Osteoblast-like Cells Research Articles

Nanostructured TiNb coating improves the bioactivity of 3D printed PEEK

Polyetheretherketone (PEEK) has mechanical properties suitable for the manufacturing of osseointegration implants. However, its biological properties can be significantly improved by surface modification. In this work, the macroscopic surface topography was obtained by 3D printing and then the 3D printed surface was coated with a TiNb layer. The coating made it possible to apply anodic oxidation, by which a surface with defined nanoscale topography was obtained. Thus, a material with a gradient surface topography and suitable chemical composition was prepared. The properties of the surface were analyzed using optical and scanning electron microscopy. The chemical composition was then evaluated by photoelectron spectroscopy. The roughness, the wettability, and the wear resistance of the surface were observed. The adhesion of the coating and the oxide layer was evaluated. The biological activity of samples was analyzed by seeding them with human osteoblast-like Saos-2 cells. The cell adhesion, proliferation, metabolic activity and viability was observed and was statistically evaluated. After this, the functionality of this surface modification was tested by applying it to an implant with a real geometry. The main conclusions show that the nanostructured TiNb layer on 3D printed PEEK significantly improves biological activity.

Comparison of the Biological Behavior and Topographical Surface Assessment of a Minimally Invasive Dental Implant and a Standard Implant: An In Vitro Study.

The current study aimed to assess the topographical and physical properties of a minimally invasive implant (MagiCore®: MC®, InnosBioSurg, IBS) and to evaluate its biological behavior compared to a gold standard implant (NobelParallel™: NB™, Nobel Biocare™). After surface characterization, the biological behavior assessment was conducted regarding human gingival fibroblasts (hGF) and osteoblast-like cells (MG63). Roughness values for NBTM were Ra = 1.28 µm and for MC® they were Ra = 2.02 µm. Alamar BlueTM assay LIVE/DEADTM staining results indicated equivalent biological development regarding both cell types for the two implants. Significant enhancement was found for hGF ALP activity in the presence of the two tested implants in a time-dependent manner from day 7 to day 14 (** p < 0.01). Alizarin red staining demonstrated significant calcium deposition enhancement when cells were interfaced with the NB™ compared to the MC® implant (** p < 0.05). Moreover, SEM and confocal imaging revealed good cell adhesion with a denser cellular layer on the MC® than the NB™ surface. The MC® cytocompatibility was ranked as equivalent to the gold standard implant despite the surface properties differences. These findings provide new insights about the minimally invasive implant's biological behavior and its potential clinical implication in different implantology situations.

Surface Modification of PHBV Fibrous Scaffold via Lithium Borohydride Reduction.

In this study, lithium borohydride (LiBH4) reduction was used to modify the surface chemistry of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) fibers. Although the most common reaction employed in the surface treatment of polyester materials is hydrolysis, it is not suitable for fiber modification of bacterial polyesters, which are highly resistant to this type of reaction. The use of LiBH4 allowed the formation of surface hydroxyl groups under very mild conditions, which was crucial for maintaining the fibers' integrity. The presence of these groups resulted in a noticeable improvement in the surface hydrophilicity of PHBV, as revealed by contact angle measurements. After the treatment with a LiBH4 solution, the electrospun PHBV fibrous mat had a significantly greater number of viable osteoblast-like cells (SaOS-2 cell line) than the untreated mat. Moreover, the results of the cell proliferation measurements correlated well with the observed cell morphology. The most flattened SaOS-2 cells were found on the surface that supported the best cell attachment. Most importantly, the results of our study indicated that the degree of surface modification could be controlled by changing the degradation time and concentration of the borohydride solution. This was of great importance since it allowed optimization of the surface properties to achieve the highest cell-proliferation capacity.

Dual pulsed laser deposition of Ag nanoparticles on calcium phosphate coatings for biomedical applications

