• MeSH
• artróza kolenních kloubů farmakoterapie   MeSH
• chondrogeneze účinky léků   MeSH
• glukokortikoidy aplikace a dávkování   farmakologie   terapeutické užití   MeSH
• hodnotící studie jako téma MeSH
• kloubní chrupavka účinky léků   MeSH
• osteoartróza * etiologie   farmakoterapie   MeSH
• triamcinolonacetonid aplikace a dávkování   farmakologie   terapeutické užití   MeSH

Caspase-1, as the main pro-inflammatory cysteine protease, was investigated mostly with respect to inflammation-related processes. Interestingly, caspase-1 was identified as being involved in lipid metabolism, which is extremely important for the proper differentiation of chondrocytes. Based on a screening investigation, general caspase inhibition impacts the expression of Cd36 in chondrocytes, the fatty acid translocase with a significant impact on lipid metabolism. However, the engagement of individual caspases in the effect has not yet been identified. Therefore, the hypothesis that caspase-1 might be a candidate here appears challenging. The primary aim of this study thus was to find out whether the inhibition of caspase-1 activity would affect Cd36 expression in a chondrogenic micromass model. The expression of Pparg, a regulator Cd36, was examined as well. In the caspase-1 inhibited samples, both molecules were significantly downregulated. Notably, in the treated group, the formation of the chondrogenic nodules was apparently disrupted, and the subcellular deposition of lipids and polysaccharides showed an abnormal pattern. To further investigate this observation, the samples were subjected to an osteogenic PCR array containing selected markers related to cartilage/bone cell differentiation. Among affected molecules, Bmp7 and Gdf10 showed a significantly increased expression, while Itgam, Mmp9, Vdr, and Rankl decreased. Notably, Rankl is a key marker in bone remodeling/homeostasis and thus is a target in several treatment strategies, including a variety of fatty acids, and is balanced by its decoy receptor Opg (osteoprotegerin). To evaluate the effect of Cd36 downregulation on Rankl and Opg, Cd36 silencing was performed using micromass cultures. After Cd36 silencing, the expression of Rankl was downregulated and Opg upregulated, which was an inverse effect to caspase-1 inhibition (and Cd36 upregulation). These results demonstrate new functions of caspase-1 in chondrocyte differentiation and lipid metabolism-related pathways. The effect on the Rankl/Opg ratio, critical for bone maintenance and pathology, including osteoarthritis, is particularly important here as well.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• chondrocyty metabolismus   MeSH
• chondrogeneze účinky léků   MeSH
• diferenciační antigeny biosyntéza   MeSH
• inhibitory kaspas farmakologie   MeSH
• kaspasa 1 metabolismus   MeSH
• metabolismus lipidů účinky léků   MeSH
• myši MeSH
• osteogeneze účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: Statins are widely used drugs for cholesterol lowering, which were recently found to counteract the effects of aberrant fibroblast growth factor receptor (FGFR3) signaling in cell and animal models of FGFR3-related chondrodysplasia. This opened an intriguing therapeutic possibility for human dwarfing conditions caused by gain-of-function mutations in FGFR3, although the mechanism of statin action on FGFR3 remains unclear. Here, we determine the effect of statins on FGFR signaling in chondrocytes. DESIGN: Cultured chondrocyte cell lines, mouse embryonic tibia cultures and limb bud micromasses were treated with FGF2 to activate FGFR signaling. The effects of atorvastatin, fluvastatin, lovastatin and pravastatin on FGFR3 protein stability and on FGFR-mediated chondrocyte growth-arrest, loss of extracellular matrix (ECM), induction of premature senescence and hypertrophic differentiation were evaluated. RESULTS: Statins did not alter the level of FGFR3 protein expression nor produce any effect on FGFR-mediated inhibition of chondrocyte proliferation and hypertrophic differentiation in cultured chondrocyte cell lines, mouse tibia cultures or limb bud micromasses. CONCLUSION: We conclude that statins do not inhibit the FGFR signaling in chondrocytes. Therefore the statin-mediated rescue of FGFR3-related chondrodysplasia, described before, is likely not intrinsic to the growth plate cartilage.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčné linie MeSH
• chondrocyty účinky léků   metabolismus   MeSH
• chondrogeneze účinky léků   MeSH
• končetinové pupeny účinky léků   metabolismus   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• myši MeSH
• receptor fibroblastových růstových faktorů, typ 3 antagonisté a inhibitory   metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• statiny farmakologie   MeSH
• techniky tkáňových kultur MeSH
• tibie účinky léků   embryologie   růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Caspases, well-known players in apoptosis or inflammation, appear to have roles also in other processes such as cell differentiation. Caspase-3, in particular, was recently demonstrated to have non-apoptotic functions in osteogenesis. However, the molecular pathways involved are not yet known. Therefore, we used osteogenic PCR arrays to provide a comprehensive screening of possible interactions of caspases in general and specifically of caspase-3 in osteogenic networks. Embryonic micromass cultures derived from mouse forelimbs were established and pharmacological fluoromethylketone (FMK) inhibitors applied. Alterations were observed in expression of several genes after caspase inhibition (Bmp1, Bmp5, Bmp6, Col10a1, Col2a1, Comp, Egf, Fgfr2, Gli1, Igf1, Nog, Phex, Sox9, Spp1). The list suggests molecular interactions of caspases and osteogenic molecules and creates a background for further temporospatial and functional studies.

