• Klíčová slova
• Aging, Calpain, Hemidesmosome, Matreotype, Skin,
• MeSH
• epidermis * účinky záření   metabolismus   MeSH
• kalpain * metabolismus   antagonisté a inhibitory   MeSH
• keratinocyty účinky záření   metabolismus   MeSH
• lidé MeSH
• škára účinky záření   patologie   metabolismus   cytologie   MeSH
• ultrafialové záření * škodlivé účinky   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• Názvy látek
• kalpain * MeSH

Background: Repairs to deep skin wounds continue to be a difficult issue in clinical practice. A promising approach is to fabricate full-thickness skin substitutes with functions closely similar to those of the natural tissue. For many years, a three-dimensional (3D) collagen hydrogel has been considered to provide a physiological 3D environment for co-cultivation of skin fibroblasts and keratinocytes. This collagen hydrogel is frequently used for fabricating tissue-engineered skin analogues with fibroblasts embedded inside the hydrogel and keratinocytes cultivated on its surface. Despite its unique biological properties, the collagen hydrogel has insufficient stiffness, with a tendency to collapse under the traction forces generated by the embedded cells. Methods: The aim of our study was to develop a two-layer skin construct consisting of a collagen hydrogel reinforced by a nanofibrous poly-L-lactide (PLLA) membrane pre-seeded with fibroblasts. The attractiveness of the membrane for dermal fibroblasts was enhanced by coating it with a thin nanofibrous fibrin mesh. Results: The fibrin mesh promoted the adhesion, proliferation and migration of the fibroblasts upwards into the collagen hydrogel. Moreover, the fibroblasts spontaneously migrating into the collagen hydrogel showed a lower tendency to contract and shrink the hydrogel by their traction forces. The surface of the collagen was seeded with human dermal keratinocytes. The keratinocytes were able to form a basal layer of highly mitotically-active cells, and a suprabasal layer. Conclusion: The two-layer skin construct based on collagen hydrogel with spontaneously immigrated fibroblasts and reinforced by a fibrin-coated nanofibrous membrane seems to be promising for the construction of full-thickness skin substitute.

• Klíčová slova
• collagen hydrogel, fibrin, fibroblast and keratinocyte co-cultivation, full-thickness skin substitutes, nanostructure,
• MeSH
• fibrin farmakologie   MeSH
• fibroblasty cytologie   účinky léků   MeSH
• hydrogely farmakologie   MeSH
• keratinocyty cytologie   účinky léků   MeSH
• kolagen farmakologie   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• membrány umělé * MeSH
• mitochondrie účinky léků   metabolismus   MeSH
• nanovlákna chemie   MeSH
• novorozenec MeSH
• pohyb buněk účinky léků   MeSH
• polyestery farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• škára cytologie   MeSH
• umělá kůže * MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• novorozenec MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fibrin MeSH
• hydrogely MeSH
• kolagen MeSH
• membrány umělé * MeSH
• poly(lactide) MeSH Prohlížeč
• polyestery MeSH

Surface modification is an important step in making a synthetic polymer cytocompatible. We have previously reported improved cytocompatibility of immortalized human keratinocytes (HaCaT) with the otherwise bioinert fluorinated ethylene propylene (FEP) upon treatment with argon plasma discharge. In this article, we show that FEP modified with Ar plasma with the power of 3 and 8 W for 40 and 240 s served as a suitable material for cultivation of primary human dermal fibroblasts (HDF), which showed significantly improved proliferation and spreading comparable to standard tissue culture polystyrene. We also evaluated focal adhesions formed by HDF cells on modified FEP, which were far more numerous compared to pristine FEP. Moreover, we attempted spontaneous osteogenic differentiation of adipose-derived mesenchymal stem cells modified with human telomerase reverse transcriptase on Ar plasma-modified FEP. While the spontaneous osteogenic differentiation was unsuccessful, the cells were able to adhere and differentiated on tested matrices upon the administration of osteodifferentiation medium. These combined findings suggest that the treatment of FEP with Ar plasma comprises and efficient method to enable the adhesion and proliferation of various cell types on an otherwise largely bioinert material.

