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

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

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