Number of found documents: 80
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Differentiation of mesenchymal stem cells on fibrin assemblies supported by immobilized growth factors FGF2 and VEGF
Musílková, Jana; Filová, Elena; Kaplan, Ondřej; Bačáková, Lucie
2021 - English
Bioartificial heart valves and vascular grafts prepared from decellularized tissues could be recellularized with bone marrow-derived mesenchymal stem cells (MSCs) that are able to differentiate into both smooth muscle cells and endothelial cells. MSCs differentiation is facilitated by sustained release of growth factors. In our study assemblies based on fibrin, fibrin with heparin, fibrin with adsorbed or covalently-immobilized vascular endothelial growth factor A165 (VEGF) or basic fibroblast growth factor (FGF-2) via binding to heparin attached to fibrin have been prepared and were evaluated for their stimulation of MSCs differentiation. We estimated the mRNA expression of endothelial marker CD31 (PECAM1), smooth muscle marker α-actin (ACTA2), osteoblast markers osteocalcin (BGLAP) and alkaline phosphatase (ALP). The gene expression was estimated using RT-PCR on days 1, 7 and 21 after seeding. The cell morphology and viability was evaluated by LIVE/DEAD staining. VEGF, both adsorbed and covalently bound, increased significantly the expression of smooth muscle marker α-actin. The mRNA expression of ACTA2 on day 7 and 21 raised more than 200 times in comparison to control samples (undifferentiated cells before seeding). The ACTA2 gene expression significantly exceeded the expression of all other evaluated genes at all time intervals. Moreover, on day 21, the late smooth muscle marker desmin (DES) was steeply rising in cells cultivated on assemblies containing heparin and covalently bound VEGF. The expression of osteocalcin was minimal. We conclude that fibrin assembly containing covalently bound VEGF is the most convenient for MSCs differentiation towards smooth muscle cells. Keywords: stem cells; differentiation; growth factors; smooth muscle cells; fibrin assemblies Fulltext is available at external website.
Differentiation of mesenchymal stem cells on fibrin assemblies supported by immobilized growth factors FGF2 and VEGF

Bioartificial heart valves and vascular grafts prepared from decellularized tissues could be recellularized with bone marrow-derived mesenchymal stem cells (MSCs) that are able to differentiate into ...

Musílková, Jana; Filová, Elena; Kaplan, Ondřej; Bačáková, Lucie
Fyziologický ústav, 2021

The co-culture of keratinocytes and fibroblasts on a multi-layered polyester nanofibrous membrane enriched with platelet lysate
Blanquer, Andreu; Filová, Elena; Jenčová, V.; Brož, Antonín; Kuželová Košťáková, E.; Lisnenko, M.; Procházková, R.; Bačáková, Lucie
2021 - English
The prevalence of chronic wounds is increasing due to the population ageing and specific illnesses like diabetes mellitus and vascular diseases. Nanofibrous membranes fabricated using synthetic polymers are promising materials to enhance skin wound healing. PCL and PVA membranes are being studied to be used as scaffolds for skin tissue engineering and hydrogels for controlled drug delivery, respectively. The present study considers the development of a multi-layered membrane made of PCL and PVA loaded with platelet lysate (PL). PCL nanofibers allowed cell adhesion and growth, whereas PVA acted as a hydrogel that releases the bioactive compounds of platelet lysate. The cytocompatibility of the membranes containing PL and without it was demonstrated on two cell types involved in wound healing, i.e. keratinocytes and fibroblasts. Both cell types were able to adhere and proliferate on the membranes. In addition, the membrane containing PL enhanced the proliferation of fibroblasts. A co-culture study was also performed by seeding each cell type on one side of the membrane. The cells were co-cultured for 7 days and the results showed that PL increased the proliferation of cells achieving a monolayer of keratinocytes or fibroblasts on each side of the membrane. Thus, the beneficial effect of PCL-PVA+PL membranes on monocultures and co-cultures of skin cells was demonstrated, and these membranes can be considered potential scaffolds for treatment of chronic wounds. Keywords: skin wound healing; electrospun nanofibers; platelet lysate; keratinocytes; fibroblasts Fulltext is available at external website.
The co-culture of keratinocytes and fibroblasts on a multi-layered polyester nanofibrous membrane enriched with platelet lysate

The prevalence of chronic wounds is increasing due to the population ageing and specific illnesses like diabetes mellitus and vascular diseases. Nanofibrous membranes fabricated using synthetic ...

Blanquer, Andreu; Filová, Elena; Jenčová, V.; Brož, Antonín; Kuželová Košťáková, E.; Lisnenko, M.; Procházková, R.; Bačáková, Lucie
Fyziologický ústav, 2021

Mineralised polylactide and polycaprolactone soft foams with hierarchical micro-macro porous structure for tissue engineering
Beran, M.; Berková, E.; Musílková, Jana; Sedlář, Antonín; Slepička, P.; Fajstavr, D.
2021 - English
The purpose of the study was development of soft foams from resorbable polymers with unique micro-macro porous interconnected hierarchical structure specially designed as scaffold for engineering of soft tissues. The foams have been prepared by freeze-drying of solutions of polylactide (PLA) and polycaprolactone (PCL) in 1,4-dioxane. The foams prepared by freeze-drying had interconnected porous aerogel characteristics. The hierarchical structure with bimodal micro-macro pore size distribution were obtained after addition of sucrose or NaCl porogens with defined crystal size distributions to the solutions before freeze-drying and leaching the porogen crystals from the freeze-dried foams with demineralised water. Polyethyleneimine was chemically conjugated to the alkali-treated foams followed by conjugation of citric acid using carbodiimide chemistry. Finally, they were mineralised by immersing and incubating in a simulated body fluid with ionic concentration similar to that of human blood plasma, to obtain tissue engineering scaffolds. To verify their biocompatibility, the scaffolds were seeded with adipose-derived stem cells (ASC) and sarcoma osteogenic-2 (SaOs-2) human osteoblast-like cells. Morphology of the cells attached to the scaffolds was evaluated and their viability was verified by a metabolic test. Biocompatibility and usability of the scaffolds was successfully verified by incubation with adipose-derived stem cells and SaOs-2 human osteosarcoma cell line. Mineralised scaffolds are more suitable growth supports for both the cell types than unmineralized collagen scaffolds. The scaffolds have been specially designed for engineering of soft tissues, but they can be used in other categories of tissue engineering, too. Keywords: tissue engineering; polylactide; polycarolactone; soft foams; hierarchical structure Fulltext is available at external website.
Mineralised polylactide and polycaprolactone soft foams with hierarchical micro-macro porous structure for tissue engineering

The purpose of the study was development of soft foams from resorbable polymers with unique micro-macro porous interconnected hierarchical structure specially designed as scaffold for engineering of ...

Beran, M.; Berková, E.; Musílková, Jana; Sedlář, Antonín; Slepička, P.; Fajstavr, D.
Fyziologický ústav, 2021

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 - English
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. Keywords: adipose tissue-derived stem cells; fibrin assemblies; growth factors; heart valves Available at various institutes of the ASCR
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 ...

Filová, Elena; Trávníčková, Martina; Knitlová, Jarmila; Matějka, Roman; Kučerová, Johanka; Riedelová, Zuzana; Brynda, Eduard; Bačáková, Lucie
Fyziologický ústav, 2019

Annual report on the progress and achievements for the years 2014 - 2017 within the project TA04011345
Hrůzová, D.; Matějka, R.; Rosina, J.; Zárubová, Jana; Filová, Elena
2017 - Czech
Annual report sums the progress and achievements for the years 2014 - 2017 within the project 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 projektu. Keywords: annual report TACR Available at various institutes of the ASCR
Annual report on the progress and achievements for the years 2014 - 2017 within the project TA04011345

Annual report sums the progress and achievements for the years 2014 - 2017 within the project TA04011345

Hrůzová, D.; Matějka, R.; Rosina, J.; Zárubová, Jana; Filová, Elena
Fyziologický ústav, 2017

Annual report on the progress and achievements for the year 2016 within the project TA04011345
Hrůzová, D.; Matějka, R.; Rosina, J.; Zárubová, Jana; Filová, Elena
2016 - Czech
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. 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. Keywords: annual report; TACR Available at various institutes of the ASCR
Annual report on the progress and achievements for the year 2016 within the project TA04011345

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, ...

Hrůzová, D.; Matějka, R.; Rosina, J.; Zárubová, Jana; Filová, Elena
Fyziologický ústav, 2016

Annual report on the progress and achievements for the year 2015 within the project TA04011345
Hrůzová, D.; Matějka, R.; Filová, Elena
2015 - Czech
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 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 Keywords: annual report; TACR Available at various institutes of the ASCR
Annual report on the progress and achievements for the year 2015 within the project TA04011345

Report sums progress of project TA04011345. Main topics are: optimization of porcine artery harvesting, optimization of decellularization process, optimization of dynamic cultivation of endothelium, ...

Hrůzová, D.; Matějka, R.; Filová, Elena
Fyziologický ústav, 2015

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 - English
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. Keywords: material nanostructure; nanofibers; osteogenic cells; bone implants; regenerative medicine Available at various institutes of the ASCR
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) ...

Bačáková, Lucie; Pařízek, Martin; Staňková, Ľubica; Novotná, Katarína; Douglas, T.E.L.; Brady, M. A.; Kromka, Alexander; Potocký, Štěpán; Stránská, D.
Fyziologický ústav, 2015

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 - English
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. Keywords: Ti6-Al4-V; SAOS-2 cells; anodization; bone implants; collagen I; mineralization Available at various institutes of the ASCR
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 ...

Doubková, Martina; Bačáková, Lucie; Pařízek, Martin; Vandrovcová, Marta; Musílková, Jana; Lisá, Věra; Gabor, R.; Marvan, J.
Fyziologický ústav, 2015

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 - English
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. Keywords: intracranial EEG; focal cortical dysplasia; IED; high frequency oscillations; delta brush Available at various institutes of the ASCR
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 ...

Balach, J.; Ježdík, P.; Čmejla, R.; Kršek, P.; Jiruška, Přemysl
Fyziologický ústav, 2015

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