scholarly journals An on-chip wound healing assay fabricated by xurography for evaluation of dermal fibroblast cell migration and wound closure

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ghazal Shabestani Monfared ◽  
Peter Ertl ◽  
Mario Rothbauer
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Wound Closure ◽  
Dermal Fibroblast ◽  
Lab On A Chip ◽  
Fibroblast Cell ◽  
Serum Starvation ◽  
Circular Membrane ◽  
Wound Healing Assay ◽  
On Chip

Abstract Dermal fibroblast cell migration is a key process in a physiological wound healing. Therefore, the analysis of cell migration is crucial for wound healing research. In this study, lab-on-a-chip technology was used to investigate the effects of basic fibroblast growth factor (bFGF), mitomycin C (MMC), MEK1/2 inhibitor (U0126) and fetal calf serum (FCS) on human dermal fibroblast cell migration. The microdevice was fabricated consisting of microchannels, pneumatic lines and pneumatically-activated actuators by xurographic rapid prototyping. In contrast to current approaches in in vitro wound healing such as scratch assays and silicone inserts in wellplate format, which show high variability and poor reproducibility, the current system aims to automate the wounding procedure at high precision and reproducibility using lab-on-a-chip. Traumatic wounding was simulated on-chip on fibroblast cell monolayers by applying air pressure on the flexible circular membrane actuator. Wound closure was monitored using light microscopy and cell migration was evaluated using image analysis. The pneumatically controlled system generates highly reproducible wound sizes compared to the conventional wound healing assay. As proof-of-principle study wound healing was investigated in the presence of several stimulatory and inhibitory substances and culture including bFGF, MMC, U0126 MEK1/2 inhibitor as well as serum starvation to demonstrate the broad applicability of the proposed miniaturized culture microsystem.

A Method for Quantitative Analysis of the Kinematics of Fibroblast Migration in a Monolayer Wound Model

2011 ◽  
Author(s):  
Gil Topman ◽  
Orna Sharabani-Yosef ◽  
Amit Gefen
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Wound Healing Assay ◽  
Complete Coverage ◽  
Time Intervals ◽  
Fibroblast Migration ◽  
Wound Model ◽  
Regular Time

A wound healing assay is simple but effective method to study cell migration in vitro. Cell migration in vitro was found to mimic migration in vivo to some extent [1,2]. In wound healing assays, a “wound” is created by either scraping or mechanically crushing cells in a monolayer, thereby forming a denuded area. Cells migrate into the denuded area to complete coverage, and thereby “heal” the wound. Micrographs at regular time intervals are captured during such experiments for analysis of the process of migration.

Wound Healing Assay for Melanoma Cell Migration

2021 ◽  
pp. 65-71
Author(s):  
Juliano T. Freitas ◽  
Ivan Jozic ◽  
Barbara Bedogni
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Melanoma Cell ◽  
Wound Healing Assay

scholarly journals Wound Healing Potential of Spirulina Protein on CCD-986sk Cells

Marine Drugs ◽  
2019 ◽  
Vol 17 (2) ◽  
pp. 130
Author(s):  
Ping Liu ◽  
Jeong-Wook Choi ◽  
Min-Kyeong Lee ◽  
Youn-Hee Choi ◽  
Taek-Jeong Nam
Keyword(s):  
Wound Healing ◽  
Dermal Fibroblast ◽  
Fibroblast Cell ◽  
Underlying Mechanism ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Cyclin Dependent Kinase ◽  
Pi3k Inhibitor Ly294002 ◽  
And Migration ◽  
Proliferation And Migration

Wound healing is a dynamic and complex process. The proliferation and migration of dermal fibroblasts are crucial for wound healing. Recent studies have indicated that the extracts from Spirulina platensis have a positive potential for wound healing. However, its underlying mechanism is not fully understood. Our previous study showed that spirulina crude protein (SPCP) promoted the viability of human dermal fibroblast cell line (CCD-986sk cells). In this study, we further investigated the wound healing effect and corresponding mechanisms of SPCP on CCD-986sk cells. Bromodeoxyuridine (BrdU) assay showed that SPCP promoted the proliferation of CCD-986sk cells. The wound healing assay showed that SPCP promoted the migration of CCD-986sk cells. Furthermore, cell cycle analysis demonstrated that SPCP promoted CCD-986sk cells to enter S and G2/M phases from G0/G1 phase. Western blot results showed that SPCP significantly upregulated the expression of cyclin D1, cyclin E, cyclin-dependent kinase 2 (Cdk2), cyclin-dependent kinase 4 (Cdk4), and cyclin-dependent kinase 6 (Cdk6), as well as inhibited the expression of CDK inhibitors p21 and p27 in CCD-986sk cells. In the meanwhile, SPCP promoted the phosphorylation and activation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt). However, the phosphorylation of Akt was significantly blocked by PI3K inhibitor (LY294002), which in turn reduced the SPCP-induced proliferation and migration of CCD-986sk cells. Therefore, the results presenting in this study suggested that SPCP can promote the proliferation and migration of CCD-986sk cells; the PI3K/Akt signaling pathway play a positive and important role in these processes.

