Osteoblast-like Cell Line MG63 Research Articles

Mussel shell-derived biogenic hydroxyapatite as reinforcement on chitosan-loaded gentamicin composite for antibacterial activity and bone regeneration

In this study, hydroxyapatite (HAp) was synthesized from natural biowaste materials, specifically mussel shells, and combined with chitosan (CS) and gentamicin sulfate antibiotic (GA) using an in-situ method. The resulting composite material, designated HAp/CS-GA, has its physicochemical and structural properties characterized by Fourier transform infrared spectroscopy (FTIR) analysis. The drug-loaded structure was confirmed by UV–visible absorption spectroscopy (UV–Vis) and X-ray diffraction (XRD) analysis. Additionally, field emission scanning electron microscopy (FE-SEM) equipped with the energy dispersive X-ray spectroscopic (EDX) technique was used to determine the surface topography and main components. The composite of HAp/CS-GA was analyzed using a drug release profile and UV–visible spectroscopy (UV–Vis). The fabricated composites antimicrobial behavior was examined against bone infection-causing Gram-positive and Gram-negative bacteria, showing potential activity against Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus compared to Escherichia coli, respectively. Simultaneously, the cytotoxicity of the composite was evaluated by MTT assay using an MG-63 osteoblast-like cell line that exhibited no toxicity in the prepared composite. After a 24 h incubation period, the MG-63 cells on the HAp/CS-GA composite showed good proliferation, according to Hoechst 33258 fluorescence staining results. The results suggested that the composite had excellent biocompatibility and antibacterial activity and enhanced the osteoblast cell proliferation. Therefore, the designed HAp/CS-GA composite would be a promising candidate for bone tissue engineering.

Revealing bactericidal events on graphene oxide nano films deposited on metal implant surfaces.

At the time when pathogens are developing robust resistance to antibiotics, the demand for implant surfaces with microbe-killing capabilities has significantly risen. To achieve this goal, profound understanding of the underlying mechanisms is crucial. Our study demonstrates that graphene oxide (GO) nano films deposited on stainless steel (SS316L) exhibit superior antibacterial features. The physicochemical properties of GO itself play a pivotal role in influencing biological events and their diversity may account for the contradictory results reported elsewhere. However, essential properties of GO coatings, such as oxygen content and the resulting electrical conductivity, have been overlooked so far. We hypothesize that the surface potential and electrical resistance of the oxygen content in the GO-nano films may induce bacteria-killing events on conductive metallic substrates. In our study, the GO applied contains 52 wt% of oxygen, and thus exhibits insulating properties. When deposited as a nano film on an electrically conducting steel substrate, GO flakes generate a Schottky barrier at the interface. This barrier, consequently, impedes the transfer of electrons to the underlying conductive substrate. As a result, this creates reactive oxygen species (ROS), leading to bacterial death. We confirmed the presence of GO coatings and their hydrolytic stability by using X-ray photoelectron spectroscopy (XPS), μRaman spectroscopy, scanning electron microscopy (SEM), and Kelvin probe force microscopy (KPFM) measurements. The biological evaluation was performed on the MG63 osteoblast-like cell line and two selected bacteria species: S. aureus and P. aeruginosa, demonstrating both the cytocompatibility and antibacterial behavior of GO-coated SS316L substrates. We propose a two-step bactericidal mechanism: electron transfer from the bacteria membrane to the substrate, followed by ROS generation. This mechanism finds support in changes observed in contact angle, surface potential, and work function, identified as decisive factors. By addressing overlooked factors and effectively bridging the gap between understanding and practicality, we present a transformative approach for implant surfaces, combating microbial resistance, and offering new application possibilitie.

