• Title/Summary/Keyword: skin vivo and vitro

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Recent Progress on Skin-Derived Mesenchymal Stem Cells in Pigs

  • Kumar, B. Mohana;Patil, Rajreddy;Lee, Sung-Lim;Rho, Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.283-290
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    • 2012
  • Skin serves as an easily accessible source of multipotent stem cells with potential for cellular therapies. In pigs, stem cells from skin tissues of fetal and adult origins have been demonstrated as either floating spheres (cell aggregates) or adherent spindle-shaped mesenchymal stem cell (MSC)-like cells depending on culture conditions. The cells isolated from the epidermis and dermis of porcine skin showed plastic adherent growth in the presence of serum and positively expressed a range of surface and intracellular markers that are considered to be specific for MSCs. The properties of primitive stem cells have been observed with the expression of alkaline phosphatase and markers related to pluripotency. Further, studies have shown the ability of skin-derived MSCs to differentiate in vitro along mesodermal, neuronal and germ-line lineages. Moreover, preclinical studies have also been performed to assess their in vivo potential, and the findings appear to be effective in tissue regeneration at the defected site after transplantation. The present review describes the recent progress on the biological features of porcine skin-derived MSCs as adherent cells, and summarizes their potential in advancing stem cell based therapies.

Immunotherapeutic Effects of CTLA4Ig Fusion Protein on Murine EAE and GVHD (마우스 EAE, GVHD 질환에서 CTLA4Ig 융합단백의 면역치료 효과)

  • Jang, Seong-Ok;Hong, Soo-Jong;Cho, Hoon-Sik;Chung, Yong-Hoon
    • IMMUNE NETWORK
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    • v.3 no.4
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    • pp.302-309
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    • 2003
  • Background: CTLA4 (CD152), which is expressed on the surface of T cells following activation, has a much higher affinity for B7 molecules comparing to CD28, and is a negative regulator of T cell activation. In contrast to stimulating and agonistic capabilities of monoclonal antibodies specific to CTLA-4, CTLA4Ig fusion protein appears to act as CD28 antagonist and inhibits in vitro and in vivo T cell priming in variety of immunological conditions. We've set out to confirm whether inhibition of the CD28-B7 costimulatory response using a soluble form of human CTLA4Ig fusion protein would lead to persistent inhibition of alloreactive T cell activation. Methods: We have used CHO-$dhfr^-$ cell-line to produce CTLA4Ig fusion protein. After serum free culture of transfected cell line we purified this recombinant molecule by using protein A column. To confirm characterization of fusion protein, we carried out a series of Western blot, SDS-PAGE and silver staining analyses. We have also investigated the efficacy of CTLA4Ig in vitro such as mixed lymphocyte reaction (MLR) & cytotoxic T lymphocyte (CTL) response and in vivo such as experimental autoimmune encephalomyelitis (EAE), graft versus host disease (GVHD) and skin-graft whether this fusion protein could inhibit alloreactive T cell activation and lead to immunosuppression of activated T cell. Results: In vitro assay, CTLA4Ig fusion protein inhibited immune response in T cell-specific manner: 1) Human CTLA4Ig inhibited allogeneic stimulation in murine MLR; 2) CTLA4Ig prevented the specific killing activity of CTL. In vivo assay, human CTLA4Ig revealed the capacities to induce alloantigen-specific hyporesponsiveness in mouse model: 1) GVHD was efficiently blocked by dose-dependent manner; 2) Clinical score of EAE was significantly decreased compared to nomal control; 3) The time of skin-graft rejection was not different between CTLA4Ig treated and control group. Conclusion: Human CTLA4Ig suppress the T cell-mediated immune response and efficiently inhibit the EAE, GVHD in mouse model. The mechanism of T cell suppression by human CTLA4Ig fusion protein may be originated from the suppression of activity of cytotoxic T cell. Human CTLA4Ig could not suppress the rejection in mouse skin-graft, this finding suggests that other mechanism except the suppression of cytotoxic T cell may exist on the suppression of graft rejection.

