• Title/Summary/Keyword: Wound healing assay

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Cell Migratory Induction by Expression of Angiogenin and Vascular Endothelial Growth Factor in Resveratrol Treated HeLa Cells (Resveratrol 처리한 HeLa세포에서 angiogenin과 vascular endothelial growth factor 발현유도에 따른 세포이동촉진)

  • Joe, I-Seul;Jeong, Sin-Gu;Cho, Goang-Won
    • Journal of Life Science
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    • v.24 no.4
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    • pp.337-342
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    • 2014
  • Resveratrol (RSV), a natural polyphenolic compound, is a modulator for cell division and cell migration, and has diverse beneficial properties. Angiogenin (ANG) and vascular endothelial growth factor (VEGF) are considered to be important mechanisms for cell proliferation, angiogenesis, the formation of tubular structures, and migration. In this study, we investigated whether RSV has a migratory effect in HeLa cells. When cells were treated with $0{\sim}50{\mu}M$ of RSV for 24 hr, the expression of ANG and VEGF was significantly increased in a dose dependent manner measured by real-time PCR. Similarly, we performed time dependent experiments for $50{\mu}M$ RSV treated cells and identified the optimized time at 24 hr. The increased expression in RSV treated cells was confirmed by Western blot analysis. To examine the toxic effects of RSV at the determined conditions, MTT assays were performed. The viabilities were unchanged for $0{\sim}50{\mu}M$ RSV treated cells, while they decreased at $100{\mu}M$ RSV. To examine the effect of migration in RSV treated cells, we performed a wound-healing assay. The migratory rates were significantly enhanced in the RSV treated group. In this study, we found that RSV induces an increase in the expression of migration factors ANG, VEGF, and enhances cell migration for the determined conditions.

Optimal Concentration of Thrombin to Activate Platelet for Wound Healing (창상치유 목적의 혈소판 치료를 위한 Thrombin의 최적 농도)

  • Eum, Soo Jin;Han, Seung Kyu;Chun, Kyung Wook;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.36 no.1
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    • pp.19-23
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    • 2009
  • Purpose: Platelet transplantation is a novel therapeutic strategy for acceleration of wound healing. When applying platelets, efficacy of adding thrombin to stimulate growth factor release from platelets has already been proved. However, no quantitative data of the thrombin treatment has been reported. The purpose of this study was to determine the optimal thrombin concentration to maximize growth factor release of platelets. In particular, this study was designed to quantify levels of platelet derived growth factor(PDGF)-BB, which is a major growth factor contained in the platelets, in vitro. Methods: Fresh platelets were obtained from a blood bank. They were suspended in DMEM/F - 12 and incubated with thrombin of various concentrations. The concentrations of thrombin tested were 0, 6.25, 12.5, 25, 50, 100, 200, and 400 IU/ml. After 30 minutes, 1, 3, 5, and 7 days, the levels of PDGF - BB were measured using enzyme linked immunosorbent assay. Platelets from four donors were included in this study. Each sample was tested in triplicate and the mean value was used as a data for each sample. Results: The addition of thrombin increased the level of PDGF - BB. Increases in storage time of platelets resulted in decreased levels of PDGF - BB. Higher levels of PDGF were detected in consort with increased thrombin concentrations. However, there was no significant difference between samples of 200 and 400 IU/ml concentrations. Conclusion: The results indicate that adding thrombin accelerates the release of growth factors from platelets and the optimal thrombin concentration to maximize this function is 200 IU/ml.

Effect of Progesterone on Cultured Human Dermal Fibroblast (배양된 인체진피섬유모세포의 증식에 대한 황체호르몬의 영향)

  • Kwon, Soon Sung;Oh, Myung June;Lee, Jin Hee;Park, Jong Lim;Chang, Hak;Minn, Kyung Won
    • Archives of Plastic Surgery
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    • v.34 no.4
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    • pp.420-425
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    • 2007
  • Purpose:The mechanism of scar formation is not fully understood. Fibroblast is an important cell in wound healing process. We experienced a patient who was taking progesterone orally. Upper blepharoplasty was performed on her but, wound healing was delayed. We hypothesized that progesterone was the cause of delayed wound healing and fibroblast proliferation inhibition. We investigated the effect of progesterone in vitro on human dermal fibroblasts to study the effects on fibroblast proliferation. Methods: Human dermal fibroblasts from four persons were cultured initially. Progesterone is mixed to them at various concentrations, and fibroblast cell count was measured by MTT assay method at 570 nm. We confirmed that progesterone has some inhibitory effect on fibroblast proliferation and maximal inhibitory concentration of progesterone was determined. Then fibroblasts from a total of nineteen persons were cultured and the effects of progesterone were studied. Results: The initial study showed the maximal inhibitory concentration of progesterone to be $50{\mu}g/ml$. The main study showed that progesterone had 70.9% inhibitory effect on human dermal fibroblast in vitro. Conclusion: Progesterone has inhibitory effect on cultured human dermal fibroblast proliferation in vitro.

Anti-inflammatory effects of low-level laser in burn wound models in rats

  • Kim, Eun-Jeong;Kim, Se-Hun
    • Physical Therapy Rehabilitation Science
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    • v.6 no.4
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    • pp.170-175
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    • 2017
  • Objective: The anti-inflammatory effects of low-level laser in burn wound model in rats were investigated. Design: Randomized controlled trial. Methods: The rats were assigned to three experimental groups. Group I received second-degree burn wounds; Group II received dressing film and low-level laser ($1.2J/cm^2$) treatment after a burn wound; Group III received dressing film and low-level laser ($2.3J/cm^2$) treatment after a burn wound. After inducing a deep second-degree burn wound, the wound was observed every day and the burn area diameter and retraction quantification at 1, 7, and 14 days were evaluated. Low-level laser was investigated on hematological parameters after 14 days. Effects of low-level laser on the inflammatory cytokines (tumor necrosis $factor-{\alpha}$ [$TNF-{\alpha}$] and interleukin-6 [IL-6]) concentrations in the serum were evaluated using immunosorbent assay kits. Results: Group III showed a significant difference in wound size on days 7 and 14 compared to Group I (p<0.05). Group II showed a significant difference in wound size on day 14 compared to Group I (p<0.05). For wound contraction percentage, both laser therapy treatment groups showed a significant difference compared with Group I (p<0.05). There was also a significant difference in wound contraction percentage in Group III compared to Group II (p<0.05). Compared with the model control group, decreased $TNF-{\alpha}$ and IL-6 levels in the serum was observed at 14 days after burn wound induction. Conclusions: The results of this study suggest that low-level laser therapy can assist in burn wound healing, which might be associated with decreased concentrations of $TNF-{\alpha}$ and IL-6 related proinflammatory cytokines.

Aqueous Extract of Anticancer Drug CRUEL Herbomineral Formulation Capsules Exerts Anti-proliferative Effects in Renal Cell Carcinoma Cell Lines

  • Verma, Shiv Prakash;Sisoudiya, Saumya;Das, Parimal
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8419-8423
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    • 2016
  • Purpose: Anti-cancer activity evaluation of aqueous extract of CRUEL (herbomineral formulation) capsules on renal cell carcinoma cell lines, and exploration of mechanisms of cell death. Materials and Methods: To detect the cytotoxic dose concentration in renal cell carcinoma (RCC) cells, MTT assays were performed and morphological changes after treatment were observed by inverted microscopy. Drug effects against RCC cell lines were assessed with reference to cell cycle distribution (flow cytometry), anti-metastatic potential (wound healing assay) and autophagy(RT-PCR). Results: CRUEL showed anti-proliferative effects against RCC tumor cell lines with an IC50 value of ${\approx}4mg/mL$ in vitro., while inducing cell cycle arrest at S-phase of cell cycle and inhibiting wound healing. LC3 was found to be up-regulated after drug treatment in RT-PCR resulting in an autophagy mode of cell death. Conclusions: This study provides the experimental validation for antitumor activity of CRUEL.

Variable Effect of Estrogen on Fibroblast Proliferation and Collagen Synthesis by Gender and Age (에스트로겐이 진피섬유아세포의 증식 및 교원질합성에 미치는 영향의 다양성)

  • Shin, Seung Han;Won, Chang Hoon;Han, Seung Kyu;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.32 no.3
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    • pp.363-368
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    • 2005
  • It was assumed that the effect of estrogen on wound healing would be variable according to patient's gender and age since estrogen is a sex steroid. This study was designed to determine the variability of the effect of estrogen on proliferation of human dermal fibroblasts and collagen synthesis which are most important in wound healing considering patient's gender and age. Fibroblasts were isolated from the dermis of female patients in premenstrual, menstrual, or postmenopausal age group and that of male patients. The isolated fibroblasts were cultivated in the presence of estrogen($1.0{\mu}g/ml$). The cells were seeded at $5.0{\times}10^3cell/well$ in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 5% fetal bovine serum in 96-well plates. The cells were incubated for 3 days. For fibroblast proliferation MTT assay method was used. To measure the production of collagen, the collagen type I carboxy- terminal propeptide enzyme immunoassay was carried out. Estrogen stimulated the proliferation of fibroblasts in female patients, but not in male patients. The greatest cell proliferation and collagen synthesis was seen at women in menstrual and postmenopausal age. These results demonstrated that effects of estrogen on dermal fibroblast proliferation and collagen synthesis were variable with gender and age.

Antineoplastic Effect of Several Herbal Medicine Mixtures on SNU-80 Anaplastic Thyroid Carcinoma Cell Line (수종 한약 복합물의 역형성갑상선암세포 SNU-80에 대한 항암효과)

  • Yeo, Hyun-Soo;Lee, Min-Hye;Choi, You-Kyung;Jun, Chan-Young;Park, Jong-Hyeong
    • The Journal of Internal Korean Medicine
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    • v.35 no.4
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    • pp.416-427
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    • 2014
  • Objectives: The purpose of this study was to investigate the antineoplastic effect of several herbal medicine mixtures (compositions of Astragalus membranaceu, Angelica gigas, Trichosanthes kirilowii, Panax ginseng, Rhus verniciflua Stokes) on the SNU-80 anaplastic thyroid carcinoma cell line. Methods: MTT assay was used to examine whether our herbal medicine mixtures decreased cell growth rate of SNU-80. Wound healing assay and Transwell invasion assay was performed to investigate whether our herbal medicine mixtures affect the migration and invasion of anaplastic cancer cells, SNU-80. ELISA assay was performed to know if our herbal medicine mixtures suppressed the expression of pro-invasive molecules, such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) secreted from SNU-80. Results: MTT assay demonstrated that A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas :T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 strongly suppressed the growth of SNU-80. Wound healing assay demonstrated that A. membranaceus:A. gigas=3:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the migration of SNU-80. Transwell invasion assay demonstrated that A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii =1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng :R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the invasion of SNU-80. ELISA assay demonstrated that A. membranaceus :A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 suppressed the expression of VEGF. Also, A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus :A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes =1:1:1:1:1 or 3:1:1:1:1 suppressed the expression of MMP-2. Conclusions: The results obtained in this study suggest that several herbal medicine mixtures suppresse the growth and inhibit the migration and invasion of SNU-80, which is anaplastic thyroid cancer cells. Especially, A. membranaceus:A. gigas: T. kirilowii=1:1:1 mixture had a stronger anti-cancer effect.

Antineoplastic Effect of Several Herbal Medicines on SNU-80 Anaplastic Thyroid Carcinoma Cell Line (황기, 당귀, 칠피, 천화분의 역형성갑상선암세포 SNU-80에 대한 항암효과)

  • Yeo, Hyun Soo;Lee, Min Hye;Ko, Seong Gyu;Choi, You Kyung;Jun, Chan Young;Park, Jong Hyeong
    • Journal of Society of Preventive Korean Medicine
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    • v.18 no.1
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    • pp.83-92
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    • 2014
  • Objective : This study was performed to investigate the antineoplastic effect of Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes and Trichosanthes kirilowii on SNU-80 anaplastic thyroid carcinoma cell line. Method : We examined whether our herbal medicines decreases cell growth rate of SNU-80 using MTT assay. We performed western blot analysis to verify that our herbal medicines induces apoptosis via caspase-dependent mechanism. We also performed wound healing assay and transwell invasion assay to investigate whether our herbal medicines affects the migration and invasion of anaplastic cancer cells, SNU-80. We also carried out ELISA assay to know our herbal medicines suppresses the expression of proinvasive molecules, such as VEGF and MMP-2 secreted from SNU-80. Results : MTT assay demonstrates that Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed strongly the growth of SNU-80. Western blot analysis demonstrates that Trichosanthes kirilowii induces apoptosis activating the cleavages of caspases (caspase-8, caspase-3) and PARP. Wound healing assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the migration of SNU-80. Transwell invasion assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the invasion of SNU-80. Elisa assay demonstrates that Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed the expression of VEGF and MMP-2. Conclusion : We could conclude that several herbal medicines suppresses the growth and inhibits the migration and invasion of SNU-80 which is anaplastic thyroid cancer cells. Especially, Rhus verniciflua Stokes, Trichosanthes kirilowii had stronger anti-cancer effect suggesting that we can apply them to treat anaplastic thyroid cancer.

Analysis of Immunomodulating Activities in Methanol Extracts from Several Kinds of Grapes (포도 품종별 메탄올 추출물로부터 면역활성 분석)

  • Heo, Jin-Chul;Woo, Sang-Uk;Kweon, Mi-Ae;Kim, Bo-Bae;Lee, Sook-Hee;Lee, Jin-Man;Choi, Jong-Uck;Chung, Shin-Kyo;Lee, Sang-Han
    • Food Science and Preservation
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    • v.14 no.4
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    • pp.419-424
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    • 2007
  • Fruits and vegetable extracts are well-known as healthy foods. Such foods have been used as herbal medicines or traditional therapies for centuries. To assess biological activities in grapes, we examined the immunomodulating activities of water extracts from four kinds of grapes (cultivars Kyoho, Delaware, Campbell, and Niagara). We explored possible antioxidant and anticancer activities using antioxidant assays such as the 1,1-diphenyl-2-picrylhydrazyl (DPPH) reduction assay, the ferric iron reducing ability of plasma (FRAP) assay, a cell proliferation assay, an NO inhibition assay, a wound healing assay, and an IL-4/IL-13 elicitation assay. Methanol extracts of grapes were tested. The results showed that each grape extract had potent antioxidant activities. The grape extracts increased cell proliferation and NO production activities in tumor cell lines. IL-4 and IL-13 cytokine levels were decreased in mouse primary spleen cells by treatment with any extract. These results suggest that grape extracts can be used as biomaterials with immunomodulating activities.

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.