• Title/Summary/Keyword: Ex vivo skin

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In Vivo and Ex Vivo Skin Reactions after Multiple Pulses of 1,064-nm, Microlens Array-type, Picosecond Laser Treatment

  • Lyu, Herin;Park, Jinyoung;Lee, Hee Chul;Lee, Sang Ju;Kim, Young Koo;Cho, Sung Bin
    • Medical Lasers
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    • v.9 no.2
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    • pp.142-149
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    • 2020
  • Background and Objectives A picosecond-domain laser treatment using a microlens array (MLA) or a diffractive optical element elicits therapeutic micro-injury zones in the skin. This study examined the patterns of tissue reactions after delivering multiple pulses of 1,064-nm, MLA-type, picosecond neodymium:yttrium-aluminum-garnet laser treatment. Materials and Methods Multiple pulses of picosecond laser treatment were delivered to ex vivo human or brown micropig skin and analyzed histopathologically. A high-speed cinematographic study was performed to visualize the multiple pulses of picosecond laser energy-induced skin reactions in in vivo human skin. Results In the ex vivo human skin, a picosecond laser treatment at a fluence of 0.3 J/cm2 over 100 non-stacking passes generated multiple lesions of thermally-initiated laser-induced optical breakdown (TI-LIOB) in the epidermis and dermis. In the ex vivo micropig skin, stacking pulses of 20, 40, 60, 80, and 100 at a fluence of 0.3 J/cm2 generated distinct round to oval zones of tissue coagulation in the mid to lower dermis. High-speed cinematography captured various patterns of twinkling, micro-spot reactions on the skin surface over 100 stacked pulses of a picosecond laser treatment. Conclusion Multiple pulses of 1,064-nm, MLA-type, picosecond laser treatment elicit marked TI-LIOB reactions in the epidermis and areas of round to oval thermal coagulation in the mid to deep dermis.

The penetration enhancement and the lipolystic effects of TAT-GKH, in both In vitro, Ex vivo, and In vivo.

  • Lim, J.M.;Chang, M.Y.;Park, S.G.;Kang, N.G.;Song, Y.S.;Lee, Y.H.;Yoo, Y.C.;Cho, W.G.;Han, S.G.;Kang, S.H.
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.87-107
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    • 2003
  • It was demonstrated that Transactivating transcriptional activator(TAT) protein from HIV-1 shown to enter cells when added to the surrounding media. TAT peptide chemically attached to various proteins was able to deliver these proteins to various cell and even in tissues in mice with high levels in heart and spleen. In this study, the tripeptide GKH(Glycine-Lysine-Histidine) derived from Parathyroid hormone (PTH), which was known as lipolytic peptide, is attached to 9-poly Lysine(TAT) to be used as a cosmetic ingredient for slimming products. When Glycerol release, expressed as extracellular glycerol concentration, is lipolysis index, TAT-GKH at $10^{-5}$mo1/L induces approximately 41.5% maximal lipolytic effects in epididymal adipocytes isolated from rats, compared with basal lipolysis. Epididymal adipose tissues of male rats is assessed ex vivo by microdialysis. Probes are perfused with Ringer solution in which increasing concentrations of TAT-GKH. The perfusion of TAT-GKH induces lipolytic effect. Penetration study showed that TAT-GKH efficiently elevates 36 times higher penetration into the excised hairless mice skin than GKH. in vivo study showed that TAT-GKH had a better effect upon the relative volume of eye bag after 28 days of application on twenty(+2) healthy female volunteers. It was identified that TAT-GKH increases penetration enhancement and lipolytic effects in both in vitro, ex vivo and in vivo.

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Contrast Enhancement of Laser Speckle Contrast Image in Deep Vasculature by Reduction of Tissue Scattering

  • Son, Taeyoon;Lee, Jonghwan;Jung, Byungjo
    • Journal of the Optical Society of Korea
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    • v.17 no.1
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    • pp.86-90
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    • 2013
  • Various methods have been proposed for enhancing the contrast of laser speckle contrast image (LSCI) in subcutaneous blood flow measurements. However, the LSCI still suffers from low image contrast due to tissue turbidity. Herein, a physicochemical tissue optical clearing (PCTOC) method was employed to enhance the contrast of LSCI. Ex vivo and in vivo experiments were performed with porcine skin samples and male ICR mice, respectively. The ex vivo LSCIs were obtained before and 90 min after the application of the PCTOC and in vivo LSCIs were obtained for 60 min after the application of the PCTOC. In order to obtain the skin recovery images, saline was applied for 30 min after the application of the PCTOC was completed. The visible appearance of the tubing under ex vivo samples and the in vivo vasculature gradually enhanced over time. The LSCI increased as a function of time after the application of the PCTOC in both ex vivo and in vivo experiments, and properly recovered to initial conditions after the application of saline in the in vivo experiment. The LSCI combined with the PCTOC was greatly enhanced even in deep vasculature. It is expected that similar results will be obtained in in vivo human studies.

Comparison of In Vitro, Ex Vivo, and In Vivo Antibacterial Activity Test Methods for Hand Hygiene Products (손 위생 제품에 대한 in vitro, ex vivo, in vivo 항균 시험법 비교)

  • Daeun Lee;Hyeonju Yeo;Haeyoon Jeong
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.35-43
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    • 2024
  • Numerous methods have been applied to assess the antibacterial effectiveness of hand hygiene products. However, the different results obtained through various evaluation methods have complicated our understanding of the real efficacy of the products. Few studies have compared test methods for assessing the efficacy of hand hygiene products. In particular, reports on ex vivo pig skin testing are limited. This study aimed to compare and characterize the methodologies applied for evaluating hand hygiene products, involving in vitro, ex vivo, and in vivo approaches, applicable to both leave-on sanitizers and wash-off products. Our further aim was to enhance the reliability of ex vivo test protocols by identifying influential factors. We performed an in vitro method (EN1276) and an in vivo test (EN1499 and ASTM2755) with at least 20 participants, against Serratia marcescens or Escherichia coli and Staphylococcus aureus. For the ex vivo experiment, we used pig skin squares prepared in the same way as those used in the in vivo test method and determined the optimal treated sample volumes for sanitizers and the amount of water required to wash off the product. The hand sanitizers showed at least a 5-log reduction in bacterial load in the in vitro test, while they showed little antibacterial activity in the in vivo and ex vivo tests, particularly those with a low alcohol content. For the hand wash products, the in vitro test was limited because of bubble formation or the high viscosity of the products and it showed low antibacterial activity of less than a 1-log reduction against E. coli. In contrast, significantly higher log reductions were observed in ex vivo and in vivo tests, consistently demonstrating these results across the two methods. Our findings revealed that the ex vivo and in vivo tests reflect the two different antibacterial mechanisms of leave-on and wash-off products. Our proposed optimized ex vivo test was more rapid and more precise than the in vitro test to evaluate antibacterial results.

Skin wound healing effects of (+)-syringaresinol from ginseng berry

  • Jee-hyun Hwang;Yeonsoo Kang;Heui-Jin Park;Seolyeong Kim;Su-Hyun Lee;Hangun Kim;Sang-Jip Nam;Kyung-Min Lim
    • Journal of Ginseng Research
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    • v.47 no.5
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    • pp.654-661
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    • 2023
  • Background: Ginseng has been used as a traditional medicine and functional cosmetic ingredients for many years. Recent studies have focused on the potential biological effects of the ginseng berry and its ingredients. (+)-Syringaresinol (SYR) is enriched in ginseng berry and its beneficial effects on the skin have been recently reported. However, little is known about the its effects on the wound healing process of skin. Methods: Here, we evaluated the skin wound healing effect of (+)-SYR using the human fibroblast Hs68 cell and ex vivo pig and human skin tissue model. Scratch wound test and hydrogen peroxide (HPO) induce chemical wound model were employed. Results: (+)-SYR promoted the migration and proliferation of Hs68 cells without significant cytotoxicity at the tested concentrations. Especially, in ex vivo pig and human skin tissue, HPO-induced chemical wound was recovered almost completely by (+)-SYR. In line with the finding in Hs68, the protein expression levels of TGF-β and PCNA, a proliferation marker were increased, demonstrating the beneficial effects of (+)-SYR on skin wound repair. Conclusion: Collectively, we demonstrated that (+)-SYR from ginseng berry, can enhance the wound healing effect by accelerating cell proliferation and skin regeneration, suggesting the potential utility of (+)-SYR for skin wound repair.

Study for Enhanced Skin Penetration of Cosmetics by Plasma-Sono Stimuli (플라즈마-초음파 자극에 의한 화장품의 피부 침투 증진 연구)

  • Yoonho Hwang;Hyeyoun Cho;Yujin Park;Hwijin Jang;Sanghyo Park;Jaehong Key
    • Journal of Biomedical Engineering Research
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    • v.44 no.4
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    • pp.275-283
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    • 2023
  • The demand for skincare has increased due to the end of the COVID-19 pandemic, leading to a focus on skincare devices and technologies designed to improve the delivery of cosmetics. Among these technologies, skincare medical devices that utilize plasma therapy (Plasma) and sonophoresis (Sono) are commonly used in dermatology clinics. However, there is still a lack of quantitative analysis for transdermal absorption effects of Plasma and Sono skincare medical devices. In this study, we quantified enhanced transdermal absorption effects of Plasma and Sono devices through in-silico and ex-vivo studies. The Sono treatment demonstrated an increased transdermal absorption effect, showing a 10~13% difference in penetration compared to the control group in the in-silico experiment, and 159% and 184% increase in the ex-vivo experiment. The Plasma treatment revealed increased transdermal absorption effects, with a 1.0~2.5% penetration difference in the in-silico experiment, and a 124% increase in the ex-vivo experiment compared to the control group. We also observed a synergistic effect from the combined treatment of Plasma and Sono, as indicated by the highest increases of 197% and 242% in penetration. Furthermore, we have determined the optimal device settings and treatment conditions for Plasma-Sono skincare medical devices. Notably, higher on/off durations (Intensity levels) and longer Sono treatments resulted in greater transdermal absorption effects.

Anti-melanogenic property of ginsenoside Rf from Panax ginseng via inhibition of CREB/MITF pathway in melanocytes and ex vivo human skin

  • Lee, Ha-Ri;Jung, Joon Min;Seo, Ji-Yeon;Chang, Sung Eun;Song, Youngsup
    • Journal of Ginseng Research
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    • v.45 no.5
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    • pp.555-564
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    • 2021
  • Background: Ginsenosides of Panax ginseng are used to enhance skin health and beauty. The present study aimed to investigate the potential use of ginsenoside Rf (Rf) from Panax ginseng as a new anti-pigmentation agent. Methods: The anti-melanogenic effects of Rf were explored. The transcriptional activity of the cyclic adenosine monophosphate (cAMP) response element binding protein (CREB) and the expression levels of tyrosinase, microphthalmia-associated transcription factor (MITF), and tyrosinase-related proteins (Tyrps) were evaluated in melanocytes and UV-irradiated ex vivo human skin. Results: Rf significantly inhibited Forskolin (FSK) or UV-stimulated melanogenesis. Consistently, cellular tyrosinase activity and levels of MITF, tyrosinase, and Tyrps were downregulated. Furthermore, Rf suppressed MITF promoter activity, which was stimulated by FSK or CREB-regulated transcription coactivator 3 (CRTC3) overexpression. Increased CREB phosphorylation and protein kinase A (PKA) activity induced by FSK were also mitigated in the presence of Rf. Conclusion: Rf can be used as a reliable anti-pigmentation agent, which has a scientifically confirmed and reproducible action mechanism, via inhibition of CREB/MITF pathway.

Laser-assisted Delivery of a Combined Antioxidant Formulation Enhances the Clinical Efficacy of Fractional Microneedle Radiofrequency Treatment: A Pilot Study

  • Kim, Jihee;Kim, Soo Min;Jung, Bok Ki;Oh, Sang Ho;Kim, Young-Koo;Lee, Ju Hee
    • Medical Lasers
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    • v.10 no.3
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    • pp.161-169
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    • 2021
  • Background and Objectives Fractional microneedle radiofrequency systems are popular options to increase elasticity in aging skin. Laser-assisted drug delivery is a promising method for the epidermal injection of topically applied drugs and cosmetic ingredients. This study assesses the safety and efficacy of topical delivery of L-ascorbic acid, vitamin E, and ferulic acid after fractional microneedle radiofrequency treatment for reducing photodamage. Materials and Methods In this prospective, single-center, split-face, controlled pilot study, six women (mean age, 48.0 years; range, 35-57 years; Fitzpatrick skin types III and IV) exhibiting mild to moderate photodamage, underwent a single session of fractional microneedle radiofrequency treatment. The patients were instructed to apply the antioxidant formulation to only one side of the face. Patients were evaluated 3 days, 7 days, and 4 weeks thereafter, using three-dimensional imaging and ultrasound. Ex vivo, the full-thickness human skin was used for molecular and histological evaluation. Statistical analysis was achieved by applying t-tests, Mann-Whitney U tests, and one-way analyses of variance. Results Compared to the untreated side, the antioxidant-treated side exhibited a significant increase in dermal thickness (10.32% vs. 17.54%, p < 0.05), but not in skin elasticity (4.76% vs. 4.69%, p > 0.05). The difference in erythema between the sides was statistically not significant (p > 0.05). In the ex vivo model, expression of FGF2 in the skin was significantly increased after application of the antioxidant formulation, as compared to results obtained subsequent to fractional microneedle radiofrequency treatment only (p < 0.01). Conclusion This study demonstrates that for the treatment of photodamaged skin, laser-assisted delivery of the antioxidant formulation is a safe and effective adjuvant modality following fractional microneedle radiofrequency.

Visualization of Epidermis and Dermal Cells in ex vivo Human Skin Using the Confocal and Two-photon Microscopy

  • Choi, Sang-Hoon;Kim, Wi-Han;Lee, Yong-Joong;Lee, Ho;Lee, Weon-Ju;Yang, Jung-Dug;Shim, Jong-Won;Kim, Jin-Woong
    • Journal of the Optical Society of Korea
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    • v.15 no.1
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    • pp.61-67
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    • 2011
  • The confocal laser scanning microscopy and two-photon microscopy was implemented based on a single laser source and an objective lens. We imaged and compared the morphology of identical sites of ex vivo human skin using both microscopes. The back-scattering emission from the sample provided the contrast for the confocal microscopy. The intrinsic autofluorescence and the second harmonic generation were used as the luminescence source for the two-photon microscopy. The wavelength of the Ti:Sapphire laser was tuned at 710 nm, which corresponds to the excitation peak of NADH and FAD in skin tissue. The various cell layers in the epidermis and the papillary dermis were clearly distinguished by both imaging modalities. The two-photon microscopy more clearly visualized the intercellular region and the nucleus of the cell compared to the confocal microscopy. The fibrous structures in the dermis were more clearly resolved by the confocal microscopy. Numerous cells in papillary dermal layer, as deep as $100\;{\mu}m$, were observed in both CLSM and two-photon microscopy. While most previous studies focused on fibrous structure imaging (collagen and elastin fiber) in the dermis, we demonstrated that the combined imaging with the CLSM and two-photon microscopy can be applied for the non-invasive study of the population, distribution and metabolism of papillary dermal cells in skin.

Moisturization and Transdermal Penetration Characteristics of PEGimpregnated Aloe vera Gel from DIS Processing (DIS에 의한 Polyethylene Glycol 함침 알로에 베라 겔의 보습 및 경피흡수 특성)

  • Kwon, Hye Mi;Hur, Won;Lee, Shin Young
    • KSBB Journal
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    • v.28 no.5
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    • pp.319-326
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    • 2013
  • This study was carried out to investigate the in vitro and in vivo moisturizing properties and percutaneous absorption of PEG-impregnated Aloe vera gel. The PEG-i-Aloe gel was obtained from dewatering and impregnation by soaking (DIS) of Aloe vera leaf slice. The moisturizing property of the obtained sample was evaluated by moisture determination using gravimetric method in desiccator under different RH% and by water sorption-desorption test on human skin. The transdermal penetration characteristics of PEG-i-Aloe gel was investigated by Franz diffusion cell in vitro transdermal absorption method. PEG-i-Aloe gel had high moisture retention ability and could significantly lead the enhancing skin hydration status as well as reducing the skin water loss due to the film formation as a skin barrier. The skin penetration rate of PEGi- Aloe gel at steady state was 9.76 ${\mu}g/(h{\cdot}cm^2)$ and the quantity of the transdermal absorption was 144 ${\mu}g/cm^2$ in 9 hr. The penetration mechanism was well fitted with Higuchi model ($R^2$ = 0.974-0.994). The results show that PEG-i-Aloe gel has the significant moisturizing effect and strong penetration of the animal skin. It could be used as the moisturizing additive in cosmetic skin products.