Improving mechanical, degradation and biological behavior of biodegradable Mg–2Ag alloy: effects of Y addition and heat treatment

Biodegradable Mg–Ag alloys are promising implants for bone regeneration owing to their ability to decrease inflammation and infection after implantation by the presence of Ag. However, their bioperformance requires further enhancement. In this regard, the effects of 1 wt % Y addition and solid solution heat treatment (T4) on microstructure, biocorrosion behavior, mechanical properties and biological features of a cast Mg–2Ag alloy were studied. Corrosion tests indicated the remarkable role of T4 treatment in promoting corrosion resistance by increasing microstructure homogeneity, reduction of defects and volume fraction of secondary phases as well as eliminating the dendritic microstructure of as-cast Mg–2Ag alloy. The presence of a stable Y-rich corrosion film on the surface of heat-treated Mg–2Ag–1Y alloy could effectively improve the corrosion resistance by inhibiting corrosion propagation. The results of cell culture experiments proved the good cell attachment and proliferation of osteoblast-like cell line MG63 on Mg–2Ag and Mg–2Ag–1Y alloys. Furthermore, the cell attachment and viability were significantly enhanced by the incorporation of Y in the solution state of Mg–2Ag alloy due to the low volume of evolved hydrogen bubbles and superb corrosion resistance.

Influence of Copper-Strontium Co-Doping on Bioactivity, Cytotoxicity and Antibacterial Activity of Mesoporous Bioactive Glass.

Mesoporous bioactive glass (MBG) is an extensively studied biomaterial used for the healing of bone defects. Its biological applications can be tailored by introducing metallic ions, such as strontium (Sr) and copper (Cu), which can enhance its functionalities, including osteogenetic, angiogenetic and antibacterial functionalities. In this study, Cu and Sr ions were co-doped (ratio 1:1) with x = 0.5, 1 and 2 mol% each in glass with an intended nominal composition of 80SiO2-(15-2x)CaO-5P2O5-xCuO-xSrO and synthesized with an evaporation-induced self-assembly (EISA)-based sol-gel technique. XRD confirmed the amorphous nature of the glass, while compositional analysis using ICP-OES confirmed the presence of dopant ions with the required amounts. A TEM study of the MBG powders showed fringes that corresponded to the formation of a highly ordered mesoporous structure. The Cu-Sr-doped MBG showed a positive effect on apatite formation when immersed in SBF, although the release of Cu and Sr ions was relatively slow for 1 mol% of each co-dopant, which signified a stable network structure in the glass. The impact of the Cu and Sr ions on the osteoblast-like cell line MG-63 was assessed. At the particle concentrations of 1 wt./vol.% or lower, the cell viability was above 50%. An antibacterial test was conducted against Gram-negative E. coli and Gram-positive S. aureus bacteria. With a sequential increase in the co-doped ion content in the glass, the zone of inhibition for bacteria increased. The results suggest that the doping of MBG with Cu and Sr ions at up to 2 mol% can result in tailored sustained release of ions to enhance the applicability of the studied glass as a functional biomaterial for bone regeneration applications.

Poly(acrylic acid)-grafted metal-organic framework carrying Mg ions for bone repair

In the field of biomedicine, metal-organic frameworks (MOFs) have been extensively studied. However, few studies have applied them to bone healing. In recent years, iron-based MOFs such as MIL-100(Fe) have been proven biocompatible and suitable for drug delivery applications. We used water-phase synthesis methods to prepare MIL-100(Fe) and NH2 functionalized MIL-100(Fe). We then performed post-synthesis modification to incorporate Magnesium (Mg) ions into the cages of the MIL-100(Fe) (i.e., Mg@MIL-100(Fe)). The poly(acrylic acid) (PAA) modification on the surface of Mg@MIL-100(Fe) (i.e., Mg@MIL-100(Fe)-PAA) prevented the Mg ions from leaking, thus significantly increasing the Mg loading. The cytotoxicity test of the osteoblast-like cell line MG-63 showed that Mg@MIL-100(Fe)-PAA was biocompatible and could slightly improve cell proliferation. Furthermore, the ALP assay showed that Mg@MIL-100(Fe)-PAA could alter the time course of cell differentiation, which could eventually accelerate the bone healing process. These findings revealed the potential of utilizing metal-organic frameworks as bone repairing materials.

