• Title/Summary/Keyword: scratch assay

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The effects of ATO-ALL on skin regeneration in human epidermal keratinocytes, HaCaT cells (ATO-ALL이 인간유래 피부 각질형성세포인 HaCaT 세포에서 피부재생에 미치는 효과)

  • Shin, Younmin;Lee, Won-Kil;Kim, Seung Hyung;Choi, Jeong June
    • Journal of Haehwa Medicine
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    • v.28 no.2
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    • pp.41-47
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    • 2019
  • Objectives : Atopic dermatitis is an irritable skin disease accompanying rash and itching leading to impaired skin barrier. ATO-ALL is an ethanol extract of natural products comprising 12 herbs and effective on atopic dermatitis. In this study, we aimed to propose that the effect of ATO-ALL on skin regeneration in human keratinocyte cell line, HaCaT cells. Methods : To evaluate the skin regenerating effects of ATO-ALL, scratch wound healing assay, bromodeoxyuridine (BrdU) assay, and propidum iodide (PI) assay were performed using cultured HaCaT cell line. Result : Scratch wound healing assay showed that ATO-ALL was able to enhance the gap filling activity more than 2-fold at 7 ppm concentration compared with control group. BrdU assay demonstrated that ATO-ALL treatment increased the de novo cell proliferation in a dose-dependent manner. Finally, PI assay indicated that the cell cycle of HaCaT cells was modulated by ATO-ALL treatment. Conclusions : These results suggested that ATO-ALL may have skin regenerating effects by increasing cell proliferation via cell cycle regulation. Taken together, ATO-ALL is supposed to have a potential on regeneration of damaged skin or functional disease including atopic dermatitis.

Cytocompatibility and cell proliferation evaluation of calcium phosphate-based root canal sealers

  • Mestieri, Leticia Boldrin;Zaccara, Ivana Maria;Pinheiro, Lucas Siqueira;Barletta, Fernando Branco;Kopper, Patricia Maria Polli;Grecca, Fabiana Soares
    • Restorative Dentistry and Endodontics
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    • v.45 no.1
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    • pp.2.1-2.7
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    • 2020
  • Objectives: This study aimed to evaluate the cell viability and migration of Endosequence Bioceramic Root Canal Sealer (BC Sealer) compared to MTA Fillapex and AH Plus. Materials and Methods: BC Sealer, MTA Fillapex, and AH Plus were placed in contact with culture medium to obtain sealers extracts in dilution 1:1, 1:2 and 1:4. 3T3 cells were plated and exposed to the extracts. Cell viability and migration were assessed by 3-(4,5-dimethylthiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and Scratch assay, respectively. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). Results: The MTT assay revealed greater cytotoxicity for AH Plus and MTA Fillapex at 1:1 dilution when compared to control (p < 0.05). At 1:2 and 1:4 dilutions, all sealers were similar to control (p > 0.05) and MTA Fillapex was more cytotoxic than BC Sealer (p < 0.05). Scratch assay demonstrated the continuous closure of the wound according to time. At 30 hours, the control group presented closure of the wound (p < 0.05). At 36 hours, only BC Sealer presented the closure when compared to AH Plus and MTA Fillapex (p < 0.05). At 42 hours, AH Plus and MTA Fillapex showed a wound healing (p > 0.05). Conclusions: All tested sealers demonstrated cell viability highlighting BC Sealer, which showed increased cell migration capacity suggesting that this sealer may achieve better tissue repair when compared to other tested sealers.

The Effect of Hyaluronic Acid on the Invasiveness of Malignant Glioma Cells : Comparison of Invasion Potential at Hyaluronic Acid Hydrogel and Matrigel

