Nejvíce citovaný článek - PubMed ID 15736406
In vitro testing is the first important step in the development of new biomaterials. The human fetal osteoblast cell line hFOB 1.19 is a very promising cell model; however, there are vast discrepancies in cultivation protocols, especially in the cultivation temperature and the presence of the selection reagent, geneticin (G418). We intended to use hFOB 1.19 for the testing of Zn-based degradable metallic materials. However, the sensitivity of hFOB 1.19 to zinc ions has not yet been studied. Therefore, we compared the toxicity of zinc towards hFOB 1.19 under different conditions and compared it with that of the L929 mouse fibroblast cell line. We also tested the cytotoxicity of three types of Zn-based biomaterials in two types of media. The presence of G418 used as a selection reagent decreased the sensitivity of hFOB 1.19 to Zn2+. hFOB 1.19 cell line was more sensitive to Zn2+ at elevated (restrictive) temperatures. hFOB 1.19 cell line was less sensitive to Zn2+ than L929 cell line (both as ZnCl2 and extracts of alloys). Therefore, the appropriate cultivation conditions of hFOB 1.19 during biomaterial testing should be chosen with caution.
- Klíčová slova
- hFOB 1.19 osteoblasts, in vitro cytotoxicity testing, zinc degradable materials,
- Publikační typ
- časopisecké články MeSH
Bone regeneration after injury or after surgical bone removal due to disease is a serious medical challenge. A variety of materials are being tested to replace a missing bone or tooth. Regeneration requires cells capable of proliferation and differentiation in bone tissue. Although there are many possible human cell types available for use as a model for each phase of this process, no cell type is ideal for each phase. Osteosarcoma cells are preferred for initial adhesion assays due to their easy cultivation and fast proliferation, but they are not suitable for subsequent differentiation testing due to their cancer origin and genetic differences from normal bone tissue. Mesenchymal stem cells are more suitable for biocompatibility testing, because they mimic natural conditions in healthy bone, but they proliferate more slowly, soon undergo senescence, and some subpopulations may exhibit weak osteodifferentiation. Primary human osteoblasts provide relevant results in evaluating the effect of biomaterials on cellular activity; however, their resources are limited for the same reasons, like for mesenchymal stem cells. This review article provides an overview of cell models for biocompatibility testing of materials used in bone tissue research.
- Klíčová slova
- Bone engineering, Human osteoblasts, Mesenchymal stem cells, Osteodifferentiation, Osteosarcoma cell lines,
- MeSH
- biokompatibilní materiály farmakologie MeSH
- buněčná diferenciace MeSH
- kosti a kostní tkáň * MeSH
- kultivované buňky MeSH
- lidé MeSH
- osteoblasty MeSH
- osteogeneze MeSH
- proliferace buněk MeSH
- tkáňové inženýrství * metody MeSH
- tkáňové podpůrné struktury MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- biokompatibilní materiály MeSH
The present study investigates the effect of an oxidized nanocrystalline diamond (O-NCD) coating functionalized with bone morphogenetic protein 7 (BMP-7) on human osteoblast maturation and extracellular matrix mineralization in vitro and on new bone formation in vivo. The chemical structure and the morphology of the NCD coating and the adhesion, thickness and morphology of the superimposed BMP-7 layer have also been assessed. The material analysis proved synthesis of a conformal diamond coating with a fine nanostructured morphology on the Ti6Al4V samples. The homogeneous nanostructured layer of BMP-7 on the NCD coating created by a physisorption method was confirmed by AFM. The osteogenic maturation of hFOB 1.19 cells in vitro was only slightly enhanced by the O-NCD coating alone without any increase in the mineralization of the matrix. Functionalization of the coating with BMP-7 resulted in more pronounced cell osteogenic maturation and increased extracellular matrix mineralization. Similar results were obtained in vivo from micro-CT and histological analyses of rabbit distal femurs with screws implanted for 4 or 12 weeks. While the O-NCD-coated implants alone promoted greater thickness of newly-formed bone in direct contact with the implant surface than the bare material, a further increase was induced by BMP-7. It can be therefore concluded that O-NCD coating functionalized with BMP-7 is a promising surface modification of metallic bone implants in order to improve their osseointegration.
