Adhesion Of Pre-osteoblastic Cells Research Articles

Dual Mg-Reinforced PCL Membrane with a Janus Structure for Vascularized Bone Regeneration and Bacterial Elimination.

Commercially available guided bone regeneration (GBR) membranes often exhibit limited mechanical properties or bioactivity, leading to poor performance in repairing bone defects. To surmount this limitation, we developed a Janus structural composite membrane (Mg-MgO/PCL) reinforced by dual Mg (Mg sheets and MgO NPs) by using a combined processing technique involving casting and electrospinning. Results showed that the addition of Mg sheets and MgO NPs enhanced the mechanical properties of the composite membrane for osteogenic space maintenance, specifically tensile strength (from 10.2 ± 1.2 to 50.3 ± 4.5 MPa) and compression force (from 0 to 0.94 ± 0.09 N mm-1), through Mg sheet reinforcement and improved crystallization. The dense cast side of the Janus structure membrane displayed better fibroblast barrier capacity than a single fiber structure; meanwhile, the PCL matrix protected the Mg sheet from severe corrosion due to predeformation. The porous microfibers side supported preosteoblast cell adhesion, enhanced osteogenesis, and angiogenesis in vitro, through the biomimetic extracellular matrix and sustainable Mg2+ release. Furthermore, the Mg-MgO/PCL membrane incorporating 2 wt % MgO NPs exhibited remarkable antimicrobial properties, inducing over 88.75% apoptosis in Staphylococcus aureus. An in vivo experiment using the rat skull defect model (Φ = 5 mm) confirmed that the Mg-MgO/PCL membrane significantly improved new bone formation postsurgery. Collectively, our investigation provides valuable insights into the design of multifunctional membranes for clinical oral GBR application.

Development of Super-Paramagnetic Iron Oxide Nanoparticle-Coated Melt Electrowritten Scaffolds for Biomedical Applications.

Polycaprolactone (PCL) is usually the material chosen for melt electrowriting (MEW) due to its biocompatibility, mechanical strength, and melt processability. This work first investigates the effect of different processing parameters to obtain optimum PCL-MEW scaffolds. Secondly, to increase PCL`s hydrophilicity and cell affinity, and to enable coating with superparamagnetic iron oxide nanoparticles (SPIONs) and silica-coated-SPIONs (Si-SPIONs), the scaffolds are modified with alkaline surface treatment. Finally, SPIONs and Si-SPIONs are successfully coated on MEW scaffolds. Results show that reproducible scaffolds are fabricated. Additionally, the alkaline treatment does not change the three-dimensionalmorphology of scaffolds while reducing fiber diameter. Furthermore, SEM images and ATR-FTIR results confirmed that SPIONs and Si-SPIONs-were coated on scaffolds. A cytocompatibility assay showed a non-toxic effect on MG-63 osteoblast-like cells in all scaffolds. Additionally, higher MC3T3-E1 pre-osteoblastic cell adhesion efficiency and proliferation are achieved for the alkaline-treated scaffolds and SPIONs/Si-SPIONs-coated scaffolds. All samples demonstrated the ability to generate heat, useful for hyperthermia-treatment, when subjected to an alternating magnetic field. Overall, the findings suggest that the strategy of coating PCL-MEW scaffolds with SPIONs/Si-SPIONs has the potential to improve scaffold performance for biomedical applications, especially for producing magnetically responsive MEW scaffolds.

