CHANGES OF PER2 GENE KNOCKOUT ON TISSUE REPAIR: EPIDERMAL WOUND HEALING ASSAY IN VIVO

2020 ◽  
Vol 130 (3) ◽  
pp. e254
Author(s):  
VERONICA QUISPE YUJRA ◽  
ERICKA JANINE D. DA SILVEIRA ◽  
DANIEL ARAKI RIBEIRO ◽  
ROGERIO CASTILHOS ◽  
CRISTIANE SQUARIZE
Keyword(s):  
Wound Healing ◽  
Tissue Repair ◽  
Gene Knockout ◽  
Wound Healing Assay

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.

Neuropilin 1 modulates TGF-β1-induced epithelial–mesenchymal transition in non-small cell lung cancer

2019 ◽  
Author(s):  
Zongli Ding ◽  
Wenwen Du ◽  
Zhe Lei ◽  
Yang Zhang ◽  
Jianjie Zhu ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Wound Healing ◽  
Epithelial Mesenchymal Transition ◽  
Small Cell ◽  
Migration And Invasion ◽  
Wound Healing Assay ◽  
Small Cell Lung ◽  
Mesenchymal Transition ◽  
Tgf Β1

Abstract Background: TGF-β1 signaling is a potent inducer of epithelial-mesenchymal transition (EMT) in various cancers. Our previous study has indicated that NRP1 was significantly up-regulated and acted as a vital promoter in the metastasis of non-small cell lung cancer (NSCLC). However, the function of NRP1 in regulation of TGF-β1-induced EMT and NSCLC cell migration and invasion remained unclear. Methods: The differential expression level of NRP1 was determined by RT-PCR analysis in human tissue samples with or without lymph node metastasis. Transwell assay and wound healing assay were conducted to determine cell ability of migration. Lentivirus-mediated stable knockdown and overexpression of NRP1 cell lines were constructed. Exogenous TGF-β1 stimulation, SIS3 treatment, western blot analysis and in vivo metastatic model were utilized to clarify the underlying regulatory mechanism. Results: Increased expression of NRP1 was found in metastatic NSCLC tissues and can promote NSCLC metastasis in vivo. Transwell assays, wound healing assay and western blot analysis showed that knockdown of NRP1 significantly inhibited TGF-β1-mediated EMT and migratory and invasive capabilities of A549 and H226 cells. Furthermore, overexpression of NRP1 could weak the decreased migratory and invasive capabilities with SIS3 treatment. Co-IP data showed that NRP1 can interact with TGFβRⅡ to induce EMT. Conclusion: This is the first time to report that NRP1 can modulate TGF-β1-induced EMT and cell migration and invasion in NSCLC.

Investigating Epidermal Interactions Through an In Vivo Cutaneous Wound-Healing Assay

2020 ◽  
pp. 1-11
Author(s):  
John L. Zemkewicz ◽  
Racheal G. Akwii ◽  
Constantinos M. Mikelis ◽  
Colleen L. Doçi
Keyword(s):  
Wound Healing ◽  
Cutaneous Wound Healing ◽  
Wound Healing Assay ◽  
Cutaneous Wound

scholarly journals Effect of Combination therapy of Desmopressin and Docetaxel on prostate cancer cell DU145 proliferation, migration and growth

2016 ◽  
Vol 2 (1) ◽  
Author(s):  
Azik Hoffman ◽  
Hiroshi Sasaki ◽  
Domenica Roberto ◽  
Michelle J Mayer ◽  
Laurence Klotz ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Wound Healing ◽  
Combination Therapy ◽  
Tumor Volume ◽  
Prostate Cancer Cell ◽  
Wound Healing Assay ◽  
Castrate Resistant Prostate Cancer ◽  
Castrate Resistant

Background: This study was designed to assess the efficacy of the combination of Desmopressin and Docetaxel for prostate cancer. Desmopressin has been demonstrated to inhibit tumor progression and metastasis in in vitro and in vivo models of breast cancer. Docetaxel, an anti-mitotic chemotherapeutic agent, is widely used for the treatment of castration resistant prostate cancer. However, it is associated with adverse effects and eventual drug resistance. This is the first report on the effect of combining Desmopressin and Docetaxel in prostate cancer, both in vitro and in vivo. Methods: An established castrate resistant prostate cancer cell line DU145 was used. Cellular proliferation was determined using the MTS assay. The migratory inhibition potential of Desmopressin alone and in combination with Docetaxel was accessed using the wound healing assay. In vivo evaluation was performed on a prostate cancer xenograft model using athymic nude mouse. Treatment was administered bi- weekly and tumor volume were measured throughout the treatment period. Eventually, after a six-week treatment period, tumors were excised and measured.  Results: A combination therapy of 1 µM Desmopressin with 100nM Docetaxel resulted in dramatic inhibition of proliferation of DU145 cells 72 hours post treatment compared to either agent along (p < 0.05). Wound healing assay revealed inhibition of cellular migration as well (p<0.05). The use of a xenograft mouse model followed by treatment with 5 mg/kg Docetaxel intraperitoneally with concomitant 2 µg/ml/kg Desmopressin administered intravenously 30 minutes before administering chemotherapy and 24 hours after, resulted in a significant decrease in tumor volume (P<0.05), while not impacting body weight.Conclusions: Desmopressin significantly enhanced the anti-proliferative and inhibiting the migratory potential of Docetaxel. Combination treatment had no additional effect on mice weight or mortality. These studies could enhance the efficacy of Docetaxel- based chemotherapy treatment for castrate resistant prostate cancer.

Expression of the beta 6 integrin subunit in development, neoplasia and tissue repair suggests a role in epithelial remodeling

1995 ◽  
Vol 108 (6) ◽  
pp. 2241-2251 ◽  
Author(s):  
J.M. Breuss ◽  
J. Gallo ◽  
H.M. DeLisser ◽  
I.V. Klimanskaya ◽  
H.G. Folkesson ◽  
...  
Keyword(s):  
Wound Healing ◽  
Epithelial Cells ◽  
Lung Injury ◽  
Tissue Repair ◽  
Expression Patterns ◽  
Alveolar Epithelial Cells ◽  
Integrin Subunit ◽  
Fibronectin Receptor ◽  
High Level

The alpha v beta 6 integrin was identified in cultured epithelial cells and functions as a fibronectin receptor. We have now used monoclonal antibodies to determine in vivo expression patterns of the beta 6 subunit in normal and pathological human or primate tissues, and during experimental wound healing or induced lung injury. The results indicate that beta 6 expression is restricted to epithelia and is up-regulated in parallel with morphogenetic events, tumorigenesis, and epithelial repair. During development of the kidney, lung, and skin, we found that beta 6 is expressed by specific types of epithelial cells, whereas it is mostly undetectable in normal adult kidney, lung and skin. In contrast, we detected high-level expression in several types of carcinoma. For example, beta 6 is almost invariably neo-expressed in squamous cell carcinomas derived from the oral mucosa, often focally localized at the infiltrating edges of tumor islands. Expression of beta 6 is also upregulated in migrating keratinocytes at the wound edge during experimental epidermal wound healing. Similarly, beta 6 expression is induced in type II alveolar epithelial cells during lung injury caused by injection of live bacteria. We also observed beta 6 expression in adult lungs and kidneys at focal sites of subclinical inflammation, as well as in a variety of clinical specimens from patients with chronic or acute inflammation of the lungs or kidneys. From these findings and earlier results, we hypothesize that alpha v beta 6 affects cell spreading, migration and growth during reorganization of epithelia in development, tissue repair, and neoplasia.

scholarly journals Nanobiomaterials for vascular biology and wound management: A review

2018 ◽  
Vol 7 (2) ◽  
Author(s):  
Ajay Vikram Singh ◽  
Donato Gemmati ◽  
Anurag Kanase ◽  
Ishan Pandey ◽  
Vatsala Misra ◽  
...  
Keyword(s):  
Wound Healing ◽  
Tissue Repair ◽  
Wound Care ◽  
Viable Cell ◽  
Clinical Science ◽  
Wound Management ◽  
Clinical Practices ◽  
Hydrogel Scaffold ◽  
Extracellular Matrix Components