Pulsed laser deposition (PLD) represents a promising bottom-up methodology for the synthesis and transference of nanoparticles to the surface of a biomedical device. Silver (Ag) nanoparticles directly incorporated on the metallic implant emerge as an alternative strategy for local action against prosthetic joint-associated infections. In the present research, a dual sequential PLD process is proposed to obtain a bilayer coating with (1) a bio-derived calcium phosphate (CaP) layer, to provide osteointegrative properties and (2) the controlled growth of the Ag nanoparticles over it, ranging the number of laser pulses from 100 to 500. The characterization by SEM, EDS, TEM, XPS and AFM revealed the uniform deposit of Ag rounded nanoparticles, with a narrow mean size distribution, in the original non-oxidized metallic state. Moreover, given the evidences from XPS and AFM techniques, the occurrence of a coalescence phenomenon from 400 pulses onwards was proposed together with the expected positive linear relation between the number of pulses and Ag contribution with a deposition rate of 0.05 at. % of Ag per pulse. Conversely, the decrease in roughness as the Ag content increased was also verified. Finally, the expected bacteriostatic activity for these PLD deposited metallic state Ag nanoparticles against the bacterial strain Staphylococcus aureus was confirmed. Moreover, the evaluation of the osteoblast-like MG-63 cells viability on the Ag(100–500)-CaP coatings revealed a significant increased proliferation (p > 0.05) on the Ag100-CaP coating compared to the control (Ag0-CaP). When same coating was evaluated against S. aureus the effect was not significant. The possibility of modulating the amount of nanoparticles in the bilayer coating to obtain a greater or lesser effect in combination with CaP was revealed.

In vitro and in vivo evaluation of surface functionalization of titanium with H2 O2 hydrothermal treatment and FGF-2.

The goals of the study were to investigate the effects on bone bioactivity of a titanium dioxide layer formed by hydrothermal oxidation of a titanium surface with hydrogen peroxide (H2 O2 ) and loading with fibroblast growth factor-2 (FGF-2) in vitro and in vivo. Ti-6Al-4V discs were hydrothermally oxidized with H2 O2 and then loaded with FGF-2. After cytotoxicity testing, Ti-6Al-4V mini-implants were subjected to the same treatment, and their osteogenic potential was evaluated histologically in a rat model. H2 O2 hydrothermal oxidation resulted in a dense porous network structure and hydrophilic changes, which improved retention of FGF-2. Morphologically, the cell density was higher, cell elongation was more pronounced, and the cell adhesion area was significantly higher in FGF-2-loaded cells than in those without FGF-2. In a cell proliferation assay using mouse osteoblast-like cells, absorbance tended to increase over time, especially in the FGF-2 group after 7 and 14 days, and in a bone differentiation assay based on ALP activity, there was a significant increase in the FGF-2 group after 14 days. In the rat model, H2 O2 hydrothermal oxidation and FGF-2 loading both resulted in more laminar bone tissue in the bone marrow around the mini-implant. These results suggest that titanium surface functionalization by H2 O2 hydrothermal oxidation and FGF-2 may promote initial cell adhesion, proliferation, and osteodifferentiation, and enhance bone bioactivity. These effects all contribute to early bonding of an implant with the surrounding bone.

Mechanical and morphological responses of osteoblast-like cells to two-photon polymerized microgrooved surfaces.

Microgrooved surfaces are recognized as an important strategy of tissue engineering to promote the alignment of bone cells. In this work, we have investigated the mechanical and morphological aspects of osteoblasts cells after interaction with different micro-structured polymeric surfaces. Femtosecond laser writing technique was used for the construction of circular and parallel microgrooved patterns in biocompatible polymeric surfaces based on pentaerythritol triacrylate. Additionally, we have studied the influence of the biocompatible TiO2 nanocrystals (NCs) related to the cell behavior, when incorporated to the photoresin. The atomic force microscopy technique was used to investigate the biomechanical reaction of the human osteoblast-like MG-63 cells for the different microgroove. It was demonstrated that osteoblasts grown on circular microgrooved surfaces exhibited significantly larger Young's modulus compared to cells sown on flat films. Furthermore, we could observe that TiO2 NCs improved the circular microgrooves effects, resulting in more populated sites, 34% more elongated cells, and increasing the cell stiffness by almost 160%. These results can guide the design and construction of effective scaffold surfaces with circular microgrooves for tissue engineering and bone regeneration.

Engineering collagenous analogs of connective tissue extracellular matrix.

Connective tissue extracellular matrix (ECM) consists of an interwoven network of contiguous collagen fibers that regulate cell activity, direct biological function, and guide tissue homeostasis throughout life. Recently, ECM analogs have emerged as a unique ex vivo culture platform for studying healthy and diseased tissues and in the latter, enabling the screening for and development of therapeutic regimen. Since these tissue models can mitigate the concern that observations from animal models do not always translate clinically, the design and production of a collagenous ECM analogue with relevant chemistry and nano- to micro-scale architecture remains a frontier challenge in the field. Therefore, the objectives of this study are two-fold— first, to apply green electrospinning approaches to the fabrication of an ECM analog with nanoscale mimicry and second, to systematically optimize collagen crosslinking in order to produce a stable, collagen-like substrate with continuous fibrous architecture that supports human cell culture and phenotypic expression. Specifically, the “green” electrospinning solvent acetic acid was evaluated for biofabrication of gelatin-based meshes, followed by the optimization of glutaraldehyde (GTA) crosslinking under controlled ambient conditions. These efforts led to the production of a collagen-like mesh with nano- and micro-scale cues, fibrous continuity with little batch-to-batch variability, and proven stability in both dry and wet conditions. Moreover, the as-fabricated mesh architecture and native chemistry were preserved with augmented mechanical properties. These meshes supported the in vitro expansion of stem cells and the production of a mineralized matrix by human osteoblast-like cells. Collectively these findings demonstrate the potential of green fabrication in the production of a collagen-like ECM analog with physiological relevance. Future studies will explore the potential of this high-fidelity platform for elucidating cell-matrix interactions and their relevance in connective tissue healing.

Laser-Deposited Beta Type Ti-42Nb Alloy with Anisotropic Mechanical Properties for Pioneering Biomedical Implants with a Very Low Elastic Modulus.

Present commercial titanium alloy implants have an elastic modulus higher than 100 GPa, whereas that of the cortical bone is much smaller (17–28 GPa). This elastic modulus mismatch produces a stress shielding effect and the resorption of the bone surrounding the implant. In the present work, a <100> fiber texture is developed in β type Ti-42Nb (wt%) alloy ingots generated by laser-directed energy deposition (LDED) in order to achieve anisotropic mechanical properties. In addition, we demonstrate that laser-deposited β type Ti-42Nb alloy ingots with an intense <100> fiber texture exhibit a very low elastic modulus in the building direction (Ez < 50 GPa) and high yield (σ0.2z > 700 MPa) and tensile (UTSz > 700 MPa) strengths. Laser-deposited Ti-42Nb alloy enhances the osteoinductive effect, promoting the adhesion, proliferation, and spreading of human osteoblast-like cells. Hence, we propose that laser-deposited β type Ti-42Nb alloy is a potentially promising candidate for the manufacturing of pioneering biomedical implants with a very low elastic modulus that can suppress stress shielding.

Surface characteristics and in vitro biocompatibility of surface-modified titanium foils as a regenerative barrier membrane for guided bone regeneration.

This study evaluated surface characteristics and biocompatibility of surface-modified thin titanium (Ti) foils as a regenerative barrier membrane for future application in guided bone regeneration (GBR) surgery to augment atrophic alveolar bone. Anodic oxidation and post-heat treatment were performed to prepare various Ti foil samples. Then, the in vitro soft and hard tissue compatibility of the samples was evaluated by examining the cell responses using primary human gingival fibroblasts (HGFs) and MG63 human osteoblast-like cells. Investigated Ti foil samples showed marked differences in physicochemical surface properties. Additional 400°C heat treatment applied to the anodized Ti surface led to formation of an anatase titanium dioxide structure and well-organized nanoscale protrusions, and significantly increased surface wettability. Anodization and heat treatment enhanced the growth of HGFs and MG63 cells in Ti foil samples. Additional heat treatment for 10 and 30min further significantly improved the response of HGFs including spreading and proliferation, and upregulated the mRNA expression of cell adhesion- and maturation-related genes as well as the osteoblast differentiation of MG63 cells. Ti foil sample with thin oxide coating obtained by a 30min heat treatment exhibited poor clinical plasticity as a regenerative barrier membrane, which showed complete coating failure in the bending test. Our results indicate that anatase Ti oxide coating of a specific film thickness with nanoscale surface protrusion morphology and hydrophilic characteristics obtained by anodization and post-heat treatment would be an effective approach as a biocompatible Ti regenerative membrane for inducing better regeneration of both gingival tissue and bone.