COMPARATIVE CHARACTERIZATION OF ARC-PVD ZrN AND PEO HYDROXYAPATITE COATINGS ON PURE Ti-6Al-4V SUBSTRATES FOR BIOMEDICAL APPLICATIONS

The nanocrystalline films of zirconium nitride have been synthesized using ion-plasma vacuum-arc deposition technique on pure Ti-6Al-4V, whereas the hydroxyapatite coatings were deposited by the method of plasmaelectrolytic oxidation in alkaline electrolytes (hydroxyapatite + 1 М potassium hydroxide). Structure evaluation by X-ray diffraction analysis (XRD), scanning electron microscopy (SEM) with microanalysis (EDS) – were performed to study phase and chemical composition, surface morphology, microstructure of coatings. The formed HA PEO coatings with spheroidal structure contain the phases of CaTiO3 calcium titanate, CaHPO4 anhydrous dicalcium phosphate, and Ca10(PO4)6(OH)2 hydroxyapatite. It was revealed that ZrN single-phase coatings of cubic modification with finecrystalline grains of 20 nm in size were formed. This experimental study found that coating Ti-6Al-4V with ZrN and HA positively influences preosteoblast cell adhesion, which results in the almost complete coverage of the surface, along with a less number of cells grown on uncoated titanium surface.

Disordered Protein Stabilization by Co-Assembly of Short Peptides Enables Formation of Robust Membranes

Molecular self-assembly is a spontaneous natural process resulting in highly ordered nano to microarchitectures. We report temperature-independent formation of robust stable membranes obtained by the spontaneous interaction of intrinsically disordered elastin-like polypeptides (ELPs) with short aromatic peptides at temperatures both below and above the conformational transition temperature of the ELPs. The membranes are stable over time and display durability over a wide range of parameters including temperature, pH, and ultrasound energy. The morphology and composition of the membranes were analyzed using microscopy. These robust structures support preosteoblast cell adhesion and proliferation as well as pH-dependent cargo release. Simple noncovalent interactions with short aromatic peptides can overcome conformational restrictions due to the phase transition to facilitate the formation of complex bioactive scaffolds that are stable over a wide range of environmental parameters. This approach offers novel possibilities for controlling the conformational restriction of intrinsically disordered proteins and using them in the design of new materials.

Chemical vapor deposited polyelectrolyte coatings with osteoconductive and osteoinductive activities

We report solvent-free incorporation of osteogenic coatings including polycations, polyanions, and polyampholytes on the polystyrene cell culture plates via initiated chemical vapor deposition. The osteoconductive and osteoinductive properties of coatings with different surface charges were systematically studied in vitro. Initial adhesion of preosteoblast cells was significantly improved by the negatively charged and mixed charged coatings but was inhibited by the positively charged one. Compared with the control, all the polyelectrolyte coated surfaces presented similar cell proliferation and higher differentiation after 10 days of culture, indicating their positive effects on osteoconduction and osteoinduction. The surface charge of vapor-deposited coatings helped to induce nucleation and crystallization of apatites during mineralization. Of all the coated surfaces, the Ca/P ratio in apatites formed on the coated surfaces with mixed charges was closest to that in natural apatite-mineralized bones.

Multifunctional Titanium Surfaces for Orthopedic Implants: Antimicrobial Activity and Enhanced Osseointegration.

The use of implants in orthopedics and dental practice is a widespread surgical procedure to treat diverse diseases. However, peri-implantitis due to infections and/or poor osseointegration can lead to metallic implant failure. The aim of this study was to develop a multifunctional coating on titanium (Ti) surfaces, to simultaneously deal with both issues, by combining antibacterial silver nanoparticles (AgNPs) and regenerative properties of lactoferrin (Lf). A simple and cost-effective methodology that allows the direct multifunctionalization of Ti surfaces was developed. The modified surfaces were characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy, and contact angle measurements. Additionally, in vitro preosteoblast cell adhesion, cell viability, and differentiation were evaluated. The antibacterial capability of the surfaces was tested against Staphylococcus aureus as a prosthesis infection model strain. Our results showed that Lf adsorbed on both Ti surfaces and Ti surfaces with adsorbed AgNPs. Simultaneously, the presence of Lf and AgNPs notably improved preosteoblast adhesion, proliferation, and differentiation, whereas it reduced the bacterial colonization by 97.7%. Our findings indicate that this simple method may have potential applications in medical devices to both improve osseointegration and reduce bacterial infection risk, enhancing successful implantation and patients' quality of life.