- MeSH
- biokompatibilní potahované materiály chemie farmakologie MeSH
- diamant chemie MeSH
- extracelulární matrix MeSH
- kostní morfogenetický protein 7 * farmakologie MeSH
- králíci MeSH
- osteointegrace * MeSH
- slitiny MeSH
- titan MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biokompatibilní potahované materiály MeSH
- diamant MeSH
- kostní morfogenetický protein 7 * MeSH
- slitiny MeSH
- titan MeSH
- titanium alloy (TiAl6V4) MeSH Prohlížeč
Poly(ε-caprolactone) (PCL) is a biocompatible resorbable material, but its use is limited due to the fact that it is characterized by the lack of cell adhesion to its surface. Various chemical and physical methods are described in the literature, as well as modifications with various nanoparticles aimed at giving it such surface properties that would positively affect cell adhesion. Nanomaterials, in the form of membranes, were obtained by the introduction of multi-walled carbon nanotubes (MWCNTs and functionalized nanotubes, MWCNTs-f) as well as electro-spun carbon nanofibers (ESCNFs, and functionalized nanofibers, ESCNFs-f) into a PCL matrix. Their properties were compared with that of reference, unmodified PCL membrane. Human osteoblast-like cell line, U-2 OS (expressing green fluorescent protein, GFP) was seeded on the evaluated nanomaterial membranes at relatively low confluency and cultured in the standard cell culture conditions. The attachment and the growth of the cell populations on the polymer and nanocomposite samples were monitored throughout the first week of culture with fluorescence microscopy. Simultaneously, Raman microspectroscopy was also used to track the dependence of U-2 OS cell development on the type of nanomaterial, and it has proven to be the best method for the early detection of nanomaterial/cell interactions. The differentiation of interactions depending on the type of nanoadditive is indicated by the ν(COC) vibration range, which indicates the interaction with PCL membranes with carbon nanotubes, while it is irrelevant for PCL with carbon nanofibers, for which no changes are observed. The vibration range ω(CH2) indicates the interaction for PCL with carbon nanofibers with seeded cells. The crystallinity of the area ν(C=O) increases for PCL/MWCNTs and for PCL/MWCNTs-f, while it decreases for PCL/ESCNFs and for PCL/ESCNFs-f with seeded cells. The crystallinity of the membranes, which is determined by Raman microspectroscopy, allows for the assessment of polymer structure changes and their degradability caused by the secretion of cell products into the ECM and the differentiation of interactions depending on the carbon nanostructure. The obtained nanocomposite membranes are promising bioactive materials.
A silicalite-1 film (SF) deposited on Ti-6Al-4V alloy was investigated in this study as a promising coating for metallic implants. Two forms of SFs were prepared: as-synthesized SFs (SF-RT), and SFs heated up to 500 °C (SF-500) to remove the excess of template species from the SF surface. The SFs were characterized in detail by X-ray photoelectron spectroscopy (XPS), by Fourier transform infrared spectroscopy (FTIR), by scanning electron microscopy (SEM) and water contact angle measurements (WCA). Two types of bone-derived cells (hFOB 1.19 non-tumor fetal osteoblast cell line and U-2 OS osteosarcoma cell line) were used for a biocompatibility assessment. The initial adhesion of hFOB 1.19 cells, evaluated by cell numbers and cell spreading area, was better supported by SF-500 than by SF-RT. While no increase in cell membrane damage, in ROS generation and in TNF-alpha secretion of bone-derived cells grown on both SFs was found, gamma H2AX staining revealed an elevated DNA damage response of U-2 OS cells grown on heat-treated samples (SF-500). This study also discusses differences between osteosarcoma cell lines and non-tumor osteoblastic cells, stressing the importance of choosing the right cell type model.
