• Title/Summary/Keyword: Human Fibroblast

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In vitro cytotoxicity of self-etching primers (자가 산부식 프라이머의 세포독성에 관한 실험적 연구)

  • Rhee, Chang-Hoon;Kim, In-Ryeon;Kim, Gyoo-Cheon;Kim, Seong-Sik;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.36 no.6
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    • pp.422-433
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    • 2006
  • Objective: Several ions and components are released from self-etching primers in the oral cavity. This may cause injury to the periodontal tissues throughout orthodontic treatment. The purpose of this study was to assess the cytotoxicity of self-etching primers to HGF-1, HaCaT, and RHEK cells. Method: Transbond XT Primer (3M Unitek, Monrovia, CA, USA), and self-etching primers, Clearfil SE Bond (Kuraray, Osaka, Japan), Transbond Plus SEP (3M Unitek, Monrovia, CA, USA), and Adper Prompt L-Pop (3M Unitek, Monrovia, CA, USA), were evaluated by MTT assay, and cellular changes were also observed. Results: In all cells after 72 hours with all primers, severe morphological changes such as atrophy and necrosis were observed. In the MTT assay using HGF-1, Clearfil SE Bond, Transbond XT Primer, Transbond Plus SEP, and Adper Prompt L-Pop were lined up in order of ascending cytotoxicity When using HaCaT, Clearfil SE Bond, Adper Prompt L-Pop, Transbond Plus SEP, and Transbond XT Primer were lined up in order of ascending cytotoxicity. When using RHEK, Clearfil SE Bond, Transbond XT Primer, Adper Prompt L-Pop, and Transbond Plus SEP were lined up in order of ascending cytotoxicity. Conclusion: The result of this study shows that care is needed because self-etching primers show cytotoxic properties similar to conventional primers.

Photoprotection and Anti-inflammatory Effects of Chinese Medical Plants (약용식물추출물의 광보호 효과와 항염증 효과 연구)

  • Jin-Hwa, Kim;Sung-Min, Park;Gwan-Sub, Sim;Bum-Chun , Lee;Hyeong-Bae, Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.227-233
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    • 2004
  • Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

The Effect of Photomodulation in Human Dermal Fibroblasts (피부 섬유아세포에서 광자극의 효과)

  • Kim, Mi Na;Kwak, Taek Jong;Kang, Nae Gyu;Lee, Sang Hwa;Park, Sun Gyoo;Lee, Cheon Koo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.325-331
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    • 2015
  • Skin is exposed to sunlight or artificial indoor light on a daily. The reached solar light on the earth surface consist of 50% visible light and 45% infrared (IR) except for ultra violet (UV). The negative effects of UV including UVB and UVA have been steadily investigated within the last decades. However, little is known about the effects of visible or IR light. In this study, we irradiated human dermal fibroblasts using light emitting diode (LED) to investigate the optimal parameter for enhancing cell growth and collagen synthesis. We found that red of 630 nm and green of 520 nm enhance the cell proliferation, but irradiation with purple and blue light exerts toxic effects. To examine the response of irradiation time and light intensity on the fibroblasts, cells were exposed to red or green light with intensities from 0.05 to $0.75mW/cm^2$. Procollagen secretion was increased of 1.4 fold by 10 min irradiation, while 30 min treatment decreased the collagen synthesis of dermal fibroblasts. Treatment with red of $0.3mW/cm^2$ and green of 0.15 and $0.3mW/cm^2$ resulted in enhancement of collagen mRNA. Lastly, we investigated the combinatorial effect of red and green light on dermal fibroblasts. The sequential irradiation of red and green light is an efficient way for the purpose of the increase in the number of fibroblasts than single light treatment. On the other hand, the exposure of red light alone was more effective method for enhancing of collagen secretion. Our study showed that specific light parameters accelerated cell proliferation, gene expression and collagen secretion on human dermal fibroblasts. In conclusion, we demonstrate that light exposure with specific parameter has beneficial effects on the function of dermal fibroblasts, and suggests the possibility of its cosmetically and clinical application.

