This study evaluated cell viability and cytokine release in immortalized human oral fibroblasts (hTERT-hNOFs) and keratinocytes (IHOK) exposed to a dental-impregnated gingival retraction cord. To prepare the extracts, dental gingival retraction cords impregnated with aluminum chloride hexahydrate were immersed in a cell culture medium for 24 h at 37 °C. hTERT-hNOFs and IHOK were cultured for 24 h. The cell culture medium was removed and extracts of the dental gingival retraction cords were added. After incubation with the extract solution, cell viability was evaluated using an MTT assay. The levels of the cytokines IL-1α and IL-8 were measured in the supernatants of each cell type. The cell viability after exposure to the extract solution for 10 min exceeded 70 % in both cell types. The ET50 values for hTERT-hNOF and IHOK were 35.75 and 28.98 min, respectively. For IHOK, the IL-1α level was (5.35 ± 5.22) pg/mL at 10 min, (3.58 ± 5.38) pg/mL at 20 min, and (2.85 ± 4.28) pg/mL at 60 min of exposure (p > 0.05). The IL-8 level in IHOK was (67.16 ± 18.70) pg/mL at 10 min, (78.36 ± 7.50) pg/mL at 20 min, and (111.9 ± 26.10) pg/mL at 60 min of exposure (p > 0.05). Cytokine release was not observed from hTERThNOFs. Based on these results, cell viability and cytokine release were confirmed in cells exposed to the impregnated gingival retraction cord. In addition, the application of the extracts to hTERT-hNOF and IHOK during the actual contact time and determination of ET50 may be beneficial for evaluating the biocompatibility of dental-impregnated gingival retraction cords.

쥐참외뿌리 추출물이 항산화 활성 및 피부 세포에서의 독성을 확인하여, 피부에 효과적으로 사 용할 수 있는 기능성 소재로써의 활용 가능성을 확인해 보고자 하였다. 쥐참외뿌리 추출물의 항산화 활성 의 지표가 되는 총 폴리페놀과 총 플라보노이드 함량을 확인하였고, 피부에서의 Neutral red assay를 이용 하여 세포 독성을 확인하였다. 연구 결과, 총 폴리페놀과 총 플라보노이드의 함량은 농도 의존적으로 증가 하였다. 섬유아 세포인 HDF cell에서의 높은 생존율이 확인되었으며, B16F10 melanoma cel와 RAW 264.7 cell에서는 5 μg/mL부터 세포 생존율이 유의하게 낮아지는 것이 확인되었다. 본 연구 결과는 쥐참 외뿌리 추출물의 항산화 활성 및 피부 세포에서의 기초적인 자료로 사용가능할 것으로 사료되어 진다.

The purpose of this study was to investigate the biological activity of fucoidan, a sulfur-containing polysaccharide, on cytotoxicity and apoptosis in the human HT-29 colorectal cancer cell line using cell viability, Flow cytometry, Western blot, and RT-PCR analyses. Fucoidan inhibited the proliferation of HT-29 cells by 39.6% at a concentration of 100 μg/mL for 72 h. The inhibition was dose-dependent and accompanied by apoptosis. Flow cytometric analysis showed that fucoidan increased early apoptosis and late apoptosis by 65.84% and 72.09% at concentrations of 25 and 100 μg/mL, respectively. Analysis of the mechanism of these events indicated that fucoidan-treated cells exhibited increases in the activation of caspase-3, caspase-8, and PARP in a dose-dependent manner. These results suggest that fucoidan may inhibit the growth of human colorectal cancer cells by various apoptosis-promoting effects, as well as by apoptosis itself.

