• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• ortopedie

Titanium-based alloys have established a crucial role in implantology. As material deteriorates overtime, nanoparticles of TiO2 and Ni are released. This study is focused on the impact of TiO2 and Ni nanoparticles with size of 100 nm on cytoskeletal and adhesive changes in human physiological and osteoarthritic osteoblasts. The impact of nanoparticles with concentration of 1.5 ng/mL on actin and tubulin expression and gene expression of FAK and ICAM-1 was studied. The cell size and actin expression of physiological osteoblasts decreased in presence of Ni nanoparticles, while TiO2 nanoparticles caused increase in cell size and actin expression. Both cell lines expressed more FAK as a response to TiO2 nanoparticles. ICAM-1 gene was overexpressed in both cell lines as a reaction to both types of nanoparticles. The presented study shows a crucial role of Ni and TiO2 nanoparticles in human osteoblast cytoskeletal and adhesive changes, especially connected with the osteoarthritic cells. Graphical abstract.

• MeSH
• buněčná adheze MeSH
• cytoskelet MeSH
• lidé MeSH
• nanočástice * MeSH
• osteoblasty MeSH
• titan * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Nickel(Ni)-containing materials have been widely used in a wide range of medical applications, including orthopaedics. Despite their excellent properties, there is still a problem with the release of nickel ions into the patient's body, which can cause changes in the behaviour of surrounding cells and tissues. This study aims to evaluate the effects of Ni on bone cells with an emphasis on the determination of Ni localization in cellular compartments in time. For these purposes, one of the most suitable models for studying the effects induced by metal implants was used-the patient's osteoarthritic cells. Thanks to this it was possible to simulate the pathophysiological conditions in the patient's body, as well as to evaluate the response of the cells which come into direct contact with the material after the implantation of the joint replacement. The largest differences in cell viability, proliferation and cell cycle changes occurred between Ni 0.5 mM and 1 mM concentrations. Time-dependent localization of Ni in cells showed that there is a continuous transport of Ni ions between the nucleus and the cytoplasm, as well as between the cell and the environment. Moreover, osteoarthritic osteoblasts showed faster changes in concentration and ability to accumulate more Ni, especially in the nucleus, than physiological osteoblasts. The differences in Ni accumulation process explains the higher sensitivity of patient osteoblasts to Ni and may be crucial in further studies of implant-derived cytotoxic effects.

• MeSH
• buněčný cyklus účinky léků   MeSH
• ionty metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• nikl * metabolismus   MeSH
• osteoartróza * metabolismus   patologie   MeSH
• osteoblasty * metabolismus   účinky léků   MeSH
• proliferace buněk * účinky léků   MeSH
• viabilita buněk * účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

PURPOSE: The purpose of this study was to evaluate the effect of progressively increasing concentrations of activated and nonactivated platelet-rich plasma (PRP) on proliferation of human osteoblasts in vitro. MATERIALS AND METHODS: Human osteoblasts (hFOB 1.19) obtained from the American Type Culture Collection (ATCC, Manassas, VA) were used in the experiment. PRP was obtained from a 28-year-old healthy male volunteer by means of a Haemonetics gradient density cell separator (Haemonetics, Munich, Germany). Human thrombin was used to activate PRP. Three independent experiments were conducted. Samples containing 10% (0.38x increase in platelet count), 25% (0.95x increase in platelet count), 50% (1.95x increase in platelet count), and 75% (2.86x increase in platelet count) of activated PRP and nonactivated PRP were prepared including controls. After culture periods of 24, 48, and 72 hours osteoblast proliferation was evaluated by counting the number of cells using a Multisizer 3 Coulter Counter (Beckman Coulter, Inc, Fullerton, CA). RESULTS: After 24, 48, and 72 hours of incubation, the number of cells in the control group (without PRP) was higher than that of cells in samples containing activated or nonactivated PRP. Osteoblasts with 10% activated PRP (0.38x increase in platelet count) had the highest viability of all samples containing PRP. CONCLUSIONS: Activated PRP resulted in higher proliferation of osteoblasts compared with nonactivated PRP at concentrations of 10% (0.38x increase in platelet count) and 25% (0.95x increase in platelet count) in culture. This study failed to show significant increases in proliferation of human osteoblasts treated with activated or nonactivated PRP compared with controls in vitro.

