Pulmonary artery banding is a surgical procedure performed when there is a shunt between the left and right ventricle. Its aim is to constrict the lumen of the pulmonary artery by using a band to reduce blood flow to the lungs. In this study, we report the results of investigating the mechanical properties of a composite composed of poly(L-lactide-co-ε-caprolactone) layers and a collagen matrix (PLCL-COLL). PLCL layers were obtained by electrospinning, impregnated with collagen solution, and finally cross-linked to increase the stiffness of the material. Bands of PLCL-COLL were implanted into a rat peritoneum and explanted after 1, 3, and 6 months in vivo. The mechanical properties of the material before and after implantation were determined using uniaxial tensile tests. The same was done with samples of strips prepared from GORE-TEX material. By comparing the results of tensile tests before implantation and after explantation, it was found that PLCL-COLL degrades in the rat's body and that it exhibits a mechanical response showing of elastic modulus values that correspond well to arterial biomechanics (elastic modulus measured in the initial linear region of the deformation was found to be: 4.14 MPa ± 1.11 MPa, 2.34 MPa ± 1.02 MPa, 1.11 MPa ± 0.77 MPa, and 0.88 MPa ± 0.60 MPa before implantation, and 1, 3, and 6 months after implantation respectively). Similar to the elastic modulus, the strength of the PLCL-COLL composite decreased during in vivo exposure (1.32 ± 0.32 MPa, 0.60 ± 0.26 MPa, 0.44 ± 0.11 MPa, and 0.46 ± 0.28 MPa before implantation, and 1, 3, and 6 months after implantation respectively). In our experiments, PLCL-COLL material was always more compliant than GORE-TEX (elastic modulus 34.7 MPa ± 2.06 MPa before implantation, and 9.35 MPa ± 6.80 MPa after implantation). The results suggest that PLCL-COLL could be a suitable candidate for the development of artery banding tapes, and also for further use in cardiovascular surgery.
- Klíčová slova
- Bioresorbable material, Collagen, Degradation, Elasticity, In vivo, L-lactide, Pulmonary artery banding, ε-caprolactone,
- MeSH
- arteria pulmonalis * chirurgie MeSH
- biokompatibilní materiály chemie MeSH
- biomechanika MeSH
- kolagen * chemie metabolismus MeSH
- krysa rodu Rattus MeSH
- mechanické jevy * MeSH
- peritoneum * chirurgie MeSH
- pevnost v tahu MeSH
- polyestery * chemie metabolismus MeSH
- testování materiálů MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biokompatibilní materiály MeSH
- kolagen * MeSH
- lactide-caprolactone copolymer MeSH Prohlížeč
- polyestery * MeSH
AIM: To evaluate the long-term impact of accelerated corneal cross-linking (A-CXL) on selected refractive and topographical parameters in eyes with progressive keratoconus. METHODS: 77 eyes with keratoconus in 54 patients treated with A-CXL (10 min "epi-off" protocol) were included in the analysis. Preoperative and postoperative (1, 3 and 5 years after A-CXL) values of the studied parameters were compared. RESULTS: In the cohort, there was an improvement in best corrected central visual acuity (BCCVA) 1 year (p = 0.004) and 3 years (p.
- Klíčová slova
- progressive keratoconus; accelerated corneal cross-linking; refractive and topographical parameters,
- MeSH
- dospělí MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky MeSH
- keratokonus * farmakoterapie diagnóza patofyziologie metabolismus MeSH
- kolagen metabolismus MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- reagencia zkříženě vázaná * MeSH
- refrakce oka MeSH
- riboflavin terapeutické užití MeSH
- rohovková topografie * MeSH
- zraková ostrost * MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fotosenzibilizující látky MeSH
- kolagen MeSH
- reagencia zkříženě vázaná * MeSH
- riboflavin MeSH
Fibrotic changes in pediatric clubfoot provide an opportunity to improve corrective therapy and prevent relapses with targeted drugs. This study defines the parameters of clubfoot fibrosis and presents a unique analysis of a simple pseudo-3D in vitro model for disease-specific high-throughput drug screening experiments. The model combines clubfoot-derived fibroblasts with a biomimetic cultivation environment induced by the water-soluble polymers Ficoll and Polyvinylpyrrolidone, utilizing the principle of macromolecular crowding. We achieved higher conversion of soluble collagen into insoluble collagen, accelerated formation of the extracellular matrix layer and upregulated fibrosis-related genes in the mixed Ficoll environment. To test the model, we evaluated the effect of a potential antifibrotic drug, minoxidil, emphasizing collagen content and cross-linking. While the model amplified overall collagen deposition, minoxidil effectively blocked the expression of lysyl hydroxylases, which are responsible for the increased occurrence of specific collagen cross-linking in various fibrotic tissues. This limited the formation of collagen cross-link in both the model and control environments. Our findings provide a tool for expanding preclinical research for clubfoot and similar fibroproliferative conditions.
- MeSH
- biomimetika metody MeSH
- extracelulární matrix metabolismus účinky léků MeSH
- fibroblasty * metabolismus účinky léků MeSH
- fibróza * farmakoterapie MeSH
- kolagen * metabolismus chemie MeSH
- kultivované buňky MeSH
- lidé MeSH
- pes equinovarus * metabolismus farmakoterapie patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kolagen * MeSH
Natural products have many healing effects on the skin with minimal or no adverse effects. In this study, we analyzed the regenerative properties of a waste product (hydrolate) derived from Helichrysum italicum (HH) on scratch-tested skin cell populations seeded on a fluidic culture system. Helichrysum italicum has always been recognized in the traditional medicine of Mediterranean countries for its wide pharmacological activities. We recreated skin physiology with a bioreactor that mimics skin stem cell (SSCs) and fibroblast (HFF1) communication as in vivo skin layers. Dynamic culture models represent an essential instrument for recreating and preserving the complex multicellular organization and interactions of the cellular microenvironment. Both cell types were exposed to two different concentrations of HH after the scratch assay and were compared to untreated control cells. Collagen is the constituent of many wound care products that act directly on the damaged wound environment. We analyzed the role played by HH in stimulating collagen production during tissue repair, both in static and dynamic culture conditions, by a confocal microscopic analysis. In addition, we performed a gene expression analysis that revealed the activation of a molecular program of stemness in treated skin stem cells. Altogether, our results indicate a future translational application of this natural extract to support skin regeneration and define a new protocol to recreate a dynamic process of healing.
- Klíčová slova
- Helichrysum italicum, bioreactor, dynamic cultures, fibroblasts, molecular mechanisms, stem cells, tissue regeneration, wound healing,
- MeSH
- fibroblasty metabolismus účinky léků MeSH
- Helichrysum * chemie MeSH
- hojení ran * účinky léků MeSH
- kmenové buňky metabolismus účinky léků cytologie MeSH
- kolagen * metabolismus MeSH
- kultivované buňky MeSH
- kůže * metabolismus účinky léků MeSH
- lidé MeSH
- regenerace * účinky léků MeSH
- rostlinné extrakty * farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kolagen * MeSH
- rostlinné extrakty * MeSH
Human skin is exposed to various physical and chemical stress factors, which commonly cause the oxidation of lipids and proteins. In this study, azo initiator AAPH [2,2' -azobis(2-methylpropionamidine) dihydrochloride] was employed to initiate lipid peroxidation in porcine skin as an ex vivo model for human skin. We demonstrate that malondialdehyde (MDA), a secondary product of lipid peroxidation, is covalently bound to collagen in the dermis, forming MDA-collagen adducts. The binding of MDA to collagen results in an unfolding of the collagen triple helix, formation of the dimer of α-chains of collagen, and fragmentation of the collagen α-chain. It is proposed here that the MDA is bound to the lysine residues of α-chain collagen, which are involved in electrostatic interaction and hydrogen bonding with the glutamate and aspartate of other α-chains of the triple helix. Our data provide crucial information about the MDA binding topology in the skin, which is necessary to understand better the various types of skin-related diseases and the aging process in the skin under stress.
- Klíčová slova
- Collagen, Lysine, Malondialdehyde, Reactive oxygen species, Skin,
- MeSH
- kolagen * metabolismus MeSH
- lidé MeSH
- malondialdehyd metabolismus MeSH
- oxidace-redukce MeSH
- oxidační stres * MeSH
- peroxidace lipidů MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kolagen * MeSH
- malondialdehyd MeSH
Non-alcoholic fatty liver disease (NAFLD), encompassing fatty liver and its progression into nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma (HCC), is one of the rapidly rising health concerns worldwide. SIRT6 is an essential nuclear sirtuin that regulates numerous pathological processes including insulin resistance and inflammation, and recently it has been implicated in the amelioration of NAFLD progression. SIRT6 overexpression protects from formation of fibrotic lesions. However, the underlying molecular mechanisms are not fully delineated. Moreover, new allelic variants of SIRT6 (N308K/A313S) were recently associated with the longevity in Ashkenazi Jews by improving genome maintenance and DNA repair, suppressing transposons and killing cancer cells. Whether these new SIRT6 variants play different or enhanced roles in liver diseases is currently unknown. In this study, we aimed to clarify how these new centenarian-associated SIRT6 genetic variants affect liver metabolism and associated diseases. We present evidence that overexpression of centenarian-associated SIRT6 variants dramatically altered the metabolomic and secretomic profiles of unchallenged immortalized human hepatocytes (IHH). Most amino acids were increased in the SIRT6 N308K/A313S overexpressing IHH when compared to IHH transfected with the SIRT6 wild-type sequence. Several unsaturated fatty acids and glycerophospholipids were increased, and ceramide tended to be decreased upon SIRT6 N308K/A313S overexpression. Furthermore, we found that overexpression of SIRT6 N308K/A313S in a 3D hepatic spheroid model formed by the co-culture of human immortalized hepatocytes (IHH) and hepatic stellate cells (LX2) inhibited collagen deposition and fibrotic gene expression in absence of metabolic or dietary challenges. Hence, our findings suggest that novel longevity associated SIRT6 N308K/A313S variants could favor the prevention of NASH by altering hepatocyte proteome and lipidome.
- Klíčová slova
- Aging, Hepatic stellate cells, Hepatocytes, Liver, Metabolomics, SIRT6, Spheroids,
- MeSH
- hepatocelulární karcinom * metabolismus patologie MeSH
- hepatocyty metabolismus patologie MeSH
- kolagen metabolismus MeSH
- lidé MeSH
- nádory jater * metabolismus patologie MeSH
- nealkoholová steatóza jater * genetika metabolismus patologie MeSH
- senioři nad 80 let MeSH
- sirtuiny * genetika metabolismus MeSH
- století lidé MeSH
- Check Tag
- lidé MeSH
- senioři nad 80 let MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kolagen MeSH
- SIRT6 protein, human MeSH Prohlížeč
- sirtuiny * MeSH
Resident tissue macrophages are organ-specialized phagocytes responsible for the maintenance and protection of tissue homeostasis. It is well established that tissue diversity is reflected by the heterogeneity of resident tissue macrophage origin and phenotype. However, much less is known about tissue-specific phagocytic and proteolytic macrophage functions. Here, using a quantitative proteomics approach, we identify cathepsins as key determinants of phagosome maturation in primary peritoneum-, lung-, and brain-resident macrophages. The data further uncover cathepsin K (CtsK) as a molecular marker for lung phagosomes required for intracellular protein and collagen degradation. Pharmacological blockade of CtsK activity diminished phagosomal proteolysis and collagenolysis in lung-resident macrophages. Furthermore, profibrotic TGF-β negatively regulated CtsK-mediated phagosomal collagen degradation independently from classical endocytic-proteolytic pathways. In humans, phagosomal CtsK activity was reduced in COPD lung macrophages and non-COPD lung macrophages exposed to cigarette smoke extract. Taken together, this study provides a comprehensive map of how peritoneal, lung, and brain tissue environment shapes phagosomal composition, revealing CtsK as a key molecular determinant of lung phagosomes contributing to phagocytic collagen clearance in lungs.
- MeSH
- fagozomy * metabolismus MeSH
- kathepsin K * metabolismus MeSH
- kolagen metabolismus MeSH
- lidé MeSH
- makrofágy * metabolismus MeSH
- plíce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- CTSK protein, human MeSH Prohlížeč
- kathepsin K * MeSH
- kolagen MeSH
Scaffolds made of degradable polymers, such as collagen, polyesters or polysaccharides, are promising matrices for fabrication of bioartificial vascular grafts or patches. In this study, collagen isolated from porcine skin was processed into a gel, reinforced with collagen particles and with incorporated adipose tissue-derived stem cells (ASCs). The cell-material constructs were then incubated in a DMEM medium with 2% of FS (DMEM_part), with added polyvinylalcohol nanofibers (PVA_part sample), and for ASCs differentiation towards smooth muscle cells (SMCs), the medium was supplemented either with human platelet lysate released from PVA nanofibers (PVA_PL_part) or with TGF-β1 + BMP-4 (TGF + BMP_part). The constructs were further endothelialised with human umbilical vein endothelial cells (ECs). The immunofluorescence staining of alpha-actin and calponin, and von Willebrand factor, was performed. The proteins involved in cell differentiation, the extracellular matrix (ECM) proteins, and ECM remodelling proteins were evaluated by mass spectrometry on day 12 of culture. Mechanical properties of the gels with ASCs were measured via an unconfined compression test on day 5. Gels evinced limited planar shrinkage, but it was higher in endothelialised TGF + BMP_part gel. Both PVA_PL_part samples and TGF + BMP_part samples supported ASC growth and differentiation towards SMCs, but only PVA_PL_part supported homogeneous endothelialisation. Young modulus of elasticity increased in all samples compared to day 0, and PVA_PL_part gel evinced a slightly higher ratio of elastic energy. The results suggest that PVA_PL_part collagen construct has the highest potential to remodel into a functional vascular wall.
- Klíčová slova
- adipose tissue-derived stem cells, collagen particles, endothelial cells, extracellular matrix, gel reinforcement, remodelling, stem cells differentiation, tissue engineering, vascular patches,
- MeSH
- buněčná diferenciace MeSH
- endoteliální buňky pupečníkové žíly (lidské) MeSH
- extracelulární matrix - proteiny metabolismus MeSH
- gely metabolismus MeSH
- kmenové buňky metabolismus MeSH
- kolagen * metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- myocyty hladké svaloviny metabolismus MeSH
- prasata MeSH
- tkáňové inženýrství metody MeSH
- tuková tkáň * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- extracelulární matrix - proteiny MeSH
- gely MeSH
- kolagen * MeSH
Lung fibrosis is a serious human pathology. MiR-146b-5p is down-regulated in idiopathic pulmonary fibrosis, and the Notch1/PDGFRβ/ROCK1 pathway is activated. However, the relation between miR-146b-5p and the Notch1/PDGFRβ/ROCK1 pathway in lung fibrosis remains unclear. To investigate the function of miR-146b-5p in lung fibrosis, an in vivo model of lung fibrosis was established in mice by bleomycin. The fibrosis in lung tissues of mice was observed by HE, Masson and Sirius Red staining. Lung pericytes were isolated and identified by fluorescence microscopy. Immunofluorescence staining and Western blot were used to investigate the expression of desmin, NG2, collagen I and α-SMA. CCK8 assay was used to assess the cell viability, and flow cytometry was performed to evaluate the cell cycle in pericytes. Furthermore, the correlation between miR-146b-5p and Notch1 was analysed by Spearman analysis. The mechanism by which miR-146b-5p affects pericytes and lung fibrosis via the Notch1/ PDGFRβ/ROCK1 pathway was explored by RT-qPCR, Western blot, immunofluorescence staining and dual luciferase reporter gene assay. In bleomycin-treated mice, miR-146b-5p was down-regulated, while Notch1 was up-regulated. Up-regulation of miR-146b-5p significantly inhibited the viability and induced G1 phase arrest of lung pericytes. MiR-146b-5p mimics up-regulated miR-146b-5p, desmin, and NG2 and down-regulated α-SMA and collagen I in the lung pericytes. Additionally, miR-146b-5p was negatively correlated with Notch1, and miR-146b-5p interacted with Notch1. Over-expression of miR-146b-5p inactivated the Notch1/PDGFRβ/ROCK1 pathway. Our results indicate that up-regulation of miR-146b-5p inhibits fibrosis in lung pericytes via modulation of the Notch1/PDGFRβ/ROCK1 pathway. Thus, our study might provide a novel target against lung fibrosis.
- MeSH
- bleomycin metabolismus MeSH
- desmin genetika metabolismus MeSH
- kinázy asociované s Rho genetika metabolismus MeSH
- kolagen genetika metabolismus MeSH
- lidé MeSH
- mikro RNA * genetika metabolismus MeSH
- myši MeSH
- pericyty metabolismus patologie MeSH
- plíce metabolismus patologie MeSH
- plicní fibróza * genetika metabolismus patologie MeSH
- receptor Notch1 genetika metabolismus MeSH
- upregulace genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bleomycin MeSH
- desmin MeSH
- kinázy asociované s Rho MeSH
- kolagen MeSH
- mikro RNA * MeSH
- NOTCH1 protein, human MeSH Prohlížeč
- receptor Notch1 MeSH
- ROCK1 protein, human MeSH Prohlížeč
Stem cells have emerged as promising therapeutic options for several human diseases, including pulmonary fibrosis (PF). In this study, we investigated the therapeutic effects of adipose tissue-derived mesenchymal stem cells (ADMSCs) in the bleomycin-induced PF model rats and the underlying mechanisms. The PF model rats were generated by intratracheal injections of 5 mg/kg bleomycin sulfate. The ADMSC group rats were generated by injecting 2×10(6) ADMSCs via the tail vein at 0, 12, and 24 h after bleomycin injection. The control, PF, and ADMSC group rats were sacrificed on day 21 after bleomycin injections and the changes in lung histology and the levels of pro-inflammatory cytokines, collagen I, and caveolin-1 (Cav-1), and the activity of the NF-kappaB signaling pathway in the lung tissues was assessed by hematoxylin-eosin staining, ELISA, and western blotting assays. The lung tissues of the PF model rats showed significant infiltration of neutrophils, tissue destruction, and collagen deposition, but these effects were abrogated by the ADMSCs. The levels of pro-inflammatory cytokines such as IL-6, IL-1beta, and TGF-beta1 were elevated in the lung tissues and the bronchoalveolar lavage fluid (BALF) of the bleomycin-induced PF model rats, but these effects were reversed by the ADMSCs. The lung tissues of the PF model rats showed significant downregulation of Cav-1 and significantly higher activation of the pro-inflammatory NF-kappaB pathway. However, administration of the ADMSCs restored the expression levels of Cav-1 and suppressed the NF-kappaB signaling pathway in the lungs of the bleomycin-induced PF model rats. In conclusion, this study demonstrated that the ADMSCs protected against bleomycin-induced PF in the rat model by modulating the Cav-1/NF-kappaB axis.
- MeSH
- bleomycin toxicita MeSH
- cytokiny metabolismus MeSH
- kaveolin 1 metabolismus farmakologie terapeutické užití MeSH
- kolagen metabolismus MeSH
- krysa rodu Rattus MeSH
- mezenchymální kmenové buňky * metabolismus MeSH
- NF-kappa B metabolismus MeSH
- plíce MeSH
- plicní fibróza * chemicky indukované terapie metabolismus MeSH
- pneumonie * metabolismus MeSH
- potkani Sprague-Dawley MeSH
- signální transdukce MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bleomycin MeSH
- cytokiny MeSH
- kaveolin 1 MeSH
- kolagen MeSH
- NF-kappa B MeSH