• MeSH
• apoptóza účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• chondrogeneze účinky léků   MeSH
• inhibitory kaspas aplikace a dávkování   MeSH
• kaspasa 3 genetika   metabolismus   MeSH
• mezenchymální kmenové buňky účinky léků   MeSH
• myši MeSH
• osteogeneze účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Foetal calf serum (FCS) is a standard supplement used in media for in vitro stem cell cultivation. This xenogeneic supplement remains widely used for its favourable growth-promoting properties and ease of accessibility; however, it is inherently not fit for human medicine due to its capacity to temper with the cultured cell quality. For this reason, the international community encourages research and development of allogeneic sera, which would expunge this issue. This study aims to investigate the differences in proliferative capacity, phenotype, and differentiation capacity of ecto-mesenchymal stem cells from human exfoliated deciduous teeth (SHED) cultured in vitro in media supplemented with allogeneic and xenogeneic sera. To address these aims, we cultured three lineages of stem cells in media supplemented with FCS in a concentration of 2% + growth factors; human blood plasma and platelet-rich plasma in concentrations of 2% + growth factors, and 10%. Here, the xenogeneic cultivation was considered as a basis for comparison because this serum is commonly used in studies concerning ecto-mesenchymal stem cells. The study shows that multipotent ecto-mesenchymal SHED can be feasibly cultivated in media where the xenogeneic FCS is substituted by allogeneic platelet-rich plasma, considering the cultured cell proliferative and differentiation capacities. We have also proved that different sera impact the cultured cells' phenotype differently, which has major implications for previous and future stem cell research and regenerative therapy.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• chondrogeneze účinky léků   MeSH
• dítě MeSH
• fenotyp MeSH
• kultivační média farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   MeSH
• neurogeneze účinky léků   MeSH
• osteogeneze účinky léků   MeSH
• předškolní dítě MeSH
• proliferace buněk účinky léků   MeSH
• tvar buňky účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• vývoj svalů účinky léků   MeSH
• zuby mléčné cytologie   MeSH
• zvířata MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• artróza kolenních kloubů klasifikace   terapie   MeSH
• buněčná a tkáňová terapie * metody   trendy   MeSH
• chitosan terapeutické užití   MeSH
• chondrocyty transplantace   MeSH
• chondrogeneze účinky léků   MeSH
• kloubní chrupavka patologie   MeSH
• kongresy jako téma MeSH
• lidé MeSH
• osteoartróza * terapie   MeSH
• Check Tag
• lidé MeSH

Hyaluronan-based tissue substitutes are promising materials in cartilage reconstruction surgery. Herein, the chondrogenesis of human mesenchymal stem cells (MSC) in a hydrogel based on a tyramine derivative of hyaluronan crosslinked by hydrogen peroxidase (HA-TA) was evaluated. Human MSC seeded in the scaffold were incubated in standard chondrogenic medium and medium enriched with bone morphogenetic protein-6 (BMP6). Cell viability, the gene expression of selected markers (collagen type II, aggrecan, SOX9, collagen type X, and osteopontin), and the histological characteristics were examined during three weeks of in vitro cultivation. The tissue reaction of both unseeded and MSC seeded HA-TA scaffolds were tested in vivo after subcutaneous application in rats for 12 weeks. The data showed that cells resisted the process of crosslinking and remained viable for the whole time while exhibiting changes in cell organization. Human MSC cultivated in HA-TA hydrogel expressed genes of both chondrogenic and osteogenic differentiation and the addition of BMP6 revealed a tendency to potentiate both processes. Histological analysis of HA-TA in vivo implants did not reveal a chronic inflammatory reaction. In both cases, in vivo HA-TA implants were continuously degraded and MSC-seeded hydrogels tended to form clusters similar to in vitro samples. In conclusion, MSC chondrogenic differentiation may proceed in a HA-TA scaffold that is biocompatible. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3523-3530, 2014.