본 연구에서는 국내 다빈도로 사용되고 있는 가공보조제 효소제 7종에 대한 최대 사용량 기반 세포독성을 연구하였다. 기원이나 원산지 및 제조형태가 다른 효소제가 시판되고 있는 경우 모두 시험대상에 포함하였고, 효소제의 경우 가공과정에 사용 후 최종제품에서 잔존하지 않거나 불활성화되기 때문에 native 및 inactivated 두 형태에 대한 세포독성을 확인하였다. 세포처리 최고 농도는 품목 제조보고서를 기반으로 다빈도 첨가 식품품목 및 그 최대 사용량과 국민영양통계 일일섭취량을 기반으로 설정하였 다. 그 결과, native 및 inactivated 효소제 모두 세포성장 및 세포막 손상에 의한 세포사멸을 유발하지 않는 것으로 나타났다. Native 효소제의 경우 종국 3종 모두에서 ROS 유발이 확인되었으나, inactivated 종국에서 ROS의 유의적 증가는 나타나지 않았다. 일부 native 형태의 종국 및 프로테아제에서 β-hexosaminidase 용출에 의한 탈 과립화가 유발될 수 있는 것으로 분석되었으나, 모든 inactivated 효소 제에서 탈 과립화는 유의적으로 나타나지 않았다. 이러한 결과는 효소제의 기원에 따라서 native 형태의 경우 ROS 유발에 의한 산화스트레스 및 탈 과립화와 같은 알레르기 원성 세포독성을 나타낼 수 있으나, 실제 가공제품에 잔존 하는 불활성화된 효소제에 의한 세포독성 영향은 낮음을 제시한다 할 수 있다. 이와 같은 연구결과는 안전성 정보 가 미흡한 가공보조제 효소제에 대한 구체적인 독성자료 및 향후 in vivo 독성연구를 위한 기초자료로 활용될 수 있을 것이다.

We investigated the cytotoxic potential of three different commercially available absorbent feminine hygiene products and one transdermal patch using direct contact and extract exposure methods. Two different cell lines were used – mouse fibroblast L929 and normal human skin fibroblast CCD-986sk cell lines. The test samples were extracted using three different methods in accordance to International Organization for Standardization (ISO). Viability of cells was analyzed using MTT assay and morphology of the cells were also observed using phase contrast microscopy. Overall, the direct contact method using L929 cells showed that all the test samples had no toxic effect when exposed to extracts for 1 h. For the exposure method, no toxic effect was observed in both L929 and CCD986sk cells incubated with all the test samples regardless of the extraction methods used.

Natural killer (NK) cells are innate lymphocytes that aid in the protection of the host from infectious diseases and cancer. Regulation of cytotoxicity in NK cells is mediated by inhibitory receptors that bind major histocompatibility complex class I (MHC-I) molecules on target cells. Human myelogenous leukemia K562 cells are readily attacked by NK cells, because K562 cells expressed very low levels of MHC-I molecules for inhibitory NK cell receptors. In this study, we compared the ability of γ-irradiated- or mitomycin C (MMC)-treated K562 feeder cells to support expansion and activation of canine NK cells. We isolated CD5 negative cells from canine peripheral blood mononuclear cells by immunomagnetic separation and co-cultured with γ-irradiated (100 Gy)- or MMC (20 μg/mL)-treated K562 cells in the presence of interleukin (IL)-2, IL-15 and IL-21 for 21 days. As a result, number of CD5 negative cells, co-cultured with γ-irradiated- or MMC-treated K562 cells (56.72 ± 13.77 fold or 32.99 ± 10.83 fold), was increased than those of CD5 negetive cells (2.99 ± 1.42 fold). Also, we found that gene expression markers of activated NK cells such as NKp30, NKp44, NKp46, Ly49, NKG2D, CD244, perforin, and granzyme B and production of interferon gamma were similarly upregulated in NK cells co-cultured with γ-irradiated- or MMC-treated K562 cells, respectively. Next, we observed that cytotoxicity of NK cells co-cultured with γ-irradiated K562 cells was more sensitively reacted to canine mammary carcinoma cells than those of MMC-treated K562 cells. These results revealed that γ-irradiated K562 cells are more efficient feeder cells than MMC-treated K562 cells for enhancing NK cells expansion and activation.