• MeSH
• časové faktory MeSH
• dospělí MeSH
• financování organizované MeSH
• kultivované buňky MeSH
• lidé MeSH
• osteoblasty účinky léků   MeSH
• plazma bohatá na destičky MeSH
• proliferace buněk účinky léků   MeSH
• trombin farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• klinické zkoušky kontrolované MeSH

PURPOSE OF THE STUDY A failure of total hip or knee artroplasty is associated with an increased production of joint fluid. This contains wear particles and host cells and proteins, and is assumed to be involved in the pathogenesis of aseptic loosening and periprosthetic osteolysis. This study investigated the effect of synovial fluid from patients with aseptically failed joint prostheses on osteoblast cultures. MATERIAL AND METHODS Synovial fluid samples were obtained from patients with failed total joint prostheses (TJP; n=36) and from control patient groups (n=16) involving cases without TJP and osteoarthritis, without TJP but with osteoarthritis, and with stable TJP. The samples were treated in the standard manner and then cultured with the SaOS-2 cell line which shows the characteristics and behaviour of osteoblasts. Each fluid sample was also examined for the content of proteins, cells and selected cytokines (IL-1?, TNF-&alfa;, IL-6, RANKL and OPG detected by ELISA). We tested the hypothesis assuming that the fluids from failed joints would show higher cytotoxicity to osteoblast culture and we also expected higher levels of IL-1?, TNF-&alfa;, IL-6, and RANKL in patients with TJP failure and/ or with more severe bone loss. The statistical methods used included the Kruskal-Wallis ANOVA and Mann-Whitney U test. RESULTS The fluids from failed TJPs showed the highest RANKL and the lowest OPG levels resulting in the highest RANKL/OPG ratio. However, there was no evidence suggesting that the joint fluids from failed TJPs would be more toxic to osteoblast culture than the fluids from control groups. In addition, no correlation was found between the fluid levels of molecules promoting inflammation and osteoclastic activity and the extent of bone loss in the hip (in terms of Saleh's classification) or the knee (AORI classification). In fact, the fluids from failed TJPs had higher protein levels in comparison with the controls, but the difference was not significant. DISCUSSION The finding of high RANKL levels and low OPG concentrations is in agreement with the theory of aseptic loosening and periprosthetic osteolysis. The other cytokines, particularly TNF-&alfa; and IL-1?, were found in low levels. This can be explained by the stage of particle disease at which the samples were taken for ELISA analysis. It is probable that the level of signal molecules reflects osteolytic process activity and is therefore not constant. The reason for no correlation found between cytokine levels and the extent of bone loss may also lie in the use of therapeutic classifications of bone defects that is apparently less sensitive to the biological activity of aseptic loosening and/or periprosthetic osteolysis. CONCLUSIONS Synovial fluids from failed total hip or knee joint prostheses are not toxic to osteoblast cultures. Cytotoxicity indicators and levels of pro-inflammatory and pro-osteoclastic cytokines (IL-1?, TNF-&alfa;, IL-6, RANKL and OPG) do not correlate well with the extent of periprosthetic bone loss.

• MeSH
• artróza kolenních kloubů chirurgie   metabolismus   MeSH
• artróza kyčelních kloubů chirurgie   metabolismus   MeSH
• interleukin-6 analýza   MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• ligand RANK analýza   MeSH
• náhrada kyčelního kloubu MeSH
• osteoblasty cytologie   MeSH
• osteoprotegerin analýza   MeSH
• selhání protézy MeSH
• senioři MeSH
• synoviální tekutina fyziologie   chemie   MeSH
• TNF-alfa analýza   MeSH
• totální endoprotéza kolene MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH

The excellent mechanical, tribological and biochemical properties of diamond coatings are promising for improving orthopedic or stomatology implants. A crucial prerequisite for such applications is an understanding and control of the biological response of the diamond coatings. This study concentrates on the correlation of diamond surface properties with osteoblast behavior. Nanocrystalline diamond (NCD) films (grain size up to 200 nm, surface roughness 20 nm) were deposited on silicon substrates of varying roughnesses (1, 270 and 500 nm) and treated by oxygen plasma to generate a hydrophilic surface. Atomic force microscopy was used for topographical characterization of the films. As a reference surface, tissue culture polystyrene (PS) was used. Scanning electron microscopy and immunofluorescence staining was used to visualize cell morphological features as a function of culture time. Metabolic activity, alkaline phosphatase activity, and calcium and phosphate deposition was also monitored. The results show an enhanced osteoblast adhesion as well as increased differentiation (raised alkaline phosphatase activity and mineral deposition) on NCD surfaces (most significantly on RMS 20 nm) compared to PS. This is attributed mainly to the specific surface topography as well as to the biocompatible properties of diamond. Hence the controlled (topographically structured) diamond coating of various substrates is promising for preparation of better implants, which offer faster colonization by specific cells as well as longer-term stability.

• MeSH
• biokompatibilní materiály chemie   MeSH
• buněčná diferenciace MeSH
• buněčné kultury metody   MeSH
• buněčné linie MeSH
• diamant chemie   MeSH
• lidé MeSH
• molekulární konformace MeSH
• nanostruktury chemie   ultrastruktura   MeSH
• osteoblasty cytologie   fyziologie   MeSH
• osteogeneze fyziologie   MeSH
• povrchové vlastnosti MeSH
• proliferace buněk MeSH
• tkáňové inženýrství metody   MeSH
• velikost buňky MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

BACKGROUND: This study was aimed to investigate whether osteoblasts from diabetic patients have a promoting effect on osteogenesis of human umbilical cord mesenchymal stem cells (HUMSCs). METHODS: HUMSCs were co-cultured with osteoblasts of diabetic and non-diabetic patients. Morphological appearance and cytochemical characteristics of the non-diabetic osteoblasts and diabetic osteoblasts were observed by hematoxylin-eosin staining, type I collagen protein expression, alkaline phosphatase (ALP) staining and Alizarin Red S staining. Cell viability, type I collagen protein expression, ALP activity and osteocalcin mRNA expression in HUMSCs were investigated. RESULTS: Compared with negative control group, the cell proliferation, type I collagen protein expression, ALP activity and osteocalcin mRNA were increased in HUMSCs co-cultured with diabetic and non-diabetic osteoblasts (P<0.05). There was no statistically significant difference in the HUMSCs cell proliferation, type I collagen protein expression, ALP activity and osteocalcin mRNA between the non-diabetic and diabetic group (P >0.05). CONCLUSIONS: Similar to osteoblasts from non-diabetic patients, osteoblasts from diabetic patients also have the ability to promote HUMSCs proliferation, and leading to HUMSCs exhibit some characteristic of osteoblasts.

• MeSH
• buněčná diferenciace MeSH
• diabetes mellitus metabolismus   patologie   MeSH
• kokultivační techniky MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   MeSH
• osteoblasty metabolismus   patologie   MeSH
• osteogeneze * MeSH
• proliferace buněk MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Regeneration of bone tissue defects that result from metabolic disorders, including periodontal diseases, can be supported by biomaterials based on hydroxyapatite. Despite of good biocompatibility of biomaterials they can cause oxidative stress and inflammatory processes as a result of mechanical interaction with surrounding tissues. Because osteoblasts are responsible for bone regeneration process in which gingival fibroblasts may also participate, the aim of the work was to investigate the influence of hydroxyapatite-based biomaterials (allogeneic and xenogeneic) and biomaterials combined with enamel matrix derivative (Emdogain) on osteoblast and fibroblast redox balance in the context of osteoblast proliferation and differentiation. The results showed that examined substitutes were not cytotoxic in vitro, but affected redox balance of osteoblasts and fibroblasts (ROS level increase and GSH level decrease) which led to oxidative stress (MDA and protein carbonyl groups level increase) resulting in an increase of the Nrf2 and NFκB expression. The consequence of these changes was partial inhibition of proliferation and osteoblast differentiation. Emdogain alone and combined with biomaterials decreased ROS generation and increased GSH level in both osteoblasts and fibroblasts leading to reduction of transcription factors expression especially proinflammatory NFκB, which promoted osteoblast differentiation and mineralization process.

• MeSH
• biokompatibilní materiály MeSH
• fibroblasty metabolismus   účinky léků   MeSH
• hydroxyapatity farmakologie   metabolismus   škodlivé účinky   MeSH
• kostní náhrady farmakologie   metabolismus   MeSH
• osteoblasty metabolismus   účinky léků   MeSH
• oxidace-redukce účinky léků   MeSH
• proteiny zubní skloviny farmakologie   metabolismus   MeSH
• řízená tkáňová regenerace * metody   MeSH
• techniky in vitro MeSH

The aim of this study was to investigate the acute effects of oral glucocorticoids in doses used in clinical practice on biochemical indices of the function of osteoclasts, osteoblasts, and osteocytes. In 17 adult patients suffering from various medical pathologies requiring systemic steroid therapy that were never before treated with glucocorticoids, glucocorticoid treatment was initiated (mean prednisolone equivalent dose of 23.1 ± 12.7 mg/day, range 10-50). Fasting morning serum concentrations of osteocalcin (OC), amino-terminal propeptide of type I procollagen (PINP), type 1 collagen cross-linked C-telopeptide (βCTX), soluble receptor activator of nuclear factor kappaB ligand (sRANKL), osteoprotegerin (OPG), sclerostin, Dickkopf-1 (Dkk-1), and high-sensitivity C-reactive protein (hsCRP) were measured at baseline and on three consecutive days. Significant reductions in serum OC, PINP, OPG, sclerostin, and hsCRP were observed during 96 h of glucocorticoid administration, while serum βCTX showed a significant percentual increase. A significant positive correlation was found between serum concentrations of Dkk-1 and βCTX after 96 h of treatment with glucocorticoids. A significant drop in serum sclerostin, OPG, and OC observed in this study may reflect the rapid glucocorticoid-induced apoptosis of osteocytes.

• MeSH
• biologické markery krev   MeSH
• C-reaktivní protein metabolismus   MeSH
• dospělí MeSH
• genetické markery MeSH
• glukokortikoidy farmakologie   terapeutické užití   MeSH
• kolagen typu I krev   MeSH
• kostní morfogenetické proteiny krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• ligand RANK krev   MeSH
• mezibuněčné signální peptidy a proteiny krev   MeSH
• osteoblasty účinky léků   metabolismus   MeSH
• osteocyty účinky léků   metabolismus   MeSH
• osteokalcin krev   MeSH
• osteoklasty účinky léků   metabolismus   MeSH
• osteoprotegerin krev   MeSH
• peptidové fragmenty krev   MeSH
• peptidy krev   MeSH
• prokolagen krev   MeSH
• prospektivní studie MeSH
• revmatické nemoci krev   farmakoterapie   patologie   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Pulsed laser deposition was proved as a suitable method for hydroxyapatite (HA) coating of coaxial poly-ɛ-caprolactone/polyvinylalcohol (PCL/PVA) nanofibers. The fibrous morphology of PCL/PVA nanofibers was preserved, if the nanofiber scaffold was coated with thin layers of HA (200 nm and 400 nm). Increasing thickness of HA, however, resulted in a gradual loss of fibrous character. In addition, biomechanical properties were improved after HA deposition on PCL/PVA nanofibers as the value of Young's moduli of elasticity significantly increased. Clearly, thin-layer hydroxyapatite deposition on a nanofiber surface stimulated mesenchymal stem cell viability and their differentiation into osteoblasts. The optimal depth of HA was 800 nm.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• hydroxyapatit chemie   farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   MeSH
• nanovlákna chemie   MeSH
• osteoblasty cytologie   MeSH
• polyestery chemie   MeSH
• polyvinylalkohol chemie   MeSH
• prasata MeSH
• proliferace buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH