• 제목/요약/키워드: NIH 3T3

검색결과 308건 처리시간 0.025초

Improvement of biohistological response of facial implant materials by tantalum surface treatment

  • Bakri, Mohammed Mousa;Lee, Sung Ho;Lee, Jong Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제41권
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    • pp.52.1-52.8
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    • 2019
  • Background: A compact passive oxide layer can grow on tantalum (Ta). It has been reported that this oxide layer can facilitate bone ingrowth in vivo though the development of bone-like apatite, which promotes hard and soft tissue adhesion. Thus, Ta surface treatment on facial implant materials may improve the tissue response, which could result in less fibrotic encapsulation and make the implant more stable on the bone surface. The purposes of this study were to verify whether surface treatment of facial implant materials using Ta can improve the biohistobiological response and to determine the possibility of potential clinical applications. Methods: Two different and commonly used implant materials, silicone and expanded polytetrafluoroethylene (ePTFE), were treated via Ta ion implantation using a Ta sputtering gun. Ta-treated samples were compared with untreated samples using in vitro and in vivo evaluations. Osteoblast (MG-63) and fibroblast (NIH3T3) cell viability with the Ta-treated implant material was assessed, and the tissue response was observed by placing the implants over the rat calvarium (n = 48) for two different lengths of time. Foreign body and inflammatory reactions were observed, and soft tissue thickness between the calvarium and the implant as well as the bone response was measured. Results: The treatment of facial implant materials using Ta showed a tendency toward increased fibroblast and osteoblast viability, although this result was not statistically significant. During the in vivo study, both Ta-treated and untreated implants showed similar foreign body reactions. However, the Ta-treated implant materials (silicone and ePTFE) showed a tendency toward better histological features: lower soft tissue thickness between the implant and the underlying calvarium as well as an increase in new bone activity. Conclusion: Ta surface treatment using ion implantation on silicone and ePTFE facial implant materials showed the possibility of reducing soft tissue intervention between the calvarium and the implant to make the implant more stable on the bone surface. Although no statistically significant improvement was observed, Ta treatment revealed a tendency toward an improved biohistological response of silicone and ePTFE facial implants. Conclusively, tantalum treatment is beneficial and has the potential for clinical applications.

말초신경재생을 위한 hNGF-$\beta$ recombinant Adenovirus의 제작 및 수종세포주에서 신경성장인자의 발현 (CONSTRUCTION OF HNGF-$\beta$ RECOMBINANT ADENOVIRUS & SCREENING OF ITS EXPRESSION AFTER TRANSFECTION INTO VARIOUS CELL LINES)

  • 고은봉;정헌종;안강민;김윤태;박희정;성미애;김남열;유상배;명훈;황순정;김명진;김성민;장정원;이종호
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제27권5호
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    • pp.446-456
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    • 2005
  • Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$\beta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$\beta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$\beta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$\beta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$\beta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$\beta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$\beta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$\beta$ gene transfected cells.

황백(Phellodendri Cortex)으로부터 분리한 지방세포 분화 저해물질 (Adipocyte Differentiation Inhibitor Isolated from the Barks of Phellodendron amurense)

  • 김경희;안순철;이명선;권오송;오원근;김민수;손천배;안종석
    • 한국식품과학회지
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    • 제35권3호
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    • pp.503-509
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    • 2003
  • 비만 저해 물질을 탐색하고자 전통의학에서 사용되는 200여 종류의 식용 또는 약용식물을 대상으로 지방세포 분화저해활성을 보이는 생약식물을 선별하여 용매층으로 활성이 이행되는 생약 중 황백(Phellodendri Cortex)을 최종적으로 후보식물로 선정하였다. 황백을 메탄올로 추출한 후, silica gel column chromatography, ODS RP-18 column chromatography, HPLC 등을 수행하여 지방세포 분화 저해활성을 갖는 화합물 PC-4를 분리하였다. PC-4는 노란색의 분말형태로 메탄올을 용매로 UV 최대흡수치를 조사한 결과, 222, 265, 349, 429 nm에서 최대의 UV의 흡수치를 보였으며 산이나 알카리 조건에서 최대 흡수치의 변화가 관찰되지 않았다. EI-mass spectrum을 측정한 결과, 분자량이 336.1로 예상되었으며 $^1H-NMR,\;^{13}C-NMR,\;DEPT,\;^1H-^1H\;COSY$, HMQC, HMBC NMR spectrum을 통해, PC-4는 isoquionoline alkaloide계열의 berberine으로 동정되었다. PC-4는 지방의 대사에 관여하는 중요한 효소인 fatty acid synthase와 pancreatic lipase에 대한 저해활성은 나타나지 않았으나, 지방세포 분화 저해활성은 $1\;{\mu}g/mL$의 낮은 농도에서도 관찰되었다.

인간 무세포성 진피기질 위에 배양한 가토 구강각화상피세포의 중충화와 기저막 형성에 관한 연구 (FORMATION OF BASEMENT MEMBRANE AND STRATIFICATION OF RABBIT ORAL KERATINOCYTES CULTURED ON HUMAN ACELLULAR DERMAL MATRIX)

  • 김용덕;안강민;염학렬;정헌종;김성민;장정원;성미애;박희정;황순정;이종호
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제27권6호
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    • pp.510-522
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    • 2005
  • To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

바이오센서 코팅용 Polydimethylsiloxane의 생체외 세포독성 평가 (In vitro Cytotoxicity Evaluation of Polydimethylsiloxane as a Biosensor Coating Material)

  • 박수범;이종환;나경아;정재연;김명진;박성재;현진호
    • 접착 및 계면
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    • 제10권2호
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    • pp.77-83
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    • 2009
  • 생체적용 센서 코팅 재료로서 polydimethylsiloxane (PDMS)를 선정하였으며 합성 및 성형과정에서 용출될 수 있는 잔류 독성 물질의 세포독성을 확인하고자 하였다. ISO 10993-5, Biological evaluation of medical devices-Part 5 : Tests for in vitro cytotoxicity (의료기기의 생물 안정성 평가-제5부: 세포 독성 시험-체외시험)를 통하여 세포 독성 평가를 실시하였다. 양성 대조군으로 organo-tin을 사용하였으며 음성 대조군으로 혈청이 포함되지 않은 RPMI 1640배지를 사용하였다. 고체 시료의 표면적에 대하여 $125{\mu}L/cm^2$가 되도록 혈청이 포함되지 않은 RPMI 1640 배지를 용기에 첨가하였으며 $38^{\circ}C$를 유지하며 일정시간 동안 추출하였다. 세포 독성 평가는 1) NIH 3T3 fibroblast 단일세포층을 형성한 후 추출물을 첨가하는 방법과 2) 세포와 함께 추출물을 넣어 배양하는 방법을 동시에 시행하였다. 세포 형태학적인 변화 관찰과 MTT (tetrazolium dye, 2-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) 시험법에 의한 세포 활성 측정을 병행함으로써 고체 시료로부터 추출된 물질의 세포독성 여부와 고체시료의 표면에 대한 세포의 감응성도 함께 관찰할 수 있었다.

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새로운 노화 방지 성분으로서 글루쿠로닉 애씨드의 기능과 화장품 응용 (Application of Glucuronic Acid with New Cosmetic Active Ingredient)

  • 이근수;김진화;이천일;표형배;이공주
    • 대한화장품학회지
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    • 제30권4호
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    • pp.471-477
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    • 2004
  • 피부 세포는 외부의 유해 요소 즉, 스트레스 환경에 노출되었을 때 세포 자신을 보호하기 위하여 다양한 방어 및 복구 체계를 가지고 있는데 그 중 하나가 보호단백질인 열충격단백질 70 kDa의 발현이다 Glucuronic acid를 피부세포에 다양한 농도로 전처리한 다음 유해자극(열, 활성산소)을 주었을 경우, western blottting을 통해 $0.12\%$ 농도에서 세포 내 열충격단백질이 발현됨을 알 수 있었다 그리고 confocal microscopy 및 세포생존율 실험을 이용하여 열 및 활성 산소에 대한 우수한 세포보호 효과를 확인하였다. 또한 마우스 피부를 이용하여 glucuronic acid 및 수중유(O/W)형 에멀젼에 적용하였을 때 경피 흡수 양상을 비교한 결과, glucuronic acid는 빠른 경피흡수거동을 보였다(투과속도 $0.83114 mg/cm^2/h,$ 지연시간 1.2 h, 분배계수 0.114). 에멀젼에서는 투과속도는 $0.04153{\;}{\mu}g/cm^{2}/h,$ 누적투과량은 $1456.25{\;}{\mu}g/cm^{2}$로 감소하였지만 지연시간은 2.48 h으로 증가하였고 지속적인 경피흡수(서방성)를 보였다.

Cellular Toxic Effects and Action Mechanisms Of 2,2', 4,6,6'-Pentachlorobiphenyl

  • Kim Sun-Hee;Shin Kum-Joo;Kim Dohan;Kim Yun-Hee;Ryu Sung Ho;Suh Pann-Ghill
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2004년도 학술대회지
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    • pp.1-20
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    • 2004
  • Polychlorinated biphenyls (PCBs), one a group of persistent and widespread environmental pollutants, have been considered to be involved in immunotoxicity, carcinogenesis, and apoptosis. However, the toxic effects and physical properties of a PCB congener are dependent on the structure. In the present study, we investigate the toxic effects and action mechanisms of PCBs In cells. Among the various congeners tested, 2,2',4,6,6'-PeCB-pentachlorobiphenyl (PeCB), a highly ortho-substituted congener having negligible binding affinity for aryl hydrocarbon receptor (AhR), caused the most potent toxicity and specific effects in several cell types. 2,2',4,6,6'-PeCB induced apoptotic cell death of human monocytic cells, suggesting that PCB-induced apoptosis may be linked to immunotoxicity. In addition, 2,2',4,6,6'-PeCB induced mitotic arrest by interfering with mitotic spindle assembly in NIH3T3 fibroblasts, followed by genetic instability which triggers p53 activation. Which suggests that 2,2',4,6,6'-PeCB may be involved in cancer development by causing genetic instability through mitotic spindle damage. On the other hand, 2,2',4,6,6'-PeCB increased cyclooxygenase-2 (COX-2) involved in cell survival through ERK1/2 MAPK and p53 in Rat-1 fibroblasts and mouse embryonic fibroblasts, triggering compensatory mechanism for abating its toxicity. Taken together, these results demonstrate that PCB congeners of different structure have distinct mechanism of action and 2,2',4,6,6'-PeCB causes several toxicity as well as compensatory mechanism in cells.

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Photoaging protective effects of BIOGF1K, a compound-K-rich fraction prepared from Panax ginseng

  • Hong, Yo Han;Kim, Donghyun;Nam, Gibaeg;Yoo, Sulgi;Han, Sang Yun;Jeong, Seong-Gu;Kim, Eunji;Jeong, Deok;Yoon, Keejung;Kim, Sunggyu;Park, Junseong;Cho, Jae Youl
    • Journal of Ginseng Research
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    • 제42권1호
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    • pp.81-89
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    • 2018
  • Background: BIOGF1K, a compound-K-rich fraction, has been shown to display anti-inflammatory activity. Although Panax ginseng is widely used for the prevention of photoaging events induced by UVB irradiation, the effect of BIOGF1K on photoaging has not yet been examined. In this study, we investigated the effects of BIOGF1K on UVB-induced photoaging events. Methods: We analyzed the ability of BIOGF1K to prevent UVB-induced apoptosis, enhance matrix metalloproteinase (MMP) expression, upregulate anti-inflammatory activity, reduce sirtuin 1 expression, and melanin production using reverse transcription-polymerase chain reaction, melanin content assay, tyrosinase assay, and flow cytometry. We also evaluated the effects of BIOGF1K on the activator protein-1 signaling pathway, which plays an important role in photoaging, by immunoblot analysis and luciferase reporter gene assays. Results: Treatment of UVB-irradiated NIH3T3 fibroblasts with BIOGF1K prevented UVB-induced cell death, inhibited apoptosis, suppressed morphological changes, reduced melanin secretion, restored the levels of type I procollagen and sirtuin 1, and prevented mRNA upregulation of MMP-1, MMP-2, and cyclo-oxygenase-2; these effects all occurred in a dose-dependent manner. In addition, BIOGF1K markedly reduced activator-protein-1-mediated luciferase activity and decreased the activity of mitogen-activated protein kinases (extracellular response kinase, p38, and C-Jun N-terminal kinase). Conclusion: Our results strongly suggest that BIOGF1K has anti-photoaging activity and that BIOGF1K could be used in anti-aging cosmeceutical preparations.

뽕나무버섯부치(Armillaris tabescens)의 자실체에서 추출한 조다당류의 생쥐 Sarcoma 180에 대한 항암 및 면역증강 효과 (Antitumor and Immuno-modulatory Effect Against Mouse Sarcoma 180 of Crude Polysaccharides Extracted from Fruiting Body of Armillaria tabescens)

  • 이건우;김혜영;이우윤;이태수
    • 한국균학회지
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    • 제35권2호
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    • pp.101-108
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    • 2007
  • 뽕나무버섯부치는 주름버섯목, 송이버섯과에 속하는 버섯으로 예로부터 만성간염, 담낭염 및 암의 치료에 널리 이용해온 맛이 좋은 식의약용 버섯이다. 뽕나무버섯부치의 자실체로부터 중성염용액, 열수 및 메탄올을 이용하여 조다당류를 추출하여 Sarcoma 180가 접종된 ICR mice에 주사하여 항암 및 면역증강 효과를 조사하였다. 세포독성 실험결과, 각각의 세포는 $10{\sim}2000\;{\mu}g/ml$ 추출물 농도에서 세포독성을 나타내지 않았다. 각각의 조다당류가 투여된 실험군은 대조군에 비해 수명이 각각 $28.8{\sim}46.5%$ 연장되었다. 중성염용액으로 추출한 조다당류는 B 임파구의 alkaline phosphatase 활성을 대조군에 비해 약 $1.8{\sim}2.1$배 내외의 증가율을 나타냈다. 중성염추출 조다당류를 투여한 생쥐의 총 복강 세포 수는 대조군에 비하여 최고 9배 정도 증가하였으며, 혈액 중 백혈구의 수도 대조군에 비하여 약 1.9배 증가하였다. 또한 면역에 관련된 장기인 간, 비장 및 흥선의 체중이 대조군에 비하여 증가된 것을 확인하였다.

Characterization of TNNC1 as a Novel Tumor Suppressor of Lung Adenocarcinoma

  • Kim, Suyeon;Kim, Jaewon;Jung, Yeonjoo;Jun, Yukyung;Jung, Yeonhwa;Lee, Hee-Young;Keum, Juhee;Park, Byung Jo;Lee, Jinseon;Kim, Jhingook;Lee, Sanghyuk;Kim, Jaesang
    • Molecules and Cells
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    • 제43권7호
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    • pp.619-631
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    • 2020
  • In this study, we describe a novel function of TNNC1 (Troponin C1, Slow Skeletal and Cardiac Type), a component of actin-bound troponin, as a tumor suppressor of lung adenocarcinoma (LUAD). First, the expression of TNNC1 was strongly down-regulated in cancer tissues compared to matched normal lung tissues, and down-regulation of TNNC1 was shown to be strongly correlated with increased mortality among LUAD patients. Interestingly, TNNC1 expression was enhanced by suppression of KRAS, and ectopic expression of TNNC1 in turn inhibited KRASG12D-mediated anchorage independent growth of NIH3T3 cells. Consistently, activation of KRAS pathway in LUAD patients was shown to be strongly correlated with down-regulation of TNNC1. In addition, ectopic expression of TNNC1 inhibited colony formation of multiple LUAD cell lines and induced DNA damage, cell cycle arrest and ultimately apoptosis. We further examined potential correlations between expression levels of TNNC1 and various clinical parameters and found that low-level expression is significantly associated with invasiveness of the tumor. Indeed, RNA interference-mediated down-regulation of TNNC1 led to significant enhancement of invasiveness in vitro. Collectively, our data indicate that TNNC1 has a novel function as a tumor suppressor and is targeted for down-regulation by KRAS pathway during the carcinogenesis of LUAD.