• Korean Journal of Veterinary Research
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• v.63 no.4
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• pp.40.1-40.7
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• 2023

To optimize the most efficient method for porcine in vitro maturation (IVM), we compared the effects of supplementing extracellular vesicles (EVs) derived from porcine follicular fluid (pFF). The cumulus oocyte complexes were grouped into 4 groups with different supplementations as following: pFF (G1), pFF-depleted EVs (G2), EVs (G3) and control (G4) groups. After IVM with different supplementations, maturation rates and the developmental competences of porcine oocytes and blastocyst development were investigated. Additionally, glutathione (GSH) and reactive oxygen species (ROS) levels were measured in mature oocytes. The EVs were isolated and characterized with cryo-TEM and nanoparticle tracking analysis. The pFF significantly affected the maturation rate, whereas the presence of EVs did not show notable difference in the maturation rates. Although there were numerical increases in the measured parameters in EV and pFF-depleted EVs groups, no significant differences were observed between them. The EV group showed similar oocyte maturation rate for both positive and negative control groups. The GSH was not different among the groups, but ROS levels were significantly lower in pFF-supplemented group when compared with other groups with the highest level in the control group. G2 group wasn't significantly different G1 and G3 group. G3 group wasn't significantly different from G2 and G4 group. This suggests that EVs in IVM medium which probably effected partially to protect against oxidative stress and potentially enhance the quality of oocytes. This study indicates that the EVs in pFF play a significant role in improving the efficiency of oocyte maturation in porcine.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.131-137
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• 2024

Background: Porcine embryonic development is widely utilized in the medical industry. However, the blastocyst development rate in vitro is lower compared to in vivo. To address this issue, various supplements are employed. Extracellular vesicles (EVs) play the role of communicators that carry many bioactive cargoes. Additionally, the contents of EVs can vary on the estrous cycle. Methods: We compared the effects of adding EVs derived from porcine uterine fluid (UF), categorized as non-EV (G1), EVs in estrus (G2) and EVs in diestrus (G3). After in vitro culture (IVC) was performed in three different groups, cleavage rate and blastocyst development rate were examined. In addition, glutathione (GSH) and reactive oxygen species (ROS) levels were measured 2 days after activation to assess oxidative stress. Results: Using NTA and cryo-TEM, we confirmed the presence of EVs with sizes ranging from 30 nm to 200 nm, that the particles were suitable for analysis for analysis. In IVC data, the highest cleavage rate was observed in G2, which was significantly different from G1 but not significantly different from the next highest, G3. Similarly, the highest blastocyst development rate was observed in G2, which was significantly different from G1 but not significantly different from the next highest, G3. Conclusions: These results indicate that estrus derived EVs contain biofactors beneficial for early blastocyst development, including GSH which protects the blastocyst from oxidative stress. Additionally, although diestrus-derived EVs are expected to have some effect on blastocyst development, it appeared to be less effective than estrus-derived EVs.

• Journal of the Korean Applied Science and Technology
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• v.37 no.5
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• pp.1285-1297
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• 2020

In this study, we report the results of a study on the clinical evaluation of wrinkle improvement by developing a method for liposome of high-purity Centella asiatica extract used in pharmaceuticals and cosmetics, and a cream using the same. In order to make Centellasome-10EX stabilizing centella asiatica extract in liposome lamella vesicle, it could be completed using 5% hydrogenated lecithin and 2% sucrose distearate. The appearance of Centellasome-10EX was a creamy form of low viscosity, the color was pale yellow, and the odor had the inherent odor of the raw material. The pH was about 6.12, the specific gravity was 1.09, and the acid value was about 0.35. The content of the main constituents of centella asiatica extract contained in the liposome vesicle contains 10,800 ppm of asiatic acid, 10,900 ppm of asiaticoside, 6,000 ppm of madecasic acid, and 1,600 ppm of madecassoside, and long-term storage. There was no discoloration even at the time, and it was found that the main component remained stable thermodynamically. To mechanistically analyze the structure of the liposome vesicle of Centellasome-10EX, as a result of observation with a transmission electron microscope (Cryo-TEM), the multilayer vesicles are formed and filled with moisture, and there are 10 to 60 multilayers around it. It was confirmed that the liposome lamella vesicle was formed. As a clinical trial (in-vivo) test, the moisturizing effect of centellasome cream after application for 5 weeks was 28.3%, which was significantly increased compared to placebo. The skin elasticity effect was 13.6%, which significantly increased the moisturizing power than the placebo. The effect of improving fine wrinkles around the eyes was improved by 23.52% compared to placebo cream. Through the results of this study, it was possible to study the formulation and manufacturing method for encapsulation and stabilization of the developed Centellasome-10EX in the liposome vesicle. It is expected that the results obtained through clinical research on the wrinkle improvement effect of the cream using this can be widely used to study skin science in the cosmetic industry and to develop high-quality cosmetics with high efficacy.

• Journal of the Korean Applied Science and Technology
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• v.38 no.6
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• pp.1687-1698
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• 2021

This study is to produce multiple layers of liposomes in a supercritical state and encapsulates active ingredients in order to stably encapsulate thermodynamically unstable active ingredients. In order to form a liposome in a supercritical state, a mixed surfactant development including vegetable-derived hydrogenated phosphatidyl choline and their delivative, hydrogenated sucrose distearate was synthesized as high purity. It describes a manufacturing method of injecting liquid carbon dioxide into a reactor to create a supercritical state and stirring to produce a giant liposome, and adding and loading genistein and quercetin. The HLB of the mixed lipid complex (SC-Lipid Complex) was 12.50, and multiple layers of liposome vesicles were formed even at very low concentrations. This surfactant had a specific odor with a pale yellow flake, the specific gravity was 0.972, and the acid value was 0.12, indicating that it was synthesized with high purity. As a result of the emulsifying capacity experiment using 20 wt% capric/capric triglyceride and triethylhexanoin using SC-Lipid Complex, it was found to have 96.2% emulsifying power. SC LIPOSOME GENISTEIN was confirmed that a multi-layer liposome vesicle was formed through a transmission electron microscope (Cryo-TEM) for the supercritical liposome encapsulated with genistein. The primary liposome particle size in which genistein was encapsulated was 253.9 nm, and the secondary capsule size was 18.2 ㎛. Using genistein as the standard substance, the encapsulation efficiency of supercritical liposomes was 99.5%, and general liposomes were found to have an efficiency of 93.6%. In addition, the antioxidant activity experiment in which quercetin was sealed was confirmed by the DPPH method, and it was found that the supercritical liposome significantly maintained excellent antioxidant activity. In this study, thermodynamically unstable raw materials were sealed into liposomes without organic solvents in a supercritical state. Based on these results, it is expected that it can be applied to various forms such as highly functional skincare cosmetics, makeup cosmetics, and scalp protection cosmetics.