• Title/Summary/Keyword: In-vivo

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The effect of different crystallization temperature of the hydroxyapatite coating produced by ion beam-assisted deposition on anodizing-treated titanium disks on human osteosarcoma cells (양극산화처리된 티타늄 표면에 이온빔보조증착방식을 이용한 수산화인회석 코팅시 소결온도의 차이가 조골세포에 미치는 영향)

  • Pae, Ah-Ran;Won, Hyun-Du;Lee, Richard Sung-Bok;Kim, Hyeong-Seob;Woo, Yi-Hyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.4
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    • pp.333-340
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    • 2011
  • Purpose: The aim of this study was to study the effect of hydroxyapatite (HA) coating crystallinity on the proliferation and differentiation of human osteosarcoma cells. Materials and methods: Surface roughness of the titanium disks increased by anodizing treatment and then HA was coated using ion beam-assisted deposition (IBAD). HA coating was crystallized by heat-treated at different temperature ($100^{\circ}C$, $300^{\circ}C$, $500^{\circ}C$, $800^{\circ}C$). According to the temperature, disks were divided into four groups (HA100, HA300, HA500, HA800). With the temperature, crystallinity of the HA coating was different. Anodized disks were used as control group. The physical properties of the disk surface were evaluated by surface roughness tests, XRD tests and SEM. The effect of the crystallinity of HA coating on HOS cells was studied in proliferation and differentiation. HOS cells were cultured on the disks and evaluated after 1, 3, 5, and 7 days. Growth and differentiation kinetics were subsequently investigated by evaluating cell proliferation and alkaline phosphatase activity. Results: Regardless of the heat-treated temperature, there is no difference on the surface roughness. Crystallinity of the HA was appeared in the groups of HA500, HA800. HOS cells proliferation, ALP activity were higher in HA500 and HA800 group than HA100 and HA300. Conclusion: Within the results of this limited study, heat treatment at $500^{\circ}C$ of HA coating produced by IBAD has shown greater effect on proliferation and differentiation of HOS cells. It is considered that further in vivo study will be necessary.

ROS Scavenging Effect and Cell Viability of Opuntia humifusa Extract on Osteoblastic MC3T3-E1 Cells (천년초 추출물이 조골세포의 증식과 ROS소거능에 미치는 영향)

  • Hwang, Hyun-Jung;Jung, Bok-Mi;Kim, Mi-Hyang
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1752-1760
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    • 2011
  • In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the $50{\mu}g/ml$ ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract $100{\mu}g/ml$ was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Gene Expression of Surfactant Protein B and C in Endotoxin and Thiourea Treated Rats (내독소 및 Thiourea 투여 후 Surfactant protein B와 C 유전자 발현의 비교 관찰)

  • Sohn, Dong Hyun;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.510-521
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    • 2003
  • Background : The surfactant specific proteins, SP-B and SP-C are believed to be important regulators of the surfactant function and homeostasis. Since acute respiratory distress syndrome(ARDS) is usually viewed as the functional and morphological expression of a similar underlying lung injury caused by a variety of insults, and since abnormalities in the surfactant function have been described in ARDS, the authors investigated the different effects of endotoxin and thiourea on the accumulation of mRNA encoding SP-B and SP-C. Methods : Sprague-Dawley rats were given 5 mg/kg of an intraperitoneal endotoxin from Salmonella enteritidis and 3.5 mg/kg intraperitoneal thiourea and were sacrificed at different time periods. Results : 1. The SP-B mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 26.1% and 50%, respectively(P<0.01, P<0.001). 2. The SP-B mRNA levels 24 hours after the 3.5 mg/kg thiourea treatment was reduced by 9.8% and 12.5%, respectively. 3. The SP-C mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 38.7% and 53.6%, respectively(P<0.01, P<0.001). 4. The SP-C mRNA level 6 hours after the 3.5 mg/kg thiourea treatment was reduced by 22.8%(P<0.05). Conclusion : These results indicate that the differential regulation of the hydrophobic surfactant proteins in vivo is evident, and suggest that the hydrophobic surfactant proteins might be differentially regulated during lung injury at different time periods without altering the lung wet to dry ratios. The mechanism of these alternations at the different time periods and the different kinds of etiology remain to be determined.

Enhancement of Sensitivity of Human Lung Cancer Cell Line to TRAIL and Gefitinib by IGF-1R Blockade (폐암세포주에서 IGF-1R 억제를 이용한 TRAIL 및 gefitinib에 대한 감수성 증가를 위한 연구)

  • Lee, Yoon-Jin;Park, Mi-Young;Kang, Young-Ae;Kwon, Sung-Youn;Yoon, Ho-Il;Lee, Jae-Ho;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.1
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    • pp.42-51
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    • 2007
  • Background: TRAIL is a cytokine that selectively induces apoptosis in various cancer cell lines. Gefitinib is new targeted drug applied in lung cancer that selectively inhibits EGFR tyrosine kinase. However, lung cancers have shown an initial or acquired resistance to these drugs. This study examined the effect of IGF-1R and its blockade on enhancing the sensitivity of lung cancer cell lines to TRAIL and gefitinib. Methods: Two lung cancer cell lines were used in this study. NCI H460 is very sensitive to TRAIL and gefitinib. On the other hand, A549 shows moderate resistance to TRAIL and gefitinib. The IGF-1R blockade was performed using adenoviruses expressing the dominant negative IGF-1R and shRNA to IGF-1R and AG1024 (IGF-1R tyrosine kinase inhibitor). Results: The adenovirus expressing dominant negative IGF-1R(950st) induced the increased expression of defective IGF-1R on the lung cancer cell surface, and the adenovirus-shIGF-1R effectively decreased the level of IGF-1R expression on cell surface. The genetic blockade of IGF-1R by the adenovirus-dnIGF-1R and AG1024 increased the sensitivity of A549 cells to TRAIL. The reduction of IGF-1R by transduction with ad-shIGF-1R also increased the sensitivity of the A549 cells to gefitinib. Conclusion: The blockade of IGF-1R through various mechanisms increased the sensitivity of the lung cancer cell line that was resistant to TRAIL and gefitinib. However, further studies using other cell lines showing acquired resistance as well as in vivo animal experiments will be needed.

Antifungal effects of synthetic human β-defensin 3-C15 peptide

  • Lim, Sang-Min;Ahn, Ki-Bum;Kim, Christine;Kum, Jong-Won;Perinpanayagam, Hiran;Gu, Yu;Yoo, Yeon-Jee;Chang, Seok Woo;Han, Seung Hyun;Shon, Won-Jun;Lee, Woocheol;Baek, Seung-Ho;Zhu, Qiang;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.41 no.2
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    • pp.91-97
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    • 2016
  • Objectives: The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human ${\beta}$-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. Materials and Methods: C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and $300{\mu}g/mL$), CH ($100{\mu}g/mL$), and Nys ($20{\mu}g/mL$) for 7 days at $37^{\circ}C$. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. Results: C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (${\geq}100{\mu}g/mL$). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (${\geq}100{\mu}g/mL$) showed significant fungicidal activity compared to CH group (p < 0.05). Conclusions: Synthetic HBD3-C15 peptide (${\geq}100{\mu}g/mL$) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

Protective effect of Gabjubaekmok (Diospyros kaki) extract against amyloid beta (Aβ)-induced cognitive impairment in a mouse model (아밀로이드 베타(amyloid beta)로 유도된 인지장애 마우스 모델에서 갑주백목(Diospyros kaki) 추출물의 인지기능 및 뇌 신경세포 보호 효과)

  • Yoo, Seul Ki;Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Han, Hye Ju;Park, Hyo Won;Kim, Chul-Woo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.379-392
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    • 2019
  • The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

Protective Effect of Enzymatically Modified Stevia on C2C12 Cell-based Model of Dexamethasone-induced Muscle Atrophy (덱사메타손으로 유도된 근위축 C2C12 모델에서 효소처리스테비아의 보호 효과)

  • Geon Oh;Sun-Il Choi;Xionggao Han;Xiao Men;Se-Jeong Lee;Ji-Hyun Im;Ho-Seong Lee;Hyeong-Dong Jung;Moon Jin La;Min Hee Kwon;Ok-Hwan Lee
    • Journal of Food Hygiene and Safety
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    • v.38 no.2
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    • pp.69-78
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    • 2023
  • This study aimed to investigate the protective effect of enzymatically modified stevia (EMS) on C2C12 cell-based model of dexamethasone (DEX)-induced muscle atrophy to provide baseline data for utilizing EMS in functional health products. C2C12 cells with DEX-induced muscle atrophy were treated with EMS (10, 50, and 100 ㎍/mL) for 24 h. C2C12 cells were treated with EMS and DEX to test their effects on cell viability and myotube formation (myotube diameter and fusion index), and analyze the expression of muscle strengthening or degrading protein markers. Schisandra chinensis Extract, a common functional ingredient, was used as a positive control. EMS did not show any cytotoxic effect at all treatment concentrations. Moreover, it exerted protective effects on C2C12 cell-based model of DEX-induced muscle atrophy at all concentrations. In addition, the positive effect of EMS on myotube formation was confirmed based on the measurement and comparison of the fusion index and myotube diameter when compared with myotubes treated with DEX alone. EMS treatment reduced the expression of muscle cell degradation-related proteins Fbx32 and MuRF1, and increased the expression of muscle strengthening and synthesis related proteins SIRT1 and pAkt/Akt. Thus, EMS is a potential ingredient for developing functional health foods and should be further evaluated in preclinical models.

Quantification of the Elastic Property of Normal Thigh Muscles Using MR Elastography: Our Initial Experience (자기 공명 탄성 검사를 이용한 대퇴 근육의 탄성도의 정량화: 초기 경험)

  • Junghoon Kim;Jeong Ah Ryu;Juhan Lee
    • Journal of the Korean Society of Radiology
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    • v.82 no.6
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    • pp.1556-1564
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    • 2021
  • Purpose This study aimed to apply MR elastography (MRE) to achieve in vivo evaluation of the elastic properties of thigh muscles and validate the feasibility of quantifying the elasticity of normal thigh muscles using MRE. Materials and Methods This prospective study included 10 volunteer subjects [mean age, 32.5 years, (range, 23-45 years)] who reported normal activities of daily living and underwent both T2-weighted axial images and MRE of thigh muscles on the same day. A sequence with a motion-encoding gradient was used in the MRE to map the propagating shear waves in the muscle. Elastic properties were quantified as the shear modulus of the following four thigh muscles at rest; the vastus medialis, vastus lateralis, adductor magnus, and biceps femoris. Results The mean shear modulus was 0.98 ± 0.32 kPa and 1.00 ± 0.33 kPa for the vastus medialis, 1.10 ± 0.46 kPa and 1.07 ± 0.43 kPa for the vastus lateralis, 0.91 ± 0.41 kPa and 0.93 ± 0.47 kPa for the adductor magnus, and 0.99 ± 0.37 kPa and 0.94 ± 0.32 kPa for the biceps femoris, with reader 1 and 2, respectively. No significant difference was observed in the shear modulus based on sex (p < 0.05). Aging consistently showed a statistically significant negative correlation (p < 0.05) with the shear modulus of the thigh muscles, except for the vastus medialis (p = 0.194 for reader 1 and p = 0.355 for reader 2). Conclusion MRE is a quantitative technique used to measure the elastic properties of individual muscles with excellent inter-observer agreement. Age was consistently significantly negatively correlated with the shear stiffness of muscles, except for the vastus medialis.

Analgesic Effects of Gyejibokryeong-hwan Aqueous Extracts on the Rat Model of Primary Dysmenorrhea (원발성 월경통에 대한 계지복령환(桂枝茯苓丸) 열수 추출물의 진통 효과)

  • Cho, Su-Yun;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.33 no.3
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    • pp.95-118
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    • 2020
  • Objectives: The objective of this in vivo study is to observe the analgesic effects or improvements of Gyejibokryeong-hwan aqueous extracts (GJBRHe) on the Primary dysmenorrhea (PD) in rats as compared to those of Indomethacin (IND). Methods: The rats were administered with estradiol benzoate for 10 days and oxytocin 1 hour after the last 10th administration of estradiol benzoate to make the primary dysmenorrhea rat model. Gyejibokryeong-hwan aqueous extracts 500, 250 and 125 mg/kg were orally administrated, for 10 days once a day. Then the changes on the body weights and gains during experimental periods, uterine weights and gross inspections, abdominal writhing response for analgesic activities, uterus lipid peroxidation (malondialdehyde (MDA) levels), antioxidant defense system - glutathione (GSH) contents, activities of superoxide dismutase (SOD) and catalase (CAT), Nuclear factor-κB (NF-κB) and Cyclooxygenase (COX)-2 mRNA expressions, were monitored with uterus histopathology and immunohistochemistry for tumor necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS). The results of Gyejibokryeong-hwan aqueous extracts were compared to those of Indomethacin adminstered rats. Results: As results of estradiol benzoate and oxytocin treatment, noticeable decreases of body weights and gains, uterus GSH contents, SOD and CAT activities, increases of abdominal writhing responses, uterus lipid peroxidation (MDA level), uterus weights, NF-κB and COX-2 mRNA expressions were observed with increases of TNF-α and iNOS immunolabeled cells, inflammatory cell infiltrations, congestion and enlargement of the uterus at gross and histopathological inspections. These means classic inflammatory and oxidative stress mediated primary dysmenorrhea are relatively well induced. However, these signs were favorably and dose-dependently inhibited by administration of three different dosages of Gyejibokryeong-hwan aqueous extracts, but lesser than those of Indomethacin. Conclusions: The results obtained in this study suggest that Gyejibokryeong-hwan aqueous extracts has favorable analgesic and refinement activities dose-dependently on the estradiol benzoate and oxytocin treatment-induced primary dysmenorrhea signs.

Establishment of a Hepatocellular Carcinoma Cell Line Expressing Dual Reporter Genes: Sodium Iodide Symporter (NIS) and Enhanced Green Fluorescence Protein (EGFP) (나트륨 옥소 공동수송체 유전자와 녹색 형광 유전자의 이중 리포터 유전자를 발현하는 간암세포주 확립)

  • Kwak, Won-Jung;Koo, Bon-Chul;Kwon, Mo-Sun;Lee, Yong-Jin;Lee, Hwa-Young;Yoo, Jeong-Soo;Kim, Te-Oan;Chun, Kwon-Soo;Cheon, Gi-Jeong;Lee, Sang-Woo;Ahn, Byeong-Cheol;Lee, Jae-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.3
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    • pp.226-233
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    • 2007
  • Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.