Infrared attenuated total reflection spectroscopy was used for in situ observation of the deposition of collagen I on poly(2-hydroxyethyl methacrylate-co-methacrylic acid, 2.9%) hydrogels and subsequent attachment of laminin or fibronectin on the collagen surface. While there was no adsorption of collagen dissolved in an acid solution on the hydrogel surface, it deposited on the surface at pH 6.5. The collagen layers with attached laminin or fibronectin were stable on hydrogel surface in physiological solution. The modification with collagen and particularly with collagen and laminin or fibronectin allowed the adhesion and growth of mesenchymal stromal cells and astrocytes on the hydrogel surface.

• MeSH
• extracelulární matrix - proteiny chemie   MeSH
• hydrogely chemie   MeSH
• koncentrace vodíkových iontů MeSH
• mikroskopie atomárních sil MeSH
• polyhydroxyethylmethakrylát analogy a deriváty   chemie   MeSH
• povrchové vlastnosti MeSH
• spektroskopie infračervená s Fourierovou transformací MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• extracelulární matrix - proteiny MeSH
• hydrogely MeSH
• polyhydroxyethyl methacrylate hydrogels MeSH Prohlížeč
• polyhydroxyethylmethakrylát MeSH

The cell/tissue engineering therapy of extensive or chronic skin wounds is a highly topical task of the contemporary medicine. One of possible therapeutic approaches is grafting of in vitro cultured keratinocytes directly to the wound bed, where the cells colonize the wound, proliferate and improve the re-epithelization process. Because the successful cultivation of keratinocytes needs an application of feeder cells, the exclusion of these cells from the cultivation system is highly required. In this study we show a positive influence of 2-ethoxyethyl methacrylate as a component of cultivation support on growth of keratinocytes without feeder cells. Keratinocytes cultured on these surfaces are able to migrate to the model wound bed in vitro, where they form distinct colonies and have a normal differentiation potential.

• MeSH
• biokompatibilní materiály chemie   MeSH
• buněčné kultury MeSH
• keratinocyty cytologie   fyziologie   MeSH
• kultivační média MeSH
• kultivované buňky MeSH
• lidé MeSH
• methakryláty chemie   MeSH
• polymery chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biokompatibilní materiály MeSH
• kultivační média MeSH
• methakryláty MeSH
• polymery MeSH

Four series of macroporous hydrogels based on crosslinked copolymers of 2-hydroxyethyl methacrylate (HEMA)-sodium methacrylate (MANa), copolymer HEMA-[2-(methacryloyloxy)ethyl]trimethylammonium chloride (MOETACl), terpolymer HEMA-MANa-MOETACl and on a polyelectrolyte complex were used as carriers for immobilization of proteins, chicken egg white albumin and avidin. The adsorption capacity of the hydrogels for the two proteins, kinetics and pH dependence of albumin adsorption and desorption were studied. The morphology of the hydrogels with and without immobilized albumin was studied by low-vacuum scanning electron microscopy.

• MeSH
• adsorpce MeSH
• albuminy analýza   chemie   ultrastruktura   MeSH
• avidin analýza   chemie   ultrastruktura   MeSH
• biokompatibilní potahované materiály analýza   chemie   MeSH
• hydrogely analýza   chemie   MeSH
• kinetika MeSH
• methakryláty analýza   chemie   MeSH
• poréznost MeSH
• povrchové vlastnosti MeSH
• testování materiálů MeSH
• vazba proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• albuminy MeSH
• avidin MeSH
• biokompatibilní potahované materiály MeSH
• hydrogely MeSH
• hydroxyethyl methacrylate MeSH Prohlížeč
• methakryláty MeSH

We studied the biocompatibility of the carbon composites and polyethylene materials with and without collagen or collagen and proteoglycan cover. We used the in vitro technology to study the adhesion of model cells evalution, their metabolic activity and the production of TNF-alpha as a cytokine model. Under in vivo condition, the biocompatibility of tested polymers were studied in the implantation experiment, subcutaneously in the interscapular region in the laboratory rat. We have found in the in vitro assay favorable proliferation and the smallest production of pro-inflammatory TNF-alpha cytokine in cells adherent to the hydrophobic polyethylene material coated with biological macromolecules. Using in vivo tests performed by the implantation of materials to the rat we demonstrated that the materials are not cytotoxic. The tissue capsule surrounding the implants was not significantly influenced by the type of the implant and the pre-treatment by the biological molecules. However, the foreign-body giant multinucleated cells were observed only in the vicinity of the collagen - covered hydrophobic polyethylene implant. Interestingly, while the collagen coating improved the biocompatibility of tested polymers in vitro, the inflammatory reaction against this covered materials was higher under in vivo conditions. The pre-treatment of carbon composites by both types of biological macromolecules reduced the occurrence of carbon debris in the implantation site. The tested carbon composites and polyethylene materials are not toxic. The pre-treatment of the materials by extracellular matrix components increased their biological tolerance in vitro and reduced implant wears in animal experiment, which can be important for the medical application.

• Publikační typ
• časopisecké články MeSH

This study compares subcutaneous and intracerebral biocompatibilty of two hydrogels: copolymer of 2-hydroxyethyl methacrylate with 2-(methylsulfanyl)ethyl methacrylate and poly(2-hydroxyethyl methacrylate) as reference polymer. The experimental copolymer was more biologically inert than poly(2-hydroxyethyl methacrylate) in both the studied parameters, hence the former material is a suitable candidate for biomedical application.

• Publikační typ
• časopisecké články MeSH

Interaction of organism with non-toxic implanted polymers depends on the physicochemical properties of the implant surface, which influence the adsorption of bioactive proteins and subsequently adhesion and growth of cells. The synthetic hydrogels are known as poorly adhesive surfaces. In this study we demonstrated the adsorption of albumin, fibrinogen, fibronectin, basic fibroblast growth factor, heparin-binding epidermal growth factor-like growth factor and epidermal growth factor to poly(2-hydroxyethyl methacrylate) (pHEMA) and copolymer of 2-hydroxyethyl methacrylate (HEMA) and potassium salt of 3-sulfopropyl methacrylate (SPMAK). The adhesion and growth of 3T3 cells and human keratinocytes on surface of these polymers was tested without and with pretreatment of polymers with heparin-binding epidermal growth factor-like growth factor. The adhesion of mixture of human granulocytes and monocytes to these surfaces was also tested. The strips of both polymers were subcutaneously and intracerebrally implanted into the rat and the extent of foreign body reaction and brain biocompatibility was evaluated. The results showed the extensive adsorption of basic fibroblast growth factor and heparin-binding epidermal growth factor-like growth factor to copolymer containing SPMAK. However the adhesion (and growth) of cells to this type of copolymers was very low. Preadsorption of human plasma to pHEMA clearly stimulated the leukocyte adhesion in contrary to copolymer containing SPMAK. The extent of foreign-body reaction was significantly higher against the pHEMA compared to tested copolymer p(HEMA-co-SPMAK). In conclusion, the tested copolymer was a poorly adhesive substrate that is only poorly recognized by the non-specific immunity, although the adsorption of basic growth factors to this substrate is highly significant. Both polymers were well tolerated by the brain tissue. The phenotype of surrounding neurons was more close to the control neurons in the brain tissue surrounding the p(HEMA-co-SPMAK) implants.