Effects of Amomi, Semen Extract on Synthesis of Insulin-like Growth Factor-1 and Anti-wrinkle in Skin (사인추출물의 인슐린 유사 성장인자-3의 합성과 피부 노화 개선에 대한 효과)

  • Choi Gyu Ho;Kim Su Nam;Lee So Hee;Sung Dae Seok;Son Eui Dong;Lee Chang Hoon;Lee Byeong Gon;Jang Ih Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.389-392
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    • 2004
  • We screened several materials to stimulate IGF-1 promoter activity using luciferase reporter assay and found that Amomi Semen extract (ASE) among them is the most powerful stimulator We also studied about the anti-wrinkle effect of ethanolic extract of Amoni Semen in vitro and in vivo. Semi-quantitative RT-PCR showed that the extract elevated the presence level of IGF-1 mRNA. And $[^3H]$ proline incorporation and semi-quantitative RT-PCR showed that the extract increased the expression of type-I collagen compared with vehicle in vitro and in vivo, respectively. Significant inhibition of MMP-1 expression was determined by ELISA and Western blot. Finally, topical treatment of the extract on hairless mouse's dorsal skin expanded the volume of collagen and dermal thickness. These results suggest that Amomi Semen may be a good candidate for improving extracellular matrix through the increase of collagen expression and inhibition of MMP-1 expression. Moreover, this study enables us to guess that IGF-1 stimulated by the extract may be involved in the mechanism of anti-wrinkle effect of it.

Skin Permeation Enhancement of Drugs by Lipophilic and Hydrophilic Vehicles

  • Lee, Cheon-Koo;Goto, Shigeru
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.43-51
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    • 1995
  • The in vitro skin permeability of 16 drugs with a wide span of lipophilicity (log P ranging from -0,95 to 4.40) was evaluated with an ethanol/panasate 800 (tricaprylin, P-800) (40/60) lipophilic binary vehicle and an ethanol/water (60/40) hydrophilic binary vehicle with lauric acid, The skin permeability of the drugs was enhanced by the use of the ethanol/P-800 (40/60) binary vehicle or the ethanol/water (60/40) binary vehicle with lauric acid; permeation rate was increased and lag time' was decreased. The relationship between lipophilicity and skin permeation rate of the drugs showed parabolic shapes with their peaks at much greater hydrophilic range compared with other past references. In the in vivo skin absorption of theophylline using abdominal rat skin, the ethanol/P-800 (40/60)-7% (w/w) ethycellulose gel produced a good feature as a sustained-release preparation, and the ethanol/water (60/40)-3 % (w/w) HPMC gel with lauric acid showed the highest BA value. The results suggest that the lipophilicity of a drug is a main factor for prediction of the skin permeability of the drug and that the ethanol/P-800 (40/60) binary vehicle and ethanol/water (60/40) binary vehicle with lauric acid would be good candidates for clinical transdermal application of hydrophilic drugs.

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Evaluating the Micronucleus Induction Potential for the Genotoxicity Assay Using the Human Skin Model, KeraSkinTM (인공피부모델 KeraSkinTM을 이용한 유전독성 평가)

  • Lee, Su-Hyon;Jung, Haeng-Sun;Kim, Seol-Yeong;Kim, Hye Soo;Lim, Kyung-Min;Chung, Young-Shin;Choe, Tae-Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.211-216
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    • 2016
  • Micronucleus test is genotoxicity assay for detection of micronuclei in the cytoplasm of interphase cells. The reduction and replacement of in vivo toxicity testing on animals require the development of in vitro models to predict the genotoxicity or other tests for cosmetic products. In this study, we evaluated a genotoxicity assay for topically applied chemicals using a three-dimensional human reconstructed skin model, KeraSkin$^{TM}$. Two genotoxins, mitomycin C (MMC) and methyl methanesulfonate (MMS), induced significant dose-related increases in cytotoxicity and micronuclei induction in the skin model. In contrast, two non-genotoxins, 4-nitrophenol (4-NP) and trichloroethylene (TCE), induced cytotoxicity but not micronucleus formation. In conclusion, micronucleus test using human skin model may be useful for predicting in vitro genotoxic potentials of cosmetic products.

Phototoxicity: Its Mechanism and Animal Alternative Test Methods

  • Kim, Kyuri;Park, Hyeonji;Lim, Kyung-Min
    • Toxicological Research
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    • v.31 no.2
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    • pp.97-104
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    • 2015
  • The skin exposure to solar irradiation and photoreactive xenobiotics may produce abnormal skin reaction, phototoxicity. Phototoxicity is an acute light-induced response, which occurs when photoreacive chemicals are activated by solar lights and transformed into products cytotoxic against the skin cells. Multifarious symptoms of phototoxicity are identified, skin irritation, erythema, pruritis, and edema that are similar to those of the exaggerated sunburn. Diverse organic chemicals, especially drugs, are known to induce phototoxicity, which is probably from the common possession of UV-absorbing benzene or heterocyclic rings in their molecular structures. Both UVB (290~320 nm) and UVA (320~400 nm) are responsible for the manifestation of phototoxicity. Absorption of photons and absorbed energy (hv) by photoactive chemicals results in molecular changes or generates reactive oxygen species and depending on the way how endogenous molecules are affected by phototoxicants, mechanisms of phototoxcity is categorized into two modes of action: Direct when unstable species from excited state directly react with the endogenous molecules, and indirect when endogeneous molecules react with secondary photoproducts. In order to identify phototoxic potential of a chemical, various test methods have been introduced. Focus is given to animal alternative test methods, i.e., in vitro, and in chemico assays as well as in vivo. 3T3 neutral red uptake assay, erythrocyte photohemolysis test, and phototoxicity test using human 3-dimensional (3D) epidermis model are examples of in vitro assays. In chemico methods evaluate the generation of reactive oxygen species or DNA strand break activity employing plasmid for chemicals, or drugs with phototoxic potential.

Effect of skin penetration enhancer on the drug release from indomethacin-soft hydrogel (인도메타신 소프트 하이드로겔로 부터 약물 방출에 미치는 피부투과촉진제의 영향)

  • Nam, Hyun-Gue;Lee, Chi-Ho;Shin, Young-Hee
    • Journal of Pharmaceutical Investigation
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    • v.32 no.1
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    • pp.35-40
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    • 2002
  • We prepared a novel dosage form, peel-off type soft hydrogel using poly(vinyl alcohol), and evaluated the effect of skin penetration enhancer on the indomethacin release from soft hydrogel by in vitro permeation and in vivo absorption test. In this study, we used four enhancers-urea, dimethyl urea, 1,1,3,3-tetramethyl urea, and pirotiodecane (1-[2(decylthio)ethyl]azacyclopentane-2-one, $HPE-101^{circledR}$). In addition, we evaluated the primary skin irritation test of soft hydrogel using rabbit. From these results, we could find the pirotiodecane was a prominent enhancer, and soft hydrogel seemed to be safe and have no irritancy.