EFFECT OF PERIOSTIN ON AUTOPHAGY AND FACTORS RELATED TO BONE METABOLISM IN OSTEOBLASTS

The purpose of this study was to explore the effect of periostin on autophagy and factors related to bone metabolism in osteoblasts. According to experimental design, some osteoblast-like cells (MC3T3-E1) were transfected by lentiviral vector to silence periostin gene. Celigo[Formula: see text] Image Cytometer and Real-Time PCR were used to assess target gene silencing effects. Western blotting was used to evaluate autophagy-associated factors (LC3B,Beclin1) and factors related to bone metabolism (RANKL, OPG, NF-[Formula: see text]B[Formula: see text]P65). Another part of osteoblasts was added with recombinant periostin protein at concentration of 75[Formula: see text]ng/mL. Monodansylcadaverine (MDC) detection was conducted to trace formation of autophagy vesicles. Factors associated with autophagy and bone metabolism were reassessed using western blotting. When periostin gene was silenced, western blotting analysis showed that the expression of LC3B-I, Beclin1, RANKL and NF-[Formula: see text]B[Formula: see text]P65 in experimental group was significantly increased; however, OPG expression was decreased. After adding recombinant periostin protein, MDC detection revealed significantly fewer green fluorescent particles in the experimental group. Western blotting results showed that the expression of LC3B, Beclin1, RANKL and NF-[Formula: see text]B[Formula: see text]P65 was decreased; however, OPG expression was increased. This study examined the regulatory mechanism of periostin in bone metabolism, which will provide a future reference for the treatment of primary teeth abnormal replacement caused by inflammation.

Comparison of Different Chemical and Mechanical Modalities for Implant Surface Decontamination: Activity against Biofilm and Influence on Cellular Regrowth-An In Vitro Study.

ObjectivesThe aim of this in vitro study was to compare the efficacy of chemical and mechanical methods for decontamination of titanium dental implant surfaces previously infected with polymicrobial biofilms in a model simulating a peri-implant defect. Furthermore, the effect of each decontamination protocol on MG-63 osteoblast-like cells morphology and adhesion to the treated implants was assessed.BackgroundPeri-implantitis is a growing issue in dentistry, and evidence about implant surface decontamination procedures is lacking and inconclusive.MethodsA total of 40 previously biofilm-contaminated implants were placed into a custom-made model simulating a peri-implant defect and randomly assigned to five treatment groups: (C) control (no treatment); (AW) air abrasion without any powder; (ESC) air abrasion with powder of erythritol, amorphous silica, and 0.3% chlorhexidine; (HBX) decontamination with a sulfonic/sulfuric acid solution in gel; and (HBX + ESC) a combination of HBX and ESC. Microbiological analysis was performed on five implants per treatment group, and the residual viable bacterial load measured in log 10 CFU/mL was counted for each bacterial strain and for the total number of colonies. The remaining three implants per group and three noncontaminated (NC) implants were used to assess surface biocompatibility using a scanning electron microscope and a backscattered electron microscope after seeding with MG-63 cells.ResultsA significant decontaminant effect was achieved using HBX or HBX + ESC, while no differences were observed among other groups. The percentage of implant surface covered by adherent MG-63 cells was influenced by the treatment method. Progressive increases in covered surfaces were observed in groups C, AW, ESC, HBX, HBX + ESC, and NC.ConclusionsA combination of mechanical and chemical decontamination may provide more predictable results than mechanical cleaning alone.

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.

Influence of the Components and Orientation of Hydroxyapatite Fibrous Substrates on Osteoblast Behavior.

Synthetic hydroxyapatite has good biocompatibility, bioactivity and osteoconductive ability because its chemical properties and biological properties are similar to those of bioapatite in bone tissue. Strontium-substituted hydroxyapatite has better degradability than hydroxyapatite and can both promote osteogenesis and inhibit adipogenesis in mesenchymal stem cells. Hence, hydroxyapatite and strontium-substituted hydroxyapatite are widely used as bone graft materials, cell carriers and drug/gene delivery carriers. In addition, osteoblasts cultured on aligned nanofibrous substrates had higher expression of osteogenesis-related genes than did those cultured on random nanofibrous substrates. However, to date, no study has explored the effects of the components and orientation of hydroxyapatite nanofibrous substrates on osteoblastic behavior. In this study, a random hydroxyapatite nanofibrous substrate (R-HANF), a random strontium-substituted hydroxyapatite nanofibrous substrate (R-SrHANF), an aligned hydroxyapatite nanofibrous substrate (A-HANF) and an aligned strontium-substituted hydroxyapatite nanofibrous substrate (A-SrHANF) were successfully fabricated by using the electrospinning technique. The effect of fiber composition on osteoblast-like MG63 cells was assessed by evaluating cell morphology, cell proliferation and osteogenesis-related gene expression. The results showed that MG63 cells cultured on A-SrHANF had higher osteogenesis-related gene expression than those cultured on A-HANF. Additionally, MG63 cells were cultured on R-SrHANF and A-SrHANF to evaluate the effects of fiber orientation on cell behavior. On A-SrHANF, the cells aligned along the direction of the nanofibers, with typical bipolar morphologies, and exhibited higher osteogenesis-related gene expression than cells on R-SrHANF. Hence, the components and orientation of hydroxyapatite nanofibrous substrates are critical parameters affecting the osteogenesis process.

PREFACE: A SPECIAL SELECTION ON EMERGING TECHNIQUES FOR BIOMECHANICS–PART II

PREFACE: A SPECIAL SELECTION ON EMERGING TECHNIQUES FOR BIOMECHANICS–PART II

Highly effective electrospun polycaprolactone/ layered double hydroxide nanofibrous scaffold for bone tissue engineering

Bone tissue engineering is considered a promising approach for repairing, regenerating and orthopedic implants. In this regard, polymeric supportive scaffolds reinforced with bioactive nanomaterials provide unique functions help in substitution of impaired bone tissues. In this constant, various layered double hydroxide (LDH) concentrations (ranging from 0.1 wt% to 10 wt%.) within the poly (ε-caprolactone) (PCL) scaffolds were fabricated successfully through electrospinning. The scaffolds were investigated by XRD, SEM, FTIR, and EDX analyses to study the phase identification, morphology, chemical properties, and elemental composition, respectively. The findings demonstrated that the inclusion of LDH to PCL scaffold increased mechanical tensile strength from 1.63 ± 0.4 to 1.920 ± 0.3 Mpa for the pure PCL and PCL + LDH (1 wt%) scaffolds, respectively. A decrease in the average diameter of PCL nanofibers was observed with the addition of LDH. The enrichment of PCL scaffold with LDH promotes in vitro biomineralization when subjected to simulated body fluid (SBF), indicating the bioactive features of the nanocomposite scaffolds. In addition, increased of LDH leads to improvement in protein adsorption capacity compared to the pure one. Furthermore, the cytocompatibility and cytotoxicity of nanocomposite fibers was assessed using human MG-63 osteoblast-like cells. The LDH/PCL scaffold induced mineral deposition and osteogenic differentiation of MG-63 as evident by an increase in ALP activity in comparison with the pure PCL. Overall, our results revealed that electrospun PCL-LDH nanofiber might be an ideal choice for bone tissue engineering.

Biological and Physico-Chemical Properties of Composite Layers Based on Magnesium-Doped Hydroxyapatite in Chitosan Matrix.

In the present study, we report the development and characterization of composite layers (by spin coating) based on magnesium-doped hydroxyapatite in a chitosan matrix, (Ca10−xMgx(PO4)6(OH)2; xMg = 0, 0.08 and 0.3; HApCh, 8MgHApCh and 30MgHApCh). The MgHApCh composite layers were investigated using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) techniques. The in vitro biological evaluation included the assessment of their cytotoxicity on MG63 osteoblast-like cells and antifungal activity against Candida albicans ATCC 10231 fungal cell lines. The results of the physico-chemical characterization highlighted the obtaining of uniform and homogeneous composite layers. In addition, the biological assays demonstrated that the increase in the magnesium concentration in the samples enhanced the antifungal effect but also decreased their cytocompatibility. However, for certain optimal magnesium ion concentrations, the composite layers presented both excellent biocompatibility and antifungal properties, suggesting their promising potential for biomedical applications in both implantology and dentistry.

Investigation of the influence of SrO addition on the bioactivity of CaO[sbnd]Na2O-P2O5-B2O3 glass system

In this work, the role of strontium oxide in the bioactive glasses calcium-sodium-borate was studied to evaluate the bioactivity of the system and the rate of the ion release in Simulated Body Fluid (SBF). Five samples were prepared, with concentration of strontium oxide (SrO) from 0 to 20 wt%, in order to understand how SrO change the glass structure and the ionic leaching process in SBF. Results of density and molar volume, XRD, FTIR, weight loss, pH and cytotoxicity assay were presented and discussed as a function of SrO. The measured pH values of SBF, presented lower values for samples with greater amounts of SrO, suggesting that the increase of SrO in the glasses can reduce the release of the ions in SBF. The fraction of tetracoordinate boron, N4, calculated by FTIR results, increases with the addition of SrO, which shows an increase in the glass connectivity and a decrease of the chemical reactivity. The FTIR spectra and X-ray diffraction results, after immersion in SBF, confirmed the materials bioactivity with the formation of carbonated hydroxyapatite or crystalline calcium phosphate. The samples did not show in vitro cytotoxicity against osteoblast cell line. Also, we highlight that 10SrO sample has induced proliferation of osteoblast-like cells in vitro to all concentrations of glass. These results show that SrO doped calcium-sodium-borate glasses are materials with high rate of bioactivity, low cytotoxicity and reduced dissolution rate of the glasses in an aqueous medium, besides induce cells proliferation and bone tissue growth and their biocompatibility with living tissues.

