• Title/Summary/Keyword: WST-1 assay

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Effect of acidic solutions on the microhardness of dentin and set OrthoMTA and their cytotoxicity on murine macrophage

  • Oh, Soram;Perinpanayagam, Hiran;Lee, Yoon;Kum, Jae-Won;Yoo, Yeon-Jee;Lim, Sang-Min;Chang, Seok Woo;Shon, Won-Jun;Lee, Woocheol;Baek, Seung-Ho;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.41 no.1
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    • pp.12-21
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    • 2016
  • Objectives: To evaluate the effects of three acids on the microhardness of set mineral trioxide aggregate (MTA) and root dentin, and cytotoxicity on murine macrophage. Materials and Methods: OrthoMTA (BioMTA) was mixed and packed into the human root dentin blocks of 1.5 mm diameter and 5 mm height. Four groups, each of ten roots, were exposed to 10% citric acid (CA), 5% glycolic acid (GA), 17% ethylenediaminetetraacetic acid (EDTA), and saline for five minutes after setting of the OrthoMTA. Vickers surface microhardness of set MTA and dentin was measured before and after exposure to solutions, and compared between groups using one-way ANOVA with Tukey test. The microhardness value of each group was analyzed using student t test. Acid-treated OrthoMTA and dentin was examined by scanning electron microscope (SEM). Cell viability of tested solutions was assessed using WST-8 assay and murine macrophage. Results: Three test solutions reduced microhardness of dentin. 17% EDTA demonstrated severe dentinal erosion, significantly reduced the dentinal microhardness compared to 10% CA (p = 0.034) or 5% GA (p = 0.006). 10% CA or 5% GA significantly reduced the surface microhardness of set MTA compared to 17% EDTA and saline (p < 0.001). Acid-treated OrthoMTA demonstrated microporous structure with destruction of globular crystal. EDTA exhibited significantly more cellular toxicity than the other acidic solutions at diluted concentrations (0.2, 0.5, 1.0%). Conclusions: Tested acidic solutions reduced microhardness of root dentin. Five minute's application of 10% CA and 5% GA significantly reduced the microhardness of set OrthoMTA with lower cellular cytotoxicity compared to 17% EDTA.

Effect of Sodium Hydroxide Treatment on Scaffold by Solid Freeform Fabrication (조형가공기술을 이용한 인공지지체의 수산화나트륨 개질 효과)

  • Park, SuA;Lee, JungBok;Kim, YangEun;Kim, JiEun;Kwon, IlKeun;Lee, JunHee;Kim, WanDoo;Kim, HyungKeun;Kim, MiEun;Lee, JunSik
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.815-819
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    • 2014
  • Scaffolds of tissue engineering should be biocompatible and biodegradable for cell attachment, proliferation and differentiation. In the various scaffold fabrication, 3D printing technique can make the three dimensional scaffold with interconnected pores for cell ingrowth. Polycaprolactone (PCL) is biodegradable polyester with a low melting temperature and has been approved by the Food and Drug Administration (FDA). In this study, PCL scaffold was fabricated by 3D bioprinting system and surface modification of PCL scaffold was controlled by NaOH treatment. Morphological change and wetability of NaOH-treated scaffold were observed by SEM and contact angle measurement system. The remnant of PCL treated with NaOH was measured by ATR-FTIR. In vitro study of scaffolds was evaluated with WST-1 and ALP activity assay. NaOH treatment of PCL scaffolds increased surface roughness, hydrophilicity, cell proliferation and osteogenic differentiation. These results indicate that NaOH-treated PCL scaffold made by 3D bioprinting has tissue engineered potential for the development of biocompatible material.

Neuroprotective effects of Salacca wallichiana extract against glutamate-induced oxidative stress in mouse Hippocampal HT22 cells (쥐 해마 HT22 세포에서 글루타메이트 유도 산화 스트레스에 대한 Salacca wallichiana 추출물의 신경 보호 효과)

  • Ji Hun Byeon;Ye Yeong Hong;Jungwhoi Lee;Thet Thet Mar Win;Su Su Hlaing;Song-I Han;Jae Hoon Kim
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.250-257
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    • 2023
  • Glutamate is an excitatory neurotransmitter distributed in the central nervous system of mammals. However, high concentrations of glutamate are known to cause neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and stroke by causing nerve cell death. In this study, the antioxidant activity and neuroprotective effect of subtropical natural products were analyzed. Among 11 subtropical plant extracts mainly tested, Sallacca wallichiana extract (SE) showed the greatest free radical scavenging activity. Then, we confirmed through WST-1 assay that SE protected HT22 cells against glutamate-induced cell death in a concentration-dependent manner. The protective effects of SE against glutamate-induced apoptosis in HT22 cells were also confirmed by flow cytometry analysis using Annexin V/PI double staining. We also confirmed using H2DCF-DA single staining that SE inhibits glutamate-induced intracellular reactive oxygen species. And we were confirmed through that SE inhibited glutamate-induced phosphorylation of Mitogen-activated Protein kinases. Consequently, our results propose that SE may contribute to the development of therapeutics to prevent neurodegenerative diseases.

Protective Effect of Radiation-induced New Blackberry Mutant γ-B201 on H2O2-induced Oxidative Damage in HepG2 Cells (H2O2 에 의해 유도된 HepG2 세포의 산화적 스트레스에 대한 신품종 방사선 돌연변이 블랙베리 γ-B201의 세포 보호 효과)

  • Cho, Byoung Ok;Lee, Chang-Wook;So, Yangkang;Jin, Chang-Hyun;Yook, Hong-Sun;Byun, Myung-Woo;Jeong, Yong-Wook;Park, Jong Chun;Jeong, Il-Yun
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.384-389
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    • 2014
  • The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.

Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts (괭생이 모자반 추출물의 비장세포 면역활성 증강 효과)

  • Kim, Dong-Sub;Sung, Nak-Yun;Han, In-Jun;Lee, Byung-Soo;Park, Sang-Yun;Nho, Eun Young;Eom, Ji;Kim, Geon;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.52 no.6
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    • pp.515-528
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    • 2019
  • Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

Effects of SLA surface treated with NaOH on surface characteristics and response of osteoblast-like cell (염기처리한 SLA 표면이 표면 특성 및 골모유사세포의 반응에 미치는 영향)

  • Park, Jin-Chul;Kim, Joo-Hyeun;Kang, Eun-Sook;Ryu, Jae-Jun;Huh, Jung-Bo
    • The Journal of Korean Academy of Prosthodontics
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    • v.52 no.3
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    • pp.211-221
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    • 2014
  • Purpose: The purpose of this study was to evaluate the surface characteristics and response of osteoblast-like cell at SLA surface treated with NaOH. Materials and methods: Three kinds of specimens were fabricated for the experiment groups. Control group was a machined surface, SLA group was a conventionally SLA treated surface, and SLA/NaOH gorup was SLA surface treated with NaOH. To evaluate the surface characteristics, the surface elemental composition (XPS), surface roughness and surface contact angle were evaluated in each group. And the cytotoxicity, cell adhesion, cell proliferation and ATP activity of osteoblast-like cells (MG-63 cells) were compared in each group for evaluatation of the cell responses. Statistical comparisons between groups were carried out via one-way ANOVA using the SPSS software (SPSS Inc., Chicago, USA), and then performed multiple comparisons. The differences were considered statistically significant at P<.05. Results: SLA surface treated with NaOH (SLA / NaOH group) was changed to hydrophilic surface. All groups did not show the cytotoxicity to the MG-63. In cell adhesion studies, SLA / NaOH group showed the higher degree of adhesion than anothers (P<.05), Up to 7 days of incubation, the proliferation was showed the increasing tendency in all groups but SLA / NaOH group showed the highest cell proliferation between the three groups (P<.05). At 7 days of incubation, there was no difference in ALP activities between the three groups, but at 14 days, SLA / NaOH group showed significant increase in ALP activities (P<.05). Conclusion: In this study, SLA surface treated with NaOH promoted cell adhesion, proliferation and differentiation. It means that SLA/NaOH group is possible to promote osseointegration of implants.

Effects of Microbial Fermentation on the Antioxidant Activities of Protaetia brevitarsis Larvae (미생물 발효가 흰점박이꽃무지(Protaetia brevitarsis) 유충의 항산화 활성에 미치는 영향)

  • Han Bi Kim;Hye Soo Kim;Soo Jeong Cho
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1052-1061
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    • 2023
  • This study was carried out to evaluate the effect of fermentation by B. subtilis (BPLE), L. brevis (LPLE), S. cerevisiae (SPLE) and C. militaris (CPLE) on the antioxidant activity of Protaetia brevitarsis larvae fed with mushroom substrates (king oyster mushroom). The total polyphenol content of Protaetia brevitarsis larvae (PLE), BPLE, LPLE, SPLE and CPLE were 58.07±0.67, 83.33±0.98, 79.21±1.32, 61.02±0.87 and 57.90±1.02 mg GAEs/extract g, respectively. The flavonoid contents of the PLE, BPLE, LPLE, SPLE and CPLE were 17.35±1.57, 19.49±0.95, 16.90±1.57, 18.12±0.95 and 16.99±0.95 mg QEs/extract g, respectively. The DPPH radical scavenging activity showed no significant difference between the PLE, BPLE, LPLE, SPLE and CPLE at a concentration of 0.2 mg/ml. However, at a concentration of 0.4 mg/ml or more, the DPPH radical scavenging activity of the BPLE and LPLE was higher than that of the PLE. The reducing power of the BPLE and LPLE was also higher than that of the PLE, and more than twice as high at a concentration of 0.8 mg/ml or more. The ORAC value of the BPLE (79.77±0.82 uM TEs/extract g) was higher than that of the PLE (61.34±0.97 uM TEs/extract g). A WST-1 assay of the RAW 264.7 cells indicated that the PLE, BPLE, LPLE, SPLE and CPLE showed no cytotoxicity.

Extract from Eucheuma cottonii Induces Apoptotic Cell Death on Human Osteosarcoma Saos-2 Cells via Caspase Cascade Apoptosis Pathway (Eucheuma cottonii 추출물에 의한 인체 골육종암 Saos-2 세포의 자가사멸 유도)

  • Kang, Chang-Won;Kang, Min-Jae;Kim, Kyong Rok;Kim, Nan-Hee;Seo, Yong Bae;Kang, Keon-Hee;Kim, Sang-Ho;Kim, Gun-Do
    • Journal of Life Science
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    • v.26 no.2
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    • pp.147-154
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    • 2016
  • Osteosarcoma (OS) is the most common and malignant bone tumors. Although many types of resection surgery and experimental agents were developed, median survival and clinical prognosis are poorly investigated. Recently, several researches have reported that Eucheuma cottonii has potent as protective effects of coal dust-induced lung damage via inhibition of malondialdehyde (MDA) and oxidative stress in bronchoalveolar lavage fluids (BALF). However, anti-cancer effects and specific molecular mechanism of extract from Eucheuma cottonii (EE) has not been clearly studied yet. This study evaluated that anti-cancer potential of EE in human osteosarcoma Saos-2 cells. EE indicated cytotoxicity on Saos-2 cells in a dose-dependent manner. Morphological degradation and nucleic condensation were also observed under the EE treatment. However, it did not significantly affect on non-cancerous kidney HEK-293 cells under the same concentration which is shown cytotoxicity on Saos-2 cells. The phosphorylation of Fas-Associated Death Domain (FADD) and expression of cleaved caspase-8, -7 and -3 were upregulated in a dose-dependent manner. In immunofluorescence staining, expression level of Fas and cleaved PARP were upregulated by EE treatment. Furthermore, treatment of EE induces upregulation of sub G1 phase by flow cytometry analysis. The results demonstrated that EE has a therapeutic potential against osteosarcoma via FADD mediated caspase cascade apoptosis signal pathway.

Variations in antioxidant activity in Protaetia brevitarsis larvae depending on the feeding source (먹이원에 따른 흰점박이꽃무지(Protaetia brevitarsis) 유충의 항산화활성)

  • Kim, Hye Soo;Park, Hyun-Young;Kwon, Hyun-Sook;Lee, Sang-Ho;Ha, Jun;Lee, Sang-Won;Cho, Soo-Jeong
    • Journal of Mushroom
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    • v.17 no.4
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    • pp.261-267
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    • 2019
  • The objective of this study was to evaluate the antioxidant activity of extracts of Protaetia brevitarsis larvae fed on fermented oak sawdust (FOS) or spent mushroom substrates (SMS, Pleurotus eryngii). Total polyphenol content was 32% higher in extracts of larvae fed on SMS (P. eryngii) (75.33±0.43 mg GAE/g) than in extracts of larvae fed on FOS (57.02±1.73 mg GAE/g). The flavonoid content of extracts of larvae grown on FOS and SMS (P. eryngii) was 24.6±0.28 mg/g and 25.4±0.75 mg/g, respectively. DPPH radical scavenging activity increased in an extract concentration-dependent manner, and the DPPH radical scavenging capacity of the extract of larvae produced on SMS (P. eryngii) was higher than that of the larvae produced on FOS. The reducing power of the larval extracts produced on FOS and SMS (P. eryngii) increased in an extract concentration-dependent manner, but there was no significant difference between them. The extract of larvae fed on SMS (P. eryngii) (66.55±0.99 uM TE/g) had a higher oxygen radical absorbance capacity (ORAC) than extracts of larvae grown on FOS (76.32±0.48 uM TE/g). The effect of larval extracts on cell proliferation was investigated using a WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) assay on RAW 264.7 cells. When cells were treated with larval extracts produced on FOS and SMS (P. eryngii) at concentrations of 0, 2, 4, 8, 16, 32, 40, and 64 mg/ml, RAW 264.7 cells proliferated at 90% or more. Therefore, larval extracts produced on FOS and SMS (P. eryngii) were not toxic to RAW 264.7 cells.