Chestnut Extracts as new Anti-aging agent

  • 김범준;김정하
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.24 no.1
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    • pp.113-126
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    • 1998
  • Inner nutshell of Castanea crenata has been used as an anti-aging and anti-wrinkle agent from the ancient time in east Asia. In order to develop new anti-aging and anti-wrinkle, ethanolic extract of inner nutshell of Castanea crenata was prepared and various biological activities were evaluated. Cor-285 possessed potent tree radical scavenging activity in vitro compared to gallic acid. Cor-285 showed the preventive effect against UV-induced cytotoxicity of fibroblast at concentration of 25-250mg/ml. When Cor-285 was evaluated for its anti-allergic activity, it effectively inhibited histamine release from mast cells induced by compound 48/80. The inhibitory activity was stronger than that of glycyrrhizinate. Cor-285 also showed on vivo inhibition against delayed hypersensitivity as well as croton-oil induced ear edema in mice when topically applied. These results strongly suggest that Cor-285 may reduce immunoregulatory/inflammatory skin trouble. From the attempts to isolate the scavenger, were isolated. In a clinical trial of twenty healthy volunteers with aged skin, 6 weeks application of Cor-285 decreased wrinkle about 26% and increased moisturizing 20% on the skin. All of these results indicate that Cor-285 may be an effective anti-aging and anti-wrinkle agent.

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Phototherapy in Allergic Rhinitis: From In Vitro Studies to Clinical Trials

  • Yoo, Shin Hyuk;Chung, Young-Jun;Bae, Jun-Sang;Mo, Ji-Hun
    • Medical Lasers
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    • v.9 no.2
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    • pp.95-102
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    • 2020
  • Phototherapy has a profound immunosuppressive effect and is widely used for the treatment of inflammatory skin diseases. Since allergic rhinitis (AR) shares several common pathologic features with atopic dermatitis, intranasal phototherapy is a novel approach for treating AR. Phototherapy reduces the antigen presenting capacity of dendritic cells, induces apoptosis of immune cells, and inhibits synthesis and release of pro-inflammatory mediators from inflammatory cells. The effectiveness of phototherapy for AR has been extensively researched: from in vitro studies to several clinical trials. Data indicates the potential of intranasal phototherapy as an alternative treatment for AR as well as other inflammatory mucosal diseases. In this review, we introduce the in vitro, in vivo studies, and clinical trials which demonstrated the effect of phototherapy on AR.

Effects of Aurantii Immaturus Fructus (AI) on Atopic Dermatitis (AD) Induced by DNCB in Mice (DNCB로 유발된 생쥐의 아토피 피부염에 대한 지실(枳實)의 효능)

  • Park, Jem Ma;Chae, Joong Won
    • The Journal of Pediatrics of Korean Medicine
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    • v.29 no.1
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    • pp.27-43
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    • 2015
  • Objectives The purpose of this study was to investigate the effects of AI on AD induced by DNCB in mice. AI has antiallergic property that is useful in treating allergy-related-diseases, such as asthma, anaphylactic shock, acute bronchitis and skin diseases, skin pruritus from gastrointestinal diseases. However, AI has not been studied intensively yet regarding anti-inflammatory effect on AD. Therefore, this study was conducted on 2,4-dinitrochlorobezene (DNCB)-induced mice to investigate effects of AI in AD. Methods In the experiment, we divided mice into four groups: a normal group (NOR), a control group (CON), an AI spread group (AI spread), and an AI spread and feeding group (AI spread & feeding). Then examined the changes in the body weight, weights of spleen and ear, thickness of dorsum skin and ear skin, clinical aspects on dorsum skin, historical assessments, proliferation of splenocytes in vitro and in vivo, and cytokine (TNF-${\alpha}$, IL-10). Results From the experiment, the ear weight of AI spread & feeding group was significantly dropped and the ear thickness of both AI spread and AI spread & feeding were decreased significantly. Dorsum skin thickness was also decreased significantly in both AI spread and AI spread & feeding group. Also, AI treatment improved the symptoms of AD, such as coloration, erythema and desquamation and had a better effect on AI spread & feeding group. In histopathological observation, thickened epidermis, hyperkeratosis, pigmentation, hypergranulosis, parakeratosis were diminished as well in both AI spread and AI spread & feeding group. In vitro, we could observe when AI was increased as proliferation rate of splenocytes were increased, too. Conclusions In conclusion, these data suggest that AI can decrease symptoms of AD and show AI can be useful herbal therapy for AD.