Research of degradable hydrogel polymeric materials exhibiting high water content and mechanical properties resembling tissues is crucial not only in drug delivery systems but also in tissue engineering, medical devices, and biomedical-healthcare sensors. Therefore, we newly offer development of hydrogels based on poly(2-hydroxyethyl methacrylate-co-2-(acetylthio) ethyl methacrylate-co-2-methacryloyloxyethyl phosphorylcholine) [P(HEMA-ATEMA-MPC)] and optimization of their mechanical and in vitro and in vivo degradability. P(HEMA-ATEMA-MPC) hydrogels differed in chemical composition, degree of crosslinking, and starting molar mass of polymers (15, 19, and 30 kDa). Polymer precursors were synthesized by a reversible addition fragmentation chain transfer (RAFT) polymerization using 2-(acetylthio)ethyl methacrylate containing protected thiol groups, which enabled crosslinking and gel formation. Elastic modulus of hydrogels increased with the degree of crosslinking (Slaughter et al., 2009) [1]. In vitro and in vivo controlled degradation was confirmed using glutathione and subcutaneous implantation of hydrogels in rats, respectively. We proved that the hydrogels with higher degree of crosslinking retarded the degradation. Also, albumin, γ-globulin, and fibrinogen adsorption on P(HEMA-ATEMA-MPC) hydrogel surface was tested, to simulate adsorption in living organism. Rat mesenchymal stromal cell adhesion on hydrogels was improved by the presence of RGDS peptide and laminin on the hydrogels. We found that rat mesenchymal stromal cells proliferated better on laminin-coated hydrogels than on RGDS-modified ones.
Monodisperse nonmagnetic macroporous poly(glycidyl methacrylate) (PGMA) microspheres were synthesized by multistep swelling polymerization of glycidyl methacrylate, ethylene dimethacrylate and 2-[(methoxycarbonyl)methoxy]ethyl methacrylate (MCMEMA). This was followed (a) by ammonolysis to modify the microspheres with amino groups, and (b) by incorporation of iron oxide (γ-Fe2O3) into the pores to render the particles magnetic. The resulting porous and magnetic microspheres were characterized by scanning and transmission electron microscopy (SEM and TEM), atomic absorption and Fourier transform infrared spectroscopy (AAS and FTIR), elemental analysis, vibrating magnetometry, mercury porosimetry and Brunauer-Emmett-Teller adsorption/desorption isotherms. The microspheres are meso- and macroporous, typically 5 μm in diameter, contain 0.9 mM · g-1 of amino groups and 14 wt.% of iron according to elemental analysis and AAS, respectively. The particles were conjugated to p46/Myo1C protein, a potential biomarker of autoimmune diseases, to isolate specific autoantibodies in the blood of patients suffering from multiple sclerosis (MS). The p46/Myo1C loaded microspheres are shown to enable the preconcentration of minute quantities of specific immunoglobulins prior to their quantification via SDS-PAGE. The immunoglobulin M (IgM) with affinity to Myo1C was detected in MS patients. Graphical abstract Monodisperse magnetic poly(glycidyl methacrylate) microspheres were synthesized, conjugated with 46 kDa form of unconventional Myo1C protein (p46/Myo1C) via carbodiimide (DIC) chemistry, and specific autoantibodies isolated from blood of multiple sclerosis (MS) patients; immunoglobulin M (IgM) level increased in MS patients.
- MeSH
- autoimunitní nemoci imunologie MeSH
- autoprotilátky krev chemie imunologie izolace a purifikace MeSH
- kyseliny polymethakrylové chemie MeSH
- lidé MeSH
- magnety chemie MeSH
- mikrosféry * MeSH
- molekulová hmotnost MeSH
- myosin typu I chemie imunologie MeSH
- roztroušená skleróza imunologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Heat-treated polyacrylonitrile (HT-PAN), also referred to as black orlon (BO), is a promising carbon-based material used for applications in tissue engineering and regenerative medicine. To the best of our knowledge, no such complex bone morphology-mimicking three-dimensional (3D) BO structure has been reported to date. We report that BO can be easily made into 3D cryogel scaffolds with porous structures, using succinonitrile as a porogen. The cryogels possess a porous morphology, similar to bone tissue. The prepared scaffolds showed strong osteoconductive activity, providing excellent support for the adhesion, proliferation, and mitochondrial activity of human bone-derived cells. This effect was more apparent in scaffolds prepared from a matrix with a higher content of PAN (i.e., 10% rather than 5%). The scaffolds with 10% of PAN also showed enhanced mechanical properties, as revealed by higher compressive modulus and higher compressive strength. Therefore, these scaffolds have a robust potential for use in bone tissue engineering.
- MeSH
- akrylové pryskyřice chemie MeSH
- kosti a kostní tkáň MeSH
- lidé MeSH
- pevnost v tlaku MeSH
- poréznost MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury MeSH
- vysoká teplota MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The aim of the present study is to develop new magnetic polymer microspheres with functional groups available for easy protein and antibody binding. Monodisperse macroporous poly(2-hydroxyethyl methacrylate) (PHEMA-COOH) microspheres ~4 µm in diameter and containing ∼1 mmol COOH/g were synthesized by multistep swelling polymerization of 2-hydroxyethyl methacrylate (HEMA), ethylene dimethacrylate (EDMA), and 2-[(methoxycarbonyl)methoxy]ethyl methacrylate (MCMEMA), which was followed by MCMEMA hydrolysis. The microspheres were rendered magnetic by precipitation of iron oxide inside the pores, which made them easily separable in a magnetic field. Properties of the resulting magnetic poly(2-hydroxyethyl methacrylate) (mgt.PHEMA) particles with COOH functionality were examined by scanning and transmission electron microscopy (SEM and TEM), static volumetric adsorption of helium and nitrogen, mercury porosimetry, Fourier transform infrared (FTIR) and atomic absorption spectroscopy (AAS), and elemental analysis. Mgt.PHEMA microspheres were coupled with p46/Myo1C protein purified from blood serum of multiple sclerosis (MS) patients, which enabled easy isolation of monospecific anti-p46/Myo1C immunoglobulin G (IgG) antibodies from crude antibody preparations of mouse blood serum. High efficiency of this approach was confirmed by SDS/PAGE, Western blot, and dot blot analyses. The newly developed mgt.PHEMA microspheres conjugated with a potential disease biomarker, p46/Myo1C protein, are thus a promising tool for affinity purification of antibodies, which can improve diagnosis and treatment of MS patients.
- MeSH
- imobilizované proteiny chemie imunologie MeSH
- imunoglobulin G imunologie izolace a purifikace MeSH
- lidé MeSH
- magnetismus metody MeSH
- magnety chemie MeSH
- mikrosféry MeSH
- myosin typu I chemie imunologie MeSH
- myši MeSH
- polyhydroxyethylmethakrylát chemie MeSH
- protein - isoformy chemie imunologie MeSH
- roztroušená skleróza diagnóza imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Three-dimensional hydrogel supports for mesenchymal and neural stem cells (NSCs) are promising materials for tissue engineering applications such as spinal cord repair. This study involves the preparation and characterization of superporous scaffolds based on a copolymer of 2-hydroxyethyl and 2-aminoethyl methacrylate (HEMA and AEMA) crosslinked with ethylene dimethacrylate. Ammonium oxalate is chosen as a suitable porogen because it consists of needle-like crystals, allowing their parallel arrangement in the polymerization mold. The amino group of AEMA is used to immobilize RGDS and SIKVAVS peptide sequences with an N-γ-maleimidobutyryloxy succinimide ester linker. The amount of the peptide on the scaffold is determined using 125 I radiolabeled SIKVAVS. Both RGDS- and SIKVAVS-modified poly(2-hydroxyethyl methacrylate) scaffolds serve as supports for culturing human mesenchymal stem cells (MSCs) and human fetal NSCs. The RGDS sequence is found to be better for MSC and NSC proliferation and growth than SIKVAVS.
- MeSH
- buněčné linie MeSH
- lidé MeSH
- methylmetakryláty chemie farmakologie MeSH
- mezenchymální kmenové buňky cytologie MeSH
- nervové kmenové buňky cytologie metabolismus MeSH
- oligopeptidy * chemie farmakologie MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The design of favorable mechanical properties and suitable surface modifications of hydrogels in order to stimulate specific cell response is a great challenge. N-(2-Hydroxypropyl) methacryl-amide (HPMA) was utilized to form macroporous cryogel scaffolds for stem cell applications. Furthermore, one group of scaffolds was enhanced by copolymerization of HPMA with methacryloyl-GGGRGDS-OH peptide in an effort to integrate biomimetic adhesion sites. The cryogels were characterized by stiffness and equilibrium swelling measurements as well as by scanning electron microscopy. Cell culture experiments were performed with human adipose-derived stem cells and substrates were found completely non-toxic. Moreover, RGDS-enriched cryogels supported cell attachment, spreading and proliferation, so they can be considered suitable for designed aims.
- MeSH
- akrylamidy * MeSH
- biokompatibilní materiály MeSH
- biomimetika * MeSH
- buněčná adheze MeSH
- kmenové buňky * MeSH
- kryogely MeSH
- kultivované buňky MeSH
- lidé MeSH
- poréznost MeSH
- proliferace buněk MeSH
- pružnost MeSH
- tkáňové podpůrné struktury MeSH
- tukové buňky MeSH
- voda chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Identification and evaluation of small changes in β-amyloid peptide (Aβ) levels in cerebrospinal fluid is of crucial importance for early detection of Alzheimer's disease. Microfluidic detection methods enable effective preconcentration of Aβ using magnetic microparticles coated with Aβ antibodies. Poly(glycidyl methacrylate) microspheres are coated with α-amino-ω-methoxy-PEG5000 /α-amino-ω-Boc-NH-PEG5000 Boc groups deprotected and NH2 succinylated to introduce carboxyl groups. Capillary electrophoresis with laser-induced fluorescence detection confirms the efficient capture of Aβ 1-40 peptides on the microspheres with immobilized monoclonal anti-Aβ 6E10. The capture specificity is confirmed by comparing Aβ 1-40 levels on the anti-IgG-immobilized particles used as a control.