Nanobiomaterials application into tissue repair and ulcer management is experiencing its golden age due to spurring diversity of translational opportunity to clinics. Over the past years, research in clinical science has seen a dramatic increase in medicinal materials at nanoscale those significantly contributed to tissue repair. This chapter outlines the new biomaterials at nanoscale those contribute state of the art clinical practices in ulcer management and wound healing due to their superior properties over traditional dressing materials. Designing new recipes for nanobiomaterials for tissue engineering practices spanning from micro to nano-dimension provided an edge over traditional wound care materials those mimic tissue in vivo. Clinical science stepped into design of artificial skin and extracellular matrix components emulating the innate structures with higher degree of precision. Advances in materials sciences polymer chemistry have yielded an entire class of new nanobiomaterials ranging from dendrimer to novel electrospun polymer with biodegradable chemistries and controlled molecular compositions assisting wound healing adhesives, bandages and controlled of therapeutics in specialized wound care. Moreover, supportive regenerative medicine is transforming into rational, real and successful component of modern clinics providing viable cell therapy of tissue remodeling. Soft nanotechnology involving hydrogel scaffold revolutionized the wound management supplementing physicobiochemical and mechanical considerations of tissue regeneration. Moreover, this chapter also reviews the current challenges and opportunities in specialized nanobiomaterials formulations those are desirable for optimal localized wound care considering their in situ physiological microenvironment.

scholarly journals miR-126a-3p induces proliferation, migration and invasion of trophoblast cells in pre-eclampsia-like rats by inhibiting A Disintegrin and Metalloprotease 9

2019 ◽  
Vol 39 (12) ◽  
Author(s):  
Shenglong Zhao ◽  
Jiandong Wang ◽  
Zheng Cao ◽  
Lei Gao ◽  
Yuanyuan Zheng ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Wound Healing ◽  
Underlying Mechanism ◽  
Migration And Invasion ◽  
Cell Cycle Distribution ◽  
Wound Healing Assay ◽  
Trophoblast Cells ◽  
Transwell Assay ◽  
Blank Group

Abstract The present study aimed to investigate the underlying mechanism of miR-126a-3p in the proliferation, migration and invasion of trophoblast cells in pre-eclampsia-like rats by targeting A Disintegrin and Metalloprotease 9 (ADAM9). First, the interaction between miR-126a-3p and ADAM9 was confirmed via biochemical assays. Placental tissues and trophoblast cells were then obtained. RNA in situ hybridization was performed in order to detect miR-126a-3p expression in the placenta. Subsequently, a series of biological assays, including reverse transcription-quantitative PCR (RT-qPCR), Western blotting, MTT assay, apoptosis assay, cell cycle assay, wound healing assay and transwell assay were adopted in order to determine the cell proliferation, cell cycle distribution, apoptotic rate, and migration and invasion of trophoblast cells in each group. The results revealed that miR-126a-3p was down-regulated in the placenta of pre-eclampsia-like rats. In vivo experiments’ results indicated that miR-126a-3p could inhibit ADAM9 expression, and induce cyclin D1, Matrix metalloproteinase (MMP) 2 (MMP-2), MMP-9 expression. MTT, apoptosis and cell cycle assay results revealed that trophoblast cells transfected with miR-126a-3p mimic or si-ADAM9 exhibited higher proliferative activity and a lower apoptotic rate compared with the blank group (all P&lt;0.05). The wound healing assay and transwell assay results confirmed that, compared with the blank group, the migration and invasion ability of trophoblast cells in the miR-126a-3p mimic group and small interfering RNA (siRNA)-ADAM9 group were significantly increased (all P&lt;0.05). Conversely, miR-126a-3p inhibitor treatment revealed the opposite effect (all P&lt;0.05). In conclusion, the present study demonstrated that miR-126a-3p could enhance proliferation, migration and invasion, but decrease the apoptosis rate of trophoblast cells in pre-eclampsia-like rats through targeting ADAM9.

scholarly journals In vivo incisional wound healing augmented by platelet-derived growth factor and recombinant c-sis gene homodimeric proteins.

