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Biomedical subjects

J Vacík

Publications and source records attributed to J Vacík.

At least 19 recordsLinked to original sources
PubMed2006

Macroporous hydrogels based on 2-hydroxyethyl methacrylate. Part 4: growth of rat bone marrow stromal cells in three-dimensional hydrogels with positive and negative surface charges and in polyelectrolyte complexes.

The growth of bone marrow stromal cells was assessed in vitro in macroporous hydrogels based on 2-hydro- xyethyl methacrylate (HEMA) copolymers with different electric charges. Copolymers of HEMA with sodium methacrylate (MA(-)) carried a negative electric charge, copolymers of HEMA with [2-(methacryloyloxy)ethyl] trimethylammonium chloride (MOETA(-)) carried a positive electric charge and terpolymers of HEMA, MA(-) and MOETA(+) carried both, positive and negative electric charges. The charges in the polyelectrolyte complexes were shielded by counter-ions. The hydrogels had similar porosities, based on a comparison of their diffusion parameters for small cations as measured by the real-time tetramethylammonium iontophoretic method of diffusion analysis. The cell growth was studied in the peripheral and central regions of the hydrogels at 2 hours and 2, 7, 14 and 28 days after cell seeding. Image analysis revealed the highest cellular density in the HEMA-MOETA(+) copolymers; most of the cells were present in the peripheral region of the hydrogels. A lower density of cells but no difference between the peripheral and central regions was observed in the HEMA-MA(-) copolymers and in polyelectrolyte complexes. This study showed that positively charged functional groups promote the adhesion of cells.

Animals↗
PubMed2005

Macroporous hydrogels based on 2-hydroxyethyl methacrylate. Part II. Copolymers with positive and negative charges, polyelectrolyte complexes.

Crosslinked macroporous hydrogels based on 2-hydroxyethyl methacrylate (HEMA)-[2-(methacryloyloxy)ethyl]trimethylammonium chloride (MOETACl) copolymer, HEMA-MOETACl-methacrylic acid (MA) terpolymer, and on a polyelectrolyte complex of HEMA-MA copolymer with poly(MOETACl) were prepared. All the hydrogels were prepared in the presence of fractionated sodium chloride particles. The hydrogels were characterized by the number of pores and the total volume of all pores in unit volume, the average volume and the average diameter of single pore. Morphology of the hydrogels was investigated by confocal and scanning electron microscopy. The hydrogels based on polyelectrolyte complexes were also characterized by chemical composition. Homogeneous (non-porous) hydrogels with the same composition as macroporous hydrogels were prepared and characterized by their biocompatibility.

Animals↗
PubMed2002

Development of hydrogel implants for urinary incontinence treatment.

Hydrogel implants for urinary incontinence treatment based on HEMA supplemented with 10% methacrylic acid have been developed. The swelling properties of implants were tested in vitro and in vivo after implantation to laboratory mice. Biocompatibility has been determined by incubation of implants in tissue culture, by histological examination of adjacent tissues after subcutaneous application of implants to laboratory mouse in a long-term experiment, and by flow cytometry examination of blood cells. The swelling of hydrogel implants was completed in 6-24 h. There was no effect on in vitro growth of cells incubated with implants. In mice, implants were well tolerated without any sign of inflammatory reaction. The material allows an elastic compression of urethra compensating a damaged sphincter after trans-urethral sub-mucosal implantation of hydrogel cylinders.

Animals↗
PubMed2002

Polymer hydrogels usable for nervous tissue repair.

The implantation of non-resorbable biocompatible polymer hydrogels into defects in the central nervous system can reduce glial scar formation, bridge the lesion and lead to tissue regeneration within the hydrogel. We implanted hydrogels based on crosslinked poly hydroxyethyl-methacrylate (pHEMA) and poly N-(2-hydroxypropyl)-methacrylamide (pHPMA) into the rat cortex and evaluated the cellular invasion into the hydrogels by means of immunohistochemical methods and tetramethylammonium diffusion measurements. Astrocytes and NF160-positive axons grew similarly into both types of hydrogels. We found no cell types other than astrocytes in the pHEMA hydrogels. In the pHPMA hydrogels, we found a massive ingrowth of connective tissue elements. These changes were accompanied by corresponding changes in the extracellular space volume fraction and tortuosity of the hydrogels.

Animals↗
PubMed2000

[Surgical treatment of recurrent urethral strictures in males after unsuccessful operations].

BACKGROUND: Subvesical obstructions of any origin represent a frequent and serious disorder occurring predominantly in males. Often it brings incontinence and/or erectility dysfunction, infection of urinary tract. Relapses of the acute pyelonephritis can turn into chronic tubulointersticial one and terminate in the renal insufficiency. To treat strictures, dilation, intermittent catheterization and recently stent introduction were used. Most suitable appears a stent from composite polymers. The aim of our work was to test properties of stents developed in the Institute of Macromolecular Chemistry ASCR. METHODS AND RESULTS: Stents from composite polymers, which are non-toxic, not-irritable can swell in body fluids and have mechanical properties similar to that of silicone rubber. Properties of the material are functionally graded and the casting or repoussé from the material can subsequently change its shape. Ten patients (males, aged 25 to 78 years) with long urethral strictures in its bulbocavernous part (50%) were treated with this method. Strictures were caused by pelvical fractures (4 times), prostate hypertrophy surgery (4 times), prolonged catheterizations (2 times). All patients were followed for 16 to 26 month and had no severities. CONCLUSION: Our results indicate that stent from composite polymers and silicone may have long-acting effects without irritation or crust formation and beneficially effected healing of the spongio-fibrous process.

Adult↗
PubMed2000

Synthetic hydrogel capacity to induce formation of foreign-body giant multinucleate cells differs in vivo and in vitro.

The granulomatous reaction accompanied with MGC formation represents the most striking feature of the non-favourable biological tolerance of implanted devices. We compared MGC formation in the course of the granulomatous reaction in vitro and in vivo employing three types of hydrogels whose biocompatibility had been well studied earlier. The efficiency of the in vitro assay for the granulomatous reaction, including MGC formation, was verified employing the nematode Nippostrongylus brasiliensis, a well-known inductor of MGC formation in vitro. The in vitro results demonstrated a very low level of MGC formation in reaction against all three types of hydrogels without polymer-specific differences in comparison with the nematode experiment characterized by a high extent of MGC formation. On the other hand, the extent of MGC formation was implant type-specific in vivo: pHEMA-co-DMAEMA > pHEMA > pHEMA-co-NaMA. These results indicate that in the in vitro assay it was not possible to discriminate among the types of polymers used in the experiment in comparison with the animal experiment. They also indicate potential differences between granuloma formation induced by parasites and by foreign bodies.

Animals↗
PubMed2000

Development of fixable stent for long-term derivation of lower urinary tract.

The object of the envisaged project is the development of easily fixable non-irritant stent of the lower urinary tract. The project has the aim to remove the obstruction and make passage-way through urethra which, after repeated operations, remains narrowed and requires repeated dilatations or, in the patients who have a chronic epicystostomy due to the non-passable urethra. The prosthesis must be self fixable, non-irritant, easily introduceable and removable by endoscopy, enabling normal urination with urinary continence and, last but not least, at a reasonable price.

Equipment Design↗
PubMed1999

Brain lesion-induced alteration of selected phenotypic properties of spleen macrophages and their partial restoration in the course of foreign body reaction against intraperitoneally implanted polymers.

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.

Journal Article↗
PubMed1998

Reactive oxygen species (ROS) generated by xanthine oxidase in the corneal epithelium and their potential participation in the damage of the corneal epithelium after prolonged use of contact lenses in rabbits.

Prolonged use of contact lenses (for 14 days) evoked an imbalance between the activity of xanthine oxidase (an enzyme belonging to reactive oxygen species-generating oxidases) and catalase (an enzyme belonging to reactive oxygen species-scavenging oxidases) in the corneal epithelium of rabbits. The activity of catalase decreased, while xanthine oxidase activity was very high. Of other enzymes studied in the corneal epithelium, the activities of xanthine oxidoreductase, glucoso-6-phosphate dehydrogenase and succinate dehydrogenase were decreased. In contrast, the activities of lactate dehydrogenase and lysosomal hydrolases (acid beta-galactosidase, dipeptidyl peptidase II) were increased and appeared in animals sacrificed immediately after contact lens removal. In rabbits sacrificed later (after 1 h), an additional increase of lactate dehydrogenase and lysosomal hydrolase activities developed in the superficial layers of the corneal epithelium. Catalase supplementation during use of contact lenses prevented both the significant decrease of catalase activity in the corneal epithelium and the development of additional epithelial damage. In contrast, topical treatment with 3-aminotriazole (an inhibitor of catalase) resulted in the nearly complete loss of catalase activity in the corneal epithelium and the appearance of more serious epithelial damage. We conclude that ROS generated by xanthine oxidase induce additional damage of the corneal epithelium related to the use of contact lenses.

Animals↗
PubMed1998

[The histopathologic picture of experimental poisoning with ethylmethacrylates].

Toxic effect of hydroxyethylmethacrylate, acetoxyethylmethacrylate and diethylenglycomethacrylate were studied in rats surviving as long as 1 to 20 days after intramuscular administration. Conspicuous lesion were found only in calf muscles at the site of application. Muscle fibre necroses with inflammatory reaction occurred repeatedly in animals surviving 1-2 days. Newly formed connective tissue replacing impaired muscle fibres was found in rats surviving 5 days. Intoxication related other lesion in rat organs were not identified.

Animals↗
PubMed1997

Anionic polymers for implantation.

The biocompatibility of hydrogels containing carboxylate anions was studied by a panel of tests in vitro and in vivo. In comparison with other types of similar hydrogels, those with COO- anions induced a lower extent of foreign body reaction, and their systemic effect on the immune system also seems lower. The biomimetic effect of carboxylate anions on biocompatibility of synthetic materials could be explained by a similar biomimetic effect of N-acetylneuraminic acid inhibiting the non-self recognition of bacterial cells by non-specific immunity. The encouraging long-term results of clinical trials of intraocular lenses prepared from copolymer of 2-hydroxyethyl methacrylate and sodium methacrylate support this hypothesis.

Animals↗
PubMed1997

Poly (triethylenglycol monomethacrylate) and poly (glycerol monomethacrylate) cross-linked gel as potential viscoelastics for intraoperative use.

BACKGROUND: The highly swelling poly (glycerol monomethacrylate) gel (polyGLYMA) and hydrophilic polymer poly (triethylenglycol monomethacrylate (polyTEGMA) were tested as potential viscoelastics for intraoperative use in anterior segment surgery. METHODS: PolyGLYMA was implanted into the anterior chamber in 5 rabbits, and 40% polyTEGMA in 16 rabbits. The eyes were enucleated 1 week to 3 months after the operation. The corneal endothelium was examined with specular microscopy, and then the whole eye histopathologically. RESULTS: In all eyes of the polyGLYMA group, the clinical findings were characterized by a marked ciliary injection and severe secondary glaucoma, and the histologic ones by a marked inflammatory infiltration and thickening of Descemet's membrane in the anterior chamber angle. Specular microscopy revealed a decrease in the endothelial cell density and polymorphism of the endothelial cells. In the polyTEGMA group, the anterior segment and the fundus were physiologic all the time, and specular microscopy and histologic findings showed no degenerative and/or inflammatory changes. CONCLUSIONS: PolyGLYMA proved unsuitable for intracameral application in rabbits. The new polymer polyTEGMA is characterized by high biologic tolerance after its implantation into the anterior chamber of rabbits. PolyTEGMA 40% might be considered as a potential viscoelastic material in humans.

Animals↗
PubMed1997

Effect of chemical structure of hydrogels on the adhesion and phenotypic characteristics of human monocytes such as expression of galectins and other carbohydrate-binding sites.

The reactivity of diverse immune aspects to the presence of synthetic polymers represents one of the most important aspects of implantable device biocompatibility. In this report, we show the effect of the chemical structure of a synthetic polymer support on monocyte adhesion and selected phenotypic characteristics in vitro as a model for the initial steps of non-self-recognition of an implant. The extent of monocyte adhesion was significantly influenced by the support chemistry. The highest level of monocyte adhesion was observed on a surface copolymer of 2-hydroxyethyl methacrylate with dimethyl aminoethyl methacrylate relative to results of experiments in which poly(2-hydroxyethyl methacrylate) or the copolymer of 2-hydroxyethyl methacrylate with the sodium salt of methacrylic acid was used. Cell adhesion to the polymers tested and to glass was accompanied by enhanced expression of the carbohydrate-binding sites tested for asialoglycoprotein beta-galactosides such as galectins, beta-N-acetylgalactosamine, alpha-mannoside, specific lectin for heparin as well as the lymphokine-macrophage migration inhibitory factor in the monocytes tested. These results suggest the importance of monocyte adhesion to the biomaterial surface for their development into macrophages and further non-self-recognition of the implanted device.

Acetylgalactosamine↗
PubMed1997

Comparative study of human monocyte and platelet adhesion to hydrogels in vitro - effect of polymer structure.

