• 제목/요약/키워드: a wound assay

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The Anti-Migratory Effect of Cirsium japonicum Pharmacopuncture in C6 Glioma Cell (대계 약침액의 C6 신경교종 세포에 대한 이주 억제 효과)

  • Park, Juyeon;Lee, Kangpa;Chang, Haeryong;Moon, Jinyoung
    • Korean Journal of Acupuncture
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    • v.30 no.4
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    • pp.212-219
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    • 2013
  • Objectives : Cirsium japonicum is a traditional Korean medicine that has been used in the treatment of inflammatory diseases such as appendicitis, hepatitis, pulmonary abscess and tumor. The aim of study was to elucidate anti-migratory activity of CJP(Cirsium japonicum pharmacopuncture) through regulation of inflammatory mediators in C6 glioma cell. Methods : Nitric oxide(NO) production was determined by using nitrite assay. The cell migration was analyzed by wound-healing assay and Boyden chamber assay. The expression levels of iNOS, and protein kinase C(PKC)-${\alpha}$ were measured by western blotting assay. Results : CJP showed a significant decrease on NO production. Moreover, glioma cell migration was effectively suppressed by CJP. Furthermore, CJP inhibited the expressions of iNOS and PKC-${\alpha}$ in C6 glioma cells. Conclusions : These results suggest that CJP inhibits glioma cell migration and iNOS expression through regulation of PKC-${\alpha}$. Therefore, it is expected that CJP could be an effective agents for blocking malignant progression of glioma.

Visible light-cured glycol chitosan hydrogel dressing containing endothelial growth factor and basic fibroblast growth factor accelerates wound healing in vivo

  • Yoo, Youngbum;Hyun, Hoon;Yoon, Sun-Jung;Kim, So Yeon;Lee, Deok-Won;Um, Sewook;Hong, Sung Ok;Yang, Dae Hyeok
    • Journal of Industrial and Engineering Chemistry
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    • v.67
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    • pp.365-372
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    • 2018
  • Wounds that heal with excessive scar formation result in poor functional and aesthetic outcomes. To address this, in our study, visible light cured glycol chitosan (GCH) hydrogels containing endothelial growth factor (EGF) and basic fibroblast growth factor (bFGF) were prepared (GCH-EGF, GCH-FGF and GCH-EGF/FGF) and evaluated their efficacies on the improvement of wound healing in vivo. In vitro release test showed that the growth factors were released in a sustained manner along with initial burst for 24 h. In vitro cell proliferation assay of L-929 mouse fibroblast cell line resulted in the superior ability of GCH-EGF/FGF on the rate. In vivo results demonstrated that the growth factor loaded GCHs further enhanced wound healing compared with GCH. In particular, GCH-EGF/EFG showed the most remarkable wound healing effect among the samples.

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.

Keratinocyte Migration in a Three-Dimensional In Vitro Wound Healing Model Co-Cultured with Fibroblasts

  • Iyer, Kritika;Chen, Zhuo;Ganapa, Teja;Wu, Benjamin M.;Tawil, Bill;Linsley, Chase S.
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.721-733
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    • 2018
  • BACKGROUND: Because three-dimensional (3D) models more closely mimic native tissues, one of the goals of 3D in vitro tissue models is to aid in the development and toxicity screening of new drug therapies. In this study, a 3D skin wound healing model comprising of a collagen type I construct with fibrin-filled defects was developed. METHODS: Optical imaging was used to measure keratinocyte migration in the presence of fibroblasts over 7 days onto the fibrin-filled defects. Additionally, cell viability and growth of fibroblasts and keratinocytes was measured using the $alamarBlue^{(R)}$ assay and changes in the mechanical stiffness of the 3D construct was monitored using compressive indentation testing. RESULTS: Keratinocyte migration rate was significantly increased in the presence of fibroblasts with the cells reaching the center of the defect as early as day 3 in the co-culture constructs compared to day 7 for the control keratinocyte monoculture constructs. Additionally, constructs with the greatest rate of keratinocyte migration had reduced cell growth. When fibroblasts were cultured alone in the wound healing construct, there was a 1.3 to 3.4-fold increase in cell growth and a 1.2 to 1.4-fold increase in cell growth for keratinocyte monocultures. However, co-culture constructs exhibited no significant growth over 7 days. Finally, mechanical testing showed that fibroblasts and keratinocytes had varying effects on matrix stiffness with fibroblasts degrading the constructs while keratinocytes increased the construct's stiffness. CONCLUSION: This 3D in vitro wound healing model is a step towards developing a mimetic construct that recapitulates the complex microenvironment of healing wounds and could aid in the early studies of novel therapeutics that promote migration and proliferation of epithelial cells.