Three-dimensional nano-hydroxyapatite sodium silicate glass composite scaffold for bone tissue engineering - A new fabrication process at a near-room temperature

Hydroxyapatite nanoparticles (n-HAp) due to their structural similarity to the mineral part of human bone is one of the most promising materials for the preparation of highly biocompatible and osteoconductive scaffolds to be used in bone tissue regeneration. Yet, the conversion of n-HAp powder into a 3D scaffold is usually occurring by sintering at high temperatures for several hours. Such a high temperature process, strongly affects the physicochemical and biological properties of n-HAp and therefore losing their similarity to the human bone tissue. In the present study, sodium silicate solution is applied, for the first time, as a mineral binder for consolidation of n-HAp through a dehydration-drying process at a near-room temperature (37 °C). The new consolidation process preserves the low crystallinity, non-stoichiometry, and nanosize of the precursor n-HAp, so important to mimic bone tissue. Furthermore, the consolidated 3D composite scaffold presents an adequate porosity and mechanical profile required for bone-healing applications. The in vitro cytotoxicity tests proved the non-hazardous and inductive nature of the fabricated 3D composite over the MG-63 osteoblast-like cell line. In fact, the results show significantly enhanced cell proliferation. Overall, this new consolidation process can deliver a porous 3D composite scaffold with high potential for bone tissue engineering applications.

Fabrication of polycaprolactone/β-tricalciumphosphate based nano scaffolds using electrospinning method for biomedical applications

In this study, we have focused on preparation, fabrication and characterization of polycaprolactone and beta tricalcium phosphate (PCL/β-TCP) composite scaffolds that are used for bone tissue engineering applications. The electrospun composites have been characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM). The developed scaffolds are effectively simulated the morphology, mechanical property and bioactivity for load-bearing tissue engineering applications. The bioactivity of the scaffolds has been evaluated with in vitro cell adhesion and growth studies. The results confirm the nontoxic behaviour of the composite biomaterials and developed scaffolds with MG-63 osteoblast-like cell line. The synthesized scaffolds have shown promising bioactivity with the growth as well as proliferation of new bone cells with considerable osteoconductive properties. The nanoscaffolds possess better physical properties and support high cell adhesion suggesting their application in bone tissue engineering field.

Physicochemical and biological characteristics of chitosan/κ-carrageenan thin layer-by-layer films for surface modification of nitinol

Thin films based on the natural polysaccharides κ-carrageenan (Carr) and chitosan (Chit) were formed by layer-by-layer deposition technique. Surface topography and mechanical characteristics (Young's modulus, adhesion strength) of the polymer films with different number of layers were determined using various modes of atomic force microscopy (AFM). Polymer films were used to deposit on the surface of nitinol (NiTi) plates. The creation of polysaccharide coatings on nitinol led to a change in surface properties, such as hydrophilicity and root mean square roughness. in vitro cytotoxicity assay for nitinol plates with and without polymer coating by the MG-63 osteoblast-like cell line was conducted and was shown that all the studied samples are not toxic. A decrease in cytotoxicity for samples with a polymer film consisting of 4 layers of chitosan and carrageenan was shown as compared to an uncoated nitinol plates.

Bioactive hydrogel scaffolds reinforced with alkaline-phosphatase containing halloysite nanotubes for bone repair applications

Alkaline phosphatase (ALP), biomineralization promoting enzyme, was immobilized in halloysite (HAL) nanotubes and used as a bioactive component of the chitosan (CH) and chitosan-collagen (C-CH) hydrogel scaffolds for bone regeneration. The influence of HAL-ALP and collagen on the properties of the obtained materials was studied. 30 wt% of HAL-ALP increased significantly the swelling ratio of chitosan-based scaffolds. The presence of both: collagen and HAL-ALP had positive effect on the scaffolds' porosity, which was considerably higher for C-CH. The presence of HAL has improved the mechanical properties of both types of scaffolds, while the addition of 20% of collagen to the chitosan hydrogels have considerably decreased their storage modulus. Biomineralization tests showed that although mineral was formed in both CH and C-CH scaffolds with HAL-ALP, the process was more effective for collagen-containing hydrogels. Biological studies, performed using MG-63 osteoblast-like cell line showed that C-CH scaffolds, especially those after biomineralization, were a better material for cell adhesion and growth. Both types of scaffolds degraded slowly in physiological pH. C-CH scaffolds containing 30% of HAL-ALP have the highest potential as bioactive material for bone regeneration.

Strontium- and Cobalt-Doped Multicomponent Mesoporous Bioactive Glasses (MBGs) for Potential Use in Bone Tissue Engineering Applications.

Mesoporous bioactive glasses (MBGs) offer suitable platforms for drug/ion delivery in tissue engineering strategies. The main goal of this study was to prepare strontium (Sr)- and cobalt (Co)-doped MBGs; strontium is currently used in the treatment of osteoporosis, and cobalt is known to exhibit pro-angiogenic effects. Sr- and Co-doped mesoporous glasses were synthesized for the first time in a multicomponent silicate system via the sol–gel method by using P123 as a structure-directing agent. The glassy state of the Sr- and Co-doped materials was confirmed by XRD before immersion in SBF, while an apatite-like layer was detected onto the surface of samples post-immersion. The textural characteristics of MBGs were confirmed by nitrogen adsorption/desorption measurements. In vitro experiments including MTT assay, Alizarin red staining, and cell attachment and migration showed the cytocompatibility of all the samples as well as their positive effects on osteoblast-like cell line MG-63. Early experiments with human umbilical vein endothelial cells also suggested the potential of these MBGs in the context of angiogenesis. In conclusion, the prepared materials were bioactive, showed the ability to improve osteoblast cell function in vitro and could be considered as valuable delivery vehicles for therapeutics, like Co2+ and Sr2+ ions.

Relationship between the Surface Roughness of Biodegradable Mg-Based Bulk Metallic Glass and the Osteogenetic Ability of MG63 Osteoblast-Like Cells.

Mg-based bulk metallic glass materials have been investigated for their large potential for application in orthopedic implants due to their biocompatibility, low degradation rate, and osteogenetic ability. As an orthopedic implant, initial cell adhesion has been a critical issue for subsequent osteogenesis and bone formation because the first contact between cells and the implant occurs upon the implants surface. Here, we aimed to create Mg-based bulk metallic glass samples with three different surface roughness attributes in order to understand the degradation behavior of Mg-based bulk metallic glass and the adhesion ability and osteogenetic ability of the contact cells. It was found that the degradation behavior of Mg66Zn29Ca5 bulk metallic glass was not affected by surface roughness. The surface of the Mg66Zn29Ca5 bulk metallic glass samples polished via #800 grade sandpaper was found to offer a well-attached surface and to provide a good cell viability environment for Human MG63 osteoblast-like cell line. In parallel, more calcium and mineral deposition was investigated on extracellular matrix with higher surface roughness that verify the relationship between surface roughness and cell performance.

Electrophoretic deposition of Bioactive glass – Chitosan nanocomposite coatings on Ti-6Al-4V for orthopedic applications