• Klíčová slova
• Biocompatibility, Fluorinated ethylene propylene, Focal adhesion, Human primary dermal fibroblasts, Osteodifferentiation, hTERT immortalized adipose-derived mesenchymal stem cells,
• MeSH
• argon chemie   MeSH
• buněčná adheze MeSH
• buněčná diferenciace MeSH
• buněčné linie MeSH
• fibroblasty cytologie   metabolismus   MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• osteogeneze MeSH
• plazmové plyny chemie   MeSH
• polytetrafluoroethylen analogy a deriváty   chemie   MeSH
• proliferace buněk MeSH
• škára cytologie   metabolismus   MeSH
• tuková tkáň cytologie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• argon MeSH
• fluorinated ethylene propylene MeSH Prohlížeč
• plazmové plyny MeSH
• polytetrafluoroethylen MeSH

Dermal fibroblasts, which make up the major cell type in the dermis, have, historically, been considered to be relatively 'passive' cells which are responsible for the synthesis and remodeling of extracellular matrix proteins. However, the dermal fibroblast population is composed of heterogeneous and distinct cell types, and it has been established that, under the stress conditions of healing wound environments, dermal fibroblasts participate in the regulation of ongoing inflammation and cell proliferation by secreting a variety of signaling molecules that modulate the functions of immune cells, keratinocyte, endothelial cells and mast cells via both direct cell to cell communication and autocrine and paracrine interactions. This review describes the capacity of dermal fibroblasts to sense and respond to signals from the micro-environment and to communicate with surrounding cells during cutaneous wound healing. The review further emphasizes the, to date, poorly understood roles of heterogeneous dermal fibroblast populations in the wound healing process.

• Klíčová slova
• Dermal fibroblast, Intercellular communication, Papillary, Reticular, Wound healing,
• MeSH
• endoteliální buňky fyziologie   MeSH
• extracelulární matrix - proteiny fyziologie   MeSH
• fibroblasty fyziologie   MeSH
• hojení ran * MeSH
• keratinocyty fyziologie   MeSH
• lidé MeSH
• mezibuněčná komunikace * MeSH
• proliferace buněk MeSH
• signální transdukce MeSH
• škára cytologie   MeSH
• zánět imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• extracelulární matrix - proteiny MeSH

OBJECTIVES: Tribbles homologue 3 (TRB3) is a pseudokinase that modifies the activation of various intracellular signalling pathways to control fundamental processes extending from mitosis and cell activation to apoptosis and modulation of gene expression. Here, we aimed to analyse the role of TRB3 in fibroblast activation in systemic sclerosis (SSc). METHODS: The expression of TRB3 was quantified by quantitative PCR, western blot and immunohistochemistry. The role of TRB3 was analysed in cultured fibroblasts and in experimental fibrosis using small interfering RNA (siRNA)-mediated knockdown and overexpression of TRB3. RESULTS: TRB3 expression was increased in fibroblasts of patients with SSc and in murine models of SSc in a transforming growth factor-β (TGF-β)/Smad-dependent manner. Overexpression of TRB3 stimulated canonical TGF-β signalling and induced an activated phenotype in resting fibroblasts. In contrast, knockdown of TRB3 reduced the profibrotic effects of TGF-β and decreased the collagen synthesis. Moreover, siRNA-mediated knockdown of TRB3 exerted potent antifibrotic effects and ameliorated bleomycin as well as constitutively active TGF-β receptor I-induced fibrosis with reduced dermal thickening, decreased hydroxyproline content and impaired myofibroblast differentiation. CONCLUSIONS: The present study characterises TRB3 as a novel profibrotic mediator in SSc. TGF-β induces TRB3, which in turn activates canonical TGF-β/Smad signalling and stimulates the release of collagen, thereby inducing a positive feedback loop that may contribute to aberrant TGF-β signalling in SSc.

• Klíčová slova
• Fibroblasts, Systemic Sclerosis, Treatment,
• MeSH
• bleomycin toxicita   MeSH
• dospělí MeSH
• fibroblasty metabolismus   MeSH
• fibróza chemicky indukované   genetika   MeSH
• genový knockdown MeSH
• genový knockin MeSH
• imunohistochemie MeSH
• kolagen metabolismus   MeSH
• kožní nemoci chemicky indukované   genetika   MeSH
• kultivované buňky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• protein Smad3 metabolismus   MeSH
• protein-serin-threoninkinasy antagonisté a inhibitory   genetika   metabolismus   MeSH
• proteiny buněčného cyklu genetika   metabolismus   MeSH
• protinádorová antibiotika toxicita   MeSH
• receptory transformujícího růstového faktoru beta MeSH
• represorové proteiny genetika   metabolismus   MeSH
• senioři MeSH
• signální transdukce genetika   MeSH
• škára cytologie   MeSH
• studie případů a kontrol MeSH
• systémová sklerodermie genetika   metabolismus   MeSH
• TGF-beta receptor I. typu MeSH
• transformující růstový faktor beta metabolismus   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• myši MeSH
• senioři MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bleomycin MeSH
• kolagen MeSH
• protein Smad3 MeSH
• protein-serin-threoninkinasy MeSH
• proteiny buněčného cyklu MeSH
• protinádorová antibiotika MeSH
• receptory transformujícího růstového faktoru beta MeSH
• represorové proteiny MeSH
• TGF-beta receptor I. typu MeSH
• transformující růstový faktor beta MeSH
• TRB3 protein, mouse MeSH Prohlížeč
• TRIB3 protein, human MeSH Prohlížeč

MicroRNA (miRNAs) are short noncoding RNA molecules involved in many cellular processes and shown to play a key role in somatic cell induced reprogramming. We performed an array based screening to identify candidates that are differentially expressed between dermal skin fibroblasts (DFs) and induced pluripotent stem cells (iPSCs). We focused our investigations on miR-145 and showed that this candidate is highly expressed in DFs relative to iPSCs and significantly downregulated during reprogramming process. Inhibition of miR-145 in DFs led to the induction of "cellular plasticity" demonstrated by: (a) alteration of cell morphology associated with downregulation of mesenchymal and upregulation of epithelial markers; (b) upregulation of pluripotency-associated genes including SOX2, KLF4, C-MYC; (c) downregulation of miRNA let-7b known to inhibit reprogramming; and (iv) increased efficiency of reprogramming to iPSCs in the presence of reprogramming factors. Together, our results indicate a direct functional link between miR-145 and molecular pathways underlying reprogramming of somatic cells to iPSCs.

• Klíčová slova
• Induced pluripotent stem cells, KLF4, Mesenchymal-to-epithelial transition, OCT4, Reprogramming, SOX2, c-MYC, miR-145, microRNA,
• MeSH
• fibroblasty cytologie   metabolismus   MeSH
• indukované pluripotentní kmenové buňky cytologie   MeSH
• Krüppel-like faktor 4 MeSH
• lidé MeSH
• mikro RNA genetika   metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• přeprogramování buněk * genetika   MeSH
• regulace genové exprese MeSH
• reprodukovatelnost výsledků MeSH
• sekvence nukleotidů MeSH
• škára cytologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• KLF4 protein, human MeSH Prohlížeč
• Krüppel-like faktor 4 MeSH
• mikro RNA MeSH
• MIRN145 microRNA, human MeSH Prohlížeč

Complex regulation of the wound healing process involves multiple interactions among stromal tissue cells, inflammatory cells, and the extracellular matrix. Low molecular weight hyaluronan (LMW HA) derived from the degradation of high molecular weight hyaluronan (HMW HA) is suggested to activate cells involved in wound healing through interaction with HA receptors. In particular, receptor CD44 is suggested to mediate cell response to HA of different MW, being the main cell surface HA receptor in stromal tissue and immune cells. However, the response of dermal fibroblasts, the key players in granulation tissue formation within the wound healing process, to LMW HA and their importance for the activation of immune cells is unclear. In this study we show that LMW HA (4.3kDa) induced pro-inflammatory cytokine IL-6 and chemokines IL-8, CXCL1, CXCL2, CXCL6 and CCL8 gene expression in normal human dermal fibroblasts (NHDF) that was further confirmed by increased levels of IL-6 and IL-8 in cell culture supernatants. Conversely, NHDF treated by HMW HA revealed a tendency to decrease the gene expression of these cytokine and chemokines when compared to untreated control. The blockage of CD44 expression by siRNA resulted in the attenuation of IL-6 and chemokines expression in LMW HA treated NHDF suggesting the involvement of CD44 in LMW HA mediated NHDF activation. The importance of pro-inflammatory mediators produced by LMW HA triggered NHDF was evaluated by significant activation of blood leukocytes exhibited as increased production of IL-6 and TNF-α. Conclusively, we demonstrated a pro-inflammatory response of dermal fibroblasts to LMW HA that was transferred to leukocytes indicating the significance of LMW HA in the inflammatory process development during the wound healing process.

• Klíčová slova
• Chemokines, Hyaluronan, IL-6, IL-8, Inflammation,
• MeSH
• antigeny CD44 metabolismus   MeSH
• chemokiny biosyntéza   genetika   MeSH
• fibroblasty účinky léků   imunologie   MeSH
• interleukin-6 biosyntéza   genetika   metabolismus   MeSH
• interleukin-8 biosyntéza   MeSH
• kyselina hyaluronová farmakologie   MeSH
• leukocyty účinky léků   metabolismus   MeSH
• lidé MeSH
• malá interferující RNA metabolismus   MeSH
• mediátory zánětu metabolismus   MeSH
• messenger RNA genetika   metabolismus   MeSH
• molekulová hmotnost MeSH
• přirozená imunita účinky léků   genetika   MeSH
• signální transdukce účinky léků   genetika   MeSH
• škára cytologie   MeSH
• TNF-alfa biosyntéza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD44 MeSH
• chemokiny MeSH
• interleukin-6 MeSH
• interleukin-8 MeSH
• kyselina hyaluronová MeSH
• malá interferující RNA MeSH
• mediátory zánětu MeSH
• messenger RNA MeSH
• TNF-alfa MeSH

Cancer-associated fibroblasts (CAFs) play a role in the progression of malignant tumors. They are formed by conversion of fibroblasts to smooth muscle α-actin-positive (SMA-positive) myofibroblasts. Polyamines are known to change the arrangement of the actin cytoskeleton by binding to the anionic actin. We tested the effect of the synthetic polyamine BPA-C8 on the transition of human dermal fibroblasts to myofibroblasts induced either by TGF-β1 alone or by TGF-β1 together with adhesion/growth-regulatory galectin-1. Pre-existing CAFs, myofibroblasts from pancreatitis, and rat smooth muscle cells were also exposed to BPA-C8. BPA-C8 impaired myofibroblast formation from activated fibroblasts, but it had no effect on cells already expressing SMA. BPA-C8 also reduced the occurrence of an extracellular matrix around the activated fibroblasts. The reported data thus extend current insights into polyamine activity, adding interference with tumor progression to the tumor-promoting processes warranting study.

• Klíčová slova
• actin, cancer, galectin, polycations, tenascin,
• MeSH
• aktiny metabolismus   MeSH
• fibroblasty účinky léků   patologie   MeSH
• galektin 1 metabolismus   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• myofibroblasty účinky léků   patologie   MeSH
• nádorové buňky kultivované MeSH
• nádory farmakoterapie   metabolismus   patologie   MeSH
• polyaminy chemie   farmakologie   MeSH
• škára cytologie   účinky léků   MeSH
• transformující růstový faktor beta1 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• galektin 1 MeSH
• polyaminy MeSH
• transformující růstový faktor beta1 MeSH

Biological meshes are biomaterials consisting of extracellular matrix that are used in surgery particularly for hernia treatment, thoracic wall reconstruction, or silicone implant-based breast reconstruction. We hypothesized that combination of extracellular matrices with autologous mesenchymal stem cells used for hernia repair would result in increased vascularization and increased strength of incorporation. We cultured autologous adipose-derived stem cells harvested from the inguinal region of Wistar rats on cross-linked and noncross-linked porcine extracellular matrices. In 24 Wistar rats, a standardized 2×4 cm fascial defect was created and repaired with either cross-linked or noncross-linked grafts enriched with stem cells. Non-MSC-enriched grafts were used as controls. The rats were sacrificed at 3 months of age. The specimens were examined for the strength of incorporation, vascularization, cell invasion, foreign body reaction, and capsule formation. Both materials showed cellular ingrowth and neovascularization. Comparison of both tested groups with the controls showed no significant differences in the capsule thickness, foreign body reaction, cellularization, or vascularization. The strength of incorporation of the stem cell-enriched cross-linked extracellular matrix specimens was higher than in acellular specimens, but this result was statistically nonsignificant. In the noncross-linked extracellular matrix, the strength of incorporation was significantly higher in the stem cell group than in the acellular group. Seeding of biological meshes with stem cells does not significantly contribute to their increased vascularization. In cross-linked materials, it does not ensure increased strength of incorporation, in contrast to noncross-linked materials. Owing to the fact that isolation and seeding of stem cells is a very complex procedure, we do not see sufficient benefits for its use in the clinical setting.

• Klíčová slova
• Acellular matrix, Adipose-derived stem cells, Animal model, Biocompatibility, Cross-linking, Extracellular matrix, Hernia, Mesenchymal stem cells, Scaffold,
• MeSH
• buněčná diferenciace MeSH
• extracelulární matrix chemie   MeSH
• hernie terapie   MeSH
• kultivované buňky MeSH
• mezenchymální kmenové buňky cytologie   MeSH
• potkani Wistar MeSH
• prasata MeSH
• reagencia zkříženě vázaná chemie   MeSH
• škára cytologie   MeSH
• tkáňové inženýrství MeSH
• tkáňové podpůrné struktury chemie   MeSH
• tuková tkáň cytologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• reagencia zkříženě vázaná MeSH

It has been shown that fibroblasts within the stroma of malignant tumours can affect the tumour's biological character, influencing such properties as local aggressiveness and metastasis potential. This influence is asserted via paracrine secretion of multiple cell factors, including chemokines. This study demonstrates that both normal keratinocytes and cancer cells can stimulate the secretion of chemokines IL-8 and CXCL-1 from normal dermal fibroblasts and stromal fibroblasts from squamous cell carcinoma. The effect of epithelia on normal fibroblasts leads to a transient secretory change, in contrast to stromal fibroblasts which generate a more prolonged one. This observation demonstrates that stimulated expression of both IL-8 and CXCL-1 is not specific to cancer, supporting the hypothesis that similar mechanisms exist between wound healing and oncogenesis. It also shows that stromal fibroblasts isolated from a tumour have significantly different features from normal fibroblasts.

• MeSH
• buňky stromatu metabolismus   MeSH
• chemokin CXCL1 metabolismus   MeSH
• fibroblasty metabolismus   MeSH
• hojení ran fyziologie   MeSH
• interleukin-8 metabolismus   MeSH
• keratinocyty fyziologie   MeSH
• kokultivační techniky MeSH
• kultivační média speciální MeSH
• kultivované buňky MeSH
• lidé MeSH
• nádorové proteiny metabolismus   MeSH
• nádory hypofaryngu metabolismus   patologie   MeSH
• nádory kůže metabolismus   patologie   MeSH
• rychlost sekrece MeSH
• škára cytologie   MeSH
• spinocelulární karcinom metabolismus   patologie   MeSH
• upregulace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• chemokin CXCL1 MeSH
• CXCL1 protein, human MeSH Prohlížeč
• interleukin-8 MeSH
• kultivační média speciální MeSH
• nádorové proteiny MeSH