• 생명과학회지
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• 제14권2호
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• pp.362-367
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• 2004

장염비브리오균의 숙주 내 감염을 일으키는 기작을 이해하기 위하여 세포외 효소 중의 하나인 콜라겐분해효소의 유전자가 불활성화된 돌연변이체를 제작하였다. 콜라겐분해효소의 유전자인 vppC 유전자에 항생제 내성 유전자인 nptII를 삽입하여 제작된 재조합 DNA를 suicide vector인 pDMS197에 클로닝하여 pVCM03이라 명명하였다. 재조합 suicide 플라스미드 pVCM03을 E. coli 7213에 형질전환하여 접합을 통하여 원 균주인 V. varahaemolyticus 04에 전달하였다. 전달된 pVCM03 유래의 재조합 vvpC::npfII DNA는 homologous recombination에 의해 wild-type allele와 교환되어 돌연변이체를 형성하게 되고, 돌연변이체는 10% sucrose가 함유된 TCBS 배지에서 선별되었다. Allele exchange는 PCR에 의한 증폭된 DNA의 크기 비교로 확인하였다. 돌연변이체인 V. parahaemolyticus CM은 원 균주와 비교하였을 때 약 4배정도 낮은 콜라겐 분해 활성을 나타내었고, vero cell을 이용한 MTT assay에서도 원 균주에 비하여 낮은 세포독성을 보였다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권3호
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• pp.216-225
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• 2002

Hyaluronic acid (HA) is an almost essential component of extracellular matrices. Early in embryogenesis mesenchymal cells migrate, proliferate and differentiate, in part, because of the influence of HA. Since the features of embryogenesis are revisited during wound repair, including bone fracture repair, this study was initiated to evaluate whether HA has an effect on calcification and bone formation in an in vitro system of osteogenesis. Mouse calvaria Pre-osteoblast (MC3T3-E1) cells were cultured in ${\alpha}-MEM$ medium with microorganism-derivative hyaluronic acid that was produced by Strep. zooepidemicus which of molecular weight was 3 million units. The dosages were categorized in each 0.5, 1.0 and 2.0 mg/ml concentration experimental groups. After 2 and 4 days cultures in expeirmental and control groups, the tendency of cell proliferation, MTT assay, protein synthesis ability, collagen synthesis and alkaline phosphatase activity were analysed and bone nodule formation capacity were measured with Alizarin Red S stain after 29 days cultures. The cell proliferation was increased in time, especially the group of 0.5 and 1.0 mg/ml concentration of HA were showed prominent cell proliferation. After 2 and 4 days culture, experimental groups in general were greater cell activity in MTT assay. The protein synthesis was increased in all experimental groups compared to control group, especially most prominent in 1.0 mg/ml concentration group. The collagen synthesis capacity were increased in HA experimental groups, especially prominent in 1.0 mg/ml group and the activity of alkaline phosphatase were increased, especially also prominent in 1.0 mg/ml group, compared to control group. Above these, the activity of mouse carvarial pre-osteoblast cells was showed greater bone osteogenesis activity in all applied HA experimental group, especially group of 1.0 mg/ml concentration of HA.

• Korean Journal of Acupuncture
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• 제19권1호
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• pp.77-84
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• 2002

목적 : 소아의 정신적 및 신체적 발달지연에 사용되고 있는 신기환이 해마의 신경세포 생성에 미치는 영향을 관찰하고자 하였다. 방법 : 신기환이 세포생성에 미치는 영향을 알아보기 위하여 해마 세포인 HiB5에 신기환을 처리, 배양하여 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay를 통해서 세포생성을 관찰하였다. 또한 3주령의 Sprague-Dawley (S-D)계 흰쥐의 중완혈에 신기환 약침을 자침후 해마의 치상회에서 bromodeoxyuridine (BrdU) immunohistochemistry를 시행하여 세포생성을 관찰하였다. 결과 : HiB5 배양세포에서는 신기환을 처리시 배양시간과 농도별로 세포생성율이 증가하였다. 또한, 대조군의 치상회에서 BrdU 양성세포수는 $128.50{\pm}9.53$, 1 mg/kg 신기환 자침군은 $157.08{\pm}10.82$ 및 10 mg/kg 신기환 자침군은 $204.80{\pm}17.68$로 나타났다. 이러한 결과로 신기환은 어린 S-D계 흰쥐의 치상회에서 세포생성을 증가시키는 것으로 사려된다.

• Restorative Dentistry and Endodontics
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• 제38권4호
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• pp.204-209
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• 2013

Objectives: The objective of this in vitro study was to evaluate and compare the cytotoxicity of four different root canal sealers i.e. Apexit Plus (Ivoclar Vivadent), Endomethasone N (Septodont), AH-26 (Dentsply) and Pulpdent Root Canal Sealer (Pulpdent), on a mouse fibroblast cell line (L929). Materials and Methods: Thirty two discs for each sealer (5 mm in diameter and 2 mm in height) were fabricated in Teflon mould. The sealer extraction was made in cell culture medium (Dulbecco's Modified Eagle's Medium, DMEM) using the ratio 1.25 $cm^2/mL$ between the surface of the sealer samples and the volume of medium in a shaker incubator. Extraction of each sealer was obtained at 24 hr, 7th day, 14th day, and one month of interval. These extracts were incubated with L929 cell line and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay was done. Two-way ANOVA for interaction effects between sealer and time and Post-hoc multiple comparison using Tukey's test across all the 16 different groups were used for statistical analysis. Results: Apexit Plus root canal sealer was significantly less toxic than other sealers (p < 0.05) and showed higher cellular growth than control. Endomethasone N showed mild cytotoxicity. AH-26 showed severe toxicity which became mild after one month while Pulpdent Root Canal Sealer showed severe to moderate toxicity. Conclusions: Apexit Plus was relatively biocompatible sealer as compared to other three sealers which were cytotoxic at their initial stages, however, they became biocompatible with time.

• 융합정보논문지
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• 제10권7호
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• pp.249-256
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• 2020

본 연구는 피부 관리에 사용되는 천연화장품 원료로서 동과자 추출물의 활용 가능성을 조사하기 위해 항산화, 항염 그리고 미백 효능을 평가하였다. DPPH 자유라디컬 소거 활성은 농도 의존적으로 감소하였으며, 총폴리페놀 함량은 메탄올추출물(22.42±0.002 mgGAE/g)이 열수추출물(9.77±0.002 mgGAE/g)보다 더 높게 나타났다. MTT assay 결과 동과자 추출물에 의한 RAW264.7 세포와 B16F10 세포에서의 세포독성은 없었다. 또한 LPS 처리에 의해 활성화된 RAW264.7 세포에서 NO 생성과 TNF-α 분비 억제능을 확인하였다. B16F10 melanoma 세포에서 동과자 추출물은 α-MSH에 의한 멜라닌 합성을 유의하게 저해하는 것으로 나타났다. 게다가, westernblot 분석결과 동과자 메탄올추출물은 MITF, TRP-1 및 TRP-2 단백질의 발현 수준을 현저하게 감소시켰다. 이상의 결과로 동과자 추출물은 항산화, 항염 그리고 미백 효능을 가진 기능성 화장품 소재로써의 가치가 높을 것으로 사료된다.

• International Journal of Oral Biology
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• 제35권4호
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• pp.153-158
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• 2010

Angelica decursiva has been used in Korean traditional medicine as an antitussive, an analgesic, an antipyretic and a cough remedy. However, the anti-cancer properties of Angelica decursiva have not yet been well defined. In our current study the cytotoxic activity of ethanol extracts of Angelica decursiva root (EEAD) and the mechanism of cell death exhibited by EEAD were examined in FaDu human head and neck squamous cell carcinoma cells. The cytotoxic effects of EEAD upon the growth of FaDu cells were examined with an MTT assay. In addition, the mechanism of cell death induced by EEAD was evaluated by DNA fragmentation analysis, immunoblotting and caspase activation measurements. EEAD induced apoptotic cell death in FaDu cells in a concentration- and time-dependent manner, as determined by MTT assay and DNA fragmentation analysis. Furthermore, the proteolytic processing of caspase-3, -7 and -9 was increased by EEAD treatment of FaDu cells. In addition, the activation of caspase-3 and -7 was detected in living FaDu cells by fluorescence microscopy. These results suggest that EEAD can induce apoptosis and suppress cell growth in cancer cells and may have utility as a future anticancer therapy.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.147-154
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• 2005

Objective: In this study, the effects of Liriope Platyphylla against LPS-induced inflammation was investigated. Methods: Cell viability was determined using the MTT assay. To identify expressions of COX-2 and iNOS mRNA, RT-PCR was performed. Assessment of PGE2 synthesis was performed using the PGE2 immunoassay. Measurement of NO synthesis was performed using the NO detection. Result : The MTT assay revealed that Liriope Platyphylla exerted no significant cytotoxicity in the microglial BV2 cells. RT-PCR analysis revealed that the mRNA levels of COX-2 and iNOS were significantly decreased in the LPS- and 5 mg/ml Liriope Platyphylla treated group. From the PGE2 immunoassay and NO detection, PGE2 and NO synthesis was significantly suppressed in the LPS- and 5 mg/ml Liriope Platyphylla treated group. Conclusion : In these study, Liriope Platyphylla was shown to suppress PGE2 and NO production by inhibiting LPS-stimulated enhancement of COX-2 enzyme activity and iNOS expression. It is very possible that Liriope Platyphylla can offer a valuable mode of therapy for the treatment of brain inflammatory diseases.

• 대한본초학회지
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• 제21권1호
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• pp.101-107
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• 2006

Objectives : It is well known that hexavalent chromium has toxic effect on normal cells. Recently, toxic effect of hexavalent chromium is diminished by the some extracts derived from herbs or plants. But, the toxic or protective mechanism of hexavalent chromium is well unknown. This study was performed to examine the protective effect of poncirin against $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Methods : The protective effect of the cytotoxicity induced by $Na_2Cr_2O_7$ was measured by the cell viability after NIH3T3 fibroblasts were cultured with or without $Na_2Cr_2O_7$ for 48 hours. Antitoxic effects of poncirin on the cytotoxicity induced by $Na_2Cr_2O_7$ were examined by colorimetric assays such as MTT or XTT assay. Results : $Na_2Cr_2O_7$ decreased cell viability by the decreased absorbance in MTT or XTT assay, but, the poncirin increased cell viability which was decreased by $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Conclusion : These results suggest that $Na_2Cr_2O_7$ showed cytotoxicity effect on NIH3T3 fibroblasts by the decrease of cell viavility, and poncirin was effective in the protection of $Na_2Cr_2O_7$-induced cytotoxicity in these cultures.

• 한국결정성장학회지
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• 제22권6호
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• pp.279-285
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• 2012

BCP, Mg-BCP, Si-BCP 분말을 공침법(co-precipitation process)을 이용하여 합성하였다. 제조된 분말을 X-선 회절 분석(XRD), 적외선분광분석(FTIR)을 이용하여 특성을 분석하였다. Ca/P 몰 비율이 1.602인 칼슘 결손 아파타이트 공침물을 제조하여 $1000^{\circ}C$ 열처리 과정을 통하여 HAp와 ${\beta}$-TCP 상이 혼재된 BCP, Mg-BCP, Si-BCP을 합성할 수 있었다. 제조된 분말의 생체활성 거동을 평가하기 위하여 Hanks' Balanced Salt Solution(HBSS)에 침적시켜 시간에 따라 형상의 변화 및 결정상을 분석한 결과, BCP 분말에 비하여 금속 이온이 치환된 BCP 분말에서 빠른 생체활성을 관찰할 수 있었다. MTT assay를 통한 세포 성장률 평가에서모든 분말에서 시간 경과에 따라 독성을 나타내지 않았으며, 세포의 활성이 증가하였다.

• 대한화장품학회지
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• 제41권2호
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• pp.151-157
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• 2015

본 연구에서는 기능성 화장품 소재 개발을 위하여 비파엽 추출물을 제조하고 효소 및 세포 활성평가를 진행하였다. 활성평가는 tyrosinase 측정법을 이용한 미백 효능, NO 측정법을 이용한 항염 효능에 대한 활성등을 분석하였다. 미백 효능 관련 tyrosinase 활성 저해는 10% 농도의 비파엽 초임계 추출물에서 미약한 활성을 나타내었다. 항염 효능 관련 NO 활성 저해 분석 결과, 초임계 추출물, 에탄올 환류 추출물, 열수 추출물의 순서로 NO 억제 활성을 나타내었다. MTT assay를 실시한 결과, 70% 에탄올 환류 추출물 5% 처리를 제외하고는 모든 농도에서 세포독성이 관찰되지 않았다. 미백 효능 관련 멜라닌 생성 저해는 5% 처리 농도에서 30%, 70% 에탄올 환류 추출물이 강하게 멜라닌 생성을 억제하는 결과를 보였다. 이상의 연구결과들로부터 비파엽 추출물 제조를 통한 우수한 항염 효능 및 미백 효능 관련 활성을 나타내는 추출조건을 도출하였으며, 향후 기능성 화장품 소재 개발을 위한 기초 연구용 자료로서의 역할을 제시할 수 있을 것이다.