The present study investigates the effect of an oxidized nanocrystalline diamond (O-NCD) coating functionalized with bone morphogenetic protein 7 (BMP-7) on human osteoblast maturation and extracellular matrix mineralization in vitro and on new bone formation in vivo. The chemical structure and the morphology of the NCD coating and the adhesion, thickness and morphology of the superimposed BMP-7 layer have also been assessed. The material analysis proved synthesis of a conformal diamond coating with a fine nanostructured morphology on the Ti6Al4V samples. The homogeneous nanostructured layer of BMP-7 on the NCD coating created by a physisorption method was confirmed by AFM. The osteogenic maturation of hFOB 1.19 cells in vitro was only slightly enhanced by the O-NCD coating alone without any increase in the mineralization of the matrix. Functionalization of the coating with BMP-7 resulted in more pronounced cell osteogenic maturation and increased extracellular matrix mineralization. Similar results were obtained in vivo from micro-CT and histological analyses of rabbit distal femurs with screws implanted for 4 or 12 weeks. While the O-NCD-coated implants alone promoted greater thickness of newly-formed bone in direct contact with the implant surface than the bare material, a further increase was induced by BMP-7. It can be therefore concluded that O-NCD coating functionalized with BMP-7 is a promising surface modification of metallic bone implants in order to improve their osseointegration.

• MeSH
• biokompatibilní potahované materiály chemie   farmakologie   MeSH
• diamant chemie   MeSH
• extracelulární matrix MeSH
• kostní morfogenetický protein 7 * farmakologie   MeSH
• králíci MeSH
• osteointegrace * MeSH
• slitiny MeSH
• titan MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Hydrogels are suitable for osteochondral defect regeneration as they mimic the viscoelastic environment of cartilage. However, their biomechanical properties are not sufficient to withstand high mechanical forces. Therefore, we have prepared electrospun poly-ε-caprolactone-chitosan (PCL-chit) and poly(ethylene oxide)-chitosan (PEO-chit) nanofibers, and FTIR analysis confirmed successful blending of chitosan with other polymers. The biocompatibility of PCL-chit and PEO-chit scaffolds was tested; fibrochondrocytes and chondrocytes seeded on PCL-chit showed superior metabolic activity. The PCL-chit nanofibers were cryogenically grinded into microparticles (mean size of about 500 µm) and further modified by polyethylene glycol-biotin in order to bind the anti-CD44 antibody, a glycoprotein interacting with hyaluronic acid (PCL-chit-PEGb-antiCD44). The PCL-chit or PCL-chit-PEGb-antiCD44 microparticles were mixed with a composite gel (collagen/fibrin/platelet rich plasma) to improve its biomechanical properties. The storage modulus was higher in the composite gel with microparticles compared to fibrin. The Eloss of the composite gel and fibrin was higher than that of the composite gel with microparticles. The composite gel either with or without microparticles was further tested in vivo in a model of osteochondral defects in rabbits. PCL-chit-PEGb-antiCD44 significantly enhanced osteogenic regeneration, mainly by desmogenous ossification, but decreased chondrogenic differentiation in the defects. PCL-chit-PEGb showed a more homogeneous distribution of hyaline cartilage and enhanced hyaline cartilage differentiation.

• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

Stroke is despite of progressive improvements in treatment and reperfusion strategies one of the most devastating human pathology. However, as quality of acute health care improves and more people survive ischemic attack, healthcare specialists have to solve new challenges to preserve reasonable quality of life to these patients. Thus, novel approaches which prevents comorbidities of stroke and improve quality of life of stroke survivors in general has to be developed and experimentally tested. The aim of the present paper was to establish reliable rat model of middle cerebral occlusion and set of methods allowing selection of animals suitable for long-term experiments. We have compared mortality rates, cerebral blood flow and extension of ischemic lesion induced by intraluminal filament in three widely used outbred rat strains. We have additionally used an animal 18F-DG PET scans to verify its reliability in noninvasive detection of ischemic infarct in acute period (24 h after MCAO) for selecting animals eligible for long survival experiments. Our data clearly indicates that high variability between rat strains might negatively influence stroke induction by intraluminal thread occlusion of middle cerebral artery. Most reliable outbred rat strain in our hands was Sprague-Dawley where maximal reduction of cerebral blood flow and extensive ischemic lesion was observed. Contrary, Wistar rats exhibited higher mortality and Long-Evans rats significantly smaller or no ischemic region in comparison to Sprague-Dawley. Additionally, we have confirmed a positron emission tomography with 18F-fluorodeoxyglucose as suitable method to assess extension of ischemic region in acute period after the experimental arterial occlusion in rats.

• MeSH
• druhová specificita MeSH
• infarkt arteria cerebri media diagnostické zobrazování   genetika   patofyziologie   MeSH
• krysa rodu rattus MeSH
• mozkový krevní oběh fyziologie   MeSH
• potkani Long-Evans MeSH
• potkani Sprague-Dawley MeSH
• potkani Wistar MeSH
• pozitronová emisní tomografie metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Aim: This study evaluates the effect of electrospun dressings in critical sized full-thickness skin defects in rabbits. Materials & methods: Electrospun poly-ε-caprolactone (PCL) and polyvinyl alcohol (PVA) nanofibers were tested in vitro and in vivo. Results: The PCL scaffold supported the proliferation of mesenchymal stem cells, fibroblasts and keratinocytes. The PVA scaffold showed significant swelling, high elongation capacity, limited protein adsorption and stimulation of cells. Nanofibrous dressings improved wound healing compared with the control group in vivo. A change of the PCL dressing every 7 days resulted in a decreased epithelial thickness and type I collagen level in the adhesive group, indicating peeling off of the newly formed tissue. In the PVA dressings, the exchange did not affect healing. Conclusion: The results demonstrate the importance of proper dressing exchange.

• MeSH
• buňky 3T3 MeSH
• hojení ran účinky léků   MeSH
• králíci MeSH
• kůže * zranění   metabolismus   patologie   MeSH
• myši MeSH
• nanovlákna chemie   MeSH
• obvazy * MeSH
• polyestery * chemie   farmakologie   MeSH
• polyvinylalkohol chemie   farmakologie   MeSH
• prasata MeSH
• tkáňová adheziva * chemie   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The study involved the electrospinning of the copolymer poly(L-lactide-co-ε-caprolactone) (PLCL) into tubular grafts. The subsequent material characterization, including micro-computed tomography analysis, revealed a level of porosity of around 70%, with pore sizes of 9.34 ± 0.19 μm and fiber diameters of 5.58 ± 0.10 μm. Unlike fibrous polycaprolactone, the electrospun PLCL copolymer promoted fibroblast and endothelial cell adhesion and proliferation in vitro. Moreover, the regeneration of the vessel wall was detected following implantation and, after six months, the endothelialization of the lumen and the infiltration of arranged smooth muscle cells producing collagen was observed. However, the degradation rate was found to be accelerated in the rabbit animal model. The study was conducted under conditions that reflected the clinical requirements-the prostheses were sutured in the end-to-side fashion and the long-term end point of prosthesis healing was assessed. The regeneration of the vessel wall in terms of endothelialization, smooth cell infiltration and the presence of collagen fibers was observed after six months in vivo. A part of the grafts failed due to the rapid degradation rate of the PLCL copolymer.

• MeSH
• aorta patologie   MeSH
• arteriae carotides patologie   MeSH
• buněčná adheze MeSH
• buňky 3T3 MeSH
• cévní protézy * MeSH
• endoteliální buňky pupečníkové žíly (lidské) MeSH
• endoteliální buňky MeSH
• fibroblasty cytologie   MeSH
• kolagen metabolismus   MeSH
• králíci MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• myocyty hladké svaloviny cytologie   MeSH
• myši MeSH
• polyestery chemie   MeSH
• polymery chemie   MeSH
• poréznost MeSH
• prasata MeSH
• psi MeSH
• regenerace MeSH
• rentgenová mikrotomografie MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury MeSH
• transplantace cév * MeSH
• zobrazování trojrozměrné MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Epilepsy is chronic neurological disease characterized by occurrence of spontaneous recurrent seizures affecting approximately 1% of people. One of the most common causes of epilepsy in adults is a stroke. The precise mechanism of development of vascular epilepsy is not known, and thus no reliable biomarker of postischemic epileptogenesis currently exists. Blood-brain barrier (BBB) impairment is phenomenon observed in several pathologies including stroke after which epilepsy often develops. Blood components such as albumin or thrombin have been experimentally shown to possess the capacity to increase excitability which results in seizures and epileptogenesis. We hypothesize that severity of the BBB breakdown during first weeks after the stroke can be used as a biomarker of postischemic epileptogenesis. This paper describes free MATLAB based software /Gd-Tracker/ and methodology developed for assessment of BBB permeability from magnetic resonance (MR) scans based on statistical voxel to voxel comparison of sequences with and without Gd-DTPA contrast. This software allows to evaluate extent and magnitude of impaired BBB region and thus serve as a tool for visualization and quantification of BBB breakdown.

• MeSH
• cévní mozková příhoda diagnostické zobrazování   MeSH
• diethylentriaminpentaacetát gadolinia aplikace a dávkování   MeSH
• epilepsie diagnostické zobrazování   etiologie   MeSH
• hematoencefalická bariéra * diagnostické zobrazování   MeSH
• kapilární permeabilita MeSH
• kontrastní látky farmakokinetika   MeSH
• lidé MeSH
• vylepšení obrazu přístrojové vybavení   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• aktivní transport MeSH
• astrocyty MeSH
• biologický transport MeSH
• endotel MeSH
• epilepsie etiologie   MeSH
• hematoencefalická bariéra * anatomie a histologie   fyziologie   patologie   účinky léků   MeSH
• lidé MeSH
• mikroglie MeSH
• mozek anatomie a histologie   fyziologie   MeSH
• neurony MeSH
• pericyty MeSH
• systémy cílené aplikace léků MeSH
• zánět imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

A three-dimensional scaffold of type I collagen and hydroxyapatite enriched with polycaprolactone nanofibers (Coll/HA/PCL), autologous mesenchymal stem cells (MSCs) in osteogenic media, and thrombocyte-rich solution (TRS) was an optimal implant for bone regeneration in vivo in white rabbits. Nanofibers optimized the viscoelastic properties of the Coll/HA scaffold for bone regeneration. MSCs and TRS in the composite scaffold improved bone regeneration. Three types of Coll/HA/PCL scaffold were prepared: an MSC-enriched scaffold, a TRS-enriched scaffold, and a scaffold enriched with both MSCs and TRS. These scaffolds were implanted into femoral condyle defects 6 mm in diameter and 10-mm deep. Untreated defects were used as a control. Macroscopic and histological analyses of the regenerated tissue from all groups were performed 12 weeks after implantation. The highest volume and most uniform distribution of newly formed bone occurred in defects treated with scaffolds enriched with both MSCs and TRS compared with that in defects treated with scaffolds enriched by either component alone. The modulus of elasticity in compressive testing was significantly higher in the Coll/HA/PCL scaffold than those without nanofibers. The composite Coll scaffold functionalized with PCL nanofibers and enriched with MSCs and TRS appears to be a novel treatment for bone defects.

• MeSH
• hydroxyapatit chemie   MeSH
• kolagen chemie   MeSH
• králíci MeSH
• kultivované buňky MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• nanovlákna chemie   MeSH
• polyestery chemie   MeSH
• regenerace kostí * MeSH
• tkáňové podpůrné struktury chemie   MeSH
• trombocyty chemie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH