scholarly journals Fibroblast Differentiation and Matrix Remodeling Impaired under Simulated Microgravity in 3D Cell Culture Model

2021 ◽  
Vol 22 (21) ◽  
pp. 11911
Author(s):  
Jiranuwat Sapudom ◽  
Mei ElGindi ◽  
Marc Arnoux ◽  
Nizar Drou ◽  
Anna Garcia-Sabaté ◽  
...  
Keyword(s):  
Simulated Microgravity ◽  
Smooth Muscle Actin ◽  
Healing Process ◽  
3D Culture ◽  
3D Cell Culture ◽  
Wound Healing Process ◽  
Interstitial Tissue ◽  
Matrix Remodeling ◽  
Alpha Smooth Muscle Actin ◽  
Matrix Production

Exposure to microgravity affects astronauts’ health in adverse ways. However, less is known about the extent to which fibroblast differentiation during the wound healing process is affected by the lack of gravity. One of the key steps of this process is the differentiation of fibroblasts into myofibroblasts, which contribute functionally through extracellular matrix production and remodeling. In this work, we utilized collagen-based three-dimensional (3D) matrices to mimic interstitial tissue and studied fibroblast differentiation under simulated microgravity (sµG). Our results demonstrated that alpha-smooth muscle actin (αSMA) expression and translocation of Smad2/3 into the cell nucleus were reduced upon exposure to sµG compared to the 1g control, which suggests the impairment of fibroblast differentiation under sµG. Moreover, matrix remodeling and production were decreased under sµG, which is in line with the impaired fibroblast differentiation. We further investigated changes on a transcriptomic level using RNA sequencing. The results demonstrated that sµG has less effect on fibroblast transcriptomes, while sµG triggers changes in the transcriptome of myofibroblasts. Several genes and biological pathways found through transcriptome analysis have previously been reported to impair fibroblast differentiation. Overall, our data indicated that fibroblast differentiation, as well as matrix production and remodeling, are impaired in 3D culture under sµG conditions.

scholarly journals Evaluation of myofibroblasts and its association with TGF-β and IFN-γ in lesions of patients with american tegumentary leishmaniasis

2012 ◽  
Vol 87 (4) ◽  
pp. 545-549 ◽  
Author(s):  
Agostinho Gonçalves Viana ◽  
Carlos Alberto de Carvalho Fraga ◽  
Paulo Rogério Ferreti Bonan
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Actin ◽  
Healing Process ◽  
Sand Fly ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Tegumentary Leishmaniasis ◽  
American Tegumentary Leishmaniasis ◽  
Ifn Γ ◽  
Tgf Β1

BACKGROUND: Leishmaniasis is caused by protozoa of Leishmania spp. genus. It is transmitted by the bite of the sand fly insect. It is believed that 12 million people are infected with this disease and that its annual incidence is 2 million; this number is increasing. OBJECTIVES: The present study aimed to evaluate the expression of myofibroblasts through alpha smooth muscle actin labeling, and to analyze their relationship with the expression of the cytokines Interferon gama (IFN-γ) and Transforming growth factor beta (TGF-β1) in lesions of American tegumentary leishmaniasis (ATL). METHODS: For this retrospective study, we gathered 28 patients diagnosed with ATL between 2002 and 2006. We verified α-SMA positivity and performed IFN-γ and TGF-β1 immunolabeling to identify the profile of these cytokines in both positive and negative cases for myofibroblasts, via immunohistochemistry, in order to assess the presence of myofibroblasts,. RESULTS: We observed that out of the 28 cases, 17 (60.71%) were positive for alpha smooth muscle actin, while 11 (39.29%) were negative, and IFN-γ was more expressed than TGF-β1 (p=0.007). The mean percentages of expression of IFN-γ and TGF-β1 in the group negative for alpha smooth muscle actin were different, with an increased expression of IFN-γ (p=0.047). However, in the group positive for alpha smooth muscle actin, there was no difference in cytokine labeling (p>0.05). CONCLUSION: We verified the presence of positive α-SMA stromal cells in the majority of the cases of ATL, indicating the presence of myofibroblasts. In cases negative for alpha smooth muscle actin, an increased expression of IFN-γ compared to TGF-β1 was observed, revealing an inflammatory phase progressing to a healing process.

scholarly journals Immunohistological changes in skin wounds during the early periods of healing in a rat model

2012 ◽  
Vol 57 (No. 2) ◽  
pp. 77-82 ◽  
Author(s):  
F. Sabol ◽  
L. Dancakova ◽  
P. Gal ◽  
T. Vasilenko ◽  
M. Novotny ◽  
...  
Keyword(s):  
Wound Healing ◽  
Wide Spectrum ◽  
Smooth Muscle Actin ◽  
Healing Process ◽  
Skin Wound ◽  
Hair Follicles ◽  
In Vivo Model ◽  
Wound Healing Process ◽  
Rat Skin

The complexity of the wound healing process, which is still poorly understood, prompted us to perform an immunohistochemical investigation using rat skin as an in vivo model. Fifteen Sprague-Dawley rats were included in the experiment. Two round full thickness wounds, 4 mm in diameter, were made on the backs of all rats. Haematoxylin and eosin basic staining as well as antibodies against wide spectrum keratin, keratin 10, keratin 14, α-smooth muscle actin, vimentin, fibronectin, collagens Type 1 and 3, and the transcription factor Sox-2 were applied to paraffin and frozen sections of skin wound specimens two, six and fourteen days after surgery, respectively. New hair follicles with Sox-2-positive cells were present after fourteen days; keratin/vimentin positivity was restricted to specimens of day two. Collagen-3 expression prevailed over collagen-1 expression at all evaluated time intervals, except in the uninjured part of the dermis. In conclusion, rat skin wound healing is a dynamic process which can serve as a model for studying phenomena such as cell-cell interactions and transitions in vivo.

scholarly journals Stromal vascular fraction promotes migration of fibroblasts and angiogenesis through regulation of extracellular matrix in the skin wound healing process

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Hongsen Bi ◽  
Hui Li ◽  
Chen Zhang ◽  
Yiqing Mao ◽  
Fangfei Nie ◽  
...  
Keyword(s):  
Wound Healing ◽  
Endothelial Cells ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Healing Process ◽  
Stromal Vascular Fraction ◽  
Wound Healing Process ◽  
Matrix Remodeling ◽  
Skin Defect ◽  
The Impact

Abstract Background A refractory wound is a typical complication of diabetes and is a common outcome after surgery. Current approaches have difficulty in improving wound healing. Recently, non-expanded stromal vascular fraction (SVF), which is derived from mature fat, has opened up new directions for the treatment of refractory wound healing. The aim of the current study is to systematically investigate the impact of SVF on wound healing, including the rate and characteristics of wound healing, ability of fibroblasts to migrate, and blood transport reconstruction, with a special emphasis on their precise molecular mechanisms. Methods SVF was isolated by digestion, followed by filtration and centrifugation, and then validated by immunocytochemistry, a MTS proliferation assay and multilineage potential analysis. A wound model was generated by creating 6-mm-diameter wounds, which include a full skin defect, on the backs of streptozocin-induced hyperglycemic mice. SVF or human adipose-derived stem cell (hADSC) suspensions were subcutaneously injected, and the wounds were characterized over a 9-day period by photography and measurements. A scratch test was used to determine whether changes in the migratory ability of fibroblasts occurred after co-culture with hADSCs. Angiogenesis was observed with human umbilical vein endothelial cells. mRNA from fibroblasts, endotheliocyte, and skin tissue were sequenced by high-throughput RNAseq, and differentially expressed genes, and pathways, potentially regulated by SVF or hADSCs were bioinformatically analyzed. Results Our data show that hADSCs have multiple characteristics of MSC. SVF and hADSCs significantly improved wound healing in hyperglycemic mice. hADSCs improve the migratory ability of fibroblasts and capillary structure formation in HUVECs. SVF promotes wound healing by focusing on angiogenesis and matrix remodeling. Conclusions Both SVF and hADSCs improve the function of fibroblast and endothelial cells, regulate gene expression, and promote skin healing. Various mechanisms likely are involved, including migration of fibroblasts, tubulogenesis of endothelial cells through regulation of cell adhesion, and cytokine pathways.

Abstract 430: Immature Cardiac Fibroblasts With Upregulated P53 Contributed to Fibrosis in Ikke Deficient Mouse Myocardial Infarction Model

2020 ◽  
Vol 127 (Suppl_1) ◽  
Author(s):  
Yong Sook Kim ◽  
Hyang Hee Cho ◽  
Ju Hee Jun ◽  
Dong Im Cho ◽  
Meeyoung Cho ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Cardiac Fibrosis ◽  
Transforming Growth Factor ◽  
Inflammatory Responses ◽  
Cardiac Fibroblasts ◽  
Smooth Muscle Actin ◽  
Healing Process ◽  
Wound Healing Process ◽  
Coronary Artery Ligation ◽  
Artery Ligation