• MeSH
• buněčná diferenciace * účinky léků   genetika   MeSH
• chondrogeneze * účinky léků   genetika   MeSH
• experimentální implantáty MeSH
• imobilizované buňky cytologie   účinky léků   metabolismus   MeSH
• krysa rodu rattus MeSH
• kyselina hyaluronová farmakologie   MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• PEG-DMA hydrogel farmakologie   MeSH
• peroxidasa metabolismus   MeSH
• reagencia zkříženě vázaná farmakologie   MeSH
• regulace genové exprese účinky léků   MeSH
• subkutánní tkáň účinky léků   MeSH
• tyramin farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• zobrazování trojrozměrné MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The focus of this study was to compare the role of nerve growth factor (NGF) and vascular endothelial growth factor (VEGF) in the regeneration of experimental skin and cartilage trauma. The role of VEGF in this process is known since decade; the NGF participation on this process has been first discussed within the spinal cord injury repair. We hypothesized that both VEGF and NGF induce angiogenesis and take part on the repair process. The angiogenesis response and the cartilage regeneration after phVEGF(165) plasmid and rat pcNGF plasmid administration were investigated using BALB/c mice. PhVEGF(165) and pcNFG were injected into the right mice ear and plain vector injection into the left ear the day before trauma. The next day, all mice were ear-punched, resulting in 2-mm diameter puncture through the center of both pinnae. In BALB/c mouse strain, a significantly faster cartilage repair was observed after phVEGF(165) and pcNGF injection into punched ear area in comparison to the control group. It has been shown that the healing process is after VEGF and NGF injection driven differentially. In case of VEGF is the cartilage wound repaired by induction of new chondrocytes differentiation. In the case of NGF, the regeneration is supported by immature leukocytes attracted into the punched area. The leukocytes induct angiogenesis so far indirectly by inflammation. The NGF-induced inflammation environment may be a part of mosaic creating the complete picture of regeneration.

• MeSH
• chondrogeneze * účinky léků   fyziologie   MeSH
• fyziologická neovaskularizace * účinky léků   fyziologie   MeSH
• genetické vektory MeSH
• hojení ran * účinky léků   fyziologie   MeSH
• krysa rodu rattus MeSH
• kůže zranění   MeSH
• látky indukující angiogenezi aplikace a dávkování   MeSH
• modely u zvířat MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• neurotrofní faktory * aplikace a dávkování   genetika   MeSH
• plazmidy MeSH
• ušní chrupavka zranění   fyziologie   MeSH
• vaskulární endoteliální růstový faktor A * aplikace a dávkování   genetika   MeSH
• zánět metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We isolated and expanded stem cells from dental pulp from extracted third molars using an innovative culture method consisting of low serum-containing medium supplemented with epidermal growth factor and platelet-derived growth factor BB. We evaluated the differentiation potential of these cells when they were growing either adherently or as micromass/spheroid cultures in various media. Undifferentiated and differentiated cells were analyzed by flow cytometry, immunocytochemistry and immunoblotting. The flow cytometry results showed that the dental pulp stem cells (DPSCs) were positive for mesenchymal stromal cell markers, but negative for hematopoietic markers. Immunocytochemical and/or immunoblotting analyses revealed the expression of numerous stem cell markers, including nanog, Sox2, nestin, Musashi-1 and nucleostemin, whereas they were negative for markers associated with differentiated neural, vascular and hepatic cells. Surprisingly, the cells were only slightly positive for α-smooth muscle actin, and a heterogeneous expression of CD146 was observed. When cultured in osteogenic media, they expressed osteonectin, osteopontin and procollagen I, and in micromass cultures, they produced collagen I. DPSCs cultured in TGF-β1/3-supplemented media produced extracellular matrix typical of cartilaginous tissue. The addition of vascular endothelial growth factor to serum-free media resulted in the expression of endothelial markers. Interestingly, when cultured in neurogenic media, DPSCs exhibited de novo or upregulated markers of undifferentiated and differentiated neural cells. Collectively, our data show that DPSCs are self-renewing and able to express markers of bone, cartilage, vascular and neural tissues, suggesting their multipotential capacity. Their easy accessibility makes these cells a suitable source of somatic stem cells for tissue engineering.

• MeSH
• aktiny metabolismus   MeSH
• antigen CD146 metabolismus   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčný rodokmen účinky léků   MeSH
• chondrogeneze účinky léků   MeSH
• endoteliální buňky cytologie   účinky léků   metabolismus   MeSH
• imunohistochemie MeSH
• kmenové buňky účinky léků   patologie   MeSH
• kultivační média bez séra farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• molár třetí patologie   MeSH
• myocyty hladké svaloviny cytologie   účinky léků   metabolismus   MeSH
• myofibroblasty cytologie   účinky léků   metabolismus   MeSH
• neurony cytologie   účinky léků   metabolismus   MeSH
• osteogeneze účinky léků   MeSH
• průtoková cytometrie MeSH
• zaklíněný zub patologie   MeSH
• zubní dřeň patologie   MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH