Changes in cytokeratin expression in epidermal keratinocytes during wound healing

1995 ◽  
Vol 103 (6) ◽  
pp. 425-433 ◽  
Author(s):  
Shinichi Watanabe ◽  
Masayoshi Osumi ◽  
Takamitsu Ohnishi ◽  
Eiko Ichikawa ◽  
Hisashi Takahashi
Keyword(s):  
Wound Healing ◽  
Epidermal Keratinocytes ◽  
Cytokeratin Expression

scholarly journals Adipose-derived stromal/stem cells improve epidermal homeostasis

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Mariko Moriyama ◽  
Shunya Sahara ◽  
Kaori Zaiki ◽  
Ayumi Ueno ◽  
Koichi Nakaoji ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Culture System ◽  
Wound Repair ◽  
Cell Types ◽  
Skin Wound ◽  
Epidermal Keratinocytes ◽  
Stromal Stem Cells ◽  
Collagen Vitrigel

AbstractWound healing is regulated by complex interactions between the keratinocytes and other cell types including fibroblasts. Recently, adipose-derived mesenchymal stromal/stem cells (ASCs) have been reported to influence wound healing positively via paracrine involvement. However, their roles in keratinocytes are still obscure. Therefore, investigation of the precise effects of ASCs on keratinocytes in an in vitro culture system is required. Our recent data indicate that the epidermal equivalents became thicker on a collagen vitrigel membrane co-cultured with human ASCs (hASCs). Co-culturing the human primary epidermal keratinocytes (HPEK) with hASCs on a collagen vitrigel membrane enhanced their abilities for cell proliferation and adhesion to the membrane but suppressed their differentiation suggesting that hASCs could maintain the undifferentiated status of HPEK. Contrarily, the effects of co-culture using polyethylene terephthalate or polycarbonate membranes for HPEK were completely opposite. These differences may depend on the protein permeability and/or structure of the membrane. Taken together, our data demonstrate that hASCs could be used as a substitute for fibroblasts in skin wound repair, aesthetic medicine, or tissue engineering. It is also important to note that a co-culture system using the collagen vitrigel membrane allows better understanding of the interactions between the keratinocytes and ASCs.

Dissecting the roles of endothelin,TGF-β and GM-CSF on myofibroblast differentiation by keratinocytes

2004 ◽  
Vol 92 (08) ◽  
pp. 262-274 ◽  
Author(s):  
Pierre Shephard ◽  
Boris Hinz ◽  
Sigrun Smola-Hess ◽  
Jean-Jacques Meister ◽  
Thomas Krieg ◽  
...  
Keyword(s):  
Wound Healing ◽  
Smooth Muscle ◽  
Contractile Activity ◽  
Smooth Muscle Actin ◽  
Mechanical Force ◽  
Force Generation ◽  
Epidermal Keratinocytes ◽  
Myofibroblast Differentiation ◽  
Potent Inducer ◽  
Gm Csf

SummaryMyofibroblasts are specialized fibroblasts that contribute to wound healing by producing extracellular matrix and by contracting the granulation tissue.They appear in a phase of wound healing when the dermis strongly interacts with activated epidermal keratinocytes. Direct co-culture with keratinocytes upregulates TGF-β???????????activity and also induces fibroblast to differentiate into α-smooth muscle actin (αSMA)-positive myofibroblasts. TGF-βactivity alone cannot completely account for αSMA induction in these co-cultures, and here we analyze mechanical force generation, another potent inducer of myofibroblast differentiation in this model. Using deformable silicone substrates, we show that contractile activity of fibroblasts is already induced after 1-2-days of co-culture, when fibroblasts are generally αSMA negative. Endothelin-1 (ET-1), the most potent inducer of smooth muscle cell contraction, was up-regulated in co-cultures, while blocking ET-1 with the ET receptor inhibitor PD156252 inhibited contraction in these early co-cultures. In 4-5 days of co-culture, however, fibroblast contractile activity correlated with an increased expression of αSMA expression. Stimulation of fibroblast mono-cultures with ET-1 in a low serum medium did not induce αSMA expression; however, ET-1 did synergize with TGF-β. Surprisingly, GM-CSF, another mediatorstimulating myofibroblast differentiation in granulation tissue, inhibited αSMA expression in fibroblasts, costimulated with TGF-β and ET-1. GM-CSF activated NFκB, thus interfering with TGF-β signaling. Blocking TGF-β and ET-1 largely impaired αSMA induction in co-cultures at day 7 and, in combination, almost completely prevented αSMA induction. Our results dissect the roles of TGF-β and ET-1 on mechanical force generation in keratinocyte-fibroblast co-cultures, and identify GM-CSF as an inducer of myofibroblasts acting indirectly.

scholarly journals Abalone Collagen Extracts Potentiate Stem Cell Properties of Human Epidermal Keratinocytes

Marine Drugs ◽  
2019 ◽  
Vol 17 (7) ◽  
pp. 424
Author(s):  
Sajee Thaweekitphathanaphakdee ◽  
Pithi Chanvorachote ◽  
Sagaw Prateepchinda ◽  
Mattaka Khongkow ◽  
Apirada Sucontphunt
Keyword(s):  
Cell Proliferation ◽  
Wound Healing ◽  
Stem Cell ◽  
Cell Migration ◽  
Expression Level ◽  
Epidermal Keratinocytes ◽  
Stem Cell Markers ◽  
Spheroid Formation ◽  
Human Epidermal Keratinocytes ◽  
Cell Markers

Stem cell activities in human tissues are critical for tissue integrity and function. Maintaining keratinocyte stem cells (KSCs) stemness helps sustain healthy skin by supporting keratinocyte renewal, involving the formation of epidermal barriers. In this study, abalone collagen (AC) extracts with molecular weights of 3 kDa (AC 1) and 300 kDa (AC 2) were compared to the epidermal growth factor (EGF) for their effects on cell proliferation, cell migration (wound healing), spheroid formation, and the expression level of stem cell markers on human keratinocytes (HaCaT cells). Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell proliferation was quantified by ATP and DNA content analysis and Sulforhodamine B (SRB) assays. Cell migration assay was determined using the scratch wound healing test. Spheroid formation was evaluated and the expression level of stem cell markers was investigated by western blot analysis. The results showed that AC 1 at the concentration of 100 µg/mL could stimulate HaCaT cell proliferation, migration, spheroid formation, and the expression level of stem cell markers (keratin 19, β-catenin, ALDH1A1) compared to the control. In conclusion, a smaller molecular weight of abalone collagen extract exhibits a better effect on keratinocytes proliferation, migration, and stemness, which could be a potential active ingredient in cosmeceutical products.

scholarly journals A gelatin/collagen/polycaprolactone scaffold for skin regeneration

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6358 ◽  
Author(s):  
Lin-Gwei Wei ◽  
Hsin-I Chang ◽  
Yiwei Wang ◽  
Shan-hui Hsu ◽  
Lien-Guo Dai ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Dermal Fibroblasts ◽  
Skin Regeneration ◽  
Collagen Content ◽  
Wound Dressings ◽  
Skin Substitute ◽  
Epidermal Keratinocytes ◽  
Adipose Derived Stem Cells

Background A tissue-engineered skin substitute, based on gelatin (“G”), collagen (“C”), and poly(ε-caprolactone) (PCL; “P”), was developed. Method G/C/P biocomposites were fabricated by impregnation of lyophilized gelatin/collagen (GC) mats with PCL solutions, followed by solvent evaporation. Two different GC:PCL ratios (1:8 and 1:20) were used. Results Differential scanning calorimetry revealed that all G/C/P biocomposites had characteristic melting point of PCL at around 60 °C. Scanning electron microscopy showed that all biocomposites had similar fibrous structures. Good cytocompatibility was present in all G/C/P biocomposites when incubated with primary human epidermal keratinocytes (PHEK), human dermal fibroblasts (PHDF) and human adipose-derived stem cells (ASCs) in vitro. All G/C/P biocomposites exhibited similar cell growth and mechanical characteristics in comparison with C/P biocomposites. G/C/P biocomposites with a lower collagen content showed better cell proliferation than those with a higher collagen content in vitro. Due to reasonable mechanical strength and biocompatibility in vitro, G/C/P with a lower content of collagen and a higher content of PCL (GCLPH) was selected for animal wound healing studies. According to our data, a significant promotion in wound healing and skin regeneration could be observed in GCLPH seeded with adipose-derived stem cells by Gomori’s trichrome staining. Conclusion This study may provide an effective and low-cost wound dressings to assist skin regeneration for clinical use.

scholarly journals Human Epidermal Keratinocytes Are a Source of Tenascin-C during Wound Healing

1997 ◽  
Vol 108 (5) ◽  
pp. 776-783 ◽  
Author(s):  
Mieke Latijnhouwers ◽  
Mieke Bergers ◽  
Maria Ponec ◽  
Henri Dijkman ◽  
Monique Andriessen ◽  
...  
Keyword(s):  
Wound Healing ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
Tenascin C

scholarly journals Loss of Vascular Endothelial Growth Factor A Activity in Murine Epidermal Keratinocytes Delays Wound Healing and Inhibits Tumor Formation

2004 ◽  
Vol 64 (10) ◽  
pp. 3508-3516 ◽  
Author(s):  
Heidemarie Rossiter ◽  
Caterina Barresi ◽  
Johannes Pammer ◽  
Michael Rendl ◽  
Jody Haigh ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Wound Healing ◽  
Growth Factor ◽  
Tumor Formation ◽  
Epidermal Keratinocytes ◽  
Vascular Endothelial ◽  
Endothelial Growth Factor

scholarly journals Expression of vascular permeability factor (vascular endothelial growth factor) by epidermal keratinocytes during wound healing.

1992 ◽  
Vol 176 (5) ◽  
pp. 1375-1379 ◽  
Author(s):  
L F Brown ◽  
K T Yeo ◽  
B Berse ◽  
T K Yeo ◽  
D R Senger ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Wound Healing ◽  
Growth Factor ◽  
Vascular Permeability ◽  
Epidermal Keratinocytes ◽  
Vascular Endothelial ◽  
Wound Surface ◽  
Vascular Permeability Factor ◽  
Permeability Factor ◽  
Endothelial Growth Factor

Persistent microvascular hyperpermeability to plasma proteins even after the cessation of injury is a characteristic but poorly understood feature of normal wound healing. It results in extravasation of fibrinogen that clots to form fibrin, which serves as a provisional matrix and promotes angiogenesis and scar formation. We present evidence indicating that vascular permeability factor (VPF; also known as vascular endothelial growth factor) may be responsible for the hyperpermeable state, as well as the angiogenesis, that are characteristic of healing wounds. Hyperpermeable blood vessels were identified in healing split-thickness guinea pig and rat punch biopsy skin wounds by their capacity to extravasate circulating macromolecular tracers (colloidal carbon, fluoresceinated dextran). Vascular permeability was maximal at 2-3 d, but persisted as late as 7 d after wounding. Leaky vessels were found initially at the wound edges and later in the subepidermal granulation tissue as keratinocytes migrated to cover the denuded wound surface. Angiogenesis was also prominent within this 7-d interval. In situ hybridization revealed that greatly increased amounts of VPF mRNA were expressed by keratinocytes, initially those at the wound edge, and, at later intervals, keratinocytes that migrated to cover the wound surface; occasional mononuclear cells also expressed VPF mRNA. Secreted VPF was detected by immunofluoroassay of medium from cultured human keratinocytes. These data identify keratinocytes as an important source of VPF gene transcript and protein, correlate VPF expression with persistent vascular hyperpermeability and angiogenesis, and suggest that VPF is an important cytokine in wound healing.

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