scholarly journals A Novel In Vitro Wound Healing Assay to Evaluate Cell Migration

10.3791/56825 ◽  
2018 ◽  
Author(s):  
Floriana Cappiello ◽  
Bruno Casciaro ◽  
Maria Luisa Mangoni
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Wound Healing Assay

scholarly journals Creation of cloned pig embryos using contact-inhibited or serum-starved fibroblast cells analysed intravitam for apoptosis occurrence / Uzyskiwanie klonalnych zarodków świni z wykorzystaniem komórek fibroblastycznych poddanych inhibicji kontaktowej lub deprywacji troficznej oraz analizowanych przyżyciowo w kierunku apoptozy

2013 ◽  
Vol 13 (2) ◽  
pp. 275-293 ◽  
Author(s):  
Marcin Samiec ◽  
Maria Skrzyszowska ◽  
Jolanta Opiela
Keyword(s):  
Dermal Fibroblast ◽  
Somatic Cells ◽  
Fibroblast Cell ◽  
Serum Starvation ◽  
Fibroblast Cells ◽  
Blastocyst Formation ◽  
Cell Nuclei ◽  
Donor Cells ◽  
Cloned Pig

Abstract Somatic cell cloning efficiency is determined by many factors. One of the most important factors is the structure-functional quality of nuclear donor cells. Morphologic criteria that have been used to date for qualitative evaluation of somatic cells may be insufficient for practical application in the cloning. Biochemical and biophysical changes that are one of the earliest symptoms in the transduction of apoptotic signal may be not reflected in the morphologic changes of somatic cells. For this reason, adult cutaneous or foetal fibroblast cells that, in our experiments, provided the source of genomic DNA for the cloning procedure had been previously analysed for biochemical and biophysical proapoptotic alterations with the use of live-DNA (YO-PRO-1) and plasma membrane (Annexin V-eGFP) fluorescent markers. In Groups IA and IB, the generation of nucleartransferred (NT) embryos using non-apoptotic/non-necrotic contact-inhibited or serum-starved adult cutaneous fibroblast cells yielded the morula and blastocyst formation rates of 125/231 (54.1%) and 68/231 (29.4%) or 99/237 (41.8%) and 43/237 (18.1%), respectively. In Groups IIA and IIB, the frequencies of embryos reconstituted with non-apoptotic/non-necrotic contact-inhibited or serum-starved foetal fibroblast cell nuclei that reached the morula and blastocyst stages were 171/245 (69.8%) and 97/245 (39.6%) or 132/227 (58.1%) and 63/227 (27.8%), respectively. In conclusion, contact inhibition of migration and proliferative activity among the subpopulations of adult dermal fibroblast cells and foetal fibroblast cells resulted in considerably higher morula and blastocyst formation rates of in vitro cultured cloned pig embryos compared to serum starvation of either type of fibroblast cell line. Moreover, irrespective of the methods applied to artificially synchronize the mitotic cycle of nuclear donor cells at the G0/G1 phases, developmental abilities to reach the morula/blastocyst stages were significantly higher for porcine NT embryos that had been reconstructed with non-apoptotic/non-necrotic foetal fibroblast cells than those for NT embryos that had been reconstructed with non-apoptotic/non-necrotic adult dermal fibroblast cells. To our knowledge, the generation of cloned pig embryos using abattoir-derived oocytes receiving cell nuclei descended from contact-inhibited or serum-deprived somatic cells undergoing comprehensive vital diagnostics for the absence of biochemical and biophysical proapoptotic alterations within their plasmalemmas has not been reported so far.

Evaluation the Effects of Helichrysum plicatum Subsp. pseudoplicatum on an In-Vitro Wound Model Using Human Dermal Fibroblast Cells

2021 ◽  
pp. 153473462110166
Author(s):  
Fatma Demirkaya Miloglu ◽  
Abdulbaki Akpınar ◽  
Leyla Güven ◽  
Alper Kursat Demirkaya ◽  
Gulsah Gundogdu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Wound Healing ◽  
Cell Migration ◽  
Phenolic Compounds ◽  
Plant Extract ◽  
Phenolic Content ◽  
Dermal Fibroblast ◽  
Human Dermal Fibroblast ◽  
Total Phenolic ◽  
Proliferative Effect

