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PROPERTIES OF FLUORIDE-RELEASING RESIN COMPOSITE RESTORATIVE MATERIALS (불소방출성 콤포짓트 레진계 수복재의 특성)

  • Kim, Sang-Hoon;Baik, Byeong-Ju;Kim, Jae-Gon;Yang, Yeon-Mi;Park, Jeong-Yeol
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.3
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    • pp.418-426
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    • 2008
  • The objectives of this study were to examine the properties of fluoride-releasing resin composite restorative materials. Four commercially available compomer materials (Compoglass F: CF, $Dyract^{(R)}$ AP: DA, $Dyract^{(R)}$ flow: DF, F2000: FT) and one fluoride-releasing composite resin ($Tetric^{(R)}$ Ceram: TC) were selected as experimental materials. Rectangular-shaped tensile test specimens were fabricated in a teflon mold giving 5mm in gauge length and 2mm in thickness. Disk-shaped specimens were fabricated in the split teflon mold with diameter of 15mm and thickness of 1mm. After curing for an hour, specimens were immersed in deionized water at $37^{\circ}C{\pm}1^{\circ}C$ for 30 days. All specimens were thermocycled for 10,000 cycles with 15 seconds of dwelling time in each $5^{\circ}C$ and $55^{\circ}C$ water baths. Toothbrush abrasion test was conducted under a load of 1.5 N and the abraded surfaces were examined with surface roughness tester (SV-3000, Mitutoyo Co, Japan) and SEM (JSM-5800, JEOL, Japan). Fluoride recharging was done by toothbrushing for 3 min. using a fluoride toothpaste (Perio Alpine Herb, LG Household & Health Care, Korea). The results obtained were summarized as follows; 1. The highest tensile strength value of 32.3 MPa was observed in TC group and the lowest value of 16.8 MPa was observed in CF group. The tensile strength of TC group was significantly higher than those of CF and DF groups (P<0.05). 2. The lowest Ra value of 0.287 was observed in TC group and the highest value of 1.516 was observed in FT group. The Ra value of FT group was significantly higher than other groups (P<0.05). 3. The abraded surfaces revealed the increase of surface roughness due to the protrusion and missing of filler particles. 4. The release of fluoride of compomers after tooth brushing by Perio Alpine Herb was initially large and then followed by small and continuously. But it remains small and constant in fluoride-releasing composite resin of TC. 5. The highest value of fluoride release after toothbrushing by Perio Alpine Herb was $2.064{\mu}g/cm^2$ in CF group and the lowest value was $0.1119{\mu}g/cm^2$ in TC group. The amount of fluoride release of CF group was significantly higher than other groups (P<0.05).

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