Background: Inhibitor of NF-κB kinase (IKK), an upstream of nuclear factor-kappa B (NF-κB), is a critical modulator for pathophysiological inflammation. IKKε is a non-classical IKK and has been studied in infectious diseases and cancers. However, the role of IKKε in a myocardial infarction (MI) has not been addressed. Methods and Results: In this study, we used IKKε knockout (KO) mice to induce MI by coronary artery ligation. The IKKε KO group showed poor early survival rate, large cardiac fibrosis (14.7±4.8% in KO vs. 31.1±10.2% in WT, p <0.05), and low fractional shortening (13.47±1.21% in KO vs. 16.36±4.46% in WT, p <0.05) compared with WT group. Next, we investigated the inflammatory responses and found that inflammatory markers such as inducible nitric oxide synthase (iNOS) and CD80 were much higher in both cardiac macrophages and bone marrow-derived macrophages (BMDM) in the IKKε KO group than in the wild type (WT) group. To explore the responsible mediator, we performed phosphorylated protein array and found phosphorylated p38 was significantly downregulated in the IKKε knockout BMDM. Conversely, both knockdown of p38 by siRNA and inhibition of p38 by SB203580 treatment in RAW264.7 cells upregulated iNOS. More interestingly, IKKε deficient cardiac fibroblasts showed highly accumulated nuclear p53 and exhibited immature differentiation. The levels of myofibroblast markers containing α-smooth muscle actin, periostin, and transforming growth factor-β1 were lower, and functional contractility was substantially impaired in the cardiac fibroblasts isolated from IKKε KO mice. Conclusion: Our data showed excessive inflammation was associated with p38 inactivation in macrophages and pathological fibrosis was resulted from immature myofibroblast phenotype with p53 upregulation. Collectively, IKKε is involved in the control of inflammation resolution and wound healing process in the infarcted myocardium.

scholarly journals Bioengineered human acellular vessels recellularize and evolve into living blood vessels after human implantation

2019 ◽  
Vol 11 (485) ◽  
pp. eaau6934 ◽  
Author(s):  
Robert D. Kirkton ◽  
Maribel Santiago-Maysonet ◽  
Jeffrey H. Lawson ◽  
William E. Tente ◽  
Shannon L. M. Dahl ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Progenitor Cell ◽  
Smooth Muscle Actin ◽  
Small Samples ◽  
Mechanical Degradation ◽  
Matrix Remodeling ◽  
Self Healing ◽  
Alpha Smooth Muscle Actin ◽  
Surgical Interventions ◽  
Host Cell Response

Traditional vascular grafts constructed from synthetic polymers or cadaveric human or animal tissues support the clinical need for readily available blood vessels, but often come with associated risks. Histopathological evaluation of these materials has shown adverse host cellular reactions and/or mechanical degradation due to insufficient or inappropriate matrix remodeling. We developed an investigational bioengineered human acellular vessel (HAV), which is currently being studied as a hemodialysis conduit in patients with end-stage renal disease. In rare cases, small samples of HAV were recovered during routine surgical interventions and used to examine the temporal and spatial pattern of the host cell response to the HAV after implantation, from 16 to 200 weeks. We observed a substantial influx of alpha smooth muscle actin (αSMA)–expressing cells into the HAV that progressively matured and circumferentially aligned in the HAV wall. These cells were supported by microvasculature initially formed by CD34+/CD31+ cells in the neoadventitia and later maintained by CD34−/CD31+ endothelial cells in the media and lumen of the HAV. Nestin+ progenitor cells differentiated into either αSMA+ or CD31+ cells and may contribute to early recellularization and self-repair of the HAV. A mesenchymal stem cell–like CD90+ progenitor cell population increased in number with duration of implantation. Our results suggest that host myogenic, endothelial, and progenitor cell repopulation of HAVs transforms these previously acellular vessels into functional multilayered living tissues that maintain blood transport and exhibit self-healing after cannulation injury, effectively rendering these vessels like the patient’s own blood vessel.

Anisotropic Strain Fields Enhance Matrix Remodeling Through Elevated TGF-β Signaling

2011 ◽  
Author(s):  
Russell Gould ◽  
Karen Chin ◽  
Puifai Santisakultam ◽  
Amanda Dropkin ◽  
Jennifer Richards ◽  
...  
Keyword(s):  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Nuclear Translocation ◽  
Smooth Muscle Actin ◽  
Matrix Remodeling ◽  
Alpha Smooth Muscle Actin ◽  
Anisotropic Strain ◽  
Unique Effect ◽  
Engineered Tissue ◽  
Growth Factor Beta

In this work, we demonstrate the unique effect of controlled anisotropic strain on fibroblast behavior in 3D engineered tissue environments. Anisotropy of biaxial strain resulted in increased cellular orientation and collagen fiber alignment. Transforming growth factor beta-1 (TGFβ1) gene expression and pSmad2 nuclear translocation increased with biaxial directionality. Myofibroblastic alpha-smooth muscle actin (α-SMA) decreased with applied strain similar to mechanically unloaded hydrogels. Collectively, these results demonstrate a novel mechanobiological mechanism by which fibroblasts develop rapid anisotropic matrix striation while maintaining phenotype quiescence.

scholarly journals The Role of MSC in Wound Healing, Scarring and Regeneration

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1729
Author(s):  
Raquel Guillamat-Prats
Keyword(s):  
Wound Healing ◽  
Wound Repair ◽  
Therapeutic Potential ◽  
Healing Process ◽  
Repair Process ◽  
Skin Wound ◽  
Wound Healing Process ◽  
Matrix Remodeling ◽  
Skin Wound Healing ◽  
Tissue Repair And Regeneration

Tissue repair and regeneration after damage is not completely understood, and current therapies to support this process are limited. The wound healing process is associated with cell migration and proliferation, extracellular matrix remodeling, angiogenesis and re-epithelialization. In normal conditions, a wound will lead to healing, resulting in reparation of the tissue. Several risk factors, chronic inflammation, and some diseases lead to a deficient wound closure, producing a scar that can finish with a pathological fibrosis. Mesenchymal stem/stromal cells (MSCs) are widely used for their regenerative capacity and their possible therapeutically potential. Derived products of MSCs, such as exosomes or extravesicles, have shown a therapeutic potential similar to MSCs, and these cell-free products may be interesting in clinics. MSCs or their derivative products have shown paracrine beneficial effects, regulating inflammation, modifying the fibroblast activation and production of collagen and promoting neovascularization and re-epithelialization. This review describes the effects of MSCs and their derived products in each step of the wound repair process. As well, it reviews the pre-clinical and clinical use of MSCs to benefit in skin wound healing in diabetic associated wounds and in pathophysiological fibrosis.

scholarly journals A Preliminary Study on the Potential of Manuka Honey and Platelet-Rich Plasma in Wound Healing

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Scott A. Sell ◽  
Patricia S. Wolfe ◽  
Andrew J. Spence ◽  
Isaac A. Rodriguez ◽  
Jennifer M. McCool ◽  
...  
Keyword(s):  
Wound Healing ◽  
Culture Media ◽  
Platelet Rich Plasma ◽  
Human Fibroblasts ◽  
Healing Process ◽  
Collagen Matrix ◽  
Wound Healing Process ◽  
Manuka Honey ◽  
Matrix Production

Aim. The purpose of this study was to determine thein vitroresponse of cells critical to the wound healing process in culture media supplemented with a lyophilized preparation rich in growth factors (PRGF) and Manuka honey.Materials and Methods. This study utilized cell culture media supplemented with PRGF, as well as whole Manuka honey and the medical-grade Medihoney (MH), a Manuka honey product. The response of human fibroblasts (hDF), macrophages, and endothelial cells (hPMEC) was evaluated, with respect to cell proliferation, chemotaxis, collagen matrix production, and angiogenic potential, when subjected to culture with media containing PRGF, MH, Manuka honey, and a combination of PRGF and MH.Results. All three cell types demonstrated increases in cellular activity in the presence of PRGF, with further increases in activity seen in the presence of PRGF+MH. hDFs proved to be the most positively responsive cells, as they experienced enhanced proliferation, collagen matrix production, and migration into anin vitrowound healing model with the PRGF+MH-supplemented media.Conclusion. This preliminaryin vitrostudy is the first to evaluate the combination of PRGF and Manuka honey, two products with the potential to increase regeneration individually, as a combined product to enhance dermal regeneration.

scholarly journals Green Peptide–nanomaterials; A Friendly Healing Touch for Skin Wound Regeneration

2019 ◽  
Vol 2 (1) ◽  
pp. 14-31 ◽  
Author(s):  
Debjani Nath ◽  
Pratyusha Banerjee ◽  
Anugrah Ray ◽  
Baishakhi Bairagi
Keyword(s):  
Wound Healing ◽  
Wound Care ◽  
Healing Process ◽  
Skin Wound ◽  
The Body ◽  
Wound Healing Process ◽  
Present Moment ◽  
Developmental Study ◽  
Matrix Remodeling ◽  
Natural Peptides

The complex phenomenon by which the body responds to any injury of skin or tissue is known as wound healing. A number of phases like exudative, proliferative, and extracellular matrix remodeling are orchestrated events to be occurred involving blood cells, parenchymal cells, and different soluble mediators. Different internal, as well as external factors, regulate the speed and quality of healing. The delay in wound healing process causes the chronic wound or scar formation. At the present moment, the upscale research for identification of agents causing accelerated healing is important. Moreover, the biocompatibility of the accelerators needs to be investigated. Recent biomedical researches for wound care target to provide antimicrobial protection as well as matrix scaffolding for quick repairing of the skin tissue. In recent studies with natural peptides have shown that they are important components in developing the nano-medicines for their usefulness and therapeutic efficiency. New therapeutic formulations can be developed using these natural peptides utilizing different nanoparticle delivery system. This review deals with the developmental study on efficient wound care system where the possible use of natural peptides in combination with nanomaterials has been explored. A trial has also been made on the findings made over the past few years on the use of peptides as tissue regenerating agents through effective wound healing pathway.

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