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.

• Klíčová slova
• Adhesion, Bone, Cytoskeleton, Ni nanoparticles, Osteoblasts, TiO2 nanoparticles,
• MeSH
• buněčná adheze MeSH
• cytoskelet MeSH
• lidé MeSH
• nanočástice * MeSH
• osteoblasty MeSH
• titan * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• titan * MeSH
• titanium dioxide MeSH Prohlížeč

Osteoblasts and osteoclasts are the unique cells occurring in bone marrow smears in situations with high bone metabolic turnover (children, trauma, rachitis, Paget disease or tumors). The collection of 2706 sternal or iliac crest aspirates from patients with hematologic malignancies and solid tumors are presented. We demonstrated significantly higher positivity for osteoblasts-osteoclasts presentation in bone marrow smears for hematological malignancies (p < 0.05), solid tumors (p < 0.01), and especially breast cancer (p < 0.001). We found a significant association between osteoblast-osteoclast positivity and dissemination of breast cancer (p < 0.05). None of the breast cancer patients without signs of dissemination (X-ray, sonography or scintigraphy) had positivity for osteoblasts or osteoclasts. We suppose that the osteoblast-osteoclast positivity in bone marrow smears can serve as a cheap marker for breast cancer dissemination.

• MeSH
• kostní dřeň patologie   MeSH
• lidé MeSH
• nádory patologie   MeSH
• osteoblasty patologie   MeSH
• osteoklasty patologie   MeSH
• retrospektivní studie MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články 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.

• Klíčová slova
• bone healing, diabetic, human umbilical cord, mesenchymal stem cells, 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

Nanostructured titanium has become a useful material for biomedical applications such as dental implants. Certain surface properties (grain size, roughness, wettability) are highly expected to promote cell adhesion and osseointegration. The aim of this study was to compare the biocompatibilities of several titanium materials using human osteoblast cell line hFOB 1.19. Eight different types of specimens were examined: machined commercially pure grade 2 (cpTi2) and 4 (cpTi4) titanium, nanostructured titanium of the same grades (nTi2, nTi4), and corresponding specimens with laser-treated surfaces (cpTi2L, cpTi4L, nTi2L, nTi4L). Their surface topography was evaluated by means of scanning electron microscopy. Surface roughness was measured using a mechanical contact profilometer. Specimens with laser-treated surfaces had significantly higher surface roughness. Wettability was measured by the drop contact angle method. Nanostructured samples had significantly higher wettability. Cell proliferation after 48 hours from plating was assessed by viability and proliferation assay. The highest proliferation of osteoblasts was found in nTi4 specimens. The analysis of cell proliferation revealed a difference between machined and laser-treated specimens. The mean proliferation was lower on the laser-treated titanium materials. Although plain laser treatment increases surface roughness and wettability, it does not seem to lead to improved biocompatibility.

• Klíčová slova
• biocompatibility, laser treatment, nanostructured titanium, osteoblasts proliferation,
• 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.

• Klíčová slova
• Implant debris, Laser ablation, Metal distribution, Metal uptake, Nickel, Osteoblasts,
• 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
• Názvy látek
• ionty MeSH
• nikl * MeSH

MiR-21 is being gradually more and more recognized as a molecule regulating bone tissue homeostasis. However, its function is not fully understood due to the dual role of miR-21 on bone-forming and bone-resorbing cells. In this study, we investigated the impact of miR-21 inhibition on pre-osteoblastic cells differentiation and paracrine signaling towards pre-osteoclasts using indirect co-culture model of mouse pre-osteoblast (MC3T3) and pre-osteoclast (4B12) cell lines. The inhibition of miR-21 in MC3T3 cells (MC3T3inh21) modulated expression of genes encoding osteogenic markers including collagen type I (Coll-1), osteocalcin (Ocl), osteopontin (Opn), and runt-related transcription factor 2 (Runx-2). Inhibition of miR-21 in osteogenic cultures of MC3T3 also inflected the synthesis of OPN protein which is essential for proper mineralization of extracellular matrix (ECM) and anchoring osteoclasts to the bones. Furthermore, it was shown that in osteoblasts miR-21 regulates expression of factors that are vital for survival of pre-osteoclast, such as receptor activator of nuclear factor κB ligand (RANKL). The pre-osteoclast cultured with MC3T3inh21 cells was characterized by lowered expression of several markers associated with osteoclasts' differentiation, foremost tartrate-resistant acid phosphatase (Trap) but also receptor activator of nuclear factor-κB ligand (Rank), cathepsin K (Ctsk), carbonic anhydrase II (CaII), and matrix metalloproteinase (Mmp-9). Collectively, our data indicate that the inhibition of miR-21 in MC3T3 cells impairs the differentiation and ECM mineralization as well as influences paracrine signaling leading to decreased viability of pre-osteoclasts.

• Klíčová slova
• differentiation, miR-21-5p, osteoblasts, osteoclasts, osteogenesis, precursor cells,
• MeSH
• buněčná diferenciace genetika   MeSH
• buněčné linie MeSH
• extracelulární matrix metabolismus   MeSH
• kokultivační techniky MeSH
• kyselá fosfatasa rezistentní k tartarátu metabolismus   MeSH
• messenger RNA genetika   MeSH
• mikro RNA genetika   metabolismus   MeSH
• myši MeSH
• osteoblasty metabolismus   MeSH
• osteogeneze genetika   MeSH
• osteoklasty metabolismus   MeSH
• osteopontin genetika   metabolismus   MeSH
• parakrinní signalizace genetika   MeSH
• protein PEBP2alfaA genetika   metabolismus   MeSH
• resorpce kosti metabolismus   MeSH
• signální transdukce genetika   MeSH
• transfekce MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Acp5 protein, mouse MeSH Prohlížeč
• kyselá fosfatasa rezistentní k tartarátu MeSH
• messenger RNA MeSH
• mikro RNA MeSH
• MIRN21 microRNA, mouse MeSH Prohlížeč
• osteopontin MeSH
• protein PEBP2alfaA MeSH
• Runx2 protein, mouse MeSH Prohlížeč
• Spp1 protein, mouse MeSH Prohlížeč

Osteogenesis is an important process of bone metabolism, and abnormal osteogenesis leads to various skeletal system diseases. Osteoblasts, the main cells involved in bone formation, are central elements in the study of bone metabolic diseases. Single-cell RNA sequencing is an important tool for studying the transcriptome of cells and can help to elucidate various cellular and molecular functions at the single-cell level, providing new avenues for life science research. Here we explore the heterogeneity of osteoblasts and try to reveal the developmental trajectory of osteoblasts, thereby contributing to efforts to describe the mechanism of osteogenesis. In this study, single-cell sequencing data of murine bone marrow cells were used to identify osteoblasts. Finally, osteoblasts were divided into four groups, each differing in characteristic genes and signal pathways. We also identify clues of the changes of some genes in the process of osteoclast formation, providing directions for further study. Collectively, our findings suggest that bone marrow osteoblasts can be divided into several subgroups, which represent different stages of cells, and that the specific genes of each subgroup respond to the molecular mechanisms of cell development. This data will likely be of great help in resolving diseases of the skeletal system.