Multi-walled carbon nanotubes (MWNTs) are suitable for delivering large biomolecules with lower cytotoxicity values and low prime cost. Surface modifications of MWNTs affect interaction with cells and proteins. Oxidation with strong acids decreases cytotoxicity of CNTs and increases protein-loading capacity. Here, after oxidation, TEM images revealed more aligned structure and carboxylated groups at the surface which decreases toxicity. Functionalized MWNTs showed more gradual degradation than the pristine MWNTs and mass loss increased by 2% in the same temperature range. Raman spectroscopy corrected graphitic structure with characteristic D and G bands at 1330 and 1579 cm−1 and increased intensity after oxidation. FTIR spectroscopy peaks at 1443 cm−1, 1560, 1640 cm−1, 2100–2200 cm−1 and 3426 cm−1 are ascribed to C–O–C vibrational stretch, C=C bonds, vibration of C≡C bonds and stretch of hydroxyl groups, respectively. The sonicationdriven dispersion of in phosphate-buffered saline, distilled water and cell culture medium were detected by UV–vis–NIR spectroscopy, water-dispersed functionalized MWNTs revealed the highest absorbance value. Cytotoxicity of MWNTs was investigated before and after functionalization in breast cancer (MDA-MB-231) and human vein endothelial (HUVEC) cells. Relatively low-toxicity results were obtained in functionalized MWNTs and cellular uptake of MWNTs were corrected with fluorescent imaging of cells and cell lysates. Protein-loading capacity of fsMWNTs (functionalized short-length multi-walled carbon nanotubes) was evaluated by using bovine serum albumin (BSA) and with an equal amount of fsMWNTs and BSA; 36% binding yield was obtained. Protein corona after covalent functionalization potentially lowered cytotoxicity up to 6%.

본 연구에서는 고구마(Ipomoea batatas L.) 수확 후 버려지는 잔재물의 활용가치를 위해 이들로부터 추출한 발효 추출물과 열수 추출물의 생리활성과 세포독성을 분석 하였다. 추출물의 pH는 모두 산성을 나타내었고, 유기물 함량은 발효 추출물과 열수 추출물에서 각각 0.98%와 0.97%로 비슷하게 나타내었다. 다량원소 중 인산, 칼륨, 칼슘, 마그네슘 성분의 함량은 열수 추출물에서 발효 추출물보다 모두 높게 나왔고, 질소 함량만 두 추출물에서 동일하게 나왔다. 미량원소 함량은 아연을 제외하고 발효 추출물보다 열수 추출물에서 높게 나타내었다. 총 폴리페놀 함량은 열수 추출물에서 60.5±2.7 mg/g로서 발효 추출물의 총 폴리페놀 함량의 22.7±4.2 mg/g 보다 37.8 mg/g 높은 함량을 나타내었다(p<0.05). 총 플라보노이드 함량은 열수 추출물에서 50.7±2.7 mg/g로서 발효 추출물의 함량인 14.0±2.1 mg/g 보다 36.7 mg/g 높은 함량을 나타내어 총 폴리페놀 함량과 마찬가지로 열수 추출물이 발효 추출물보다 높은 함량을 나타내었다(p<0.05). DPPH와 ABTS radical 소거능력은 모두 열수 추출물에서 발효 추출물 보다 높은 항산화력을 보였다(p<0.05). MTT assay를 이용한 추출물의 세포독성 실험에서는 두 추출물의 모든 농도에서 세포독성이 미약한 것으로 확인되었다. 따라서 고구마 수확 후 잔재물의 추출물이 향후 각종 바이오 소재로 이용 시에도 큰 문제가 없을 것이라 판단된다.

목적: 항균제 polyhexamethylene Biguanide(PHMB), epigallocatechin gallate(EGCG) 및 PHMB/EGCG 혼합물의 각막상피세포(primary human corneal epithelial cells, HCEpiCs)에 대한 급성독성을 평가하고자 하였다. 방법: 각막상피세포를 0.00001~0.005% PHMB, 0.001~5% EGCG 및 0.00005% PHMB/0.05% EGCG 혼합물이 각각 포함된 배양액에서 30분, 60분, 120분 및 240분 동안 배양하였다. 배양한 각막상피세포를 고정한 다음 Draq 5로 염색하고 공초점현미경과 ImageXpress UltraTM를 이용하여 세포형태를 관찰하여 세포 생존율과 세포자살(apoptosis)을 비교하였다. 결과: 배양된 각막상피세포는 0.00005% 이하의 PHMB 농도 및 0.05% 이하의 EGCG 농도에서는 세포 독성이 나타나지 않았다. 0.00005% PHMB/0.05% EGCG 혼합물이 포함된 배양액에서 급성 세포독성은 관찰되지 않았으나 240분 배양시킨 경우에는 손상된 각막상피세포 수가 증가하고 생존 세포의 수는 감소하였다. 결론: 항균 시너지 효과를 갖는다고 보고된 0.00005% PHMB/0.05% EGCG 혼합물의 경우 각막상피세포에 대한 급성독성은 없었으나 만성효과에 대해서는 추가적인 연구가 필요할 것으로 생각된다.

Nanostructured lipid carriers (NLC) are getting attention as delivery system for nutraceuticals due to its low toxicity and higher loading efficiency of active ingredients. However, the cytotoxicity of NLC had not fully evaluated especially on neuroblastoma. In this study, cytotoxicity of NLC and curcumin-loaded NLC (C-NLC) were evaluated on SH-SY5Y neuroblastoma cells investigating cell morphology, mitochondrial activity, and reactive oxygen species (ROS) production compared to H2O2 treatment as a positive control. As a result, the metabolic activity was inhibited about 40% by 250ppm of NLC along with morphological change. C-NLC exhibited 50% inhibitory effect on mitochondrial activity at 500ppm, which was lower than NLC itself. Moreover, NLCs significantly induced ROS production which was recognized as one of the indicators of cytotoxicity generated by NLCs. In conclusion, lower cytotoxic effect was observed with NLC on SH-SY5Y neuroblastoma based on ROS production and these investigation could be used for further application of NLC in food industry.

Xenotransplantation is proposed as a solution to the problem of organ shortage. However, transplantation of xenogeneic organs induces an antigen-antibody reaction in α-1,3-gal structure that are not present in humans and primates, and thus complement is also activated and organs die within minutes or hours. In this study, we used FasL gene, which is involved in the immune response of NK cell, and US11, which suppresses MHC Class I cell membrane surface expression, to inhibit cell mediated rejection in the interspecific immunity rejection, and also hDAF(CD55) was introduced to confirm the response to C3 complement. These genes were tranfeced into Korean native pig fetal fibroblasts using pCAGGS vector. And cytotoxicity of NK cell and human complement was confirmed in each cell line. The US11 inhibited the cytotoxicity of NK cell and, in addition, the simultaneous expression of US11 and Fas ligand showed excellent suppress to T-lymphocyte cytotoxicity, hDAF showed weak resistance to cytotoxicity of natural killer cell but not in CD8+ CTLs. Cytotoxicity study with human complement showed that hDAF was effective for reducing complement reaction. In this studies have demonstrated that each gene is effective in reducing immune rejection.

Fludarabine, a chain terminating anti-cancer drug, is a purine analogue that causes DNA strand breaks in normal cells. In this study, we determined if A. melanocarpa and Korean red ginseng extract mixture reduce cytotoxicity of fludarabine. Treatment of HaCaT cells with 10 μM of fludarabine for 24 hours decreased cell viability and increased DNA strand breaks. Treatment of A. melanocarpa and Korean red ginseng extract mixture for 24 hours increased cell viability as compared with single extract treatment. The protective effect of these extracts on cell activity increased in a concentration-dependent manner. DNA strand breaks induced by fludarabine decreased as concentration of extract mixture increased. p-H2AX level, a marker of DNA strand breakage, decreased depending on the concentration of extract mixture. The effect of mixed extract of A. melanocarpa and Korean red ginseng on DNA damage is due to the anti-oxidative effect of A. melanocarpa and signal transmission through glucocorticoid receptor upon binding of saponin of Korean red ginseng.

Ganoderma lucidum has been traditionally used as a medicine for treatment of bronchitis, arthritis, and high blood pressure, and it has been reported to display many biological activities including anticancer and immune activities. Since mushroom mycelium is known to have excellent biological activities together with mushroom fruiting body, studies on biological activities of mushroom mycelium have been actively conducted. Thus, the present study compared the biological activities before and after the cultivation of Ganoderma lucidum mycelium on Atractylodes rhizoma. When the radical scavenging activity was assessed by the DPPH assay, ARGL (ethanol extract of Atractylodes rhizoma mycelium fermented with Ganoderma lucidum) showed radical scavenging activity of 5.58~82.56% at concentrations of 10~500 μg/assay, while AR (ethanol extract of Atractylodes rhizoma) showed radical scavenging activity of 5.27~72.08% at the same concentrations. When measured by using the ABTS assay, ARGL showed higher radical scavenging activity than AR, which was consistent with the result obtained by the DPPH assay. In the MTT assay, the cytotoxicity of ARGL against all cell lines was higher than that of AR. In particular, the cytotoxicities of AR and ARGL against Hep3B at a concentration of 400 μg/assay were 71.81% and 86.40%, respectively. In addition, the result obtained by the SRB assay was consistent with the result obtained by the MTT assay. According to the results mentioned above, there is a high probability that medicinal herb cultures using mycelium can be used as sources of functional foods since the cytotoxicities against cancer cells and antioxidant activities increased when the mycelium was fermented with Atractylodes rhizoma.

The red ginseng is known to have effects on antioxidativity and cytotoxicity. Nanoscale active substances have various advantages such as improved bioavailability and permeation ability into the cell. However, few studies conducted with the nanoparticles of red ginseng due to its low yield rate and difficulty of manufacturing the product in pilot scale. This study, therefore, investigated the size effects of ultra-fine powder of red ginseng on antioxidativity and cytotoxicity. Red ginseng powder (6, 8, or 158 μm) prepared using a pilot scale was provided by a local company. Antioxidativity was measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assays, and cytotoxicity was tested by methylthiazolyl tetrazolium (MTT) assay. The results of DPPH and ABTS radical electron donating ability IC50 of red ginseng were ranged from 2.27 to 3.34 mg/ml and 2.94 to 3.09 mg/ml, respectively, which were not significantly different between all samples. However, the results of cytotoxicity clearly showed a pattern of decreased toxicity in 6 and 8 μm power compared to 158 μm powder. Unexpectedly, particle sizes of red ginseng did not significantly affect antioxidativity. It is believed that these were related to the process of pilot scale production. These phenomena are also believed to be caused by aggregation of low size power particle that increases water holding capacity. From our result, it is concluded that this range of particle size of red ginseng affected the reduction of cytotoxicity.

본 연구는 우엉 뿌리 추출물의 항산화 활성 및 피부에 대한 안전성을 평가하기 위하여 총 폴 리페놀 함량, 총 플라보노이드 함량, DPPH radical 소거 활성을 통하여 항산화 활성을 살펴보고, B16F10 melanoma 세포에 대한 세포 독성 및 자외선 A에 대한 피부 세포 보호 효과를 확인하였다. 또 한 화장품 소재로서의 활용을 검증하기 위하여 1차 피부 첩포 테스트를 실시하였다. 본 실험 결과 우엉 뿌리 추출물의 함량이 증가됨에 따라 높은 폴리페놀과 플라보노이드의 함량이 확인되었으며, DPPH radical 소거 활성을 확인하였다. B16F10세포에 대한 세포 독성을 확인한 결과 B16F10 melanoma 세포 에 대해 독성이 낮고, 자외선 A에 대해 80% 이상의 세포 보호 효과를 확인하였다. 또한 1차 피부 첩포 테스트를 통해 우엉 뿌리 추출물이 피부에 자극이 거의 없음을 확인하였다. 이러한 결과를 통하여 우엉 뿌리 추출물이 항산화 활성과 자외선에 대한 피부 보호 효과가 뛰어나고 피부 세포에 대한 독성이 낮으 며, 피부에 대한 안전성이 확인됨에 따라 화장품 소재로서의 가능성을 확인하였다.

Acetylcholine receptors (AChR) including muscarinic and nicotinic AChR are widely expressed and mediate a variety of physiological cellular responses in neuronal and non-neuronal cells. Notably, a functional cholinergic system exists in oral epithelial cells, and nicotinic AChR (nAChR) mediates cholinergic anti-inflammatory responses. However, the pathophysiological roles of AChR in periodontitis are unclear. Here, we show that activation of AChR elicits increased cytosolic Ca2+ ([Ca2+]i), transient cytotoxicity, and induction of receptor activator of nuclear factor kappa-B ligand (RANKL) expression. Intracellular Ca2+ mobilization in human gingival fibroblast-1 (hGF-1) cells was measured using the fluorescent Ca2+ indicator, fura-2/AM. Cytotoxicity and induction of gene expression were evaluated by measuring the release of glucose-6-phosphate dehydrogenase and RT-PCR. Activation of AChR in hGF-1 cells by carbachol (Cch) induced [Ca2+]i increase in a dose-dependent manner. Treatment with a high concentration of Cch on hGF-1 cells caused transient cytotoxicity. Notably, treatment of hGF-1 cells with Cch resulted in upregulated RANKL expression. The findings may indicate potential roles of AChR in gingival fibroblast cells in bone remodeling.

산업화를 위한 지구자 및 12가지 식물성 원료 추출물의 생리학적 효과를 평가하기 위하여, 만성 에탄 올을 투여한 마우스 혈청 내에서의 생리학적 지표 및 간과 뇌 조직 내에서의 산화적 스트레스에 대한 보호효과를 확인하였다. 에탄올을 투여한 마우스의 혈당은 정상 대조군 그룹(NG)과 에탄올 투여 그룹 (EG)에서 각각 119.43mg/dL, 305.25mg/dL로 나타났고, 에탄올과 혼합 추출물을 동시에 투여한 그룹 (100, 200mg/kg body weight + 25% ethanol 5g/kg body weight, ME100, ME200)에서 각각 272.76mg/dL, 234.60mg/dL로 감소하였다. 혈중 에탄올 함량은 EG에서 4.08mg/dL를 나타내었고 ME100, 200에서 각각 3.85mg/dL, 3.08mg/dL로 감소하였으며, 혈중 아세트알데하이드 함량은 18.72mg/dL에서 각각 15.76mg/dL, 15.16mg/dL로 감소하였다. 또한 ME100, ME200은 혈청 내의 생 리학적 지표에서 간 독성 지표인 glutamine pyruvic transaminase(GPT), glutamic oxaloacetic transaminase(GOT)와 신장 독성 지표인 blood urea nitrogen(BUN), creatine(CRE), 혈중 total cholesterol(TCHO), triglyceride(TG)의 함량이 유의하게 감소하였다. 뇌 조직에서 에탄올에 의해 acetylcholinesterase(AChE)가 EG(116.10%)에서 NG(100.00%)와 비교하였을 때 증가된 활성을 나타냈 으나, ME에서 각각 109.00%와 108.47%로 유의적으로 감소하였다. ME에서 EG에 비해 간과 뇌 조직에 서 superoxide dismutase(SOD)의 함량이 증가하였고, oxidized glutathione(GSH)/total GSH 비율과 malondialdehyde(MDA)의 함량이 감소하였다. 이러한 결과들을 통해 지구자를 포함한 혼합 추출물은 간, 뇌 조직 및 혈액 등에서 만성 에탄올 섭취에 의해 유발된 산화적 스트레스를 효과적으로 보호할 수 있는 제품으로의 개발이 가능할 것으로 판단된다.