Tuning Myoblast and Preosteoblast Cell Adhesion Site, Orientation, and Elongation through Electroactive Micropatterned Scaffolds.

Electroactive polymers are being increasingly used in tissue engineering applications. Together with the electromechanical clues, morphological ones have been demonstrated to determine cell proliferation and differentiation. This work reports on the micropatterning of poly(vinylidene fluoride-co-trifluoroethylene), P(VDF-TrFE) scaffolds, and their interaction with myoblast and preosteoblasts cell lines, selected based on their different functional morphology. The scaffolds were obtained by soft lithography and obtained in the form of arrays of lines, intermittent lines, hexagons, linear zigzags, and curved zigzags with dimensions of 25, 75, and 150 μm. Moreover, the scaffolds were tested in cell adhesion assays of myoblasts and preosteoblasts cell lines. The results show that more linear surface topographies and dense morphology have a large potential in the regeneration of musculoskeletal tissue, while nonpatterned scaffolds or more anisotropic surface microstructures present largest potential to promote the growth and regeneration of bone tissue. In this way, cell adhesion site, orientation, and elongation can be controlled by choosing properly the topography and morphology of the scaffolds, indicating their suitability and potential for further proliferation and differentiation assays.

Adhesive and oxidative response of stem cell and pre-osteoblasts on titanium and zirconia surfaces in vitro.

The aim of the present study was to investigate the initial stem cell and pre-osteoblast cell adhesion and oxidative response on zirconia in comparison with titanium. Human dental pulp stem cells (DPSC) and murine pre-osteoblasts (MC3T3-E1) cells were cultured on zirconia and titanium surfaces, and at 3-, 12-, and 24-hour intervals, cell viability and morphology were determined with tetrazolium based colorimetric assay, scanning electron microscopy, and immunofluorescence analysis. The in situ reactive oxygen species level of both cells on each material surface was examined after 24-hour culture. Both DPSC and MC3T3-E1 cells revealed comparable morphological features during 24-hour cell adhesion processes, with cells continued expanding of cell size and increasing of cell viability on titanium and zirconia surfaces during 24-hour culture. Zirconia demonstrated relatively higher mean cell viability compared to titanium within 24-hour culture, with significantly higher DPSC viability at 12hours after seeding (P<0.05). Relatively higher mean reactive oxygen species levels in both DPSC and MC3T3E1 were found on zirconia surfaces after 24-hour culture compared to titanium. From the results, zirconia as a potential dental implant substrate demonstrated equivalent or better initial cellular responses compared to titanium.

Fabrication of a nanoparticle-containing 3D porous bone scaffold with proangiogenic and antibacterial properties

3D porous scaffolds based on agarose and nanocrystalline apatite, two structural components that act as a temporary mineralized extracellular matrix, were prepared by the GELPOR3D method. This shaping technology allows the introduction of thermally-labile molecules within the scaffolds during the fabrication procedure. An angiogenic protein, Vascular Endothelial Growth Factor, and an antibiotic, cephalexin, loaded in mesoporous silica nanoparticles, were included to design multifunctional scaffolds for bone reconstruction. The dual release of both molecules showed a marked increase in the number of blood vessels on embryonic day 14 in chicken embryos grown ex ovo, while, at the same time providing an antibiotic local concentration capable of inhibiting Staphylococcus aureus bacterial growth. In this sense, different release patterns, monitored by UV-spectroscopy, could be tailored as a function of the cephalexin loading strategy, either releasing all the loaded cephalexin in the first 4 h or less than 50% after 24 h. The scaffold surface was characterized by a high hydrophilicity, with contact angles between 50° and 63°, which enabled the adhesion and proliferation of preosteoblastic cells. Statement of SignificanceThe localized delivery of bioactive molecules has attracted significant attention due to the potential for dose reduction as well as reduced side effects compared to systemic delivery. In this article multifunctional 3D porous scaffolds with a designed porosity have been fabricated. The method also enables the controlled loading of an antibiotic drug and an angiogenic protein into the scaffold. These scaffolds, whose composition resembles the extracellular matrix are suitable for the adhesion of preosteoblast cells, exhibit a sustained cephalexin delivery adequate for inhibiting bacterial growth as well as release the proangiogenic molecule which induces blood vessel formation in chicken embryos grown ex ovo.

Controlling pre-osteoblastic cell adhesion and spreading on glycopolymer brushes of variable film thickness.

Controlling the cell behavior on biocompatible polymer surfaces is critical for the development of suitable medical implant coatings as well as in anti-adhesive applications. Synthetic glycopolymer brushes, based on sugar methacrylate monomers have been reported as robust surfaces to resist protein adsorption and cell adhesion. In this study, poly(D-gluconamidoethyl methacrylate) (PGAMA) brushes of various chain lengths were synthesized directly from initiator functionalized glass substrates using surface-initiated atom transfer radical polymerization. The glycopolymer film thicknesses were determined by ellipsometry, whereas the wettability and the morphology of the surfaces were characterized by static water contact angle measurements and atomic force microscopy, respectively. Stable, grafted films with thicknesses in the dry state between 4 and 20 nm and of low roughness (~1 nm) were obtained by varying the polymerization time. Cell experiments with MC3T3-E1 pre-osteoblasts cultured on the PGAMA brushes were performed to examine the effect of film thickness on the cell morphology, cytoskeleton organization and growth. The results revealed good cell spreading and proliferation on PGAMA layers of low film thickness, whereas cell adhesion was prevented on polymer films with thickness higher than ~10 nm, indicating their potential use in medical implants and anti-adhesive surfaces, respectively.

Multifunctional Copper-Containing Carboxymethyl Chitosan/Alginate Scaffolds for Eradicating Clinical Bacterial Infection and Promoting Bone Formation.

Repairing infected bone defects relies on a scaffold that can not only fill the defects to promote bone formation but also kill clinically present bacterial pathogens such as Staphylococcus aureus (S.aureus). To meet this demand, here, we develop a new copper (Cu) containing natural polymeric scaffold with a full potential for repairing infected bone defects. Instead of directly adding antibacterial Cu2+ ions to the polymer mixtures, which caused uncontrolled polymer cross-linking, we added Cu nanoparticles to the mixture of anionic carboxymethyl chitosan (CMC) and alginate (Alg). Then, the Cu2+ ions released from the Cu nanoparticles gradually cross-linked the polymer mixtures, which was further turned into a scaffold (CMC/Alg/Cu) with an interconnected porous structure by freeze-drying. We found that the CMC/Alg/Cu scaffolds showed significantly improved capabilities of osteogenesis and killing clinical bacteria compared to CMC/Alg scaffolds fabricated by the same procedure but without adding Cu nanoparticles. Specifically, in vitro studies showed that the CMC/Alg/Cu scaffolds with excellent biocompatibility could enhance preosteoblastic cell adhesion by upregulating the expression level of adhesion-related genes (focal adhesion kinase (FAK), paxillin (PXN), and vinculin (VCL)), promoting osteogenic differentiation and mineralization by upregulating the osteogenesis-related gene expression and extracellular calcium deposition. In vivo studies further demonstrated that CMC/Alg/Cu scaffolds could induce the formation of vascularized new bone tissue in 4 weeks while avoiding clinical bacterial infection even when the implantation sites were challenged with the clinically collected S.aureus bacteria. This work represents a facile and innovative approach to the fabrication of Cu containing polymer scaffolds that can potentially be used to repair infected bone defects.

Electrochemical & osteoblast adhesion study of engineered TiO2 nanotubular surfaces on titanium alloys.

TiO2 nanotubes were grafted on the surface of cpTi, Ti6Al4V and Ti6Al4V-ELI with the aim to provide a new podium for human pre-osteoblast cell (MC3T3) adhesion and proliferation. The surface morphology and chemistry of these alloys were examined with scanning electron microscopy and energy dispersive x-ray spectroscopy. TiO2 nanotubes were further characterized by cyclic potentiodynamic polarization tests and electrochemical impedance spectroscopy. The vertically aligned nanotubes were subjected to pre-osteoblast cell proliferation in order to better understand cell-material interaction. The study demonstrated that these cells interact differently with nanotubes of different titanium alloys. The significant acceleration in the growth rate of pre-osteoblast cell adhesion and proliferation is also witnessed. Additionally, the cytotoxicity of the leached metal ions was evaluated by using a tetrazolium-based bio-assay, MTS. Each group of data was operated for p<0.05, concluded one way ANOVA to investigate the significance difference.

Incubator proof miniaturized Holomonitor to in situ monitor cancer cells exposed to green tea polyphenol and preosteoblast cells adhering on nanostructured titanate surfaces: validity of the measured parameters and their corrections.

The in situ observation of cell movements and morphological parameters over longer periods of time under physiological conditions is critical in basic cell research and biomedical applications. The quantitative phase-contrast microscope applied in this study has a remarkably small size, therefore it can be placed directly into a humidified incubator. Here, we report on the successful application of this M4 Holomonitor to observe cancer cell motility, motility speed, and migration in the presence of the green tea polyphenol, epigallocatechin gallate, as well as to monitor the adhesion of preosteoblast cells on nanostructured titanate coatings, relevant for biomedical applications. A special mechanical stage was developed to position the sample into that range of the optical arrangement where digital autofocusing works with high reproducibility and precision. By in-depth analyzing the obtained single cell morphological parameters, we show that the limited vertical resolution of the optical setup results in underestimated single cell contact area and volume and overestimated single cell averaged thickness. We propose a simple model to correct the recorded data to obtain more precise single cell parameters. We compare the results with the kinetic data recorded by a surface sensitive optical biosensor, optical waveguide lightmode spectroscopy.

Toward smart implant synthesis: bonding bioceramics of different resorbability to match bone growth rates.

Craniofacial reconstructive surgery requires a bioactive bone implant capable to provide a gradual resorbability and to adjust to the kinetics of new bone formation during healing. Biomaterials made of calcium phosphate or bioactive glasses are currently available, mainly as bone defect fillers, but it is still required a versatile processing technique to fabricate composition-gradient bioceramics for application as controlled resorption implants. Here it is reported the application of rapid prototyping based on laser cladding to produce three-dimensional bioceramic implants comprising of a calcium phosphate inner core, with moderate in vitro degradation at physiological pH, surrounded by a bioactive glass outer layer of higher degradability. Each component of the implant is validated in terms of chemical and physical properties, and absence of toxicity. Pre–osteoblastic cell adhesion and proliferation assays reveal the adherence and growth of new bone cells on the material. This technique affords implants with gradual-resorbability for restoration of low-load-bearing bone.

Effects of human fibronectin and human serum albumin sequential adsorption on preosteoblastic cell adhesion.

Fibronectin (Fn) is widely reported to promote cell adhesion and spreading, and recent reports attest to the synergistic effect of coadsorbed albumin (unexpected due to the passivating character of the latter protein). In this study, the sequential adsorption of fibronectin and albumin, and the morphology of cultured MC3T3-E1 preosteoblastic cells are investigated on three important biomaterial surfaces: silicon oxide, poly(styrene) (PS), and hydroxyapatite (HA). Using quartz crystal microgravimetry with dissipation analysis, the adsorbed protein composition and mechanics are determined. Interestingly, cell morphological changes correlate neither with the amount of Fn nor the rigidity of the protein layer. On the PS surface, Alb is seen to significantly diminish cell spreading, possibly due to Alb aggregation with a partially denatured initially placed Fn layer. HA appears to be a particularly favorable substrate for osteoblast adhesion, despite having low Fn adsorption and protein layer rigidity.

