• Title/Summary/Keyword: Cell toxicity

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Application of Primary Rat Corneal Epithelial Cells to Evaluate Toxicity of Particulate Matter 2.5 to the Eyes (눈에 대한 미세먼지의 독성 평가를 위한 쥐 각막 상피 세포의 적용)

  • Kim, Da Hye;Hwangbo, Hyun;Lee, Hyesook;Cheong, Jaehun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.32 no.9
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    • pp.712-720
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    • 2022
  • The purpose of this study was to investigate the efficacy of rat corneal-derived epithelial cells as an in vitro model to evaluate the harmfulness of the cornea caused by particulate matter 2.5 (PM2.5). To establish an experimental model for the effect of PM2.5 on corneal epithelial cells, it was confirmed that primary cultured cells isolated from rat eyes were corneal epithelial cells through pan-cytokeratin staining. Our results showed that PM2.5 treatment reduced cell viability of primary rat corneal epithelial (RCE) cells, which was associated with the induction of apoptosis. PM2.5 treatment also increased the generation of reactive oxygen species due to mitochondrial dysfunction. In addition, the production of nitric oxide and inflammatory cytokines was increased in PM2.5-treated RCE cells. Furthermore, through heatmap analysis showing various expression profiling between PM2.5-exposed and unexposed RCE cells, we proposed five genes, including BLNK, IL-1RA, Itga2b, ABCb1a and Ptgs2, as potential targets for clinical treatment of PM-related ocular diseases. These findings indicate that the primary RCE cell line is a useful in vitro model system for the study of PM2.5-mediated pathological mechanisms and that PM2.5-induced oxidative and inflammatory responses are key factors in PM2.5-induced ocular surface disorders.

Antioxidant, whitening and Anti-inflammatory Effects of "Geranium Maculatum extract" Water Extracts (열수추출 Geranium Maculatum extract (와일드제라늄추출물)의 항산화, 미백, 항염효과)

  • Choi, In-Jeong
    • Journal of Convergence for Information Technology
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    • v.12 no.3
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    • pp.244-251
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    • 2022
  • This study attempted to investigate the applicability of Geranium maculatum extract as a cosmeceutical ingredient. For this, DPPH, ABTS and FRAP assays were performed to assess radical scavenging activities. To evaluate antioxidant substances, in addition, polyphenol and flavonoid concentrations were measured. Furthermore, cytotoxicity, whitening and anti-inflammatory tests were conducted, using B16F10 and RAW 264.7 cells, and the results found the followings: In the DPPH and ABTS assays, 265.8 mg ascorbic acid/g and 168.5 mg ascorbic acid/g of antioxidant capacities were found respectively. According to the FRAP assay, 1 mg Geranium maculatum extract was same with ascorbic acid 229±9 ㎍ in terms of reducing power. In polyphenol and flavonoid concentrations, 32.989±1.610 mg/g and 11.098±0.261 mg/g were observed each. The above results show that cells survived in the test concentrations more than 80 percent, confirming the low toxicity of Geranium maculatum extract. According to whitening testing, melanin synthesis was reduced depending on concentration, and at the same time, 40.62±2.07% of melanin production inhibition was found at 100 ㎍/mL. In anti-inflammatory testing, inflammation was reduced depending on concentration, and 27.86±2.82% of inhibition of inflammation was detected simultaneously, confirming the applicability of Geranium maculatum extract as a cosmeceutical ingredient.

Study of Plant Growth Inhibition with Edible Sweetener Saccharin and Acesulfame Potassium (식용 감미료 사카린과 에이스셜팜 칼륨의 식물 성장 저해 연구)

  • Donggiun Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.5
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    • pp.661-667
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    • 2023
  • Five commercially available edible sweeteners are used as diet products because they can replace sucrose. In studies on the effects on animals and the human body, stability has been proven by excreting-oriented studies with characteristics of animal cells, and accumulation in small amounts has been ignored. On the other hand, plants can absorb, degrade, and accumulate foreign substances, so the effect of degradability and accumulation potential can be studied using plants. Metabolic effects in plants of commercially available saccharin and acesulfame potassium (Ace K) were tested using germinated barley and bean sprouts. In germinated barley and bean sprouts, saccharin and ace K showed inhibitory effects on plant growth in all organs from low concentrations in leaves, stems and roots. In addition, it can be observed that the symptoms of death appear clearly over time, so it can be seen that they are accumulated in the body of the plant. As the accumulated amount increases, the toxic effect increases and the plant reaches a state where it is unable to metabolize, turning black from the tip of the leaf and reaching a state of death. In order to remove the accumulated artificial sweetener, recovery was attempted by culturing in distilled water, but it acts as a substance that is not degraded and dies without avoiding toxicity. Saccharin and ace K cannot be excreted from the cell. Its toxic effects are thought to be persistent, inhibiting growth and eventually leading to cell death.