Impact of Cross-Linked Hyaluronic Acid on Osteogenic Differentiation of SAOS-2 Cells in an Air-Lift Model.

The aim of this study was to investigate the impact of cross-linked hyaluronic acid on osteoblast-like cells seeded on top of two collagen substrates, native porcine pericardium membrane (substrate A) and ribose cross-linked collagen membranes (substrate B), in an air-lift model. Substrates A or B, saturated with three hyaluronic acid concentrations, served as membranes for SAOS-2 cells seeded on top. Cultivation followed for 7 and 14 days in the air-lift model. Controls used the same substrates without hyaluronic pre-treatment. Cells were harvested, and four (Runx2, BGLAP, IBSP, Cx43) different osteogenic differentiation markers were assessed by qPCR. Triplicated experiment outcomes were statistically analyzed (ANOVA, t-test; SPSS). Supplementary histologic analysis confirmed the cells’ vitality. After seven days, only few markers were overexpressed on both substrates. After 14 days, targeted genes were highly expressed on substrate A. The same substrate treated with 1:100 diluted xHyA disclosed statistically significant different expression level vs. substrate B (p = 0.032). Time (p = 0.0001), experimental condition as a function of time (p = 0.022), and substrate (p = 0.028) were statistically significant factors. Histological imaging demonstrated vitality and visualized nuclei. We conclude that the impact of hyaluronic acid resulted in a higher expression profile of SAOS-2 cells on substrate A compared to substrate B in an air-lift culture after two weeks.

Effect of tantalum interlayer on hydroxyapatite biointerface for orthopedic applications

Various attempts have been made during recent decades to improve the functionality, biocompatibility and osseointegration of metallic implants. With technological advancement, modifying superficial properties of orthopedic implants has undergone great evolution. Current efforts of scientists and the orthopedic industry have focused on modifying surface properties to achieve a specific biointerface that can reduce patient's healing time and prevent aseptic loosening. This study investigates the contribution of a protective tantalum interlayer deposited on titanium substrates on the growth of hydroxyapatite (HAp) crystals with hydrothermal synthesis method. We investigated the morphology and structure of the produced coatings, as well as in vitro bioactivity in simulated body fluid and biocompatibility in osteoblast-like cell culture. The hydrothermal method allows to synthesize coating with micro-sized HAp crystallites that exhibit rod-like hexagonal shape. The diameter of HAp crystallites may be controlled and driven by nature of the interlayer (tantalum, tantalum oxide or titanium oxide) previously formed on the titanium substrate. Coatings with smaller diameters of HAp crystals (between 0,3 μm and 0,6 μm) synthesized on tantalum-based films were found to be more favorable for cell viability and attachment. High bioactivity and positive effect on the osteoblast-like cells of the HAp biointerface with a tantalum intermediate layer was observed.

A study of the interactions between human osteoblast-like cells and polymer composites with functionalized graphene derivatives using 2D correlation spectroscopy (2D-COS)

In response to the growing need for development of modern biomaterials for applications in regenerative medicine strategies, the research presented here investigated the biological potential of two types of polymer nanocomposites. Graphene oxide (GO) and partially reduced graphene oxide (rGO) were incorporated into a poly(ε-caprolactone) (PCL) matrix, creating PCL/GO and PCL/rGO nanocomposites in the form of membranes. Proliferation of osteoblast-like cells (human U-2 OS cell line) on the surface of the studied materials confirmed their biological activity. Fluorescence microscopy was able to distinguish the different patterns of interaction between cells (depending on the type of material) after 15 days of the test run. Raman micro-spectroscopy and two-dimensional correlation spectroscopy (2D-COS) applied to Raman spectra distinguished the nature of cell-material interactions after only 8 days. Combination of these two techniques (Raman micro-spectroscopy and 2D-COS analysis) facilitated identification of a much more complex cellular response (especially from proteins) on the surface of PCL/GO. The presented approach can be regarded as a method for early study of the bioactivity of membrane materials.