Nanomechanical properties of hybrid coatings for bone tissue engineering

Bone tissue engineering has emerged as a promising alternative approach in the treatment of bone injuries and defects arising from malformation, osteoporosis, and tumours. In this approach, a temporary scaffold possessing mechanical properties resembling those of natural bone is needed to serve as a substrate enhancing cell adhesion and growth, and a physical support to guide the formation of the new bone. In this regard, the scaffold should be biocompatible, biodegradable, malleable and mechanically strong. Herein, we investigate the mechanical properties of three coatings of different chemical compositions onto silanized glass substrates; a hybrid material consisting of methacryloxypropyl trimethoxysilane and zirconium propoxide, a type of a hybrid organic–inorganic material of the above containing also 50mol% 2-(dimethylamino)ethyl methacrylate (DMAEMA) moieties and a pure organic material, based on PDMAEMA. This study investigates the variations in the measured hardness and reduced modulus values, wear resistance and plastic behaviour before and after samples' submersion in cell culture medium. Through this analysis we aim to explain how hybrid materials behave under applied stresses (pile-up formations), how water uptake changes this behaviour, and estimate how these materials will react while interaction with cells in tissue engineering applications. Finally, we report on the pre-osteoblastic cell adhesion and proliferation on three-dimensional structures of the hybrid materials within the first hour and up to 7 days in culture. It was evident that hybrid structure, consisting of 50mol% organic–inorganic material, reveals good mechanical behaviour, wear resistance and cell adhesion and proliferation, suggesting a possible candidate in bone tissue engineering.

On the biodegradability, mechanical behavior, and cytocompatibility of amorphous Mg72Zn23Ca5 and crystalline Mg70Zn23Ca5Pd2 alloys as temporary implant materials

The evolution of microstructure and mechanical properties of almost fully amorphous Mg(72) Zn(23) Ca(5) and crystalline Mg(70) Zn(23) Ca(5) Pd(2) alloys during immersion in Hank's balanced salt solution (HBSS), as well as their cytocompatibility, are investigated in order to assess the feasibility of both materials as biodegradable implants. Though the crystalline Mg(70) Zn(23) Ca(5) Pd(2) sample shows lower wettability and more positive corrosion potential, this sample degrades much faster upon incubation in HBSS as a consequence of the formation of micro-galvanic couples between the nobler Pd-rich dendrites and the surrounding phases. After 22-h immersion, the concentration of Mg ions in the HBSS medium containing the Mg(70) Zn(23) Ca(5) Pd(2) sample is six times larger than for Mg(72) Zn(23) Ca(5) . Due to the Zn enrichment and the incipient porosity, the mechanical properties of the Mg(72) Zn(23) Ca(5) sample improve within the first stages of biodegradation (i.e., hardness increases while the Young's modulus decreases, thus rendering an enhanced wear resistance). Cytocompatibility studies reveal that neither Mg(72) Zn(23) Ca(5) nor Mg(70) Zn(23) Ca(5) Pd(2) are cytotoxic, although preosteoblast cell adhesion is to some extent precluded, particularly onto the surface of Mg(70) Zn(23) Ca(5) Pd(2) , because of the relatively high hydrophobicity. Because of their outstanding properties and their time-evolution, the use of the Pd-free alloy in temporary implants such as screws, stents, and sutures is envisioned.

