• Title/Summary/Keyword: Adhesion assay

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Evaluation of Physical Properties and Biocompatibility of HA-Dex Fusion Hydrogel Patch for Atopic Healing Ability (HA-Dex 융복합 하이드로겔 패치의 아토피 치유 능력에 대한 물리적 특성 및 생체 적합성 평가)

  • Hong, Gyeong Sik;Choi, Jeong Yeon;Choi, Jin Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.3
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    • pp.219-229
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    • 2020
  • Recently, since atopic dermatitis is sensitive to skin irritation, it has been suggested that the development of a patch that can effectively exhibit adhesion and absorption to a specific local area while minimizing skin irritation, and capable of appropriate drug release should be given priority. In this study, we tried to develop a hydrogel patch that minimizes skin irritation, adheres effectively to a specific area, and promotes absorption. The atopic patch was formulated into a super-absorbent hydrogel sheet using a freeze drying method. Cell viability assay was carried out using keratinocytes (HaCaT cell) and fibroblasts (L929 cells). In order to investigate the physical properties, FT-IR, FE-SEM, porosity analysis and swelling behavior were investigated. As a result, the newly prepared HA-Dex hydrogel patch was verified by biocompatibility and physical evaluation. In addition, the manufactured hydrogel patch has sufficient moisture absorption capacity and can relieve itching of atopic skin, and is expected to be applied to various drug delivery products for the treatment of atopic dermatitis in the future.

Interaction between odontoblast and bio-calcium phosphate cement reinforced with chitosan (치아모세포와 키토산으로 강화된 생체 적합 칼슘인산시멘트와의 상호작용)

  • Chun, Byung-Do;Kim, Sung-Won;Lee, Sung-Tak;Kim, Tae-Hoon;Lee, Jung-Han;Kim, Gyoo-Cheon;Kim, Yong-Deok;Kim, Uk-Kyu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.5
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    • pp.415-420
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    • 2011
  • Purpose: Calcium phosphate cement (CPC) is one of many useful materials for restoring tooth defects, periodontium and maxillofacial area. Chitosan is a biodegradable material that has been shown to promote the growth and differentiation of osteoblasts in culture. This study examined the interaction between odontoblasts and bio-calcium phosphate cement reinforced with chitosan. Materials and Methods: $5{\times}10^3$ odontoblastic cells were seeded into each well. Various concentrations of bio-calcium phosphate cement reinforced with chitosan (10, 20, 50, 100, 200, 500 ${\mu}g$/ml, 1, 2, 4 mg/ml) were diluted and added to the wells. The well was incubated for 24 h, 48 h and 72 h. After incubation, the number of cells was assessed to determine the cell viability. A cytokinesis-block micronucleus assay and chromosomal aberration test were carried out to estimate the extent of chromosomal abnormalities. Microscopic photographs and RT-PCR were performed to examine the adhesion potential of bio-calcium phosphate cement reinforced with chitosan. Results: Bio-CPC-reinforced chitosan did not show significant cytotoxicity. The number of damaged chromosomes in the cells treated with Bio-CPC-reinforced chitosan was similar to that in the control cells. There was no significant increase in the number of chromosomal aberrations in the Bio-CPC reinforced chitosan exposed cells. Microscopic photographs and RT-PCR confirmed the adhesive potential of bio-CPC reinforced chitosan to odontoblasts. Conclusion: Bio-CPC-reinforced chitosan did not affect the odontoblastic cell viability, and had no significant cytotoxic effect. Bio-CPC-reinforced chitosan showed adhesive potential to odontoblasts. These results are expected form the basis of future studies on the effectiveness of dental restorative materials in Bio-CPC reinforced with chitosan.

Preparation and Characterization of Silk Fibroin/Gelatin Hybrid Scaffolds (실크 피브로인/젤라틴 하이브리드 지지체의 제조 및 특성분석)

  • Kim, Hye-Lin;Hong, Min-Sung;Kim, Su-Jin;Jo, Han-Su;Yoo, Il-Sou;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.5
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    • pp.378-384
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    • 2011
  • Silk fibroin is a biocompatible and slowly biodegradable natural polymer. This natural polymer has excellent mechanical properties, non-toxicity, and non-immunogenic properties and has been demonstrated to support tissue regeneration. Also, gelatin is a natural material derived from collagen by hydrolysis and has an almost identical composition as that of collagen. Silk fibroin/gelatin scaffolds have been fabricated by using the freeze-drying method. To establish the scaffold manufacturing condition for silk fibroin and gelatin, we made scaffolds with various compositions of gelatin, glutaldehyde and silk fibroin. The silk fibroin/gelatin scaffolds were characterized using SEM, DSC, and water absorption ability tests. The cellular proliferation was evaluated by WST assay. These results suggested that a scaffold containing 8% of gelatin, 1% of glutaldehyde and 0.3 g of silk fibroin provided suitable characterstics for cell adhesion and proliferation. In conclusion, the silk fibroin/gelatin scaffold may serve as a potential cell delivery vehicle and a structural basis for tissue engineering.

Fibroblastic Reticular Cell Derived from Lymph Node Is Involved in the Assistance of Antigen Process (림프절 유래 fibroblastic reticular cell의 효율적 항원처리 관련성에 대한 연구)

  • Kim, Min Hwan;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1027-1032
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    • 2016
  • Antigen is substance causing disease derived from pathogen. Living organism has the immune system in terms of defense mechanism against antigen. Antigen is processed through several pathways such as phagocytosis, antibody action, complement activation, and cytotoxins by NK or cytotoxic T lymphocyte via MHC molecule. Lymph node (LN) is comprised of the complicated 3 dimensional network and several stromal cells. Fibroblastic reticular cells (FRC) are distributed in T zone for interaction with T cells. FRC produces the extra cellular matrix (ECM) into LN for ECM reorganization against pathogen infections and secretes homing chemokines. However, it has not so much been known about the involvement of the antigen process of FRC. The present report is for the function of FRC on antigen process. For this, FRC was positioned with several infected situations such as co-culture with macrophage, T cell, lipopolysaccharide (LPS) and TNFα stimulation. When co-culture between FRC with macrophage and T cells was performed, morphological change of FRC was observed and empty space between FRCs was made by morphological change. The matrix metallo-proteinase (MMP) activity was up-regulated by Y27632 and T cells onto FRC. Furthermore, inflammatory cytokine, TNFα regulated the expression of adhesion molecules and MHC I antigen transporter in FRC by gene chip assay. NO production was elevated by FRC monolayer co-cultured with macrophage stimulated by LPS. GFP antigen was up-taken by macrophage co-cultured with FRC. Collectively, it suggests that FRC assists of the facilitation of antigen process and LN stroma is implicated into antigen process pathway.

Behavior of Retinal Pigment Epithelial Cells Cultured on Silk Films (실크필름에 배양한 망막색소상피세포의 거동)

  • Lee, So Jin;Kim, Hye Yun;Kim, Seul Ji;Yang, Jaewon;Lee, Seon Ui;Park, Chan Hum;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.3
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    • pp.364-370
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    • 2014
  • The retinal pigment epithelium (RPE) plays an important role in maintaining a healthy retina and the degeneration of RPE caused a number of retinal diseases. The transplantation of RPE has recently become a possible therapeutic modality for retinal degeneration. To transplant RPE cells securely, substrates are essential, and then as a substrate, we fabricated films using silk that has unique mechanical properties and biocompatibility. After the FTIR spectra, contact angle and biodegradation of silk films were confirmed, RPE cells were seeded and the influence of RPE cells on silk films was examined. We measured the cell adhesion, cell viability, morphology and specific mRNA expression by MTT assay, SEM, immunofluorescence and RT-PCR. In this study, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells cultured on silk films were great, and thereby we were able to confirm the potential applications of silk films as tissue engineering carrier for regeneration of retina.

