• Title/Summary/Keyword: in-vitro physical model

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Extracts of Grifola frondosa inhibit the MAPK signaling pathways involved in keratinocyte inflammation and ameliorate atopic dermatitis

  • Eun-Ju Choi;Jin Kyeong Choi
    • Nutrition Research and Practice
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    • v.17 no.6
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    • pp.1056-1069
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    • 2023
  • BACKGROUND/OBJECTIVES: Grifola frondosa, commonly referred to as the maitake mushroom, has been studied extensively to explore its potential health benefits. However, its anti-inflammatory effects in skin disorders have not been sufficiently elucidated. This study aimed to elucidate the anti-inflammatory role of the ethanol extract of G. frondosa in atopic dermatitis (AD) using in vivo and in vitro models. MATERIALS/METHODS: We investigated its impact on skin and spleen inflammatory responses in Dermatophagoides farinae extract (DFE)/1-chloro-2,4 dinitrochlorobenzene (DNCB)-induced AD-like skin lesions in a mouse model. Additionally, we determined the immunosuppressive response and mechanism of G. frondosa by inducing atopic-like immune reactions in keratinocytes through tumor necrosis factor (TNF)-α/interferon (IFN)-γ stimulation. RESULTS: Our study revealed that G. frondosa ameliorates clinical symptoms in an AD-like mouse model. These effects contributed to the suppression of Th1, Th2, Th17, and Th22 immune responses in the skin and spleen, leading to protection against cutaneous inflammation. Furthermore, G. frondosa inhibited the production of antibodies immunoglobulin (Ig)E and IgG2a in the serum of AD mice. Importantly, the inhibitory effect of G. frondosa on inflammatory cytokines in TNF-α/IFN-γ-stimulated AD-like keratinocytes was associated with the suppression of MAPK (Mitogen Activated Protein Kinase) pathway activation. CONCLUSIONS: Collectively, these findings highlight the potential of G. frondosa as a novel therapeutic agent for AD treatment and prevention.

Reduction of Migration and Invasion Ability of nm23-H1 Transfected U87MG (nm23-H1 유전자가 주입된 U87MG 세포의 이동능과 침윤능의 감소)

  • Paek, Yun-Woong
    • Journal of Korean Biological Nursing Science
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    • v.7 no.1
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    • pp.47-56
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    • 2005
  • nm23-H1 gene expression has been inversely correlated with tumor metastatic potential in certain tumors including melanomas, breast carcinomas, and hepatocellular carcinomas. However, its role with respect to the invasive behavior of central nervous system tumors has scarcely been addressed Because cell motility and invasion plays an essential role in metastatic dissemination, we have studied whether motile human glioma cell(U87MG) transfected with nm23-H1 complementary DNA have any alterations in their ability to migrate and invade. There was no significant changes in the shape and size of the cells following nm23-H1 transfection. The role of nm23-H1 in glioma migration and invasion have been evaluated by in vitro simple scratch technique and brain slice invasion model Basal migration ability of nm23-H1 transfectants cell(U87MG-pEGFP-nm23) were lesser than U87MG. Accordingly, U87MG-pEGFP-nm23 didn't migrate away apparently from the tumors implanted site comparing U87MG in brain slice invasion model. These results suggest that nm23-H1 may play an important role in suppressing the human glioma migration and invasion.

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Development of a Dynamic System Simulating Pig Gastric Digestion

  • Chiang, C.-C.;Croom, J.;Chuang, S.-T.;Chiou, P.W.S.;Yu, B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1522-1528
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    • 2008
  • The objective of this study was to develop a model for simulating gastric digestion in the pig. The model was constructed to include the chemical and physical changes associated with gastric digestion such as enzyme release, digestion product removal and gastric emptying. Digesta was collected from the stomach cannula of pigs to establish system parameters and to document the ability of the model to simulate gastric digestion. The results showed that the average pH of gastric digesta increased significantly from 2.47 to 4.97 after feed consumption and then decreased 140 min postprandial. The model described the decrease in pH within the pigs' stomach as $pH_t=5.182e^{-0.0014t}$, where t represents the postprandial time in minutes. The cumulative distribution function of liquid digesta was $V_t=64.509e^{0.0109t}$. The average pepsin activity in the liquid digesta was 317Anson units/mL. There was significant gastric emptying 220 min after feed consumption. The cybernetic dynamic system of gastric digestion was set according to the above data in order to compare with in vivo changes. The time course of crude protein digestion predicted by the model was highly correlated with observed in vivo digestion (r = 0.97; p = 0.0001), Model prediction for protein digestion was higher than that observed for a traditional static in vitro method (r = 0.89; p = 0.0001).

Scutellaria baicalensis Inhibits Coxsackievirus B3-Induced Myocarditis Via AKT and p38 Pathways

  • Fu, Qiang;Gao, Lu;Fu, Xiao;Meng, Qinghua;Lu, Zhihong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1230-1239
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    • 2019
  • Scutellaria baicalensis Georgi has been widely used in China for treatment of various diseases. This study investigated the effect of Scutellaria baicalensis Georgi extracts (SBE) against Coxsackievirus B3 (CVB3)-induced myocarditis in vitro and in vivo. In vitro, Hela cells and primary myocardial cells were infected with CVB3 and treated with SBE ($50-800{\mu}g/ml$) and ribavirin ($200{\mu}M$) for 48 h and then determined by CCK8 assay. Real-time PCR and western blotting assays were performed. In vivo, a myocarditis model was induced in male BALB/c mice by injecting CVB3 suspension intraperitoneally for three times, followed by treatment with SBE (400 and 200 mg/kg) and ribavirin (100 mg/kg) for 28 days. SBE ameliorated the cytotoxicity of CVB3 in Hela cells, especially at $400{\mu}g/ml$ (39.93% vs 65.67%, p < 0.05) without influencing cell growth and also significantly reduced CVB3 replication in primary myocardial cells. The levels of AKT, ERK, and p38 were increased after CVB3 infection. SBE could downregulate the expressions of AKT and p38. In vivo, the mortality rate from CVB3 reached to 66.67%, while 10.00% and 23.33% of this came after 400 and 200 mg/kg SBE treatment, respectively (p < 0.05). The CVB3 replication was obviously reduced after SBE administration from day 5. Similarly, the levels of AKT, ERK, and p38 mRNAs and proteins were increased, and SBE suppressed the expression of AKT and p38. Our study indicates that SBE is a promising potent antiviral agent against CVB3-induced myocarditis by inhibition of virus replication via depressing AKT and p38 expressions.

Enhanced Liver Targeting by Synthesis of $N_{\b{1}}-Stearyl-5-Fu$ and Incorporation into Solid lipid Nanoparticles

  • Yu, Bo-Tao;Xun-Sun;Zhang, Zhi-Rong
    • Archives of Pharmacal Research
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    • v.26 no.12
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    • pp.1096-1101
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    • 2003
  • To enhance the liver targeting and reduce the side effects of 5-fluorouracil (5-Fu), it was acylated by stearyl chloride to obtain .$\b{N}_{\b{1}}$stearyl-5-Fu (5-FuS). The chemical structure of the prodrug was confirmed by Nuclear Magnetic Resonance and Infrared Spectrometry. 5-FuS was incorporated into solid lipid nanoparticles (SLN), which were prepared by the physical agglomeration method. The mean diameter of 5-FuS-SLN was 240.19 nm and the drug loading was 20.53%. The release characteristics in vitro of 5-FuS-SLN were fitted to the first-order pharmacokinetic model. Compared with 5-Fu injection, a study on the distribution of 5-FuS-SLN in mice showed that 5-FuS-SLN could double 5-Fu concentration in mice livers. The main pharmacokinetic parameters of 5-FuS-SLN in rabbits is shown as follows: $V_d$=0.04336L/kg, $T_{1/2} \beta$=1.2834h, CL=0.1632 L/h. In conclusion, 5-FuS-SLN has significant liver targeting properties. The employment of a prodrug to enhance drug liposoluble properties and the preparation method presented in this paper, seem to be an alternative strategy to the traditional colloidal delivery system.

Experimental studies of Glycine max Merr. (black bean), Triticum aestivum L. (wheat) and Oryza sativa L. (rice bran) extracts on the effects of hair growth activity and physical properties (검은콩, 밀, 쌀겨 추출물이 모발의 성장과 물리적 특성에 미치는 효과)

  • Park, Hye-Yoon;Kim, Su-Na;Kang, Byung-Ha;Lee, John-Hwan
    • Korean Journal of Oriental Medicine
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    • v.16 no.3
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    • pp.167-173
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    • 2010
  • Objects : Glycine max Merr. (black bean), Triticum aestivum L. (wheat) and Oryza sativa L. (rice bran) have been widely used for treatment of relaxion of smooth muscle, gastrointestinal hemorrhage and alopecia in Korean Traditional Medicine. In this research, we examined the effect of the extracts, obtained from EtOH extracts of 3 kinds of traditional plants, on hair growing activity of the DP6 and C3H10T1 cell and physical properties. Materials and Methods : On the basis of previous studies, three traditional plants were selected and we extracted them with ethanol. We evaluated their hairy dermal papillar cell proliferation activity and mouse mesenchymal stem cell in vitro model. Also, 3 herbal extracts were added to the normal shampoo formulation in ranges of 0.1% and we validated tensile properties and physical changes using aged hair. In this research, we compared the tensile strength, shine and color appearance between the hair (general formulation) and the hair after applying shampoo with natural extracts. To analyze the luster and color image, we use the SAMBA hardware and software made by Bossa Nova Technologies. Results : In the comparative test for tensile characteristic between the hair treated general formulation(control) and the hair applying special formulation including 3 kinds of extracts, tensile distance and energy of the latter are larger than control on average. The shine and color appearance were also increased after using shampoo including natural extracts(shine : 10.9%, color appearance: 24.12%). We observed the enhancement of hair growth activity in the DP6 and C3H10T1 cell. Especially black bean extracts had the most powerful effect in the dermal papillar cell proliferation. Conclusion : These experiments suggest that extracts of Glycine max Merr. (black bean), Triticum aestivum L. (wheat) and Oryza sativa L. (rice bran) stimulate the hair growth activity and can improve physical activities of aged hair. Shampoo product, which contains 3 kinds of natural extracts, would be used for the treatment for aged hair.

TISSUE-ENGINEERED RECONSTITUTION OF ORAL MUCOSA USING POLYDIOXANONE MESH (Polydioxanone mesh를 이용한 구강점막의 조직 공학적 재건)

  • Moon, Seon-Jae;Joo, So-Yeon;Kim, Jin;Kim, Hak-Yong;Park, Jung-Keug;Cha, In-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.4
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    • pp.249-256
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    • 2003
  • The lack of sufficient oral mucosa available for intra-oral reconstruction has been dealt with by the use of skin or oral mucosa grafts harvested from donor sites but grafts requires more than one surgical procedures and could cause donor site morbidity. Many investigators have attempted to increase available soft tissue by tissue engineered skin or oral mucosa replacements for clinical applications. But, reconstructed mucosa by several methods have low physical properties such as rolling and contraction. The aims of this study were to develope an in vitro experimental model that maintains an epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally cultured oral mucosa embedded with Polydioxanone mesh by histological and immunohistochemical analysis. The results were as follows; 1. Oral mucosa reconstructed by three-dimensional organotypic culture revealed similar morphologic characteristics to equvalent normal oral mucosa in the point that they show stratification and differentiation. 2. The expression of cytokeratin 10/13 and involucrin in the cultured tissue showed the same pattern with normal oral mucosa suggesting that organotypic co-culture condition is able to induce cellular differentiation. 3. After insertion of polydioxanone mesh, increased tensile strength were observed. These results suggest that three-dimensional organotypic co-culture of the oral mucosa cell lines with the dermal equvalent consisting type I collagen and fibroblasts reproduce the morphologic and immunohistochemical characteristics similar to those in vivo condition. And increased physical properties by use of polydioxanone mesh will helpful for clinical applications.