Surface modifications by gas plasma control osteogenic differentiation of MC3T3-E1 cells

Numerous studies have shown that the physicochemical properties of biomaterials can control cell activity. Cell adhesion, proliferation, differentiation as well as tissue formation in vivo can be tuned by properties such as the porosity, surface micro- and nanoscale topography and chemical composition of biomaterials. This concept is very appealing for tissue engineering since instructive properties in bioactive materials can be more economical and time efficient than traditional strategies of cell pre-differentiation in vitro prior to implantation. The biomaterial surface, which is easy to modify due to its accessibility, may provide the necessary signals to elicit a certain cellular behavior. Here, we used gas plasma technology at atmospheric pressure to modify the physicochemical properties of polylactic acid and analyzed how this influenced pre-osteoblast proliferation and differentiation. Tetramethylsilane and 3-aminopropyl-trimethoxysilane with helium as a carrier gas or a mixture of nitrogen and hydrogen were discharged to polylactic acid discs to create different surface chemical compositions, hydrophobicity and microscale topographies. Such modifications influenced protein adsorption and pre-osteoblast cell adhesion, proliferation and osteogenic differentiation. Furthermore polylactic acid treated with tetramethylsilane enhanced osteogenic differentiation compared to the other surfaces. This promising surface modification could be further explored for potential development of bone graft substitutes.

The effect of unlocking RGD-motifs in collagen I on pre-osteoblast adhesion and differentiation

Denaturation of extracellular matrix proteins exposes cryptic binding sites. It is hypothesized that binding of cell adhesion receptors to these cryptic binding sites regulates cellular behaviour during tissue repair and regeneration. To test this hypothesis, we quantify the adhesion of pre-osteoblastic cells to native (Col) and partially-denatured (pdCol) collagen I using single-cell force spectroscopy. During early stages of cell attachment (≤180 s) pre-osteoblasts (MC3T3-E1) adhered significantly stronger to pdCol compared to Col. RGD (Arg-Gly-Asp)-containing peptides suppressed this elevated cell adhesion. We show that the RGD-binding α 5β 1- and α v-integrins mediated pre-osteoblast adhesion to pdCol, but not to Col. On pdCol pre-osteoblasts had a higher focal adhesion kinase tyrosine-phosphorylation level that correlated with enhanced spreading and motility. Moreover, pre-osteoblasts cultured on pdCol showed a pronounced matrix mineralization activity. Our data suggest that partially-denatured collagen exposes RGD-motifs that trigger binding of α 5β 1- and α v-integrins. These integrins initiate cellular processes that stimulate osteoblast adhesion, spreading, motility and differentiation. Taken together, these quantitative insights reveal an approach for the development of alternative collagen I- based surfaces for tissue engineering applications.

In vitro degradation, hemolysis and MC3T3-E1 cell adhesion of biodegradable Mg–Zn alloy

In this study a kind of patent binary Mg–6 wt.%Zn magnesium alloy was investigated as degradable biomedical material. The results of in vitro degradation including electrochemical measurements and immersion tests in simulated body fluid (SBF) revealed that zinc could elevate both the corrosion potential and Faraday charge transfer resistance of magnesium and thus improve the corrosion resistance. XRD and EDS analysis proved that the corrosion products on the surface of Mg–Zn contained hydroxyapatite (HA), Mg(OH) 2 and other Mg/Ca phosphates, which could reduce the degradation rate. The degradation process of magnesium alloy and the mechanism of corrosion layer formation were also discussed in this work, i.e. the byproducts of degradation of magnesium, Mg 2+ and OH −, reacted with the phosphate and Ca 2+ in the SBF, thus the corrosion layer containing HA, Mg(OH) 2 and other magnesium-substituted apatite precipitated in corrosion pits and covered the surface of magnesium alloy. The hemolysis test found that the hemolysis rate of Mg–Zn was 3.4%, which is lower than the safe value of 5% according to ISO 10993-4. For the cell culture experiments, after 2 h incubation the pre-osteoblastic cell MC3T3-E1 was able to adhere and spread on the corrosion layer of Mg–Zn alloy, indicating that despite the fluctuation of pH value of DMEM culture solution, Mg–Zn alloy could still support the earlier adhesion of pre-osteoblastic cells on the surface. Hemolysis and adhesion of cells display good biocompatibility of Mg–Zn alloy in vitro.