Effect of Extracellular Matrix on the Growth Behavior of Corneal Endothelial Cells to Poly(lactic-co-glycolic acid) Film (각막 내피세포 성장 거동에 대한 락타이드 글리콜라이드 공중합체 필름과 세포외 기질의 효과)

  • Kim, Eun Young;Kim, Hye Min;Song, Jeong Eun;Lee, Hyun Soo;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.702-707
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    • 2014
  • Corneal endothelium is mono-inner cell layer of cornea and lay on Descmet's membrane which comprised of various proteins called extracellular matrix such as fibronectin, collagen, laminin, and proteoglycan, etc. In this study, we fabricated transparent poly(lactic-co-glycolic acid) (PLGA) film because PLGA is widely used for tissue engineering based on their properties. We investigated the behaviors of rabbit corneal endothelial cells (rCEnCs) on PLGA film surfaces coated with various cell-adhesive molecules like fibronectin, laminin, collagen type I and IV and FNC coating mix. The morphologic images, proliferation and adhesion assay, immunofluorescence for ZO-1 and $Na^+/K^+-ATPase$ and RT-PCR for expression of specific markers were conducted. These results showed that PLGA film plays a role as CEnC carriers in vitro and the cell-adhesive molecules give positive effects on the behaviors of rCEnC.

Antifungal Activity of Rheum undulatum on Candida albicans by the Changes in Membrane Permeability (막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성)

  • Lee, Heung-Shick;Kim, Younhee
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.360-367
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    • 2014
  • Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

Inhibitory Effect of Shiitake Mushroom Extracts Cultured in Aloe-Supplement on Invasion/Migration of TNF-α-Induced MDA-MB-231 Breast Cancer Cells (알로에 배지에서 재배한 표고버섯 추출물의 종양괴사인자로 유도된 유방암세포 MDA-MB231에 대한 전이 저해 활성)

  • Suh, Seok-Jong;Kim, Cheorl-Ho;Baek, Jin-Hong;Lee, Kyoung-Hae;Shin, Dong-Bum;Park, Sung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.1
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    • pp.44-48
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    • 2015
  • To investigate the anti-cancer effect of shiitake mushrooms (Lentinus edodes) cultured in aloe-supplement, we treated extract of shiitake mushroom cultured in aloe-supplement (ESA) to MDA-MB-231 human breast cancer cells. ESA-treated MDA-MB-231 cells showed decreased growth rate in XTT assay. In addition migration/invasion was noticeably inhibited by ESA in TNF-${\alpha}$-treated MDA-MB-231 cells. Western blot analysis showed that the molecular mechanism of cell migration/invasion was mediated by reduced intercellular adhesion molecule-1 expression via p-ERK signal transduction pathways. We found ESA had inhibition activity against cellular growth and migration/invasion. Taken together, ESA has putative anti-cancer activity against human breast cancer.

Embryotoxicity and Teratogenicity of Excess Zinc on Xenopus laevis (과량의 아연에 의한 아프리카 발톱개구리 (Xenopus laevis)의 배발생 이상과 독성)

  • Yoon, Chun-Sik;Jin, Jung-Hyo;Cheong, Seon-Woo
    • Korean Journal of Ecology and Environment
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    • v.36 no.1 s.102
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    • pp.83-94
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    • 2003
  • Concentrated releases of zinc into water usually results from discharges associated with industrial purpose. The released zinc into soil is corroded and released into water. In aquatic environment, exess zinc is toxic to the organisms and causes the growth inhibition and malformation of them as a heavy metal. In this study, excess zinc toxicity was tested by FETAX (frog embryo teratogenetic assay with Xenopus)as in vivo system. Xenopus embryos at st.9 were exposed to $100{\sim}900\;{\mu}M$ of zinc for 7 days and 81% of individuals were survived in 100 ${\mu}M$, and 25% were survived in 1000M of zinc solution. In external malformations, swelled belly and intestinal dysplasia were common, and all of tested individuals showed these malformations in 200 ${\mu}M$ or higher concentration of zinc. In 400 ${\mu}M$ or higher concentration, all of tested tadpoles showed faded heart. Also, hypo-pigmentation, lens hernia and loose digestive track were very frequently found in 100 ${\mu}M$ of zinc. The histological study with paraffin section of zinc treated tadpoles showed following abnormalities; regeneration of photoreceptor on retina, reduced vitreous chamber in eye, reduction of red blood cells in heart, abnormal liver, swelling of pronephric cell, muscle dysplasia and palatal papilloma. These abnormalities may be caused by the degeneration of mitochondria, inhibition of cell adhesion, and the formation of leghemoglobin by zinc due to the substitution of $Ca^{2+}$ by $Zn^{2+}$. The body length was reduced due to the excess zinc. From a statistical result, body lengths of 300 ${\mu}M$ or higher concentrative g개ups was significantly reduced comparing that of control group. Recently, many spontaneous malformations and reduction of amphibians are reported, From the results of present study, excess zinc mi호t be a factor of amphibian reduction, and the control of zinc discharges is very important.

Cellular activities of osteoblast-like cells on alkali-treated titanium surface (알칼리 처리된 타이타늄 표면에 대한 골아 유사세포의 세포 활성도)

  • Park, Jin-Woo;Lee, Deog-Hye;Yeo, Shin-Il;Park, Kwang-Bum;Choi, Seok-Kyu;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.427-445
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    • 2007
  • To improve osseointegration at the boneto-implant interface, several studies have been carried out to modify titanium surface. Variations in surface texture or microtopography may affect the cellular response to an implant. Osteoblast-like cells attach more readily to a rougher titanium surface, and synthesis of extracellular matrix and subsequent mineralization were found to be enhanced on rough or porous coated titanium. However, regarding the effect of roughened surface by physical and mechanical methods, most studies carried out on the reactions of cells to micrometric topography, little work has been performed on the reaction of cells to nanotopography. The purpose of this study was to examme the response of osteoblast-like cell cultured on blasted surfaces and alkali treated surfaces, and to evaluate the influence of surface texture or submicro-scaled surface topography on the cell attachment, cell proliferation and the gene expression of osteoblastic phenotype using ROS 17/2.8 cell lines. In scanning electron micrographs, the blasted, alkali treated and machined surfaces demonstrated microscopic differences in the surface topography. The specimens of alkali treatment had a submicro-scaled porous sur-face with pore size about 200 nm. The blasted surfaces showed irregularities in morphology with small(<10 ${\mu}m$) depression and indentation among flatter-appearing areas of various sizes. Based on profilometry, the blasted surfaces was significantly rougher than the machined and the alkali treated surfaces (p$TiO_2$) were observed on alkali treated surfaces, whereas not observed on machined and blasted surfaces. The attachment morphology of cells according to time was observed by the scanning electron microscope. After 1 hour incubation, the cells were in the process of adhesion and spreading on the prepared surfaces. After 3 hours, the cells on all prepared surfaces were further spreaded and flattened, however on the blasted and alkali treated surfaces, the cells exhibited slightly irregular shapes and some gaps or spaces were seen. After 24 hours incubation, most cells of the all groups had a flattened and polygonal shape, but the cells were more spreaded on the machined surfaces than the blasted and alkali treated surfaces. The MTT assay indicated the increase on machined, alkali treated and blasted surfaces according to time, and the alkali treated and blasted surfaces showed significantly increased in optical density comparing with machined surfaces at 1 day (p<0.01). Gene expression study showed that mRNA expression level of ${\alpha}\;1(I)$ collagen, alkaline phosphatase and osteopontin of the osteoblast-like cells showed a tendency to be higher on blasted and alkali treated surfaces than on the machined surfaces, although no siginificant difference in the mRNA expression level of ${\alpha}\;1(I)$ collagen, alkaline phosphatase and osteopontin was observed among all groups. In conclusion, we suggest that submicroscaled surfaces on osteoblast-like cell response do not over-ride the one of the surface with micro-scaled topography produced by blasting method, although the microscaled and submicro-scaled surfaces can accelerate osteogenic cell attachment and function compared with the machined surfaces.