Selection of model viruses for foot-and-mouth disease virus-related-experiments (구제역 바이러스를 대체할 모델 바이러스 선별)

  • Kim, Tae-Hwan;Herath, Thilina U. B.;Kim, Jae-Hoon;Lee, Kwang-Nyeong;Park, Jong-Hyeon;Kim, Chul-Joong;Lee, Jong-Soo
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.304-308
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    • 2017
  • Researchers have comparatively fewer opportunities to conduct experiments on foot-and-mouth disease virus (FMDV), owing to the limited availability of biosafety level 3 facilities. Bovine rhinovirus (BRV) and human rhinovirus (HRV), which are genetically closely related to FMDV, have been evaluated in this study as model viruses for FMDV. To discover whether BRV and HRV have similar physicochemical properties as FMDV, virus susceptibility tests have been performed in different physical (pH and heat) and chemical (acidic/alkaline solutions and commercial disinfectants) conditions in vitro. Our data revealed that the physicochemical characteristics of BRV and HRV were nearly similar to those of FMDV.

Alleviation of ascorbic acid-induced gastric high acidity by calcium ascorbate in vitro and in vivo

  • Lee, Joon-Kyung;Jung, Sang-Hyuk;Lee, Sang-Eun;Han, Joo-Hui;Jo, Eunji;Park, Hyun-Soo;Heo, Kyung-Sun;Kim, Deasun;Park, Jeong-Sook;Myung, Chang-Seon
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.1
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    • pp.35-42
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    • 2018
  • Ascorbic acid is one of the most well-known nutritional supplement and antioxidant found in fruits and vegetables. Calcium ascorbate has been developed to mitigate the gastric irritation caused by the acidity of ascorbic acid. The aim of this study was to compare calcium ascorbate and ascorbic acid, focusing on their antioxidant activity and effects on gastric juice pH, total acid output, and pepsin secretion in an in vivo rat model, as well as pharmacokinetic parameters. Calcium ascorbate and ascorbic acid had similar antioxidant activity. However, the gastric fluid pH was increased by calcium ascorbate, whereas total acid output was increased by ascorbic acid. In the rat pylorus ligation-induced ulcer model, calcium ascorbate increased the gastric fluid pH without changing the total acid output. Administration of calcium ascorbate to rats given a single oral dose of 100 mg/kg as ascorbic acid resulted in higher plasma concentrations than that from ascorbic acid alone. The area under the curve (AUC) values of calcium ascorbate were 1.5-fold higher than those of ascorbic acid, and the $C_{max}$ value of calcium ascorbate (91.0 ng/ml) was higher than that of ascorbic acid (74.8 ng/ml). However, their $T_{max}$ values were similar. Thus, although calcium ascorbate showed equivalent antioxidant activity to ascorbic acid, it could attenuate the gastric high acidity caused by ascorbic acid, making it suitable for consideration of use to improve the side effects of ascorbic acid. Furthermore, calcium ascorbate could be an appropriate antioxidant substrate, with increased oral bioavailability, for patients with gastrointestinal disorders.

Kinetic Characterization of Swelling of Liquid Crystalline Phases of Glyceryl Monooleate

  • Lee, Jae-Hwi;Choi, Sung-Up;Yoon, Mi-Kyeong;Choi, Young-Wook
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.880-885
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    • 2003
  • Research in this paper focuses on the kinetic evaluation of swelling of the liquid crystalline phases of glyceryl monooleate (GMO). Swelling of the lamellar and cubic liquid crystalline phases of GMO was studied using two in vitro methods, a total immersion method and a Franz cell method. The swelling of the lamellar phase and GMO having 0 %w/w initial water content was temperature dependent. The swelling ratio was greater at $20^{\circ}^C than 37^{\circ}^C$ . The water uptake increased dramatically with decreasing initial water content of the liquid crystalline phases. The swelling rates obtained using the Franz cell method with a moist nylon membrane to mimic buccal drug delivery situation were slower than the total immersion method. The swelling was studied by employing first-order and second-order swelling kinetics. The swelling of the liquid crystalline phases of GMO could be described by second-order swelling kinetics. The initial stage of the swelling (t < 4 h) followed the square root of time relationship, indicating that this model is also suitable for describing the water uptake by the liquid crystalline matrices. These results obtained from the current study demonstrate that the swelling strongly depends on temperature, the initial water content of the liquid crystalline phases and the methodology employed for measuring the swelling of GMO.