Wound is tissue damage that occurs in the skin. Helichrysum species (Altınotu) are rich in phenolic compounds used in traditional medicine for wound healing. The main component in their flower head (capitulum) is phenolic compounds. The present study investigates the proliferative, oxidative stress, and wound healing properties of the methanolic extract of Helichrysum plicatum subsp. pseudoplicatum capitulum on a human dermal fibroblast (HDF) cell line in this study. H plicatum subsp. pseudoplicatum capitulums were collected in Erzurum, Turkey (altitude 1950 m), dried, pulverized, and extracted with methanol. Firstly, total phenolic contents were determined and secondly, the proliferative effect, oxidative stress activities, and wound healing effects on HDF cells were evaluated by the cell proliferation kit (XTT) test, total antioxidant status (TAS), and total oxidant status (TOS) commercial kits, and the scratch experiment by taking microscopic images of the cells at 0, 12, 18, and 24 h, respectively. Total phenolic content was found to be 142.00 ± 0.73 mg gallic acid equivalent per gram (GAE/g) extract. The capitulum extract has a proliferative effect at 0.5 to 10 µg/mL concentrations according to the XTT test results. It was observed that TAS levels significantly increased in the plant extract at the concentration ranges 1 to 10 µg/mL ( P < .01). About 1 to 5 µg/mL plant extract started to increase cell migration at the 12 h and significantly closed the wound area at the 24 h. At the doses between 1 to 5 μg/mL, it has the most substantial effect on both cell viability and antioxidant effect, and wound healing was found to be in this concentration range. These findings suggested that the H plicatum subsp. pseudoplicatum capitulum is a valuable source of phenolic content with important antioxidant activity at wound healing and it was concluded that the capitulum extract accelerates wound healing by increasing cell migration in low doses.

scholarly journals Anti-cancer effect of dihydrokaempferol in human malignant melanoma cell is mediated via inhibition of cell migration and invasion and up-regulation of NF-kB/MAPK signalling pathways

2016 ◽  
Vol 11 (4) ◽  
pp. 810
Author(s):  
Ning Zeng ◽  
Hong Qiu ◽  
Min Wu ◽  
Yi Xu ◽  
Hai-Ping Wang ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Malignant Melanoma ◽  
Signalling Pathways ◽  
Migration And Invasion ◽  
Wound Healing Assay ◽  
Human Malignant Melanoma ◽  
Cell Migration And Invasion ◽  
Mapk Signalling

<p class="Abstract">The purpose of the present research work was to demonstrate the antitumor activity of dihydrokaempferol in SK-Mel-28 human malignant melanoma cells. MTT assay was used to study the cytotoxic effects induced by dihydrokaempferol in these cells. In vitro wound healing assay and invasion assay were used to examine its effects on cell migration and invasion. Fluorescence microscopy using acridine orange/propidium iodide was used to study effects on cell morphology and apoptosis. Western blot assay revealed its effects on NF-kB/mitogen-activated protein kinase (MAPK) protein expression levels. The results indicated that dihydrokaempferol significantly inhibited the growth of these cells and the cytotoxicity pattern was shown to follow the drug dose and incubation times. Dihydrokaempferol led to onset of red fluorescence in these cells indicating that its treatment with different doses leads to induction of apoptosis. Dihydrokaempferol also led to inhibition of cell migration and invasion in a dose-dependent manner. It was also shown to up-regulate NF-kB/MAPK signalling pathways.</p><p class="Abstract"><strong>Video Clip:</strong></p><p class="Abstract"><a href="https://youtube.com/v/g8vkXiPHG4A"><em>In vitro</em> wound healing assay:</a> 4 min 25 sec</p><p> </p>

scholarly journals Zyxin Mediates Vascular Repair via Endothelial Migration Promoted by Forskolin in Mice

2021 ◽  
Vol 12 ◽  
Author(s):  
Xuya Kang ◽  
Yanan Deng ◽  
Yang Cao ◽  
Yingqing Huo ◽  
Jincai Luo
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Carotid Artery ◽  
Vascular Injury ◽  
Camp Signaling ◽  
Blue Staining ◽  
Wild Type ◽  
Wound Healing Assay ◽  
Endothelial Repair ◽  
Endothelial Migration

Background and Purpose: Endothelial repair upon vascular injury is critical for the protection of vessel integrity and prevention of the development of vascular disorders, but the underlying mechanisms remain poorly understood. In this study, we investigated the role of zyxin and its associated cyclic adenosine monophosphate (cAMP) signaling in the regulation of re-endothelialization after vascular injury.Experimental Approach: In zyxin-/- and wild-type mice, wire injury of the carotid artery was carried out, followed by Evans blue staining, to evaluate the re-endothelialization. Mice with endothelium-specific zyxin knockout were used to further determine its role. An in vitro wound-healing assay was performed in primary human endothelial cells (ECs) expressing zyxin-specific short-hairpin RNAs (shRNAs) or scrambled controls by measuring cell migration and proliferation. The effects of the cAMP signaling agonist forskolin were assessed.Key Results: The re-endothelialization of the injured carotid artery was impaired in zyxin-deficient mice, whereas the rate of cell proliferation was comparable with that in wild-type controls. Furthermore, endothelium-specific deletion of zyxin led to similar phenotypes. Knockdown of zyxin by shRNAs in primary human ECs significantly reduced cell migration in the wound-healing assay. Notably, forskolin enhanced endothelial migration in a dose-dependent manner, and this was dependent on zyxin through its interaction with vasodilator-stimulated phosphoprotein. In addition, forskolin promoted the re-endothelialization of the injured carotid artery, and this was compromised by zyxin deficiency.Conclusion and Implications: This study reveals zyxin as a new player in endothelial repair, which is promoted by forskolin, after vascular injury. Thus, zyxin-mediated signaling might be a potential treatment target for diseases involving vascular injury.

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