Antimicrobial Effect on the Periodontal Pathogens and Anti-inflammatory Effect of Artemisiae Iwayomogii Herba (한인진(韓茵蔯)의 치주염세균에 대한 항균효과 및 항염효가)

  • Kim, Young-Hong;Jeong, Mi-Young;Lee, Na-Kyung;Lee, Jin-Yong;Herr, Yeek;Lee, Je-Hyun;Lim, Sabina
    • The Korea Journal of Herbology
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    • v.23 no.2
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    • pp.1-8
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    • 2008
  • Objectives : The purpose of this study was to evaluate on the antimicrobial effect on the periodontal pathogens and anti-inflammatory effect of Artemisiae Iwayomogii Herba. Artemisiae Iwayomogii Herba has been used for treating as Artemisiae Capilaris Herba in Korea. Methods : Artemisiae Iwayomogii Herba was prepared by extracting medicinal herb with water. We investigated antimicrobial activity by the minimun inhibitory concentration (MIC) test. We also investigated inhibition of IL-$1{\beta}$-induced collagenase-l(MMP-l), stromelysin-1(MMP-3), interleukin-6 gene expression in human gingival fibroblasts. Results : The antimicrobial effect of Artemisiae Iwayomogii Herba was evaluated with MIC against periodontopathogens; Porphyromonas gingivalis 2561, W50, A7A1-28, 9-14K-1, Prevotella intermedia28, and Actinobacillus actinomycetemcomitans Y4, MICs of Artemisiae Iwayomogii Herba were 0.156 mg/ml, 0.625 mg/ml, 0.313 mg/ml, 1.25 mg/ml, 10 mg/ml and 10 mg/ml. The anti-inflammatory effect of Artemisiae Iwayomogii Herba was evaluated with Influence of herbs on the IL-$1{\beta}$-induced expression of MMP-1, MMP-3, interleukin-6, IL-$1{\beta}$ increased MMP-1, MMP-3, interleukin-6 mRNA levels. Artemisiae Iwayomogii Herba significantly inhibited IL-$1{\beta}$-induced MMP-1, MMP-3, interleukin-6 gene expressions in a dose-dependent manner. Conclusions : These results suggested that Artemisiae Iwayomogii Herba might reduce the excessive proteolytic capacity of the gingival fibroblast during inflammation and could be developed a new drug in periodontitis.

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IL-23 P19 Expression Induced by IL-17 and $IL-1{\beta}$ in Rheumatoid Arthritis Synovial Mononuclear Cells (류마티스관절염 환자의 활액 세포에서 IL-17과 $IL-1{\beta}$에 의한 IL-23p19의 발현 증가)

  • Cho, Mi-La;Heo, Yu-Jung;Oh, Hye-Jwa;Kang, Chang-Min;Lee, Seon-Yeong;Hong, Yeon-Sik;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.29-37
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    • 2008
  • Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.

BONE REGENERATION WITH ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELL AND HA/TCP (HA/TCP 골이식재상에 이식된 지방유래 줄기세포의 골모세포로의 분화 및 골형성에 대한 연구)

  • Rim, Jae-Suk;Gwon, Jong-Jin;Jang, Hyon-Seok;Lee, Eui-Seok;Jeong, You-Min;Lee, Tai-Hyung;Park, Jeong-Kyun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.2
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    • pp.97-106
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    • 2010
  • Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

Physiological Activities of Cudrania tricuspidata Extracts on the Skin (꾸지뽕나무 추출물의 피부 생리 활성)

  • Choi, Hak Joo;Kim, Cheong Taek;Do, Min Yeon;Rang, Moon Jeong
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.2
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    • pp.260-274
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    • 2015
  • This paper has shown the experimental results about the physiological activities of water-, ethanol-, ethyl acetate-soluble fractions from ethanolic extracts of leaves, stems and roots of Cudrania tricuspidata on the skin, which has been used for a long time as a traditional herb medicine in Korea and China. The effects of these fractions on the secretion of nitric oxide and cytokines from macrophage(RAW 264.7 cell) exhibited that the ethyl acetate and water fractions from leaves inhibited the release of nitric oxide, all fractions inhibited thoses of inflammatory cytokine $IL-1{\alpha}$, and the ethyl acetate fractions of leaves, stems and roots inhibited thoses of inflammatory cytokine IL-6. The only ethylacetate fraction of leaves demonstrated significantly the reduction of melanin synthesis in melanoma cells. In order to evaluate the efficacy of collagen synthesis, the treatment with extracts on the human normal fibroblast cell(CCD-986sk cell) resulted in finding that the water fractions of leaves, stems and roots and the ethanol fractions of leaves and stems showed the increased synthesis of collagen.

Anti-wrinkle Effect of Mycelial Culture Broth of Paecilomyces japonica in the Mixture of Cucumber and Grape Extracts (포도와 오이즙액을 이용한 동충하초(Paecilomyces japonica) 균사체 배양액의 피부 주름개선효과)

  • Lee Young-Hoon;Choi U-Sic;Park Ki-Hoon;Choi Young-Ju;Gal Sang-Wan
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.516-521
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    • 2006
  • The possibility of usage as cosmetic resource of the mycelial culture broth of P. japonica in the mixture of cucumber and grape extracts was investigated. In the effect of collagen synthesis promotion in human fibroblast cells, the culture broth of P. japonica of 0.1, 0.5 and 1.0% concentration increased the amount of collagen synthesis than that of control cells. The culture broth increased the SOD activity in the concentration dependant manner of 0.01% to 1.0% in the antioxydation activity test. About 90% of superoxide radical was eliminated by 0.5% concentration of the culture supernatant in the antioxydation test. Anticoagulant quercetin in the course of mycelial growth in the mixture of cucumber and grape extract was accumulated to 15 folds than that of pre-culture. In the skin safety test of the culture broth, there is no any skin damage signal in the tested 30 people. Taken together, we concluded that the culture broth of P. japonica in the mixture of grape and cucumber extracts can be used as a cosmetic resource.

In vitro cytotoxicity of four kinds orthodontic band cements (수종 치과 교정용 밴드 시멘트의 세포독성에 관한 실험적 연구)

  • Lee, Won-Chul;Park, Soo-Byung
    • The korean journal of orthodontics
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    • v.34 no.4 s.105
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    • pp.351-362
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    • 2004
  • Orthodontic band cements are widely used in the fields of orthodontics, but they are commonly known as cytotoxic material. Within an oral cavity several ions and components are released from orthodontic band cements, thus causing inflammation or injury to the Periodontal tissue. Therefore, it is very important to estimate the biocompatibility of orthodontic band cements. The purpose of this study was to assess the cytotoxic effect of orthodontic band cements to HGF cells. A zinc phosphate cement, a glass ionomer, a resin modified glass ionomer, and compomer were used to evaluate three cytotoxicity assays: cell proliferation assay, MTT assay, and agar ovelay assay The results were as follows: 1. In the cell proliferation assay, Gl>ZPC, RMGI, RMGI24, GI24>compomer24, ZPC24, compomer>metal ring lined up in order of cytotoxicity 2. In the MTT assay, GI>ZPC, RMGI>GI24>ZPC24, compomer, metal ring, RMGI24, compomer24 lined up in order of cytotoxicity. 3. In the agar overlay test, GI>GI24, ZPC, ZPC24, RMGI>RMGI24, compomer, compomer24, metal ring lined up in order of cytotoxicity.

Efficient Production of Cloned Bovine Embryos from Transformed Somatic Cells (형질전환 체세포로부터 소 복제수정란의 효율적인 생산)

  • Wee G.;B. H Sohn;Park, J. S.;D. B. Koo;Lee, K. K.;Y. M. Han
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.25-34
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    • 2003
  • Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis. To direct hTPO expression in the mammary gland, an expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycin resistance gene (pBT-L neo). Fibroblast cells derived from cow's ear skin tissue were transfected with the expression vector (pBT-L neo) using Lipofectamine. Transfected cells resistant to G418 trea?nt were cultured to form the colonies for more than 2 weeks. The transformed colonies identified by PCR were further expanded prior to nuclear transfer. Reconstructed oocytes with transformed cells were electrofused, activated using calcium ionophore and 6-DMAP, and cultured in vitro for 7 days. Of 35 cell colonies analyzed by PCR, 29 colonies (82.9%) were positive for the hTPO gene. Cleavage and developmental rates to the blastocyst stage of reconstructed embryos with the transformed cells were 65.1% and 23.8%, respectively Of 29 blastocysts that developed from reconstructed embryos with the transformed cells, 27 embryos (93.1%) were transgenic. These results indicate that transgenic bovine embryos can be efficiently produced by somatic cell nuclear transfer using transformed cells.