  • Jin, Shu-Guang;Jeong, Young-Il;Jung, Shin;Ryu, Hyang-Hwa;Jin, Yong-Hao;Kim, In-Young
    • Journal of Korean Neurosurgical Society
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    • v.46 no.5
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    • pp.472-478
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    • 2009
  • Objective : Hyaluronidase (HAse), a degrading enzyme of hyaluronic acid (HA), is highly expressed in patients with malignant glioma. The purpose of this study was to verify whether HAse is related to the invasion of glioma cells. We also investigated if glioma cells with higher mobility in 2-dimensioal (2-D) method have also higher mobility at 3-dimensional (3-D) environment. Methods : Malignant glioma cell lines (U87MG, U251MG, U343MG-A, and U373MG) were used, and their HAse expressions were evaluated by HA zymography. The migration ability was evaluated by simple scratch technique. The invasiveness of each cell lines was evaluated by Matrigel invasion assay and HA hydrogel invasion assay. In HA hydrogel invasion assay, colonies larger than $150\;{\mu}m$ were regarded as positive ones and counted. Statistical analysis of migration ability and invasion properties of each cell lines was performed using t-test. Results : In scratch test to examine migration ability of each cell lines, U87MG cells were most motile than others, and U343MG-A least motile. The HAse was expressed in U251MG and U343MG-A cell lines. However, U87MG and U373MG cell lines did not express HAse activity. In Matrigel invasion assay, the cell lines expressing HAse (U251MG and U343MG-A) were more invasive in the presence of HA than HAse deficient cell lines (U87MG and U373MG). In HA hydrogel invasion assay, the HAse-expressing cell lines formed colonies more invasively than HAse-deficient ones. Conclusion : Malignant Glioma cells expressing HAse were more invasive than HAse-deficient ones in 3-dimensional environment. Therefore, it might be suggested that invasion of malignant gliomas is suppressed by inhibition of HAse expression or HA secretion. Additionally, the ability of 2-D migration and 3-D invasion might not be always coincident to each other in malignant glioma cells.

Effects of CTHRC1 on odontogenic differentiation and angiogenesis in human dental pulp stem cells

  • Jong-soon Kim;Bin-Na Lee;Hoon-Sang Chang;In-Nam Hwang;Won-Mann Oh;Yun-Chan Hwang
    • Restorative Dentistry and Endodontics
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    • v.48 no.2
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    • pp.18.1-18.10
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    • 2023
  • Objectives: This study aimed to determine whether collagen triple helix repeat containing-1 (CTHRC1), which is involved in vascular remodeling and bone formation, can stimulate odontogenic differentiation and angiogenesis when administered to human dental pulp stem cells (hDPSCs). Materials and Methods: The viability of hDPSCs upon exposure to CTHRC1 was assessed with the WST-1 assay. CTHRC1 doses of 5, 10, and 20 ㎍/mL were administered to hDPSCs. Reverse-transcription polymerase reaction was used to detect dentin sialophosphoprotein, dentin matrix protein 1, vascular endothelial growth factor, and fibroblast growth factor 2. The formation of mineralization nodules was evaluated using Alizarin red. A scratch wound assay was conducted to evaluate the effect of CTHRC1 on cell migration. Data were analyzed using 1-way analysis of variance followed by the Tukey post hoc test. The threshold for statistical significance was set at p < 0.05. Results: CTHRC1 doses of 5, 10, and 20 ㎍/mL had no significant effect on the viability of hDPSCs. Mineralized nodules were formed and odontogenic markers were upregulated, indicating that CTHRC1 promoted odontogenic differentiation. Scratch wound assays demonstrated that CTHRC1 significantly enhanced the migration of hDPSCs. Conclusions: CTHRC1 promoted odontogenic differentiation and mineralization in hDPSCs.

The Effect of the Butanol Fraction from Hydnocarpi Semen Extract on Activation of Keratinocyte and Fibroblast (각질형성세포와 섬유모세포 활성화에 미치는 대풍자 부탄올 분획의 효능)

  • Lee, Geum Seon;Lee, Ki Man;Yim, Dongsool;Cheong, Jae Hoon;Kang, Tae Jin
    • Korean Journal of Pharmacognosy
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    • v.46 no.1
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    • pp.59-64
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    • 2015
  • Wound healing is a complex process that includes inflammation, granulation tissue formation, re-epithelialization, and remodeling. We reported previously that BuOH fraction from Hydnocarpi Semen (HS) crude extract exhibited wound healing activity in animal ulcer models. In this study, we investigated whether BuOH fraction activates keratinocyte and fibroblast via wound closure test and migration assay. In the scratch test, BuOH fraction accelerated the closure of a monolayer wound scratch at $100{\mu}g/mL$. After treatment with BuOH fraction for 18 h, keratinocytes showed a increase in migration at $25{\mu}g/mL$, whereas the migration of fibroblast increased significantly at $100{\mu}g/mL$ of BuOH fraction compared to control. The mechanism that the BuOH fraction of HS helps to promote healing by inflammation is possibly associated with the migration of keratinocyte and fibroblast.

Therapeutic Effects of Amnion-Conjugated Chitosan-Alginate Membranes on Diabetic Wounds in an Induced Diabetic Swine Model: An In Vitro and In Vivo Study

  • Jeong, Woonhyeok;Hong, Jamin;Jung, Minho;Jang, Mijin;An, Sanghyun;Jo, Taehee;Kwon, Sunyoung;Son, Daegu
    • Archives of Plastic Surgery
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    • v.49 no.2
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    • pp.258-265
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    • 2022
  • Background Chitosan (CS) is a well-known antimicrobial dressing material. Moreover, widely used amniotic membranes contain growth factors beneficial for wound healing. Herein, we created a novel amnion-conjugated CS-alginate membrane dressing and tested its wound healing potency in a diabetic swine model. Methods The bovine amniotic powder growth factor contents were evaluated by protein assay, and the powder's wound healing effects were assessed in vitro by HaCaT cell scratch closure. In vivo, two minipigs developed streptozotocin-induced diabetes. Serial serum glucose measurements and intravenous glucose tolerance tests were performed to confirm their diabetic status. Twelve square-shaped wounds created on each pig's back were randomly divided into control (n = 4), CS (n = 4), and amnion-CS (AC; n = 4) groups and treated accordingly with different dressings. Wound healing in each group was assessed by measuring wound contraction over time, capturing wound perfusion with indocyanine green (ICG) angiography, and histologically analyzing inflammatory markers. Results Amniotic powder elution promoted HaCaT cell migration in the scratch wound model, suggesting its beneficial in vitro wound healing effects. In vivo, the CS and AC groups showed earlier wound contraction initiation and reepithelialization and earlier wound perfusion improvement by ICG angiography than the control group. Additionally, the wound size of the AC group at week 3 was significantly smaller than those in the control group. There was no significant difference in the numbers of acute and chronic inflammatory cells between the groups. Conclusion The amnion-conjugated CS-alginate membrane, as well as CS dressing alone, could be a favorable dressing option for diabetic wounds.

An In Vitro Study on the Effect of Fermented Citrus junos as a Cosmetic Material (화장품소재로서 유자발효물의 In Vitro 효능 연구)

  • Yeom, Hyun Sook;Cho, Seong Mi;Park, Jin Oh;Lee, Hae Kwang;Lee, Hye Ja
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.1
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    • pp.71-76
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    • 2022
  • In this study, we investigated the antioxidant activity, wound healing, moisturizing and anti-pollution effects of fermented Citrus junos (FCJ) with Lactobacillus rhamnosus as a cosmetic material. For the anti-oxidative activities, the FCJ showed potent DPPH radical scavenging activities. In addition, FCJ increased cell migration compared to the untreated group in scratch-induced wound healing assay. It was confirmed that the amount of filagrin (FLG), a moisturizing factor, increased in FCJ. FCJ prevented the decrease in cell viability, stimulated by PM10 at in vitro. Based on these results, it is believed that various effects of FCJ can provide a scientific basis for the development of cosmetic raw materials.

Experimental Studies on the Skin Barrier Improvement and Anti-inflammatory Activity based on a Bibliometric Network Map

  • Eunsoo Sohn;Sung Hyeok Kim;Chang Woo Ha;Sohee Jang;Jung Hun Choi;Hyo Yeon Son;Cheol-Joo Chae;Hyun Jung Koo;Eun-Hwa Sohn
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2023.04a
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    • pp.40-40
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    • 2023
  • Atopic dermatitis is a chronic inflammatory skin diseases caused by skin barrier dysfunction. Allium victoralis var. Platyphyllum (AVP) is a perennial plant used as vegetable and herbal medicine. The purpose of this study was to suggest that AVP is a new cosmetic material by examining the effects of AVP on the skin barrier and inflammatory response. A bibliometric network analysis was performed through keyword co-occurrence analysis by extracting author keyword from 69 articles retrieved from SCOPUS. We noted the anti-inflammatory activity shown by the results of clustering and mapping from network visualization analysis using VOSviewer software tool. HPLC-UV analysis showed that AVP contains 0.12 ± 0.02 mg/g of chlorogenic acid and 0.10 ± 0.01 mg/g of gallic acid. AVP at 100 ㎍/mL was shown to increase the mRNA levels of filaggrin and involucrin related to skin barrier function by 1.50-fold and 1.43-fold, respectively. In the scratch assay, AVP at concentrations of 100 ㎍/mL and 200 ㎍/mL significantly increased the cell migration rate and narrowed the scratch area. In addition, AVP suppressed the increase of inflammation-related factors COX-2 and NO and decreased the release of β-hexosaminidase. This study suggests that AVP can be developed as a functional cosmetic material for atopy management through skin barrier protection effects, anti-inflammatory and anti-itch effects.

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Artemisinin attenuates platelet-derived growth factor BB-induced migration of vascular smooth muscle cells

  • Lee, Kang Pa;Park, Eun-Seok;Kim, Dae-Eun;Park, In-Sik;Kim, Jin Tack;Hong, Heeok
    • Nutrition Research and Practice
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    • v.8 no.5
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    • pp.521-525
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    • 2014
  • BACKGROUND/OBJECTIVES: Artemisinin (AT), an active compound in Arternisia annua, is well known as an anti-malaria drug. It is also known to have several effects including anti-oxidant, anti-inflammation, and anti-cancer activities. To date, the effect of AT on vascular disorders has not been studied. In this study, we investigated the effects of AT on the migration and proliferation of vascular smooth muscle cells (VSMC) stimulated by platelet-derived growth factor BB (PDGF-BB). MATERIALS/METHODS: Aortic smooth muscle cells were isolated from Sprague-Dawley rats. PDGF-BB stimulated VSMC migration was measured by the scratch wound healing assay and the Boyden chamber assay. Cell viability was determined by using an EZ-Cytox Cell Viability Assay Kit. The production of reactive oxygen species (ROS) in PDGF-BB stimulated VSMC was measured through $H_2DCF$-DA staining. We also determined the expression levels of signal proteins relevant to ROS, including measures of extracellular signal-regulated kinase (ERK) 1/2 measured by western blot analysis and matrix metalloproteinase (MMP) 9 measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: AT ($10{\mu}M$ and $30{\mu}M$) significantly reduced the proliferation and migration of PDGF-BB stimulated VSMC in a dose-dependent manner. The production of ROS, normally induced by PDGF-BB, is reduced by treatment with AT at both concentrations. PDGF-BB stimulated VSMC treated with AT ($10{\mu}M$ and $30{\mu}M$) have reduced phosphorylation of ERK1/2 and inhibited MMP9 expression compared to untreated PDGF-BB stimulated VSMC. CONCLUSIONS: We suggest, based on these results, that AT may exert an anti-atherosclerotic effect on PDGF-BB stimulated VSMCs by inhibiting their proliferation and migration through down-regulation of ERK1/2 and MMP9 phosphorylation.

The Effects of Gokgisaeng on Anti-inflammation and Rat C6 Glioma Cell Migration (곡기생(槲寄生)의 항염증 효능 및 암세포 이주저해에 미치는 영향)

  • Kim, Hyun-Young;Jang, Soo-Young;Jeong, Ji-Cheon;Shin, Hyeon-Cheol
    • The Journal of Internal Korean Medicine
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    • v.34 no.1
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    • pp.31-45
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    • 2013
  • Objectives : Gokgisaeng (Korean mistletoe) is used for the treatment of inflammatory and cancer diseases in traditional Korean medicine and its major component lectins have been reported to induce nitric oxide (NO) in RAW 264.7 macrophages, and also induce apoptosis of various types of cancer cells, although its modulatory effects on cancer cell migration and macrophage activation is poorly understood. The aim of this study is to clarify molecular mechanisms of action responsible for the anti-inflammatory and antitumor migration potentials of Korean mistletoe extract (KME). Methods : We investigated the anti-inflammatory activity of KME on NO production and inducible nitric oxide synthase (iNOS) expression by lipopolysaccharide (LPS) in both RAW 264.7 macrophages and rat C6 glioma cells, and also evaluated inhibitory efficacy on glioma cell growth and migration. For assessment, XTT assay, nitrite assay, RT-PCR, scratch-wound and Boyden chamber assay, and western blot analysis were performed. Results : Previously reported, unlike the efficacy of Gokgisaeng lectin, KME inhibited NO production and iNOS expression, and suppressed pro-inflammatory mediators including IL-$1{\beta}$, IL-6, COX-2, iNOS in LPS-stimulated RAW 264.7 cells. Furthermore, KME suppressed tumor cell growth and migration, and it also inhibited LPS-induced NO release and iNOS activation by down-regulating expression of protein kinase C (PKC) and phosphorylation of ERK in C6 glioma cells. Conclusions : Our research findings provide evidence that KME can play a significant role in blocking pro-inflammatory reaction and malignant progression of tumors through the suppression of NO/iNOS by down-regulating of inflammatory signaling pathways, PKC/ERK.