scholarly journals Human keratinocyte (HaCaT) stimulation and healing effect of the methanol fraction from the decoction from leaf from Sideroxylon obtusifolium (Roem. & Schult.) T.D. Penn on experimental burn wound model

2020 ◽  
Author(s):  
Tamiris de Fátima Goebel de Souza ◽  
Taiana Magalhães Pierdoná ◽  
Fernanda Soares Macedo ◽  
Pedro Everson Alexandre de Aquino ◽  
Gisele de Fátima Pinheiro Rangel ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Human Keratinocytes ◽  
Northeastern Brazil ◽  
Easy Access ◽  
Hacat Cells ◽  
Burn Wound ◽  
Methanol Fraction ◽  
Wound Model ◽  
Human Keratinocyte ◽  
Experimental Burn

AbstractThe larger number of plants, with therapeutic potential, popularly used in Northeastern Brazil is due to their easy access and the great Brazilian biodiversity. Previously, was demonstrated that the methanol fraction from Sideroxylon obtusifolium (MFSOL) promoted an anti-inflammatory and healing activity in excisional wounds. Thus, this work aimed to investigate the healing effects of MFSOL on human keratinocytes cells (HaCaT) and experimental burn model injuries. HaCaT cells were used to investigate migration and proliferation of cell rates. Female Swiss mice were subjected to second-degree superficial burn protocol and divided into four treatment groups: Vehicle (cream-base), 1.0% Silver Sulfadiazine (Sulfa), and 0.5% or 1.0% MFSOL cream (CrMFSOL). Samples were collected for quantification of the inflammatory mediators and histological analyses after 3, 7 and 14 days on evaluation. As result, MFSOL (50 μg/ml) stimulated HaCaT cells by increasing proliferation and migration rates. Moreover, CrMFSOL 0.5% attenuated myeloperoxidase (MPO) activity and also stimulated the release of IL-1β and IL-10, after 3 days with treatment. CrMFSOL 0.5% enhanced wound contraction, promoted tissue remodeling improvement and highest collagen production after 7 days, and VEGF release after 14 days. Therefore, MFSOL evidenced the stimulation of human keratinocyte (HaCaT) cells and improvements on wound healing via inflammatory modulation on burn injuries.

scholarly journals An integrated systems biology approach to understanding the rules of keratinocyte colony formation

2007 ◽  
Vol 4 (17) ◽  
pp. 1077-1092 ◽  
Author(s):  
Tao Sun ◽  
Phil McMinn ◽  
Simon Coakley ◽  
Mike Holcombe ◽  
Rod Smallwood ◽  
...  
Keyword(s):  
Human Keratinocytes ◽  
Self Organization ◽  
Hacat Cells ◽  
Agent Based ◽  
Wound Model ◽  
Cell Substrate ◽  
Scratch Wound ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell

Closely coupled in vitro and in virtuo models have been used to explore the self-organization of normal human keratinocytes (NHK). Although it can be observed experimentally, we lack the tools to explore many biological rules that govern NHK self-organization. An agent-based computational model was developed, based on rules derived from literature, which predicts the dynamic multicellular morphogenesis of NHK and of a keratinocyte cell line (HaCat cells) under varying extracellular Ca ++ concentrations. The model enables in virtuo exploration of the relative importance of biological rules and was used to test hypotheses in virtuo which were subsequently examined in vitro . Results indicated that cell–cell and cell–substrate adhesions were critically important to NHK self-organization. In contrast, cell cycle length and the number of divisions that transit-amplifying cells could undergo proved non-critical to the final organization. Two further hypotheses, to explain the growth behaviour of HaCat cells, were explored in virtuo —an inability to differentiate and a differing sensitivity to extracellular calcium. In vitro experimentation provided some support for both hypotheses. For NHKs, the prediction was made that the position of stem cells would influence the pattern of cell migration post-wounding. This was then confirmed experimentally using a scratch wound model.

scholarly journals Microgravity Induces Transient EMT in Human Keratinocytes by Early Down-Regulation of E-Cadherin and Cell-Adhesion Remodeling

2020 ◽  
Vol 11 (1) ◽  
pp. 110
Author(s):  
Giulia Ricci ◽  
Alessandra Cucina ◽  
Sara Proietti ◽  
Simona Dinicola ◽  
Francesca Ferranti ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Human Keratinocytes ◽  
Short Exposure ◽  
Migration And Invasion ◽  
Hacat Cells ◽  
Invasive Phenotype ◽  
Adhesion Properties ◽  
Mesenchymal Transition ◽  
Cell Matrix ◽  
E Cadherin

Changes in cell–matrix and cell-to-cell adhesion patterns are dramatically fostered by the microgravity exposure of living cells. The modification of adhesion properties could promote the emergence of a migrating and invasive phenotype. We previously demonstrated that short exposure to the simulated microgravity of human keratinocytes (HaCaT) promotes an early epithelial–mesenchymal transition (EMT). Herein, we developed this investigation to verify if the cells maintain the acquired invasive phenotype after an extended period of weightlessness exposure. We also evaluated cells’ capability in recovering epithelial characteristics when seeded again into a normal gravitational field after short microgravity exposure. We evaluated the ultra-structural junctional features of HaCaT cells by Transmission Electron Microscopy and the distribution pattern of vinculin and E-cadherin by confocal microscopy, observing a rearrangement in cell–cell and cell–matrix interactions. These results are mirrored by data provided by migration and invasion biological assay. Overall, our studies demonstrate that after extended periods of microgravity, HaCaT cells recover an epithelial phenotype by re-establishing E-cadherin-based junctions and cytoskeleton remodeling, both being instrumental in promoting a mesenchymal–epithelial transition (MET). Those findings suggest that cytoskeletal changes noticed during the first weightlessness period have a transitory character, given that they are later reversed and followed by adaptive modifications through which cells miss the acquired mesenchymal phenotype.

scholarly journals Wound Healing Composite Materials of Bacterial Cellulose and Zinc Oxide Nanoparticles with Immobilized Betulin Diphosphate

Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 713
Author(s):  
Nina Melnikova ◽  
Alexander Knyazev ◽  
Viktor Nikolskiy ◽  
Peter Peretyagin ◽  
Kseniia Belyaeva ◽  
...  
Keyword(s):  
Wound Healing ◽  
Zinc Oxide ◽  
Bacterial Cellulose ◽  
Zinc Oxide Nanoparticles ◽  
Wound Dressings ◽  
Oxide Nanoparticles ◽  
Burn Wound ◽  
Zno Nps ◽  
Wound Model ◽  
And Oxidative Stress

A design of new nanocomposites of bacterial cellulose (BC) and betulin diphosphate (BDP) pre-impregnated into the surface of zinc oxide nanoparticles (ZnO NPs) for the production of wound dressings is proposed. The sizes of crystalline BC and ZnO NPs (5–25%) corresponded to 5–6 nm and 10–18 nm, respectively (powder X-ray diffractometry (PXRD), Fourier-infrared (FTIR), ultraviolet (UV), atomic absorption (AAS) and photoluminescence (PL) spectroscopies). The biological activity of the wound dressings “BC-ZnO NPs-BDP” was investigated in rats using a burn wound model. Morpho-histological studies have shown that more intensive healing was observed during treatment with hydrophilic nanocomposites than the oleophilic standard (ZnO NPs-BDP oleogel; p < 0.001). Treatment by both hydrophilic and lipophilic agents led to increases in antioxidant enzyme activity (superoxide dismutase (SOD), catalase) in erythrocytes and decreases in the malondialdehyde (MDA) concentration by 7, 10 and 21 days (p < 0.001). The microcirculation index was restored on the 3rd day after burn under treatment with BC-ZnO NPs-BDP wound dressings. The results of effective wound healing with BC-ZnO NPs-BDP nanocomposites can be explained by the synergistic effect of all nanocomposite components, which regulate oxygenation and microcirculation, reducing hypoxia and oxidative stress in a burn wound.

scholarly journals Skullcapflavone II Suppresses TNF-α/IFN-γ-Induced TARC, MDC, and CTSS Production in HaCaT Cells

2021 ◽  
Vol 22 (12) ◽  
pp. 6428
Author(s):  
Hanon Lee ◽  
Dong Hun Lee ◽  
Jang-Hee Oh ◽  
Jin Ho Chung
Keyword(s):  
P38 Mapk ◽  
Therapeutic Potential ◽  
Inflammatory Diseases ◽  
Mapk Signaling ◽  
Hacat Cells ◽  
Protein Levels ◽  
Tnf Α ◽  
Ifn Γ ◽  
Mapk Signaling Pathways ◽  
Pro Inflammatory Cytokines

Skullcapflavone II (SFII), a flavonoid derived from Scutellaria baicalensis, has been reported to have anti-inflammatory properties. However, its therapeutic potential for skin inflammatory diseases and its mechanism are unknown. Therefore, this study aimed to investigate the effect of SFII on TNF-α/IFN-γ-induced atopic dermatitis (AD)-associated cytokines, such as thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC). Co-stimulation with TNF-α/IFN-γ in HaCaT cells is a well-established model for induction of pro-inflammatory cytokines. We treated cells with SFII prior to TNF-α/IFN-γ-stimulation and confirmed that it significantly inhibited TARC and MDC expression at the mRNA and protein levels. Additionally, SFII also inhibited the expression of cathepsin S (CTSS), which is associated with itching in patients with AD. Using specific inhibitors, we demonstrated that STAT1, NF-κB, and p38 MAPK mediate TNF-α/IFN-γ-induced TARC and MDC, as well as CTSS expression. Finally, we confirmed that SFII significantly suppressed TNF-α/IFN-γ-induced phosphorylation of STAT1, NF-κB, and p38 MAPK. Taken together, our study indicates that SFII inhibits TNF-α/IFN-γ-induced TARC, MDC, and CTSS expression by regulating STAT1, NF-κB, and p38 MAPK signaling pathways.

Conditioned medium from the human umbilical cord-mesenchymal stem cells stimulate the proliferation of human keratinocytes

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Sushmitha Sriramulu ◽  
Antara Banerjee ◽  
Ganesan Jothimani ◽  
Surajit Pathak
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Umbilical Cord ◽  
Human Keratinocytes ◽  
Human Umbilical Cord ◽  
Hacat Cells ◽  
And Migration

AbstractObjectivesWound healing is a complex process with a sequence of restoring and inhibition events such as cell proliferation, differentiation, migration as well as adhesion. Mesenchymal stem cells (MSC) derived conditioned medium (CM) has potent therapeutic functions and promotes cell proliferation, anti-oxidant, immunosuppressive, and anti-apoptotic effects. The main aim of this research is to study the role of human umbilical cord-mesenchymal stem cells (UC-MSCs) derived CM in stimulating the proliferation of human keratinocytes (HaCaT).MethodsFirstly, MSC were isolated from human umbilical cords (UC) and the cells were then cultured in proliferative medium. We prepared and collected the CM after 72 h. Morphological changes were observed after the treatment of HaCaT cells with CM. To validate the findings, proliferation rate, clonal efficiency and also gene expression studies were performed.ResultsIncreased proliferation rate was observed and confirmed with the expression of Proliferating Cell Nuclear Antigen (PCNA) after treatment with HaCaT cells. Cell-cell strap formation was also observed when HaCaT cells were treated with CM for a period of 5–6 days which was confirmed by the increased expression of Collagen Type 1 Alpha 1 chain (Col1A1).ConclusionsOur results from present study depicts that the secretory components in the CM might play a significant role by interacting with keratinocytes to promote proliferation and migration. Thus, the CM stimulates cellular proliferation, epithelialization and migration of skin cells which might be the future promising application in wound healing.

Evaluation of Potential Application of Wharton’s Jelly-Derived Human Mesenchymal Stromal Cells and its Conditioned Media for Dermal Regeneration using Rat Wound Healing Model

2021 ◽  
pp. 1-14
Author(s):  
Caroline Mathen ◽  
Mrunal Ghag Sawant ◽  
Raghubansh Gupta ◽  
Wilfrid Dsouza ◽  
Shilpa G. Krishna
Keyword(s):  
Wound Healing ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Therapeutic Potential ◽  
Wound Care ◽  
Free Cell ◽  
Conditioned Media ◽  
Human Umbilical Cord ◽  
Human Mscs ◽  
Wound Model

Mesenchymal stromal cells and the derived conditioned media represent an area of tremendous medical interest and, among other clinical applications, are currently being extensively explored for wound healing. The aim of this study was to comparatively evaluate the wound healing potential of xeno-free human umbilical cord-derived mesenchymal stromal cells (MSCs) and the conditioned media (CM) in a full-thickness excision wound model in rats. The evaluation parameters included rate of wound healing, serum cytokine analyses, collagen content, histopathology, and hyperspectral imaging as an independent qualitative and quantitative tool. Both the cell-based and cell-free approaches scored better in lower inflammation, as evidenced in lower IL-10 and stable IL-6 levels, and improved rate of wound healing (<i>p</i> &#x3c; 0.0001). More importantly, no adverse reaction or rejection was observed although human MSCs and CM were used in a xenogeneic model. The presence of hFGF, hHGF, hGCSF, hIL-1Ra, hVEGF, and hIL-6 in the secretome may elucidate the regenerative potential of the xeno-free cell-based and cell-free approaches which have translational value for advanced wound care. The results revealed the therapeutic potential of both the cell-based and cell-free approaches for wound healing.

scholarly journals A heparin sulfate-regulated human keratinocyte autocrine factor is similar or identical to amphiregulin.

1991 ◽  
Vol 11 (5) ◽  
pp. 2547-2557 ◽  
Author(s):  
P W Cook ◽  
P A Mattox ◽  
W W Keeble ◽  
M R Pittelkow ◽  
G D Plowman ◽  
...  
Keyword(s):  
Human Fibroblasts ◽  
Human Keratinocytes ◽  
Mitogenic Activity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Surface Binding ◽  
Mammary Tumor Cell ◽  
Human Keratinocyte ◽  
Heparin Sulfate ◽  
Normal Human ◽  
Autocrine Factor

A novel human keratinocyte-derived autocrine factor (KAF) was purified from conditioned medium by using heparin affinity chromatography as the first step. Purified KAF stimulated the growth of normal human keratinocytes, mouse AKR-2B cells, and a mouse keratinocyte cell line (BALB/MK). Heparin sulfate inhibited KAF mitogenic activity on all cell types tested and inhibited the ability of KAF to compete with epidermal growth factor for cell surface binding. Interestingly, KAF stimulated the growth of BALB/MK cells at high cell density but failed to stimulate these cells at clonal density. Protein microsequencing of the first 20 NH2-terminal amino acid residues of purified KAF revealed identity to the NH2 terminus of human amphiregulin (AR). Northern (RNA) blot analysis with AR-specific cRNA demonstrated that human keratinocytes, as well as mammary epithelial cell cultures, expressed high levels of AR mRNA. In contrast, AR mRNA was not detected in normal human fibroblasts or melanocytes and was present at reduced levels in several mammary tumor cell lines. The mitogenic activity of purified AR was also shown to be inhibited by heparin sulfate, and an AR-specific enzyme-linked immunosorbent assay (ELISA) revealed that KAF and AR are antigenically related. We have previously shown that human keratinocytes can grow in an autocrine manner. Our present study demonstrates that one of the growth factors responsible for this autocrine growth (KAF) is similar or identical to AR and that KAF and AR bioactivity can be negatively regulated by heparin sulfate.

scholarly journals Citrus peel polymethoxyflavones, sudachitin and nobiletin, induce distinct cellular responses in human keratinocyte HaCaT cells

2018 ◽  
Vol 82 (12) ◽  
pp. 2064-2071 ◽  
Author(s):  
Shogo Abe ◽  
Saki Hirose ◽  
Mami Nishitani ◽  
Ichiro Yoshida ◽  
Masao Tsukayama ◽  
...  
Keyword(s):  
Cellular Responses ◽  
Hacat Cells ◽  
Citrus Peel ◽  
Human Keratinocyte
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