• Food Science and Preservation
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• v.23 no.7
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• pp.1026-1032
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• 2016

Citrus platymamma hort. ex Tanaka is widely used in traditional Korean medicine because of its medicinal benefits including an anti-inflammatory effect. This study aimed to evaluate changes in the flavonoid content and anti-inflammatory activities of C. platymamma during its harvest period. Fruit peel samples were obtained between September 2015 and February 2016. The results indicate that C. platymamma peel extract (CPE) was an effective inhibitor of lipopolysaccharide (LPS)-induced NO production in RAW264.7 cells. The inhibitory effects of CPE at $100{\mu}g/mL$ concentration included dose-dependent decreases in the expression of iNOS and COX-2 proteins. In addition, CPE decreased the expression of pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. The highest anti-inflammatory activity and flavonoid content were observed in CPE of C. platymamma peel harvested during the immature fruit period in September. Further, to assess the suitability of CPE for cosmetic use, we performed MTT assays using HaCaT keratinocytes and observed that CPE did not exhibit any cytotoxicity. To test the potential application of CPE as a cosmetic material, we also performed primary skin irritation tests on normal skin of 30 volunteers and no adverse reactions were observed. The results of this study indicate that CPE may be considered as an anti-inflammatory candidate for inclusion in cosmetic materials.

• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.690-695
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• 2003

Physiological activities of 40% ethanol extracts of Phellinus ribis were studied by employing several biological and biochemical assays. The extracts of Phellinus ribis displayed nitrate-scavenging activities (NSA) at pH 1.2 as with 64% NSA with 1.0 mg/mL of the extracts. They also had 91% 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities at the concentration of 1.0 mg/mL. Antioxidant activities of the extracts (at 0.5 mg/mL) on the autoxidation of linoleic acid (p<0.001) were also observed.. The inhibitory effect of the extracts on angiotensin converting enzyme was 11%. Cytotoxic effects of Phellinus ribis extracts against human cancer cell tines were also examined using MTT assay. The extracts (at 50 mg/mL) had severe growth inhibitory effects on A549, Hela, AGS, and SK-Hep-1, which were 8, 44, 76 and 42%, respectively. Ames test indicated that the extracts had no mutating effects on Salmonella typhimurium TA98 and TA100.

• The Journal of Korean Academy of Prosthodontics
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• v.44 no.2
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• pp.250-263
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• 2006

Statement of problem: Ti-alloy has been used widely since it was produced in the United States in 1947 because it has high biocompatibility and anticorrosive characteristics. Purpose: The pure titanium, however, was used limitedly due to insufficient mechanical charateristics and difficult manufacturing process. Our previous study was focused on the development of a new titanium alloy. In the previous study we found that the Ti-Ta-Nb alloy had better mechanical characteristics and similar anticorrosive characteristics to Ti-6Al-4V Material and methods: In this study, the cytotoxicity of the Ti-Ta-Nb alloy was evaluated by MTT assay using MSCs(Mesenchaimal stem cells) and L929 cells(fibroblast cell line). The biocompatibility of the Ti-Ta-Nb alloy was performed by inserting the alloy into the femur of the rabbits and observing the radiological and histological changes surrounding the alloy implant. Results: 1. In the cytotoxicity test using MSCs, the 60% survival rate was observed in pure titanium, 84% in Ti-6Al-4V alloy and 95% in Ti-10Ta-10Nb alloy. 2. In the animal study, the serial follow-up of the radiographs showed no separation or migration revealing gradual bone ingrowth surrounding the implants. Similar radiographic results were obtained among three implant groups pure titanium, Ti-6Al-4V alloy and Ti-10Ta-10Nb alloy. 3. In the histologic examination of the bone block containing the implants. the bone ingrowth was prominent around the implants with the lapse of time. There was no signs of any tissue rejection, degeneration, or inflammation. Active bone ingrowth was observed around the implants. In the comparison of the three groups, the rate of bone ingrowth was better in the Ti-10Ta-10Nb alloy group than those in pure titanium group or Ti-6Al-4V alloy group. In conclusion, Ti-10Ta-10Nb alloy revealed better biocompatibility in survival rate of the cells and bone ingrowth around the implants. Therefore we believe a newly developed Ti-10Ta-10Nb alloy can replace currently used Ti-6Al-4V alloy to increase biocompatibility and to decrease side effects. Conclusion: In conclusion, Ti-10Ta-10Nb alloy revealed better biocompatibility in survival rate of the cells and bone ingrowth around the implants. Therefore we believe a newly developed Ti-10Ta-10Nb alloy can replace currently used Ti-6Al-4V alloy to increase biocompatibility and to decrease side effects.

• Polymer(Korea)
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• v.32 no.6
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• pp.516-522
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• 2008

Biodegradable polymers have been used extensively as scaffolding materials to regenerate new tissues and the ingrowth of tissue have been reported to be dependent directly of the porosity, pore diameter, pore shape, and porous structure of the scaffold. In this study, porous poly (L-lactide-co-glycolide) (PLGA) scaffolds with five different pore sizes were fabricated to investigate the effect of pore sizes for AF tissue regeneration. Cellular viability and proliferation were assayed by MTT test. Hydroxyproline/DNA content of AF cells on each scaffold was measured. sGAG analyses were performed at each time point of 2 and 6 weeks. Scaffold seeded AF cells were implanted into the back of athymic nude mouse to observe the difference of formation of disc-like tissue depending on pore size in vivo. We confirmed that scaffold with $180{\sim}250{\mu}m$ pores displayed high cell viability in vitro and produced higher ECM than scaffold with other pore sizes in vivo.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.2
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• pp.131-140
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• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.

• Restorative Dentistry and Endodontics
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• v.45 no.4
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• pp.54.1-54.16
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• 2020

Objectives: This study aimed to synthesize nanocrystals (NCs) of zinc oxide (ZnO) and calcium ion (Ca²⁺)-doped ZnO with different percentages of calcium oxide (CaO), to evaluate cytotoxicity and to assess the effects of the most promising NCs on cytotoxicity depending on lipopolysaccharide (LPS) stimulation. Materials and Methods: Nanomaterials were synthesized (ZnO and ZnO:xCa, x = 0.7; 1.0; 5.0; 9.0) and characterized using X-ray diffractometry, scanning electron microscopy, and methylene blue degradation. SAOS-2 and RAW 264.7 were treated with NCs, and evaluated for viability using the MTT assay. NCs with lower cytotoxicity were maintained in contact with LPS-stimulated (+LPS) and nonstimulated (-LPS) human dental pulp cells (hDPCs). Cell viability, nitric oxide (NO), and reactive oxygen species (ROS) production were evaluated. Cells kept in culture medium or LPS served as negative and positive controls, respectively. One-way analysis of variance and the Dunnett test (α = 0.05) were used for statistical testing. Results: ZnO:0.7Ca and ZnO:1.0Ca at 10 ㎍/mL were not cytotoxic to SAOS-2 and RAW 264.7. +LPS and -LPS hDPCs treated with ZnO, ZnO:0.7Ca, and ZnO:1.0Ca presented similar NO production to negative control (p > 0.05) and lower production compared to positive control (p < 0.05). All NCs showed reduced ROS production compared with the positive control group both in +LPS and -LPS cells (p < 0.05). Conclusions: NCs were successfully synthesized. ZnO, ZnO:0.7Ca and ZnO:1.0Ca presented the highest percentages of cell viability, decreased ROS and NO production in +LPS cells, and maintenance of NO production at basal levels.

• Restorative Dentistry and Endodontics
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• v.47 no.4
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• pp.38.1-38.15
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• 2022

Objectives: This study investigated the cytotoxicity, radiopacity, pH, and dentinal tubule penetration of a paste of 1.0% calcium-doped zinc oxide nanocrystals (ZnO:1.0Ca) combined with propylene glycol (PRG) or polyethylene glycol and propylene glycol (PEG-PRG). Materials and Methods: The pastes were prepared by mixing calcium hydroxide [Ca(OH)₂] or ZnO:1.0Ca with PRG or a PEG-PRG mixture. The pH was evaluated after 24 and 96 hours of storage in deionized water. Digital radiographs were acquired for radiopacity analysis and bubble counting of each material. The materials were labeled with 0.1% fluorescein and applied to root canals, and images of their dentinal tubule penetration were obtained using confocal laser scanning microscopy. RAW264.7 macrophages were placed in different dilutions of culture media previously exposed to the materials for 24 and 96 hours and tested for cell viability using the MTT assay. Analysis of variance and the Tukey test (α = 0.05) were performed. Results: ZnO:1.0Ca materials showed lower viability at 1:1 and 1:2 dilutions than Ca(OH)₂ materials (p < 0.0001). Ca(OH)₂ had higher pH values than ZnO:1.0Ca at 24 and 96 hours, regardless of the vehicle (p < 0.05). ZnO:1.0Ca pastes showed higher radiopacity than Ca(OH)₂ pastes (p < 0.01). No between-material differences were found in bubble counting (p = 0.0902). The ZnO:1.0Ca pastes had a greater penetration depth than Ca(OH)₂ in the apical third (p < 0.0001). Conclusions: ZnO:1.0Ca medicaments presented higher penetrability, cell viability, and radiopacity than Ca(OH)₂. Higher values of cell viability and pH were present in Ca(OH)₂ than in ZnO:1.0Ca.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.10
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• pp.1397-1403
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• 2011

Epimedium koreanum Nakai is a wild medicinal plant commonly consumed in South Korea due to its health beneficial effects. In the present study, the antioxidative, antimutagenic and immunological activities of E. koreanum Nakai extracts were investigated for their use in food. The yields of icariin compounds from the ethanol extract as well as the ethyl acetate, butanol, hexane, water, and chloroform fractions of E. koreanum were 27.9, 2.5, 1.7, 1.4, and 1.3 ${\mu}g/g$, respectively. The icariin components (295.5 ${\mu}g/g$) were collected from the ethyl acetate fraction by thin layer chromatography (TLC) and analyzed via high performance liquid chromatography (HPLC). The antioxidant activities of each fraction were as follows: ethyl acetate (49.0 ${\mu}g/mL$), butanol (59.2 ${\mu}g/mL$), hexane (119.8 ${\mu}g/mL$), water (122.0 ${\mu}g/mL$), and chloroform (138.5 ${\mu}g/mL$), based on $RC_{50}$ ${\mu}g/mL$. Icariin, isolated and identified as the main component, showed strong antioxidant activity with a $RC_{50}$ value of 15.3 ${\mu}g/mL$, which was higher than those of ascorbic acid (19.5 ${\mu}g/mL$) and ${\alpha}$-tocopherol (18.2 ${\mu}g/mL$). In an Ames test, none of the fractions produced mutagenic effects on Salmonella Typhimurium TA98 and TA100. In an immunomodulating activity test, the effects of E. koreanum Nakai on B cells (Rhamos) and T cells (Jurkat) were investigated. These results show that the growth and viability of B and T cells were increased by isolated icariin components for 1.27 and 1.28 fold, respectively. These results also provide preliminary data for the development of E. koreanum Nakai as an edible food material.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.4773-4780
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• 2014

Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

• Journal of Acupuncture Research
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• v.24 no.6
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• pp.1-14
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• 2007

Backgrounds : Rheumatoid Arthritis(RA) is known as the chronic inflammatory diseasethat induces persistent inflammation in the joint cavity. The destruction of cartilage occurs as the result of bones destoyed by pannus, several influential cytokines induced by the synovial capsulitis, varieties of proteinases, $O_2$ radicals, and the secondary degenerative changes of articular cartilage. The type 2 collagen-induced arthritis model is used in recent experimental research on rheumatoid arthritis. Cervus elaphus sibiricus (Nockyong) has the effect of relieving pain by nourishing the muscles, joints, and bones. It is also known to be efficacious in promoting and enhancing the immune system. The objective of this study was to investigate the effect of Cervus elaphus sibiricus herbal acupuncture to inhibit the generation of proinflammatory enzyme on type 2 collagen-induced arthritis. I investigated the inhibition of mRNA transcription of MIF(macrophage migration inhibitory factor), $TNF-{\alpha}$(Tumor necrosis $factor-{\alpha}$) and MMP-9 (matrix metalloproteinase-9) of Cervus elaphus sibiricus herbal acupuncture using an in vitro test. Also investigated was the inhibition of differentiation of Th 1 cells and activation of cytokines(MIF, $TNF-{\alpha}$, IL-6, MMP-9), which are known to cause initial RA ,and are also related to the morphology of the synovial membranes of the joint capsule, by an in vivo test, using CIA(collagen induced arthritis) model mice. Materials & methods : The laboratory animals used in this experiment were 4 week-old DBA female mice, weighing approximately 20 grams, and adjusted to the laboratory environment. The experiment was divided into the normal group(NOR)-no treated group, control group(CON)-CIA induced group, and sample group(SAM)-Cervus elaphus sibiricus herbal acupuncture treated group. RA was induced in the mice via injection of $50{\mu}{\ell}$ C II mixed CFA. The Cervus elaphus sibiricus herbal acupuncture solution was applied on $GB_{35}$(陽陵泉) for 26 days from the 3rd day of RA inducement. The concentration of the solution was determined via a MTT assay. To research the effect on the expression of MIF, $TNF-{\alpha}$ and MMP-9 mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. C II induced RA knee joint's histo-chemical synovial membrane was observed using a specimen model via the Hematoxilin and Eosin dying technique. Results : The expression of mRNA of RA-related cytokines such as MIF, $TNF-{\alpha}$, and MMP-9 dosedependently decreased in the cell from the synovial membranes of the joint, which is treated with Cervus elaphus sibiricus herbal acupuncture solution. In mice treated with Cervus elaphus sibiricusherbal acupuncture, the damage of synovial membranes of the joint was lessened, and differentiation of Th 1 cells was suppressed. The activation of RA-related cytokines such as MIF was suppressed, and the generation of $TNF-{\alpha}$ and MMP-9 showed a statistically significant decreas. Conclusions : It is speculated that Cervus elaphus sibiricus herbal acupuncture has the therapeutic effect of palliating the damage of the tissue impaired by RA by inhibition of the initial RA progression and by regulating excessive differentiation of Th 1 cell as it suppresses the generation of RA-related cytokines during the highest stage of RA by acting on pro-inflammatory enzymes.