In this study, fibrous scaffolds based on poly(γ-benzyl-l-glutamate) (PBLG) were investigated in terms of the chondrogenic differentiation potential of human tooth germ stem cells (HTGSCs). Through the solution-assisted bonding of the fibres, fully connected scaffolds with pore sizes in the range 20-400 µm were prepared. Biomimetic modification of the PBLG scaffolds was achieved by a two-step reaction procedure: first, aminolysis of the PBLG fibres' surface layers was performed, which resulted in an increase in the hydrophilicity of the fibrous scaffolds after the introduction of N5 -hydroxyethyl-l-glutamine units; and second, modification with the short peptide sequence azidopentanoyl-GGGRGDSGGGY-NH2 , using the 'click' reaction on the previously modified scaffold with 2-propynyl side-chains, was performed. Radio-assay of the 125 I-labelled peptide was used to evaluate the RGD density in the fibrous scaffolds (which varied in the range 10-3 -10 pm/cm2 ). All the PBLG scaffolds, especially with density 90 ± 20 fm/cm2 and 200 ± 100 fm/cm2 RGD, were found to be potentially suitable for growth and chondrogenic differentiation of HTGSCs. Copyright © 2015 John Wiley & Sons, Ltd.
- Klíčová slova
- HTGSC, adhesion peptide, fibrous scaffolds, poly(amino acid), radio-assay,
- MeSH
- benzylové sloučeniny chemická syntéza chemie farmakologie MeSH
- chrupavka účinky léků fyziologie MeSH
- click chemie MeSH
- dítě MeSH
- glutamáty chemická syntéza chemie farmakologie MeSH
- glykosaminoglykany metabolismus MeSH
- kmenové buňky cytologie účinky léků MeSH
- kultivované buňky MeSH
- lidé MeSH
- magnetická rezonanční spektroskopie MeSH
- mladiství MeSH
- peptidy farmakologie MeSH
- povrchové vlastnosti MeSH
- proliferace buněk účinky léků MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- viabilita buněk účinky léků MeSH
- zubní zárodek cytologie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- benzylové sloučeniny MeSH
- glutamáty MeSH
- glykosaminoglykany MeSH
- peptidy MeSH
- poly-(gamma-benzyl-L-glutamate) MeSH Prohlížeč
Activating mutations in the fibroblast growth factor receptor 3 (FGFR3) cause the most common genetic form of human dwarfism, achondroplasia (ACH). Small chemical inhibitors of FGFR tyrosine kinase activity are considered to be viable option for treating ACH, but little experimental evidence supports this claim. We evaluated five FGFR tyrosine kinase inhibitors (TKIs) (SU5402, PD173074, AZD1480, AZD4547 and BGJ398) for their activity against FGFR signaling in chondrocytes. All five TKIs strongly inhibited FGFR activation in cultured chondrocytes and limb rudiment cultures, completely relieving FGFR-mediated inhibition of chondrocyte proliferation and maturation. In contrast, TKI treatment of newborn mice did not improve skeletal growth and had lethal toxic effects on the liver, lungs and kidneys. In cell-free kinase assays as well as in vitro and in vivo cell assays, none of the tested TKIs demonstrated selectivity for FGFR3 over three other FGFR tyrosine kinases. In addition, the TKIs exhibited significant off-target activity when screened against a panel of 14 unrelated tyrosine kinases. This was most extensive in SU5402 and AZD1480, which inhibited DDR2, IGF1R, FLT3, TRKA, FLT4, ABL and JAK3 with efficiencies similar to or greater than those for FGFR. Low target specificity and toxicity of FGFR TKIs thus compromise their use for treatment of ACH. Conceptually, different avenues of therapeutic FGFR3 targeting should be investigated.
- MeSH
- achondroplazie farmakoterapie MeSH
- benzamidy farmakologie MeSH
- chondrocyty metabolismus MeSH
- chrupavka účinky léků metabolismus MeSH
- fenylmočovinové sloučeniny farmakologie MeSH
- katalýza účinky léků MeSH
- kultivované buňky MeSH
- kuřecí embryo MeSH
- lidé MeSH
- myši MeSH
- piperaziny farmakologie MeSH
- pyrazoly farmakologie MeSH
- pyrimidiny farmakologie MeSH
- pyrroly farmakologie MeSH
- receptory fibroblastových růstových faktorů antagonisté a inhibitory MeSH
- signální transdukce účinky léků MeSH
- syndrom MeSH
- tyrosinkinasové receptory antagonisté a inhibitory MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- AZD 1480 MeSH Prohlížeč
- AZD4547 MeSH Prohlížeč
- benzamidy MeSH
- fenylmočovinové sloučeniny MeSH
- infigratinib MeSH Prohlížeč
- PD 173074 MeSH Prohlížeč
- piperaziny MeSH
- pyrazoly MeSH
- pyrimidiny MeSH
- pyrroly MeSH
- receptory fibroblastových růstových faktorů MeSH
- SU 5402 MeSH Prohlížeč
- tyrosinkinasové receptory MeSH
Aberrant fibroblast growth factor (FGF) signaling disturbs chondrocyte differentiation in skeletal dysplasia, but the mechanisms underlying this process remain unclear. Recently, FGF was found to activate canonical WNT/β-catenin pathway in chondrocytes via Erk MAP kinase-mediated phosphorylation of WNT co-receptor Lrp6. Here, we explore the cellular consequences of such a signaling interaction. WNT enhanced the FGF-mediated suppression of chondrocyte differentiation in mouse limb bud micromass and limb organ cultures, leading to inhibition of cartilage nodule formation in micromass cultures, and suppression of growth in cultured limbs. Simultaneous activation of the FGF and WNT/β-catenin pathways resulted in loss of chondrocyte extracellular matrix, expression of genes typical for mineralized tissues and alteration of cellular shape. WNT enhanced the FGF-mediated downregulation of chondrocyte proteoglycan and collagen extracellular matrix via inhibition of matrix synthesis and induction of proteinases involved in matrix degradation. Expression of genes regulating RhoA GTPase pathway was induced by FGF in cooperation with WNT, and inhibition of the RhoA signaling rescued the FGF/WNT-mediated changes in chondrocyte cellular shape. Our results suggest that aberrant FGF signaling cooperates with WNT/β-catenin in suppression of chondrocyte differentiation.
- Klíčová slova
- Cartilage, Chondrocyte, Differentiation, FGFR3, Fibroblast growth factor receptor, WNT,
- MeSH
- beta-katenin genetika metabolismus MeSH
- biologické modely MeSH
- buněčná diferenciace účinky léků genetika MeSH
- chondrocyty účinky léků metabolismus MeSH
- chrupavka cytologie účinky léků metabolismus MeSH
- fibroblastové růstové faktory farmakologie MeSH
- fibroblastový růstový faktor 2 farmakologie MeSH
- HEK293 buňky MeSH
- končetinové pupeny účinky léků embryologie metabolismus MeSH
- konfokální mikroskopie MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- LDL receptor related protein 6 genetika metabolismus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- protein Wnt3A farmakologie MeSH
- proteiny Wnt genetika metabolismus farmakologie MeSH
- receptory fibroblastových růstových faktorů genetika metabolismus MeSH
- signální transdukce účinky léků genetika MeSH
- synergismus léků MeSH
- transkriptom účinky léků genetika MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- beta-katenin MeSH
- fibroblastové růstové faktory MeSH
- fibroblastový růstový faktor 2 MeSH
- LDL receptor related protein 6 MeSH
- Lrp6 protein, mouse MeSH Prohlížeč
- protein Wnt3A MeSH
- proteiny Wnt MeSH
- receptory fibroblastových růstových faktorů MeSH
Under certain conditions chondrocytes form lattices with cartilage collagens, which may serve as cartilage implants. It is necessary to find the optimal conditions for culturing chondrocytes. Three different supports are compared: (a) plastic; (b) cartilage collagens; and (c) insoluble skin collagen solubilized under denaturing conditions (ISC-40). The effect of culture medium supplementation with the tripeptide (Gly-His-Lys)2.Cu.2H2O.2NaCl (GHK) on chondrocyte proliferation and synthetic activity is studied, with particular attention paid to collagen types I, II and III. The collagen supports stimulated chondrocyte proliferation, but on the ISC-40 support they started to dedifferentiate rather early. In the primary culture, chondrocytes on all three supports synthesized mainly collagen type II, and only small amounts of types I and III. In the first passage the synthesis of these two collagen types increased, relative to collagen type II, at least on the cartilage collagen support. Supplementation of culture medium with GHK stimulated chondrocyte proliferation in the primary structure mostly on the ISC-40 support. On the other two types of supports the stimulatory effect of GHK was expressed mostly in the first passages. The collagen synthetic rate was increased by GHK on both of the collagen supports; on the cartilage collagen support collagen type II was synthesized predominantly and on the ISC-40 support types I and III were mostly formed. It is suggested that supplementation of culture medium with GHK may be useful in the preparation of cartilage implants.
- MeSH
- buněčné dělení účinky léků MeSH
- chrupavka cytologie účinky léků metabolismus MeSH
- denaturace proteinů MeSH
- DNA analýza metabolismus MeSH
- ELISA MeSH
- kolagen biosyntéza chemie MeSH
- kultivační média MeSH
- kultivované buňky MeSH
- kuřecí embryo MeSH
- kůže metabolismus MeSH
- molekulární sekvence - údaje MeSH
- oligopeptidy farmakologie MeSH
- plastické hmoty chemie metabolismus MeSH
- protézy a implantáty normy MeSH
- růstové látky farmakologie MeSH
- sekvence aminokyselin MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA MeSH
- glycyl-histidyl-lysine MeSH Prohlížeč
- kolagen MeSH
- kultivační média MeSH
- oligopeptidy MeSH
- plastické hmoty MeSH
- růstové látky MeSH
Administration into mouse shank muscles of various immunomodulators which directly or indirectly activate lymphocytes (BCG, Con A, Carrageenan IV, Dextran, PHA-M, PWM, lipopolysaccharides from Corynebacterium) have a dual effect on local bones, stimulating both periosteal bone formation and bone resorption, the former being dominant. These effects vary in frequency and magnitude, the most potent stimulation of periosteal bone formation being observed after administration of Con A and BCG in complete Freund adjuvant. BCG was also a strong inducer of bone resorption in vivo. Concanavalin A and, to a lesser degree, other immunomodulators applied, when administered subcutaneously into the pinna, also have induced perichondrial chondrogenesis. These novel effects of immunodulators could be applied in the field of skeletal tissues regeneration.
- MeSH
- adjuvancia imunologická farmakologie MeSH
- chrupavka cytologie účinky léků MeSH
- inbrední kmeny myší MeSH
- kosti a kostní tkáň účinky léků fyziologie MeSH
- myši MeSH
- osteoblasty účinky léků MeSH
- osteogeneze účinky léků fyziologie MeSH
- resorpce kosti MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adjuvancia imunologická MeSH
The effects of several antirheumatic drugs on the activity of degradative enzymes in normal and pathologic knee joint cartilage and on the proliferative activity of synovial tissue cells were studied. Inflammatory arthropathy was induced in rabbits by intraarticular papain administration. Elevated contents of proteoglycanase and collagenase, together with an increase in serine and cysteine proteinase inhibitors, were found in animals with papain-induced arthropathy. Inflammation also accelerated the rate of proliferation of cells present in the synovial tissue. In the treated animals, the reduction in enzyme activity, decrease in inhibitor content and decreased DNA proliferation rate were registered to a different degree. The suppression of protein synthesis by nonsteroidal antiinflammatory drugs may explain our findings. The best therapeutic results were achieved with glycosaminoglycan polysulphate (Arteparon).
- MeSH
- antiflogistika nesteroidní terapeutické užití MeSH
- antiflogistika MeSH
- buněčné dělení MeSH
- chrupavka účinky léků metabolismus MeSH
- cytofotometrie MeSH
- DNA metabolismus MeSH
- endopeptidasy metabolismus MeSH
- králíci MeSH
- metaloendopeptidasy * MeSH
- mikrobiální kolagenasa metabolismus MeSH
- osteoartróza farmakoterapie enzymologie MeSH
- ploidie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antiflogistika nesteroidní MeSH
- antiflogistika MeSH
- DNA MeSH
- endopeptidasy MeSH
- metaloendopeptidasy * MeSH
- mikrobiální kolagenasa MeSH
- proteoglycan-degrading metalloendopeptidases MeSH Prohlížeč
- MeSH
- chrupavka účinky léků embryologie MeSH
- končetiny účinky léků embryologie MeSH
- kyselina askorbová farmakologie MeSH
- morfogeneze účinky léků MeSH
- myši MeSH
- orgánové kultury - kultivační techniky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kyselina askorbová MeSH
- MeSH
- cholekalciferol farmakologie MeSH
- chrupavka účinky léků MeSH
- kalcitonin farmakologie MeSH
- králíci MeSH
- parathormon farmakologie MeSH
- processus xiphoideus MeSH
- ucho MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- cholekalciferol MeSH
- kalcitonin MeSH
- parathormon MeSH