Blood platelet and monocyte adhesion was studied in vitro with respect to the influence of hydrophilic polymer chemical functional groups and their charge. The results showed that the strongest adhesion of human monocytes was to coverslips covered with cationic polymer. Platelet adhesion to all tested polymers proved to be negligible; no differences related to the charge of the polymers used were observed. These results show the obvious difference between the biocompatibility and haemocompatibility in vitro which must be taken into consideration during polymer biological properties testing before clinical trials.

Journal Article↗
PubMed1997

Phenotypic characterization of keratinocytes migrated from polymer support - in vitro study.

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.

Journal Article↗
PubMed1997

[New approaches in clinical ophthalmology: UV absorbers and hindered amines covalently bound to contact lenses. Importance of enzyme histochemistry in studying their effectiveness in vivo].

UV absorbers developed by us are covalently bound in hydrophilic contact lenses. This is great advantage because UV absorbers cannot be diluted from contact lenses. In laboratory investigations and "in vivo" experiments it was found that contact lenses containing UV absorbents prevent the eye against the damaging effect of UV irradiation (UVA, UVB, UVC rays). The second new approach (mentioned in this paper) is the use of Hindered Amine Light Stabilizer (4-methakryloylamino-2,2,6,6-tetramethylpiperidine) covalently bound in the hydrophilic contact or therapeutic lenses--for the healing of the damaged eye. In laboratory measurements the cleavage of hydrogen peroxide (of various concentrations) by hindered amine light stabilizer was confirmed. In "in vivo" experiments it was examined histochemically and morphologically that Hindered Amines Light Stabilizer bound in contact lenses favourable influenced the healing of the damaged anterior eye segment of rabbits. The amount as well as enzyme activation (lysosomal hydrolases) of inflammatory cells was significantly reduced in the anterior eye segment. It is suggested that Hindered Amine Light Stabilizer reacts with reactive oxygen species (oxyradicals and hydrogen peroxide) present in the damaged anterior eye segment (e.g. released from inflammatory cells). During this reaction intermediate products appear and the resulting product is a nitroxide radical. All reaction products as well as resulting product are covalently bound in hydrophilic material. For the development and also evaluation of the efficacy of both new approaches perspective for the use in clinical ophthalmology, biochemical investigations and histochemical methods are employed. Enzyme histochemistry is very important for the evaluation of the efficacy of new therapeutic approaches "in vivo" and therefore the particular attention is devoted to it in this communication.

Acrylates↗
PubMed1996

Influence of intraperitoneal injection of three types of hydrogel beads on expression of carbohydrate-binding sites in spleen macrophages.

The influence of the chemical structure of polymer implants on selected characteristics of macrophages was studied to improve our understanding of the mechanisms of non-self recognition of synthetic materials. Three types of polymers differing in net charge were prepared to compare in vivo responses. Beads from preparation of poly(2-hydroxyethyl methacrylate), a copolymer of 2-hydroxyethyl methacrylate with sodium methacrylate, and a copolymer of 2-hydroxyethyl methacrylate and dimethylaminoethyl methacrylate were injected intraperitoneally into rats and harvested 48 h later. The effects of these polymers on the presence of inflammatory cells in the peritoneal exudate, on the adhesion of macrophages to individual batches of the different types of beads and on distinct molecular aspects of macrophages in the red pulp of spleen were evaluated. Beads from both types of copolymer caused an elevation in the number of macrophages in the exudate, in contrast to the situation in rats treated with poly(2-hydroxyethyl methacrylate) beads and physiological saline solution as control. The molecular design of the implant had no significant influence on the extent of macrophage adhesion to beads or on the expression of selected carbohydrate-binding sites. Since important cellular functions such as cell adhesion and glycoprotein routing depend on the sugar part of glycoconjugates, labelled neoglycoproteins were employed to analyse this aspect of macrophages in the tested animals. The beads of the copolymer of 2-hydroxyethyl methacrylate with dimethylaminoethyl methacrylate clearly led to an elevation of the expression of specific binding sites for beta-galactoside-terminating structures which are presented by asialofetuin, for mannose, fucose, sialic acid and N-acetylgalactosamine, which had been used as the ligand parts of biotinylated neoglycoproteins, in spleen macrophages whereas the levels of sites which recognize mannose-6-phosphate were unaffected. Expression of sites with specificity to N-acetylglucosamine was lessened. The effect of beads from the copolymer of 2-hydroxyethyl methacrylate with sodium methacrylate on the measured glycobiological features in the splenic macrophages was only negligible. These results suggest the possibility of systemic effects of implanted polymers on the distinct recognitive functions of macrophages.

Animals↗