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Adhesion and growth of adipose tissue-derived stem cells on fibrin assemblies with attached growth factors for tissue engineering of heart valves
Filová, Elena; Trávníčková, Martina; Knitlová, Jarmila; Matějka, Roman; Kučerová, Johanka; Riedelová, Zuzana; Brynda, Eduard; Bačáková, Lucie
2019 - anglický
Currently used xenogeneic biological heart valve prostheses are decellularized and crosslinked with glutaraldehyde. These grafts usually undergo degeneration and calcification. Pericardium-based heart valve prostheses, re-seeded with autologous cells, i.e. Adipose tissue-derived cells (ASCs) and endothelial cells, could have longer durability and biocompatibility. In order to improve the adhesion of cells and their ingrowth into decellularized pericardium, various fibrin (Fb) layers were developed, i.e. Fb, Fb with covalently bound heparin (H), Fb with either vascular endothelial growth factor (VEGF) or fibroblast growth factor 2 (FGF) in various concentrations (1 ng/ml, 10 ng/ml, 100 ng/ml) or with both VEGF and FGF (100 ng/ml). Growth factors were attached onto Fb via heparin or were adsorbed. ASCs were seeded on theses layers in a DMEM medium supplemented with 2 % of fetal bovine serum, TGFβ1 and BMP-4 (both 2.5 ng/ml), and with ascorbic acid. Cell adhesion and growth/viability was assessed by counted cell number/MTS evaluation. ASCs were stained for differentiation markers of smooth muscle cells, such as alpha-actin, calponin, and myosin heavy chain. On day 7, ASCs on Fb-H-VEGF layers produced both calponin and alpha-actin. An increased FGF concentration caused reduced calponin staining of ASCs. Lack of heparin in fibrin assemblies with growth factors inhibited the production of both alpha-actin and calponin in ASCs. The highest ASCs density/viability was found on Fb-H-VEGF-FGF layer. The proper formulation of fibrin coatings could be favorable for ASCs growth and differentiation and could subsequently support endothelialization of cardiovascular prostheses with endothelial cells.
Klíčová slova:
adipose tissue-derived stem cells; fibrin assemblies; growth factors; heart valves
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Adhesion and growth of adipose tissue-derived stem cells on fibrin assemblies with attached growth factors for tissue engineering of heart valves
Currently used xenogeneic biological heart valve prostheses are decellularized and crosslinked with glutaraldehyde. These grafts usually undergo degeneration and calcification. Pericardium-based heart ...
Odborná zpráva o postupu prací a dosažených výsledcích za období 2014 - 2017 v rámci projektu TA04011345
Hrůzová, D.; Matějka, R.; Rosina, J.; Zárubová, Jana; Filová, Elena
2017 - český
Zpráva shrnuje postupy realizace projektu TA04011345 Cévní protézy o malém průměru osídlované endotelovými a kmenovými buňkami kostní dřeně v bioreaktoru v rámci etap 2014 - 2017. Závěrečná zpráva k projektu. Annual report sums the progress and achievements for the years 2014 - 2017 within the project TA04011345
Klíčová slova:
annual report TACR
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Odborná zpráva o postupu prací a dosažených výsledcích za období 2014 - 2017 v rámci projektu TA04011345
Zpráva shrnuje postupy realizace projektu TA04011345 Cévní protézy o malém průměru osídlované endotelovými a kmenovými buňkami kostní dřeně v bioreaktoru v rámci etap 2014 - 2017. Závěrečná zpráva k ...
Odborná zpráva o postupu prací a dosažených výsledcích za rok 2016 v rámci projektu TA04011345
Hrůzová, D.; Matějka, R.; Rosina, J.; Zárubová, Jana; Filová, Elena
2016 - český
Zpráva shrnuje postupy realizace projektu TA04011345 Cévní protézy o malém průměru osídlované endotelovými a kmenovými buňkami kostní dřeně v bioreaktoru v rámci etapy 2016. Hlavními body jsou: optimalizace decelularizačního postupu pro získání standardizovaného materiálu pro buněčné osídlení, postup trojrozměrného osídlování cévní protézy - endotelizace, vývoj systému bioreaktoru, mechanické zkoušky, chemické potahování protézy, získání buněk od pacientů a jejich charakterizace. Report sums progress of project TA04011345 in 2016 phase. Main topics are: optimization of automated decellularization, bioreactor design for 3D re-endothelisation, mechanical evaluation tests, chemical modifications, cell harvesting from patients and their characterisation.
Klíčová slova:
annual report; TACR
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Odborná zpráva o postupu prací a dosažených výsledcích za rok 2016 v rámci projektu TA04011345
Zpráva shrnuje postupy realizace projektu TA04011345 Cévní protézy o malém průměru osídlované endotelovými a kmenovými buňkami kostní dřeně v bioreaktoru v rámci etapy 2016. Hlavními body jsou: ...
Odborná zpráva o postupu prací a dosažených výsledcích za rok 2015 v rámci projektu TA04011345
Hrůzová, D.; Matějka, R.; Filová, Elena
2015 - český
Zpráva shrnuje postupy realizace projektu TA04011345 Cévní protézy o malém průměru osídlované endotelovými a kmenovými buňkami kostní dřeně v bioreaktoru v rámci etapy 2014. Hlavními body jsou: optimalizace odběru prasečích cév vhodných pro deceluarizaci; optimalizace postupů deceluarizace; optimalizace podmínek při dynamické kultivaci endotelu, kmenových buněk a jejich kokultur; návrh komory a perfuzního systému pro 3D dynamickou kultivaci v deceluarizovaných nosičích Report sums progress of project TA04011345. Main topics are: optimization of porcine artery harvesting, optimization of decellularization process, optimization of dynamic cultivation of endothelium, stem cells and their co-cultures, design of cultivation chamber and perfusion system used for 3D dynamic cultivation in decellularized tubular scaffolds
Klíčová slova:
annual report; TACR
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Odborná zpráva o postupu prací a dosažených výsledcích za rok 2015 v rámci projektu TA04011345
Zpráva shrnuje postupy realizace projektu TA04011345 Cévní protézy o malém průměru osídlované endotelovými a kmenovými buňkami kostní dřeně v bioreaktoru v rámci etapy 2014. Hlavními body jsou: ...
Polymeric nanofibrous scaffolds reinforced with diamond and ceramic nanoparticles for bone tissue engineering
Bačáková, Lucie; Pařízek, Martin; Staňková, Ľubica; Novotná, Katarína; Douglas, T.E.L.; Brady, M. A.; Kromka, Alexander; Potocký, Štěpán; Stránská, D.
2015 - anglický
Three types of nanofibrous scaffolds were prepared by electrospining: (1) poly(lactide-co-glycoside) (PLGA) scaffolds reinforced with 23 wt.% of diamond nanoparticles (DNPs), (2) poly(L-lactide) (PLLA) scaffolds with DNPs in concentration ranging from from 0.4 wt.% to 12.3 wt.%, and (3) PLLA scaffolds with 5 wt.% or 15 wt.% of hydroxyapatite (HAp) nanoparticles. The diameter of the nanofibers ranged between 160 and 729 nm. The nanofibers with nanoparticles were thicker and the void spaces among them were smaller. Mechanical properties of the nanoparticle-loaded scaffolds were better, as demonstrated by a rupture test in scaffolds with DNPs and by a creep behavior test in scaffolds with HAp. On PLGA scaffolds with DNPs, the human osteoblast-like MG-63 cells adhered in similar numbers and grew with similar kinetics as on pure PLGA scaffolds. Human bone marrow mesenchymal stem cells grew faster and reached higher population densities on PLGA-DNP scaffolds. However, on PLLA-based scaffolds, the activity of mitochondrial enzymes and concentration of osteocalcin in MG-63 cells decreased with increasing DNP concentration. On the other hand, the metabolic activity of MG-63 cells and content of osteocalcin in these cells were positively correlated with the HAp concentration in PLLA scaffolds. Thus, PLGA nanofibers with 23 wt% of DNPs and PLLA nanofibers with 5 and particularly 15 wt.% of HAp seem to be promising for bone tissue engineering.
Klíčová slova:
material nanostructure; nanofibers; osteogenic cells; bone implants; regenerative medicine
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Polymeric nanofibrous scaffolds reinforced with diamond and ceramic nanoparticles for bone tissue engineering
Three types of nanofibrous scaffolds were prepared by electrospining: (1) poly(lactide-co-glycoside) (PLGA) scaffolds reinforced with 23 wt.% of diamond nanoparticles (DNPs), (2) poly(L-lactide) ...
Growth and differentiation of human osteoblast-like cells on TI-6AL-4V alloy modified with anodization
Doubková, Martina; Bačáková, Lucie; Pařízek, Martin; Vandrovcová, Marta; Musílková, Jana; Lisá, Věra; Gabor, R.; Marvan, J.
2015 - anglický
Titanium and its alloys are widely used as hard tissue implants for their high biocompatibility and suitable physical properties. Along with other surface treatment methods, anodization technique is known to enhance osseointegration. The aim of this study is to evaluate the adhesion, growth and osteogenic differentiation of human osteoblast-like SAOS-2 cells on Ti-6Al-4V samples anodized in electrolytes composed of a stable volume of KOH (336.48 g/l) and variable volumes of liquid glass (124.47 g/l, 84,12 g/l and 37.38 g/l; samples s5, s6 and s7, respectively). Non-anodized Ti-6Al-4V samples, cell culture polystyrene (PS) and microscopic glass coverslips served as control materials. On days 2 and 4 after seeding, the cell number did not differ significantly among the tested samples. However, on day 7, the cell number on s6 samples reached the lowest values, which could be attributed to a non-homogeneous TiO2 film on s6 samples formed during anodization. Nevertheless, the osteogenic differentiation, estimated by the intensity of fluorescence of collagen I in cells grown in a differentiation medium, was the highest on s6 samples. On s5 samples, coated with homogeneous TiO2 films, both cell numbers and intensity of fluorescence of collagen I was relatively high. The bone matrix mineralization, evaluated by Alizarin Red staining, was the highest on s5 samples in standard culture medium, and similar on all tested samples in differentiation medium. Thus, the surface modification of s5 samples could be considered the most suitable for application in bone implants.
Klíčová slova:
Ti6-Al4-V; SAOS-2 cells; anodization; bone implants; collagen I; mineralization
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Growth and differentiation of human osteoblast-like cells on TI-6AL-4V alloy modified with anodization
Titanium and its alloys are widely used as hard tissue implants for their high biocompatibility and suitable physical properties. Along with other surface treatment methods, anodization technique is ...
Case study of intracranial EEG records of patients with focal cortical dysplasia type I and II
Balach, J.; Ježdík, P.; Čmejla, R.; Kršek, P.; Jiruška, Přemysl
2015 - anglický
In this study we try to find out if it is possible to differentiate type of focal cortical dysplasia by features obtained from intracranial EEG. We compare occurrence and rates of three biomarkers present in epilepsy in patients with focal cortical dysplasia type I and II. Case study is made on long term night records of 6 pediatric patients. Detection of interictal epileptiform discharges and high-frequency oscillations is made by automated algorithms, delta brush are marked visually. Position of lesion and electrodes inside were obtained from MRI. In individual rates were not found difference on significant level. No major significance were found, but as promising seem to be ratio inside to outside rates of high-frequency oscillations and presence of delta brush, which were found only in patients with focal cortical dysplasia type II.
Klíčová slova:
intracranial EEG; focal cortical dysplasia; IED; high frequency oscillations; delta brush
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Case study of intracranial EEG records of patients with focal cortical dysplasia type I and II
In this study we try to find out if it is possible to differentiate type of focal cortical dysplasia by features obtained from intracranial EEG. We compare occurrence and rates of three biomarkers ...
Growth of human adipose-derived stem cells on Ti-6Al-4V alloy with various surface modifications
Kročilová, Nikola; Bačáková, Lucie; Pařízek, Martin; Havlíková, Jana; Motarjemi, H.; Molitor, M.; Gabor, R.; Marvan, J.
2015 - anglický
Adipose-derived stem cells (ASCs) are of a great importance for bone tissue engineering. We compared ASCs obtained by liposuction under two pressures (-200 mmHg and -700 mmHg). The number and proliferation activity of cells isolated under -700 mmHg (ASC-H) were higher than in cells isolated under -200 mmHg (ASC-L). However, the ASC-L was more active in osteogenic differentiation, as manifested by a higher intensity of fluorescence of alkaline phosphatase and osteocalcin in these cells. The adhesion and growth of ASCs were then studied on Ti-6Al-4V samples either unmodified (K, roughness parameter R-a = 280 nm) or modified by shot blasting and tarnishing A, R-a = 200 nm), vibratory finishing (B, R-a = 100 nm) and vibratory finishing, shot blasting and polishing (C, R-a = 80 nm). All modified samples were more wettable than the K samples. On day 1 after seeding, the size of cell spreading area on some modified samples was lower than on K samples, namely on B samples in ASC-L, on B and C in ASC-H and on A in control MG-63 cells. On day 3, the cell number on some modified samples became higher than on K samples, namely on C samples (ASC-L), on B (ASC-H) and on B and C s (MG-63 cells). On day 7, the numbers of ASC-H and MG-63 cells on all modified samples evened out and became significantly higher than on K samples. On ASC-L samples, the highest cell numbers were obtained on A samples. Thus, all studied modifications of Ti-6Al-4V enhanced growth of ASCs and human osteoblast-like MG-63 cells.
Klíčová slova:
bone implants; bone tissue engineering; surface roughness; stem cells; liposuction; cell proliferation
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Growth of human adipose-derived stem cells on Ti-6Al-4V alloy with various surface modifications
Adipose-derived stem cells (ASCs) are of a great importance for bone tissue engineering. We compared ASCs obtained by liposuction under two pressures (-200 mmHg and -700 mmHg). The number and ...
Analysis of high-frequency activity in electroencephalogram of animal epilepsy model
Havel, T.; Kudláček, J.; Jiruška, Přemysl; Ježdík, P.; Čmejla, R.
2015 - anglický
High frequency oscillations (HFO) are believed to be a new specific biomarker of epileptogenic tissue. According to the reported findings, we hypothesized that HFOs occur more specific to the epileptogenic tissue. To test this hypothesis, we analyzed intracranial electroencephalograms (iEEG) of three subjects of animal epilepsy model. Overall 180 minutes of iEEG records were processed by the automatic high frequency activity detector and further analyzed. The dominant frequency of each segment was determined and categorized as ripples (80-200 Hz) or fast ripples (200-1000 Hz). Even though overall number of HFO detections predominate in the hemisphere where epileptic focus is located, in comparison of each brain structure separately our hypothesis cannot be fully confirmed.
Klíčová slova:
epilepsy; animal model; electroencephalogram (EEG); high-frequency oscillations (HFO)
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Analysis of high-frequency activity in electroencephalogram of animal epilepsy model
High frequency oscillations (HFO) are believed to be a new specific biomarker of epileptogenic tissue. According to the reported findings, we hypothesized that HFOs occur more specific to the ...
Průmyslová mikrofluidizace liposomálních a hydrofobních lékových forem
Vlk, M.; Poučková, P.; Ježek, Petr
2014 - český
Zavedení a validace sterilní mikrofluidizace lecitinových liposomů v čistých prostorách firmy Wake s. r. o. Bude sloužit k výrobě liposomálního přípravku s ftalocyaninem pro fotodynamickou léčbu tumorů Development and validation of sterile microfluidization of lecithin liposomes in the clean facilities of company Wake s. r. o. It will serve for manufacturing of liposomal preparate with phtalocyanine for photodynamic treatment of tumors
Klíčová slova:
phytodynamic therapy; phtalocyanine; microfluidization
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Průmyslová mikrofluidizace liposomálních a hydrofobních lékových forem
Zavedení a validace sterilní mikrofluidizace lecitinových liposomů v čistých prostorách firmy Wake s. r. o. Bude sloužit k výrobě liposomálního přípravku s ftalocyaninem pro fotodynamickou léčbu ...
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