1988 ◽  
Vol 167 (3) ◽  
pp. 974-987 ◽  
Author(s):  
G F Pierce ◽  
T A Mustoe ◽  
R M Senior ◽  
J Reed ◽  
G L Griffin ◽  
...  
Keyword(s):  
Wound Healing ◽  
Growth Factor ◽  
Tissue Repair ◽  
Inflammatory Cells ◽  
Cell Types ◽  
Platelet Derived Growth Factor ◽  
Inflammatory Cell Infiltrate ◽  
In Vitro Tests

Human platelet-derived growth factor (hPDGF) is likely to be important in stimulating tissue repair, based upon its in vivo chemotactic and stimulatory activities for inflammatory cells and fibroblasts and upon the presence of PDGF and related proteins in platelets, macrophages, and activated fibroblasts, cell types that make up the milieu of the healing wound. Recombinant human c-sis (rPDGF-B), homodimers of the B chain of PDGF, were compared with hPDGF in vitro. rPDGF-B was immunologically similar to hPDGF and, at identical concentrations, similar to hPDGF in stimulating fibroblast mitogenesis and chemotaxis of polymorphonuclear leukocytes, monocytes, and fibroblasts. Purified hPDGF and rPDGF-B were also tested in vivo for potency in a model of tissue repair using a linear incision wound through rat dermis. A single application of hPDGF or rPDGF-B (2-20 micrograms/wound) in a slow release vehicle at the time of wounding resulted in a dose-dependent, statistically highly significant increase of breaking strength of treated wounds. Wound healing in animals treated with rPDGF-B was 170% stronger and accelerated by 2 d during the first week over control wounds and by 4-6 d over the next 2 wk. Histologic evaluation of growth factor-treated wounds correlated the in vitro chemotactic activity and the accelerated healing of wounds with a striking inflammatory cell infiltrate early after wounding, markedly increased formation of granulation tissue by 4-d, and increased fibrosis by 14 d in comparison to control wounds. The results thus demonstrate that rPDGF-B is fully active in in vitro tests of mitogenesis and chemotaxis and, for the first time, demonstrate directly that PDGF significantly advances wound healing in incisional wounds of experimental animals.

scholarly journals A dynamic spectrum of monocytes arising from the in situ reprogramming of CCR2+ monocytes at a site of sterile injury

2015 ◽  
Vol 212 (4) ◽  
pp. 447-456 ◽  
Author(s):  
Daniela Dal-Secco ◽  
Jing Wang ◽  
Zhutian Zeng ◽  
Elzbieta Kolaczkowska ◽  
Connie H.Y. Wong ◽  
...  
Keyword(s):  
Wound Healing ◽  
Tissue Repair ◽  
Hepatic Injury ◽  
Dynamic Spectrum ◽  
Fluorescent Reporters ◽  
Injured Area ◽  
A Site ◽  
Local Cytokine

Monocytes are recruited from the blood to sites of inflammation, where they contribute to wound healing and tissue repair. There are at least two subsets of monocytes: classical or proinflammatory (CCR2hiCX3CR1low) and nonclassical, patrolling, or alternative (CCR2lowCX3CR1hi) monocytes. Using spinning-disk confocal intravital microscopy and mice with fluorescent reporters for each of these subsets, we were able to track the dynamic spectrum of monocytes that enter a site of sterile hepatic injury in vivo. We observed that the CCR2hiCX3CR1low monocytes were recruited early and persisted for at least 48 h, forming a ringlike structure around the injured area. These monocytes transitioned, in situ, from CCR2hiCx3CR1low to CX3CR1hiCCR2low within the ringlike structure and then entered the injury site. This phenotypic conversion was essential for optimal repair. These results demonstrate a local, cytokine driven reprogramming of classic, proinflammatory monocytes into nonclassical or alternative monocytes to facilitate proper wound-healing.

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