• MeSH
• buněčná diferenciace MeSH
• kostní dřeň * MeSH
• myši MeSH
• osteoblasty * metabolismus   MeSH
• osteogeneze genetika   MeSH
• osteoklasty metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE OF THE STUDY Nitinol (NiTi) is a biomaterial widely used in medicine based on super-elastic and shape memory properties. miR-124 has a key role in inflammatory process, osteoblasts differentiation, and mineralization. The aim of study was evaluating the differences in gene expression of miR-124 of human physiological osteoblasts (HOB) and human osteoarthritic osteoblasts (OSBA) as a response to NiTi alloy in different heat treatments. MATERIAL AND METHODS The cells were cultivated with NiTi discs with/without addition of bacterial lipopolysaccharide (LPS) for 72 hours. MicroRNAs were isolated, underwent reverse transcription and were analyzed by RT-PCR. RESULTS As a response to LPS, HOB overexpressed miR-124, while in OSBA expression change did not occur. Overexpression was also observed in both cell lines as a response to hydrogen and helium treated NiTi discs. HOB expressed significantly higher amount of miR-124 than OSBA as a response to hydrogen treatment of NiTi discs. In addition, hydrogen treatment caused significantly higher expression in HOB than LPS. The combination of NiTi disc and LPS treatment in HOB didn't cause any expression changes. Comparing to LPS-only treatment, the expression in HOB with combination of LPS and alloy was significantly lower. In OSBA, the expression was increased by the combination of LPS and hydrogen disc, in case of helium disc, the expression was decreased. CONCLUSIONS In conclusion, human physiological and osteoarthritic osteoblasts respond to NiTi alloy with both surface (hydrogen and helium atmosphere) treatment by overexpression of miR-124. The effect of LPS as inflammatory modulator suggests the presence of an "anti-inflammatory preconditioning" in osteoarthritic osteoblasts, as physiological osteoblasts overexpression was significantly higher. Key words: nitinol, osteoblast, miR-124, lipopolysaccharide.

• MeSH
• helium metabolismus   farmakologie   MeSH
• lidé MeSH
• lipopolysacharidy * farmakologie   metabolismus   MeSH
• mikro RNA * genetika   metabolismus   farmakologie   MeSH
• osteoartróza genetika   MeSH
• osteoblasty metabolismus   MeSH
• slitiny metabolismus   farmakologie   MeSH
• titan MeSH
• vodík metabolismus   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• helium MeSH
• lipopolysacharidy * MeSH
• mikro RNA * MeSH
• MIRN124 microRNA, human MeSH Prohlížeč
• nitinol MeSH Prohlížeč
• slitiny MeSH
• titan MeSH
• vodík MeSH

Titanium offers excellent biocompatibility and extraordinary mechanical properties. As a result, it is used as a material for dental implants. Implants infected by peri-implantitis can be cleaned for successful re-osseointegration. Optimal surface properties, such as roughness and wettability, have a significant impact on cell adhesion. The aim of this study was to evaluate the adhesion and proliferation of osteoblasts on the surface of repeatedly cleaned nanostructured titanium samples. Human osteoblast-like cells MG-63 were seeded on nanostructured titanium specimens manufactured from rods produced by the equal channel angular pressing. For surface characterization, roughness and wettability were measured. Cell adhesion after 2 h as well as cell proliferation after 48 h from plating was assessed. We have found that this repeated cleaning of titanium surface reduced cell adhesion as well as proliferation. These events depend on interplay of surface properties, such as wettability, roughness and topography. It is difficult to distinguish which factors are responsible for these events and further investigations will be required. However, even after the several rounds of repeated cleaning, there was a certain rate of adhesion and proliferation recorded. Therefore the attempts to save failing implants by using in situ cleaning are promising.

• Klíčová slova
• biocompatibility, nanostructured titanium, osteoblasts proliferation, peri-implantitis,
• Publikační typ
• časopisecké články MeSH

Microscopic chemical patterning of diamond surfaces by hydrogen and oxygen surface atoms is used for self-assembly of human osteoblastic cells into micro-arrays. The cell adhesion and assembly is further controlled by concentration of cells (2,500-10,000 cells/cm(2)) and fetal bovine serum (0-15%). The cells are characterized by fluorescence microscopy of actin fibers and nuclei. The serum protein adsorption is studied by atomic force microscopy (AFM). The cells are arranged selectively on O-terminated patterns into 30-200 μm wide arrays. Higher cell concentrations allow colonization of unfavorable H-terminated regions due to mutual cell communication. There is no cell selectivity without the proteins in the medium. Based on the AFM, the proteins are present on both H- and O-terminated surfaces. Pronounced differences in their thickness, surface roughness, morphology, and phase images indicate different conformation of the proteins and explain the cell selectivity.

• Klíčová slova
• atomic force microscopy, biosensors, biotechnology, cell adhesion, diamond, osteoblasts, proteins, tissue engineering,
• Publikační typ
• časopisecké články MeSH