Effect of LEDs Light of 633 nm Wavelength in Skin of Organism (633 nm 파장의 LED 광원이 생체 피부에 미치는 영향)

  • Cheon, Min-Woo
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.21 no.8
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    • pp.760-765
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    • 2008
  • Low power laser therapy is internationally certified and is known to be effective in stimulating DNA in living organisms, increasing protein synthesis and activating cell division, smoothing blood circulation, promoting cell activation, cell regeneration and function. It also has anti-inflammatory, anti-edemic, anti-fibrous dysplastic and neuralogic hyperfunctional effects. This study was intended to verify the effect of LED irradiation therapy on wound healing in cell and animal tests by applying LED irradiator using a laser and laser diode, which was independently designed and developed to emit beams of similar wavelength to that of a laser. This equipment was fabricated using a micro-controller and a high brightness LED, and designed to enable us to control light irradiation time, intensity and reservation. In case of cell proliferation experiment, each experiment was performed to irradiation group and non-irradiation group for tissue cells. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590 nm transmittance of micro-plate reader. In the wound healing experiment, 1$cm^2$ wounds on the skin wound of SD-Rat(Sprague-Dawley Rat) were made. Light irradiation group and none light irradiation group divided, each group was irradiated one hour a day for 9 days. As a result, the cell increase of tissue cells was verified in irradiation group as compared to non-irradiation group. And, compared with none light irradiation group, the lower incidence of inflammation and faster recovery was shown in light irradiation group.

Effect of Oncostatin M on Proliferation and Matrix Synthesis of Dermal Fibroblasts (Oncostatin M이 피부섬유모세포의 증식과 기질생성에 미치는 영향)

  • Chun, Kyung Wook;Lim, Hyung Woo;Han, Seung-Kyu;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.35 no.2
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    • pp.115-120
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    • 2008
  • Purpose: Oncostatin M(OSM) is a multifunctional cytokine that belongs to the interleukin(IL)-6 family. Although there have been a number of studies that focused on the role and mechanism of OSM in various organs and tissues, there are few reports on its effect on wound healing. The final purpose of this project is to evaluate the effect of OSM on wound healing. This pilot study was designed to investigate the effect of OSM on proliferation and matrix synthesis of human dermal fibroblasts, which are the major components of the wound healing. Methods: Excess skin that was obtained from patients who underwent skin grafts, was used for this study. From this material, fibroblasts were isolated and cultured. The cultured fibroblasts were treated with one of four concentrations of OSM. The OSM concentrations used were 0, 50, 100, and 200 ng/ml, respectively. After the OSM treatment, cell proliferation was determined by the MTT assay, collagen synthesis by the C1CP method, GAG levels by the Blyscan Dye method. The parameter levels of each group were compared. Results: OSM treatment increased all the components tested in the study. In particular, cell proliferation, GAG synthesis demonstrated statistically significant increases(p<0.05 in the Mann-Whitney U-test). The highest increase in all the components was obtained at a 100 ng/ml concentration of OSM.Conclusion: The results of the present study indicate that OSM stimulates proliferation and matrix synthesis of human dermal fibroblast and the optimal concentration for wound healing is 100 ng/mL.

Wound Healing Activity of Gamma-Aminobutyric Acid (GABA) in Rats

  • Han, Dong-Oh;Kim, Hee-Young;Lee, Hye-Jung;Shim, In-Sop;Hahm, Dae-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1661-1669
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    • 2007
  • Gamma-aminobutyric acid (GABA) is a non-protein amino acid. It is well known for its role as an inhibitory neurotransmitter of developing and operating nervous systems in brains. In this study, a novel function of GABA in the healing process of cutaneous wounds was presented regarding anti-inflammation and fibroblast cell proliferation. The cell proliferation activity of GABA was verified through an MTT assay using murine fibroblast NIH3T3 cells. It was observed that GABA significantly inhibited the mRNA expression of iNOS, IL-$1{\beta}$, and TNF-${\alpha}$ in LPS-stimulated RAW 264.7 cells. To evaluate in vivo activity of GABA in wound healing, excisional open wounds were made on the dorsal sides of Sprague-Dawley rats under anesthesia, and the healing of the wounds was apparently assessed. The molecular aspects of the healing process were also investigated by hematoxylineosin staining of the healed skin, displaying the degrees of re-epithelialization and linear alignment of the granulation tissue, and immunostaining and RT-PCR analyses of fibroblast growth factor and platelet-derived growth factor, implying extracellular matrix synthesis and remodeling of the skin. The GABA treatment was effective to accelerate the healing process by suppressing inflammation and stimulating re-epithelialization, compared with the epidermal growth factor treatment. The healing effect of GABA was remarkable at the early stage of wound healing, which resulted in significant reduction of the whole healing period.

(-)-Epigallocatechin-3-Gallate Induces Apoptosis and Inhibits Invasion and Migration of Human Cervical Cancer Cells

  • Sharma, Chhavi;Nusri, Qurrat El-Ain;Begum, Salema;Javed, Elham;Rizvi, Tahir A.;Hussain, Arif
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4815-4822
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    • 2012
  • Invasion and metastasis are the major causes of cancer-related death. Pharmacological or therapeutic interventions such as chemoprevention of the progression stages of neoplastic development could result in substantial reduction in the incidence of cancer mortality. (-)-Epigallocatechin-3-gallate (EGCG), a promising chemopreventive agent, has attracted extensive interest for cancer therapy utilizing its antioxidant, anti-proliferative and inhibitory effects on angiogenesis and tumor cell invasion. In this study, we assessed the influence of EGCG on the proliferative potential of HeLa cells by cell viability assay and authenticated the results by nuclear morphological examination, DNA laddering assay and cell cycle analysis. Further we analyzed the anti-invasive properties of EGCG by wound migration assay and gene expression of MMP-9 and TIMP-1 in HeLa cells. Our results indicated that EGCG induced growth inhibition of HeLa cells in a dose- and time-dependent manner. It was observed that cell death mediated by EGCG was through apoptosis. Interestingly, EGCG effectively inhibited invasion and migration of HeLa cells and modulated the expression of related genes (MMP-9 and TIMP-1). These results indicate that EGCG may effectively suppress promotion and progression stages of cervical cancer development.

Apoptosis induced by water extracts of Nypa fruticans wurmb via a mitochondria-dependent pathway in human FaDu hypopharyngeal squamous carcinoma cells

  • Lee, Seul Ah;Choi, Mi Suk;Park, Bo-Ram;Kim, Jin-Soo;Kim, Chun Sung
    • International Journal of Oral Biology
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    • v.46 no.4
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    • pp.160-167
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    • 2021
  • Nypa fruticans Wurmb (NFW) contains a large amount of phenolic acid and flavonoids, and is popular as a superfood in Myanmar. NFW has various biological activities, such as anti-inflammatory, anti-oxidant, and neuroprotective properties; however, the anti-cancer effect of NFW have not been reported. In this study, we investigated the anticancer activity of water extracts of NFW (WeNFW) and the underlying mechanism in human FaDu hypopharyngeal squamous carcinoma cells. The WeNFW inhibited FaDu cell growth in a dose-dependent manner without affecting normal cells (L929), as determined by an MTT assay and Live and Dead assay. In addition, the concentrations of WeNFW without cytotoxicity (0.025, 0.05, and 0.1 mg/mL) inhibited wound healing and colony formation. Furthermore, WeNFW significantly induced apoptosis through the proteolytic cleavage of caspase-3 and -9, poly (ADP-ribose) polymerase, and downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by DAPI staining, FACS analysis, and western blot analysis. Taken together, these results suggest that WeNFW exhibits potent anti-cancer effects by suppressing the growth of oral cancer cells, wound healing and colony formation activity. Via mitrochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, WeNFW can provide a natural chemotherapeutic drug for oral cancer in humans.

Effect of lipoaspirate cell autograft on proliferation and collagen synthesis of diabetic fibroblasts in vitro (지방기질세포 치료가 당뇨섬유아세포의 증식과 교원질합성에 미치는 영향)

  • Song, Sun Ho;Han, Seung Kyu;Chun, Kyung Wook;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.36 no.6
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    • pp.679-684
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    • 2009
  • Purpose: Human lipoaspirate cells are relatively easy to obtain in large quantities without cell culture. The aim of this in vitro pilot study was to determine the effect of cell therapy using uncultured lipoaspirate cells on cell proliferation and collagen synthesis of diabetic fibroblasts, which are the major contributing factors in wound healing. Methods: In order to get diabetic fibroblasts, dermis tissues were obtained from foot skin of diabetic patients who underwent debridements or toe amputations(n = 4). In order to isolate lipoaspirate cells, the same diabetic patients' abdominal adipose tissues were obtained by liposuction. The diabetic fibroblasts were co - cultured with or without autogenous lipoaspirate cells using porous culture plate insert. Initial numbers of the lipoaspirate cells and diabetic fibroblasts seeded were 15,000 cells/well, respectively. For cell proliferation assay, two treatment groups were included. In group I, diabetic fibroblasts were cultured with the insert having no cells, which serves as a control. In group II, the lipoaspirate cells were added in the culture plate insert. For collagen synthesis assay, one additional group(group III), in which diabetic fibroblasts were not seeded in the well and only lipoaspirate cells inside the insert were incubated without diabetic fibroblasts, was included for a reference. Results: One hundred to one hundred sixty thousand lipoaspirate cells were isolated per ml of aspirated adipose tissue. After 3 - day incubation, the mean cell numbers in group I and II were 17,294/well and 22,163/well. The mean collagen level in group I, II, and III were 29, 41, and 2 ng/ml, respectively. These results imply that both cell proliferation and collagen synthesis in the lipoaspirate cell treatment group were 28 and 44 percents higher than in the control group, respectively(p < 0.05). Conclusion: Uncultured lipoaspirate cell autografts may stimulate the wound healing activity of diabetic fibroblasts.