Chitosan-Bioactive glass (CS-BG) nanocomposite coatings were developed on the Ti-6Al-4 V alloy to investigate the effect of the BG content on the adhesion strength, bioactivity, bio-corrosion, wettability and roughness. For this purpose, BG nanoparticles were synthesized using a sol-gel process. Three nanocomposite coatings with different concentrations of BG (0.5, 1 and 1.5 g/L) were fabricated through cathodic electrophoretic deposition (EPD). The surface morphology and composition of the coatings revealed the formation of compact coatings with a uniform distribution of BG nanoparticles. Increasing the BG content enhanced the deposition rate of CS-BG nanocomposite coatings and raised the coating thickness. Moreover, the CS-BG coating containing 1.5 g/L BG showed the best corrosion performance owing to the more uniform distribution of BG nanoparticles and its higher thickness. Also, increasing the BG concentration improved the adhesion strength, raised the roughness, and promoted wettability. Further, in-vitro bioactivity evaluation of the coated and uncoated specimens in SBF revealed that the formation of bone-like apatite was significantly encouraged on the surface of CS-BG coatings, as compared to the Ti-6Al-4 V uncoated sample. So, the apatite-forming ability of the coatings was improved by increasing the BG content. For in vitro investigation, osteoblast-like cell line MG63 were cultured on Ti-6Al-4 V substrate coated with CS-BG and cellular behavior was evaluated. Results demonstrated good cell attachment with no significant levels of cytotoxicity during 5 days of culture. Therefore, the electrophoretic deposition of the CS-1.5 g/L BG coating could successfully enhance the adhesion strength, bioactivity, corrosion and cellular performance of the substrate.

Design, synthesis and biological evaluation of inhibitors of cathepsin K on dedifferentiated chondrocytes.

Selective proteinase inhibitors have demonstrated utility in the investigation of cartilage degeneration mechanisms and may have clinical use in the management of osteoarthritis. The cysteine protease cathepsin K (CatK) is an attractive target for arthritis therapy. Here we report the synthesis of two cathepsin K inhibitors (CKIs): racemic azanitrile derivatives CKI-E and CKI-F, which have better inhibition properties on CatK than the commercial inhibitor odanacatib (ODN). Their IC50 values and inhibition constants (Ki) have been determined in vitro. Inhibitors demonstrate differential selectivity for CatK over cathepsin B, L and S in vitro, with Ki amounting to 1.14 and 7.21 nM respectively. We analyzed the effect of these racemic inhibitors on viability in different cell types. The human osteoblast-like cell line MG63, MOVAS cells (a murine vascular smooth muscle cell line) or murine primary chondrocytes, were treated either with CKI-E or with CKI-F, which were not toxic at doses of up to 5 µM. Primary chondrocytes subjected to several passages were used as a model of phenotypic loss of articular chondrocytes, occurring in osteoarthritic cartilage. The efficiency of CKIs regarding CatK inhibition and their specificity over other proteases were validated in primary chondrocytes subjected to several passages. Racemic CKI-E and CKI-F at 0.1 and 1 µM significantly inhibited CatK activity in dedifferentiated chondrocytes, even better than the commercial CatK inhibitor ODN. The enzymatic activity of other proteases such as matrix metalloproteinases or aggrecanases were not affected. Taken together, these findings support the possibility to design CatK inhibitors for preventing cartilage degradation in different pathologies.

Nano-hydroxyapatite bioactive glass composite scaffold with enhanced mechanical and biological performance for tissue engineering application

Composite biomaterials can improve the mechanical and biological properties of ceramic scaffolds for bone tissue engineering application. In this study hydroxyapatite (HAp), and bioactive glass ceramics were synthesized by co-precipitation and ultrasound assisted sol-gel method. The synthesized HAp (80 wt.-%) and bioglass (15 wt.-%) bioceramics were further used with aluminum oxide (3 wt.-%) and starch (2 wt%) to prepare a composite biomaterial. The composite biomaterial was finally used to fabricate scaffold materials by employing nontoxic binder mediated gel-casting approach. The synthesized composite material and developed scaffolds were characterized through different physical, chemical, and morphological analyses, mechanical study, and in vitro biological assays. The synthesized scaffold exhibited 20–25% porosity, with a high compressive strength of ~ 157 ± 2 MPa, and Tensile strength of ~ 83 ± 2 MPa after sintering at 1200 °C for 2 h. The in vitro biological study confirmed the nontoxic behavior of the composite biomaterial and developed scaffolds with MG-63 osteoblast-like cell line. The developed composite scaffold facilitated new cell attachment, growth, and proliferation on its surface, all of which correlates with good osteoconductive properties. The developed scaffolds effectively simulated the morphology, porosity, mechanical property, and bioactivity of the bone structure for load-bearing tissue engineering application.

Electrospun F18 Bioactive Glass/PCL-Poly (ε-caprolactone)-Membrane for Guided Tissue Regeneration.

Barrier membranes that are used for guided tissue regeneration (GTR) therapy usually lack bioactivity and the capability to promote new bone tissue formation. However, the incorporation of an osteogenic agent into polymeric membranes seems to be the most assertive strategy to enhance their regenerative potential. Here, the manufacturing of composite electrospun membranes made of poly (ε-caprolactone) (PCL) and particles of a novel bioactive glass composition (F18) is described. The membranes were mechanically and biologically tested with tensile strength tests and tissue culture with MG-63 osteoblast-like cell line, respectively. The PCL-F18 composite membranes demonstrated no increased cytotoxicity and an enhanced osteogenic potential when compared to pure PCL membranes. Moreover, the addition of the bioactive phase increased the membrane tensile strength. These preliminary results suggested that these new membranes can be a strong candidate for small bone injuries treatment by GTR technique.

Fabrication and characterization of highly porous barium titanate based scaffold coated by Gel/HA nanocomposite with high piezoelectric coefficient for bone tissue engineering applications.

It is well established that the piezoelectric effect plays an important physiological role in bone growth, remodeling and fracture healing. Barium titanate, as a well-known piezoelectric ceramic, is especially an attractive material as a scaffold for bone tissue engineering applications. In this regard, we tried to fabricate a highly porous barium titanate based scaffolds by foam replication method and polarize them by applying an external electric field. In order to enhance the mechanical and biological properties, polarized/non-polarized scaffolds were coated with gelatin and nanostructured HA and characterized for their morphologies, porosities, piezoelectric and mechanical properties. The results showed that the compressive strength and piezoelectric coefficient of porous scaffolds increased with the increase of sintering temperature. After being coated with Gel/HA nanocomposite, the interconnected porous structure and pore size of the scaffolds almost remain unchanged while the Gel/nHA-coated scaffolds exhibited enhanced compressive strength and elastic modulus compared with the uncoated samples. Also, the effect of polarizing and coating of optimal scaffolds on adhesion, viability, and proliferation of the MG63 osteoblast-like cell line was evaluated by scanning electron microscope (SEM) and MTT assay. The cell culture experiments revealed that developed scaffolds had good biocompatibility and cells were able to adhere, proliferate and migrate into pores of the scaffolds. Furthermore, cell density was significantly higher in the coated scaffolds at all tested time-points. These results indicated that highly porous barium titanate scaffolds coated with Gel/HA nanocomposite has great potential in tissue engineering applications for bone tissue repair and regeneration.

Silicon bioceramic loaded with vancomycin stimulates bone tissue regeneration.

Porous ceramics doped with silicon and pure β-TCP were analyzed in terms of internal microstructure, cell behavior, and the percentage of newly formed bone. Additionally the materials were tested to determine which of the two had better properties to load and release vancomycin hydrochloride. Internal pore distribution and porosity were determined through high pressure mercury porosimetry and the specific surface area was measured by the Brunauer Emmet-Teller method. The proliferation and viability of the human osteoblast-like cell line MG-63 was studied to validate both materials. The materials were tested on eight New Zealand rabbits which created defects, 10 mm in diameter, in the calvaria bone. After 8 and 12 weeks a histological and histomorphometric analysis was performed. Si-β-TCP showed a higher porosity and specific surface area. The cytocompatibility test revealed acceptable results in terms of proliferation and viability whereas the percentage of new bone was higher in Si-β-TCP with a two-time study being statistically significant with 12 weeks of healing (p < 0.05).The vancomycin loaded within the ceramic scaffolds were burst released and the material had the ability to inhibit bacterial growth. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2307-2315, 2018.

Silver-loaded chitosan coating as an integrated approach to face titanium implant-associated infections: analytical characterization and biological activity.

The present work focuses on the idea to prevent and/or inhibit the colonization of implant surfaces by microbial pathogens responsible for post-operative infections, adjusting antimicrobial properties of the implant surface prior to its insertion. An antibacterial coating based on chitosan and silver was developed by electrodeposition techniques on poly(acrylic acid)-coated titanium substrates. When a silver salt was added during the chitosan deposition step, a stable and scalable silver incorporation was achieved. The physico-chemical composition of the coating was studied by X-ray photoelectron spectroscopy (XPS), while atomic force microscopy in intermittent contact mode (ICAFM) was used to explore the coating morphology. The amount of silver released from the coating up to 21days was evaluated by inductively coupled plasma mass spectrometry (ICP-MS). The capability of the proposed coating to interact in vitro with the biological environment in terms of compatibility and antibacterial properties was assessed using MG-63 osteoblast-like cell line and S. aureus and P. aeruginosa strains, respectively. These studies revealed that a coating showing a silver surface atomic percentage equal to 0.3% can be effectively used as antibacterial system, while providing good viability of osteoblast-like cells after 7days. The antibacterial effectiveness of the prepared coating is mainly driven by a contact killing mechanism, although the low concentration of silver released (below 0.1ppm up to 21days) is enough to inhibit bacterial growth, advantaging MG-63 cells in the race for the surface.

Simplified preparation and characterization of magnetic hydroxyapatite-based nanocomposites

Authors aimed to provide a magnetic responsiveness to bone-mimicking nano-hydroxyapatite (n-HA). For this purpose, dextran-grafted iron oxide nanoarchitectures (DM) were synthesized by a green-friendly and scalable alkaline co-precipitation method at room temperature and used to functionalize n-HA crystals. Different amounts of DM hybrid structures were added into the nanocomposites (DM/n-HA 1:1, 2:1 and 3:1weight ratio) which were investigated through extensive physicochemical (XRD, ICP, TGA and Zeta-potential), microstructural (TEM and DLS), magnetic (VSM) and biological analyses (MTT proliferation assay). X-ray diffraction patterns have confirmed the n-HA formation in the presence of DM as a co-reagent. Furthermore, the addition of DM during the synthesis does not affect the primary crystallite domains of DM/n-HA nanocomposites. DM/n-HAs have shown a rising of the magnetic moment values by increasing DM content up to 2:1 ratio. However, the magnetic moment value recorded in the DM/n-HA 3:1 do not further increase showing a saturation behaviour. The cytocompatibility of the DM/n-HA was evaluated with respect to the MG63 osteoblast-like cell line. Proliferation assays revealed that viability, carried out in the absence of external magnetic field, was not affected by the amount of DM employed. Interestingly, assays also suggested that the DM/n-HA nanocomposites exhibit a possible shielding effect with respect to the anti-proliferative activity induced by the DM particles alone.

Induction of osteogenic differentiation of osteoblast-like cells MG-63 during cultivation on fibroin microcarriers

We have developed microcarriers made from silk fibroin. Microcarriers can be used as a substrate for cell cultivation and cell delivery during cell-based therapy and for the construction of bioengineered tissue. Fibroin microcarriers were mineralized, which led to the appearance of calcium phosphate crystals on their surface. The ability of mineralized and nonmineralized microcarriers to support osteogenic differentiation of the osteoblast-like cell line MG-63 was estimated by alkaline phosphatase activity, an early marker of bone formation. The experiment showed cells actively proliferating on the surface of both mineralized and nonmodified microcarriers. Culturing MG-63 on the surface of fibroin microcarriers resulted in an increase of alkaline phosphatase activity indicative of osteogenic differentiation of MG-63 cells in the absence of inductors. The level of alkaline phosphatase was higher when mineralized microcarriers were used. Alkaline phosphatase activity of MG-63 cells cultivated using traditional two-dimensional approaches were close to zero. As opposed to conventional monolayer culturing, microcarrier culture cells are in a three-dimensional environment that is closer to physiological conditions. This can have a significant impact on their morphology and functional properties. During this study, we also characterized mechanical properties of porous scaffolds used for microcarriers.