- MeSH
- biokompatibilní materiály chemie MeSH
- buněčná membrána účinky léků MeSH
- buněčné linie MeSH
- cytotoxiny chemie farmakologie MeSH
- fotoelektronová spektroskopie metody MeSH
- lidé MeSH
- mikroskopie elektronová rastrovací metody MeSH
- nádorové buněčné linie MeSH
- osteoblasty účinky léků MeSH
- osteocyty účinky léků MeSH
- povrchové vlastnosti účinky léků MeSH
- proliferace buněk účinky léků MeSH
- slitiny MeSH
- testování materiálů metody MeSH
- titan chemie MeSH
- vysoká teplota MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biokompatibilní materiály MeSH
- cytotoxiny MeSH
- slitiny MeSH
- titan MeSH
- titanium alloy (TiAl6V4) MeSH Prohlížeč
Biofabrication and maturation of bone constructs is a long-term task that requires a high degree of specialization. This specialization falls onto the hierarchy complexity of the bone tissue that limits the transfer of this technology to the clinic. This work studied the effects of the short-term cryopreservation on biofabricated osteoblast-containing structures, with the final aim to make them steadily available in biobanks. The biological responses studied include the osteoblast post-thawing metabolic activity and the recovery of the osteoblastic function of 3D-bioprinted osteoblastic structures and beta tricalcium phosphate (β-TCP) scaffolds infiltrated with osteoblasts encapsulated in a hydrogel. The obtained structures were cryopreserved at -80 °C for 7 days using dimethyl sulfoxide (DMSO) as cryoprotectant additive. After thawing the structures were cultured up to 14 days. The results revealed fundamental biological aspects for the successful cryopreservation of osteoblast constructs. In summary, immature osteoblasts take longer to recover than mature osteoblasts. The pre-cryopreservation culture period had an important effect on the metabolic activity and function maintain, faster recovering normal values when cryopreserved after longer-term culture (7 days). The use of β-TCP scaffolds further improved the osteoblast survival after cryopreservation, resulting in similar levels of alkaline phosphatase activity in comparison with the non-preserved structures. These results contribute to the understanding of the biology of cryopreserved osteoblast constructs, approaching biofabrication to the clinical practice.
- Klíčová slova
- ALP activity, biofabrication, bioprinting, bone construct, cryopreservation, metabolic activity, osteoblast,
- Publikační typ
- časopisecké články MeSH
Bone tissue is the second tissue to be replaced. Annually, over four million surgical treatments are performed. Tissue engineering constitutes an alternative to autologous grafts. Its application requires three-dimensional scaffolds, which mimic human body environment. Bone tissue has a highly organized structure and contains mostly inorganic components. The scaffolds of the latest generation should not only be biocompatible but also promote osteoconduction. Poly (lactic acid) nanofibers are commonly used for this purpose; however, they lack bioactivity and do not provide good cell adhesion. Chitosan is a commonly used biopolymer which positively affects osteoblasts' behavior. The aim of this article was to prepare novel hybrid 3D scaffolds containing nanohydroxyapatite capable of cell-response stimulation. The matrixes were successfully obtained by PLA electrospinning and microwave-assisted chitosan crosslinking, followed by doping with three types of metallic nanoparticles (Au, Pt, and TiO2). The products and semi-components were characterized over their physicochemical properties, such as chemical structure, crystallinity, and swelling degree. Nanoparticles' and ready biomaterials' morphologies were investigated by SEM and TEM methods. Finally, the scaffolds were studied over bioactivity on MG-63 and effect on current-stimulated biomineralization. Obtained results confirmed preparation of tunable biomimicking matrixes which may be used as a promising tool for bone-tissue engineering.
- Klíčová slova
- biotechnology, properties of nanoparticles–reinforced polymers, smart hybrid materials,
- Publikační typ
- časopisecké články MeSH
With the development of a wide range of new biomaterials for the sensing of different cell behaviour, it is important to consider whether the cells tested in vitro are in direct contact with the material or whether cell-biomaterial contact is mediated by an interfacial layer of proteins originating from the culture medium or from the cells themselves. Thus, this study describes the differences between the cell adhesion mediated by proteins originating from foetal bovine serum and without the presence of such proteins 2 h following cell seeding exemplarily with different cell types (an osteoblastic cell line, primary fibroblasts, and mesenchymal stem cells). Three of the examined cell types were found to react differently to differing conditions in terms of cell shape, area, and number. Nevertheless, the expression and localization of the various proteins involved in cell adhesion and signalling (CD44, vinculin, talin, actin, focal adhesion kinase, Rho-GTPases and extracellular signal-regulated kinases 1 and 2) were, in general, similar with respect to all the cell types tested, albeit varying according to the presence or absence of serum. Moreover, no classical focal adhesions were formed during cell adhesion without serum proteins, while different signalling pathways were involved in this process. The study systematically describes and discusses the cell adhesion of three different human cell types to a well-known substrate without the presence of external proteins and it is hoped that this knowledge will be subsequently applied in biomaterial applications in which the presence of external proteins is undesirable (e.g. for biosensing purposes).