- MeSH
- adsorpce MeSH
- amyloidní beta-protein izolace a purifikace MeSH
- chromatografie afinitní MeSH
- elektroforéza kapilární MeSH
- imunoglobulin G metabolismus MeSH
- kyseliny karboxylové chemie MeSH
- kyseliny polymethakrylové chemie MeSH
- magnetické jevy MeSH
- mikrosféry * MeSH
- mikroskopie atomárních sil MeSH
- peptidové fragmenty izolace a purifikace MeSH
- polyethylenglykoly chemie MeSH
- skot MeSH
- termogravimetrie MeSH
- velikost částic MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The efficient isolation and concentration of protein antigens from complex biological samples is a critical step in several analytical methods, such as mass spectrometry, flow cytometry and immunochemistry. These techniques take advantage of magnetic microspheres as immunosorbents. The focus of this study was on the development of new superparamagnetic polymer microspheres for the specific isolation of the tumor suppressor protein p53. Monodisperse macroporous poly(glycidyl methacrylate) (PGMA) microspheres measuring approximately 5 μm and containing carboxyl groups were prepared by multistep swelling polymerization of glycidyl methacrylate (GMA), 2-[(methoxycarbonyl)methoxy]ethyl methacrylate (MCMEMA) and ethylene dimethylacrylate (EDMA) as a crosslinker in the presence of cyclohexyl acetate as a porogen. To render the microspheres magnetic, iron oxide was precipitated within their pores; the Fe content in the particles received ∼18 wt%. Nonspecific interactions between the magnetic particles and biological media were minimized by coating the microspheres with poly(ethylene glycol) (PEG) terminated by carboxyl groups. The carboxyl groups of the magnetic PGMA microspheres were conjugated with primary amino groups of mouse monoclonal DO-1 antibody using conventional carbodiimide chemistry. The efficiency of protein p53 capture and the degree of nonspecific adsorption on neat and PEG-coated magnetic microspheres were determined by western blot analysis.
- MeSH
- kyseliny polymethakrylové chemie MeSH
- mikrosféry * MeSH
- myší monoklonální protilátky chemie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 chemie izolace a purifikace MeSH
- polyethylenglykoly chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
To minimize non-specific protein adsorption on macroporous poly(glycidyl methacrylate) and poly(2-hydroxyethyl methacrylate) microspheres containing amino and/or carboxyl groups, the microspheres are coated with α,ω-bis-carboxy poly(ethylene glycol) and amino-terminated poly(ethylene glycol-co-propylene glycol) or α-methoxy-ω-amino poly(ethylene glycol). Adsorption of bovine serum albumin (BSA), γ-globulin, (125) I-BSA, pepsin, and chymotrypsin on neat and PEGylated microspheres is determined by UV-VIS spectroscopy of supernatants and eluates or by measurement of radioactivity in an ionization chamber. Neat and PEGylated microspheres adsorb 0.8-70% and 0.02-44% of protein, respectively.
- MeSH
- adsorpce MeSH
- biotechnologie metody MeSH
- chymotrypsin chemie MeSH
- gama-globuliny chemie MeSH
- kyseliny polymethakrylové chemie MeSH
- mikrosféry * MeSH
- molekulární struktura MeSH
- polyethylenglykoly chemie MeSH
- polyhydroxyethylmethakrylát chemie MeSH
- radioizotopy jodu chemie MeSH
- sérový albumin hovězí chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Poly(N,N-diethylacrylamide) (PDEAAm) hydrogel scaffolds were prepared by radical copolymerization of N,N-diethylacrylamide (DEAAm), N,N'-methylenebisacrylamide and methacrylic acid in the presence of (NH₄)₂SO₄ or NaCl. The hydrogels were characterized by low-vacuum scanning electron microscopy in the water-swollen state, water and cyclohexane regain, and by mercury porosimetry. The pentapeptide, YIGSR-NH₂, was immobilized on the hydrogel. Human embryonic stem cells (hESCs) were cultured with the hydrogels to test their biocompatibility. The results suggest that the PDEAAm hydrogel scaffolds are nontoxic and support hESC attachment and proliferation, and that interconnected pores of the scaffolds are important for hESC cultivation. Immobilization of YIGSR-NH₂ pentapeptide on the PDEAAm surface improved both adhesion and growth of hESCs compared with the unmodified hydrogel. The YIGSR-NH₂-modified PDEAAm hydrogels may be a useful tool for tissue-engineering purposes.
- MeSH
- akrylamidy chemie MeSH
- buněčné linie MeSH
- embryonální kmenové buňky cytologie MeSH
- hydrogely chemie MeSH
- lidé MeSH
- myši MeSH
- oligopeptidy chemie MeSH
- polymery chemie MeSH
- proliferace buněk MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