• Publikační typ
• časopisecké články MeSH

Polyethylene (PE) foils were modified by irradiation with Ar+ and Xe+ ions to different fluences and different physico-chemical properties of the irradiated PE were studied in relation to adhesion and proliferation of keratinocytes on the modified surface. Changes in the PE surface roughness were examined using the AFM technique, the production of conjugated double bonds and oxidized structures by UV-VIS and FTIR techniques respectively. The surface polarity was determined by measuring surface contact angle and two-point technique was used for the determination of PE sheet resistance. Adhesion and proliferation of keratinocytes was characterized using the MTT-test. The ion irradiation leads to creation of conjugated double bonds which, together with progressive carbonization, contribute to the observed decrease of sheet resistance. Oxidation of the irradiated PE surface layer during the ion implantation is observed. Besides oxidation, the PE surface polarity is affected by other factors. The observed increase of the PE surface roughness due to the ion irradiation is inversely proportional to the ion size. The adhesion and proliferation of keratinocytes on the ion irradiated PE is significantly higher than on the pristine PE. Distribution of results in keratinocyte cultivation and the number of cells is related to the ion fluence applied and to ion species as well.

• Publikační typ
• časopisecké články MeSH

A lesion in the dorsoposterior part of the rat brain septum is known to exert an inhibitory effect on the delayed skin hypersensitivity and incorporation of radiolabeled thymidine into the lymphoid organs. To determine whether distinct properties of macrophages will also be modulated by this type of injury, we have focused upon the monitoring of expression of sugar receptors (lectins). In this study we show a reduction in the number of macrophages expressing carbohydrate-binding sites for asialoglycoproteins (beta-D-galactoside), alpha-D-mannoside and alpha-D-mannoside-6-phosphate in spleen macrophages after the lesion of the dorsoposterior septum of the brain in the rat. The number of ED-1+ macrophages was not influenced. The intraperitoneal injection of beads prepared from the copolymer of 2-hydroxyethyl methacrylate with dimethyl aminoethyl methacrylate (30 wt %) elevated significantly the number of ED-1+ spleen macrophages and number of macrophages with binding site(s) recognizing asialoglycoproteins and alpha-D-mannoside-6-phosphate, respectively. These results indicate that a foreign-body reaction appears to be able to mediate a phenotypic restoration of lectin expression by spleen macrophages altered by the brain lesion. It can be suggested that, for example, a probable production of cytokines by the inflammatory cells colonizing the implanted beads plays a role in this process.

• Publikační typ
• časopisecké články MeSH

The keratinocytes are able to migrate from the poly (2- hydroxyethylmethacrylate) disc if it is transferred to the new Petri-dish colonized with irradiated 3T3 mouse fibroblasts, and form a ring-shaped colony around the disc. The phenotypic characterization of human keratinocytes migrated from these discs was studied using a group of monoclonal antibodies. The keratinocytes in the external periphery of the colony of cells which migrated from the disc express the proliferating cell nuclear antigen (PCNA), alpha2, alpha3 chains and alpha5beta1 integrin receptor. A protein of the desmosome complex, desmoplakin-1, was also expressed. Involucrin and cytokeratin-10 were expressed after prolonged cultivation. These results suggest that the migrated keratinocytes are able to proliferate, recognize extracellular matrix molecules important in the process of the re-epitelization of the wound, and terminally differentiate in vitro. They are encouraging for further experiments with respect to the development of a support for keratinocyte cultivation and for grafting in clinical practice.

• Publikační typ
• časopisecké články MeSH

Structural changes of polyethylene (PE), induced by irradiation with 40 keV Ar+ ions at a fluence of 1x10(12)-1x10(15) cm(-2), are characterized by different experimental methods and physical parameters of the modified PE are related to its biocompatibility. Production of oxidized structures and conjugated double bonds in the PE surface layer modified by the ion irradiation was proved using IR, UV-VIS spectroscopies and a Rutherford backscattering technique. The fusion of macrophages onto implants made of as-irradiated and chemically doped PE was studied in vivo. It was found that the free surface energy is not a decisive factor affecting the non-self-recognition of the modified PE by macrophages. The fusion of macrophages, however, was found to be different on the as-irradiated specimens and the specimens additionally doped with acrylic acid.

• Publikační typ
• časopisecké články MeSH