Verification of the effect of Potentillae Chinensis Chinensis Herba extract and bioconversion fraction on chronic respiratory diseases (위릉채 추출물 및 생물전환 분획물의 만성호흡기 질환 효과 검증)

  • Dong-Hee Kim;Bo Ae Kim;Yun-Hwan Kang
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.6
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    • pp.1454-1463
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    • 2023
  • Evaluating the antioxidant efficacy using Potentillae Chinensis Herba extract, the anti-inflammatory efficacy was tested in respiratory mucosal epithelium, RAW264.7 cells, and zebrafish. As a result, antioxidant activity increased in a concentration-dependent manner in DPPH free radical scavenging and ABTS+ cation radical activities. As a result of MTT assay for cell experiments, the survival rate of NCI-H292 cells was reduced to less than 70% when treated at each concentration of 100 ㎍/ml, subsequent experiments were conducted at 50 ㎍/ml. Anti-inflammatory efficacy evaluation, NO production, TNF-𝛼, IL-1𝛽, and PGE2 decreased, and COX-2 also decreased significantly at 50 ㎍/ml. The mucin protein expression of Potentillae Chinensis Herba extract and bioconverted extract, it was observed that MUC5AC expression was significantly reduced. In the zebrafish toxicity evaluation, concentrations below 50 ㎍/ml did not show embryotoxicity and showed anti-inflammatory efficacy by reducing NO production due to LPS. The above results are valid to be valuable for use as a functional material that suppresses inflammation by helping the expression of Potentillae Chinensis Herba's respiratory mucus proteins.

Anti-inflammatory, Anti-bacterial and Anti-oxidant Activities for Subcritical Extract of Chamaecyparis obtusa (편백 잎 아임계 수 추출물의 항염, 항균 및 항산화 활성)

  • Jung Eun Kim;Min Jeong Kim;Ji Young Moon;Jeong Mi Kim;Tae Heon Oh;Nam Ho Lee
    • Journal of the Korean Applied Science and Technology
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    • v.41 no.3
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    • pp.543-554
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    • 2024
  • In this study, the anti-inflammatory, anti-bacterial and anti-oxidative activities of subcritical water extract from Chamaecyparis obtusa leaves was compared with hot water extract. The yield of subcritical water extract (165℃ and 180℃, 80 bar) was 39.4~48.5%, higher than 31.5% of hot water extract. In the anti-inflammatory tests using lipopolysaccaride (LPS)-induced RAW264.7 macrophages, the subcritical water extract concentration-dependently inhibited production of nitric oxide (NO) without causing cell toxicity. Upon the anti-bacterial studies using Cutibacterium acnes and Staphylococcus epidermidis, subcritical water extract showed the stronger activity than hot water extract. In addition, DPPH and ABTS cation radical scavenging activity experiments showed that the radical scavenging activity of subcritical water extract was similar to that of hot water extract. Moreover, in the study of cell protection effect using HaCaT keratinocytes damaged by hydrogen peroxide (H2O2), the subcritical water extract (165℃, 80 bar) indicated protective effect against oxidative stress. These results suggested that the subcritical water extract of C. obtusa leaves as natural ingredients could be used as anti-inflammatory, anti-bacterial and anti-oxidative ingredients in cosmetic formulations.

Concurrent Chemoradiation Therapy in Stage III Non-small Cell Lung Cancer (III 기 비소세포성 폐암에서 Cisplatin-방사선동시병합요법의 효과)

  • Kim In Ah;Choi Ihl Bhong;Kang Ki Mun;Jang Jie Young;Song Jung Sub;Lee Sun Hee;Kuak Mun Sub;Shinn Kyung Sub
    • Radiation Oncology Journal
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    • v.15 no.1
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    • pp.27-36
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    • 1997
  • Purpose : This study was tried to evaluate the Potential benefits of concurrent chemoradiation therapy (low dose daily cisplatin combined with split course radiation therapy) compared with conventional radiation therapy alone in stage III non-small cell lung cancer. The end points of analyses were response rate. overall survival, survival without locoregional failure, survival without distant metastasis, prognostic factors affecting survival and treatment related toxicities. Materials and Methods : Between April 1992 and March 1994, 32 patients who had stage III non-small cell lung cancer were treated with concurrent chemoradiation therapy. Radiation therapy for 2 weeks (300 cGy given 10 times up to 3000 cGy) followed by a 3 weeks rest period and then radiation therapy for 2 more weeks (250 cGy given 10 times up to 2500 cGy) was combined with $6mg/m^2$ of cisplatin. Follow-up period ranged from 13 months to 48 months with median of 24 months. Historical control group consisted of 32 patients who had stage III non-small cell lung cancer were received conventionally fractionated (daily 170-200 cGy) radiation therapy alone. Total radiation dose ranged from 5580 cGy to 7000 cGy with median of 5940 cGy. Follow-up Period ranged from 36 months to 105 months with median of 62 months. Result : Complete reponse rate was higher in chemoradiation therapy (CRT) group than radiation therapy (RT) group (18.8% vs. 6.3%, CRT group showed lower in-field failure rate compared with RT group(25% vs. 47%. The overall survival rate had no significant differences in between CRT group and RT group (17.5% vs. 9.4% at 2 years). The survival without locoregional failure (16.5% vs. 5.3% at 2 years) and survival without distant metastasis (17% vs. 4.6% at 2 years) also had no significant differences. In subgroup analyses for Patients with good performance status (Karnofsky performance scale 80), CRT group showed significantly higher overall survival rate compared with RT group (62.5% vs. 15.6% at 2 years). The prognostic factors affecting survival rate were performance status and pathologic subtype (squamous cell cancer vs. nonsquamous cell cancer) in CRT group. In RT alone group, performance status and stage (IIIa vs IIIb) were identified as a Prognostic factors. RTOG/EORTC grade 2-3 nausea and vomiting(22% vs 6% and bone marrow toxicities (25% vs. 15.6% were significantly higher in CRT group compared with RT alone group. The incidence of RTOG/EORTC grade 3-4 pulmonary toxicity had no significant differences in between CRT group and RT group (16% vs. 6%. The incidence of WHO grade 3-4 pulmonary fibrosis also had no significant differences in both group (38% vs. 25%. In analyses for relationship of field size and Pulmonary toxicity, the Patients who treated with field size beyond 200cm2 had significantly higher rates of pulmonary toxicities. Conclusion : The CRT group showed significantly higher local control rate than RT group. There were no significant differences of survival rate in between two groups. The subgroup of patients who had good performance status showed higher overall survival rate in CRT group than RT group. In spite of higher incidence of acute toxicities with concurrent chemoradiation therapy, the survival gain in subgroup of patients with good performance status were encouraging. CRT group showed higher rate of early death within 1 year, higher 2 year survival rate compared with RT group Therefore, to evaluate the accurate effect on survival of concurrent chemoradiation therapy, systematic follow-up for long term survivors are needed.

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Twice Daily Radiation Therapy Plus Concurrent Chemotherapy for Limited-Stage Small Cell Lung Cancer (국한성병기 소세포폐암에서 하루 두 번 분할조사와 동시 화학방사선치료)

  • Yeo Seung-Gu;Cho Moon-June;Kim Sun-Young;Kim Ki-Whan;Kim Jun-Sang
    • Radiation Oncology Journal
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    • v.24 no.2
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    • pp.96-102
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    • 2006
  • Purpose: A retrospective study was performed to evaluate the efficiency and feasibility of twice daily radiation therapy plus concurrent chemotherapy for limited-stage small cell lung cancer in terms of treatment response, survival, patterns of failure, and acute toxicities. Materials and Methods: Between February 1993 and October 2002, 76 patients of histologically proven limited-stage small cell lung cancer (LS-SCLC) were treated with twice daily radiation therapy and concurrent chemotherapy. Male was in 84% (64/76), and median age was 57 years (range, 32-75 years). Thoracic radiation therapy consisted of 120 or 150 cGy per fraction, twice a day at least 6 hours apart, 5 days a week. Median total dose was 50.4 Gy (range, 45-51 Gy). Concurrent chemotherapy consisted of CAV ($cytoxan\;1000mg/m^2,\;adriamycin\;40mg/m^2,\;vincristine\;1mg/m^2$) alternating with PE ($cisplatin\;60mg/m^2,\;etoposide\;100mg/m^2$) or PE alone, every 3 weeks. The median cycle of chemotherapy was six (range, 1-9 cycle). Prophylactic cranial irradiation (PCI) was recommended to the patients who achieved a complete response (CR). PCI scheme was 25 Gy/10 fractions. Median follow up was 18 months (range, 1-136 months). Results: Overall response rate was 86%; complete response in 39 (52%) and partial response in 26 (34%) patients. The median overall survival was 23 months. One, two, and three year overall survival rate was 72%, 50% and 30%, respectively. In univariate analysis, the treatment response was revealed as a significant favorable prognostic factor for survival (p<0.001). Grade 3 or worse acute toxicities were leukopenia in 46 (61%), anemia in 5 (6%), thrombocytopenia in 10 (13%), esophagitis in 5 (6%), and pulmonary toxicity in 2 (2%) patients. Of 73 evaluable patients, 40 (55%) patients subsequently had disease progression. The most frequent first site of distant metastasis was brain. Conclusion: Twice daily radiation therapy plus concurrent chemotherapy produced favorable response and survival for LS-SCLC patients with tolerable toxicities. To improve the treatment response, which proved as a significant prognostic factor for survival, there should be further investigations about fractionation scheme, chemotherapy regimens and compatible chemoradiotherapy schedule.

Histological Observation of Bone Reaction with Relation to Surface Treatment of Titanium (생체합금 Titanium의 표면처리에 따른 골반응의 변화에 대한 조직학적 관찰)

  • Ahn, Myun-Whan;Choi, Yong-Sug;Ahn, Jong-Chul;Seo, Jae-Sung;Lee, Dong-Chul;Jang, Jae-Hyuk
    • Journal of Yeungnam Medical Science
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    • v.13 no.1
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    • pp.116-133
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    • 1996
  • The biocompatibility of the titanium has been estabilished through various experimental studies such as cell culture toxicity test, pyrogen test, mutagen test and others. In order to confirm biocompatibility after fabrication of titanium and to clarify the difference between the bone reaction after insertion of the lathed titanium rods and the bone reaction after insertion of the finished and polished rods, both rods were implanted into the proximal femur of a rabbit. Histologic reactions in the bone were observed according to the ASTM standards at the intervals of 6 weeks, 12 weeks and 26 weeks after implantation. The result were as follows : In 6 weeks after implantation of lathed titanium rods, inflammatory reactions, such as minimal degree infiltration of polymorphonuclear leukocytes and lymphocytes were observed in all cases. This was thought to he caused by surgical trauma. However, inflammatory cell infiltration was not seen after implantation of polished and finished rods in all cases. The cellular infiltration and the histologic reaction of the hone after implantation of lathed group were significantly more pronounced than those after implantation of the finished group. In 12 weeks after implantation of lathed rods, two of four cases revealed a minimal degree of cellular infiltration. No inflammatory cell infiltration was demonstrated after implantation of the finished group. The cellular infiltration and histologic reaction seemed to be more pronounced in the lathed group, but they were not significant statistically. At 26 weeks after implantation of the lathed and finished group, there was no cellular infiltration in both groups. New bone formation was observed up 26 weeks, and no difference between lathed titanium rods and finished titanium rods were apparent. Mild bone necrosis was observed in 1 case out of 11 cases in which lathed titanium rods were implanted. Bone necrosis was not observed in the finished titanium rod group. Fibrosis was observed in both groups, but differences were not significant between the experimental groups. In the lathed titanium rods group and the shorter interval group, inflammatory cell infiltration was significantly higher. Finished titanium rods and longer interval groups had markedly decreased tendences in histologic reaction ratings. As a conclusion, although certificated titanium might be safe to use, difference of biocompatibility were observed depending on the method of surface finish. By identifying biocompatibility as a long-term standardized animal study, we can develop progressed internal fixation device that is safe for human beings.

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Antioxidative and Cytoprotective Effects of Annona muricata (Graviola) Extract for HDF Cell Damage Induced by Hydrogen Peroxide (H2O2에 의해 유도된 HDF 세포 손상에 대한 그라비올라 추출물의 항산화 및 세포 보호 효과)

  • Shin, Yun-Mi;Kim, You-Jeong;You, Seon-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.568-576
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    • 2017
  • As interest in functionality and environmentally friendly cosmetics is growing in recent years, materials that use safe and effective plant extracts have been developed. Therefore, this study also attempted to check the possibility of the graviola extract, which is known to have various efficacy mainly as a health functional material as a functional cosmetic material. In order to find out the antioxidant activity of graviola, we measured total polyphenol, total flavonoid content and DPPH radical scavenging activity and measured the ROS activity inhibition effect and cytoprotective effect on oxidative stress by treating HDF with hydrogen peroxide cells at an appropriate concentration after checking cytotoxicity in HDF cells. Based on the results of this experiment, the graviola extract was found to contain as high as 26.6 mg(CA)/100g, 14.3 mg(Q)/100g of total polyphenol and flavonoid, which are the antioxidant indexes and to have the high radical scavenging activity. The cell survival rate of the HDF cells was measured, and as a result, no significant cytotoxicity was observed at all concentrations and the experiment was carried out at a concentration of $100{\mu}g/mL$ afterwards. Inhibition of ROS activity in HDF cells induced by hydrogen peroxide was measured and the concentration-dependent inhibition of ROS activity was found and the cell protection effect of graviola was measured after hydrogen peroxide was treated for 4, 24 and 48 hours. As a result, the cell protection effect as high as 89.92% was confirmed at a $25{\mu}g/mL$ concentration up to 24 hours. As these results show that the graviola extract has excellent antioxidant activity, almost no toxicity to HDF cells, an effective activity inhibitory effect on active oxygen generated by hydrogen peroxide and excellent cytoprotective effect, the possibility as various functional materials with antioxidant and cytoprotective effects was confirmed.

Comparative Study of Toxic Effects of Anatase and Rutile Type Nanosized Titanium Dioxide Particles in vivo and in vitro

  • Numano, Takamasa;Xu, Jiegou;Futakuchi, Mitsuru;Fukamachi, Katsumi;Alexander, David B.;Furukawa, Fumio;Kanno, Jun;Hirose, Akihiko;Tsuda, Hiroyuki;Suzui, Masumi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.929-935
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    • 2014
  • Two types of nanosized titanium dioxide, anatase ($anTiO_2$) and rutile ($rnTiO_2$), are widely used in industry, commercial products and biosystems. $TiO_2$ has been evaluated as a Group 2B carcinogen. Previous reports indicated that $anTiO_2$ is less toxic than $rnTiO_2$, however, under ultraviolet irradiation $anTiO_2$ is more toxic than $rnTiO_2$ in vitro because of differences in their crystal structures. In the present study, we compared the in vivo and in vitro toxic effects induced by $anTiO_2$ and $rnTiO_2$. Female SD rats were treated with $500{\mu}g/ml$ of $anTiO_2$ or $rnTiO_2$ suspensions by intra-pulmonary spraying 8 times over a two week period. In the lung, treatment with $anTiO_2$ or $rnTiO_2$ increased alveolar macrophage numbers and levels of 8-hydroxydeoxyguanosine (8-OHdG); these increases tended to be lower in the $anTiO_2$ treated group compared to the $rnTiO_2$ treated group. Expression of $MIP1{\alpha}$ mRNA and protein in lung tissues treated with $anTiO_2$ and $rnTiO_2$ was also significantly up-regulated, with $MIP1{\alpha}$ mRNA and protein expression significantly lower in the $anTiO_2$ group than in the $rnTiO_2$ group. In cell culture of primary alveolar macrophages (PAM) treated with $anTiO_2$ and $rnTiO_2$, expression of $MIP1{\alpha}$ mRNA in the PAM and protein in the culture media was significantly higher than in control cultures. Similarly to the in vivo results, $MIP1{\alpha}$ mRNA and protein expression was significantly lower in the $anTiO_2$ treated cultures compared to the $rnTiO_2$ treated cultures. Furthermore, conditioned cell culture media from PAM cultures treated with $anTiO_2$ had less effect on A549 cell proliferation compared to conditioned media from cultures treated with $rnTiO_2$. However, no significant difference was found in the toxicological effects on cell viability of ultra violet irradiated $anTiO_2$ and $rnTiO_2$. In conclusion, our results indicate that $anTiO_2$ is less potent in induction of alveolar macrophage infiltration, 8-OHdG and $MIP1{\alpha}$ expression in the lung, and growth stimulation of A549 cells in vitro than $rnTiO_2$.