- Klíčová slova
- Cell adhesion, Cell signalling, Fluorescence microscopy, Foetal bovine serum, Protein expression,
- MeSH
- buněčná adheze účinky léků MeSH
- fibroblasty cytologie účinky léků MeSH
- krevní proteiny chemie farmakologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- mezenchymální kmenové buňky cytologie účinky léků MeSH
- osteoblasty cytologie účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- krevní proteiny MeSH
Bone and cartilage are tissues of a three-dimensional (3D) nature. Therefore, scaffolds for their regeneration should support cell infiltration and growth in all 3 dimensions. To fulfill such a requirement, the materials should possess large, open pores. Centrifugal spinning is a simple method for producing 3D fibrous scaffolds with large and interconnected pores. However, the process of bone regeneration is rather complex and requires additional stimulation by active molecules. In the current study, we introduced a simple composite scaffold based on platelet adhesion to poly-ε-caprolactone 3D fibers. Platelets were used as a natural source of growth factors and cytokines active in the tissue repair process. By immobilization in the fibrous scaffolds, their bioavailability was prolonged. The biological evaluation of the proposed system in the MG-63 model showed improved metabolic activity, proliferation and alkaline phosphatase activity in comparison to nonfunctionalized fibrous scaffold. In addition, the response of cells was dose dependent with improved biocompatibility with increasing platelet concentration. The results demonstrated the suitability of the system for bone tissue.
- Klíčová slova
- 3D scaffold, PCL, centrifugal spinning, cytokines, growth factors, platelets,
- MeSH
- adhezivita trombocytů účinky léků MeSH
- alkalická fosfatasa metabolismus MeSH
- kinetika MeSH
- lékové transportní systémy metody MeSH
- lidé MeSH
- mezibuněčné signální peptidy a proteiny aplikace a dávkování farmakologie MeSH
- nádorové buněčné linie MeSH
- osteoblasty cytologie účinky léků ultrastruktura MeSH
- osteogeneze účinky léků MeSH
- polyestery chemie farmakologie MeSH
- proliferace buněk účinky léků MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- trombocyty účinky léků metabolismus ultrastruktura MeSH
- tvar buňky účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alkalická fosfatasa MeSH
- mezibuněčné signální peptidy a proteiny MeSH
- polycaprolactone MeSH Prohlížeč
- polyestery MeSH
Ti-6Al-4V-based nanotubes were prepared on a Ti-6Al-4V surface by anodic oxidation on 10 V, 20 V, and 30 V samples. The 10 V, 20 V, and 30 V samples and a control smooth Ti-6Al-4V sample were evaluated in terms of their chemical composition, diameter distribution, and cellular response. The surfaces of the 10 V, 20 V, and 30 V samples consisted of nanotubes of a relatively wide range of diameters that increased with the voltage. Saos-2 cells had a similar initial adhesion on all nanotube samples to the control Ti-6Al-4V sample, but it was lower than on glass. On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes. On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples. The final cellular densities on 10 V, 20 V, and 30 V samples were higher than on glass. Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties.
- Klíčová slova
- Saos-2 cells, cell adhesion, nanostructure, osteogenic differentiation, titanium nanotubes,
- MeSH
- aktiny metabolismus MeSH
- biologické markery metabolismus MeSH
- buněčná adheze účinky léků MeSH
- buněčná diferenciace účinky léků MeSH
- fluorescenční protilátková technika MeSH
- fotoelektronová spektroskopie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nanotrubičky chemie MeSH
- osteoblasty cytologie účinky léků MeSH
- osteogeneze účinky léků MeSH
- povrchové vlastnosti MeSH
- proliferace buněk účinky léků MeSH
- slitiny MeSH
- titan farmakologie MeSH
- velikost částic * MeSH
- viabilita buněk účinky léků MeSH
- vinkulin metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aktiny MeSH
- biologické markery MeSH
- slitiny MeSH
- titan MeSH
- titanium alloy (TiAl6V4) MeSH Prohlížeč
- vinkulin MeSH
