Concentrated Simulated Body Fluid Research Articles

A Study on Tailoring Silver Release from Micro‐Arc Oxidation Coating Fabricated on Titanium

This study is initiated with the aim of regulating the release of silver (Ag) as an antibacterial agent from the micro‐arc oxidation (MAO) coating. Herein, an external 5 wt% Tin(II) chloride (SnCl2) containing biodegradable polycaprolactone (PCL) layer is formed on the 0.8 wt% Ag‐incorporated MAO coating by the dip coating method. 5 wt% SnCl2 addition into PCL provides a steady release of Ag into concentrated simulated body fluid (1.5X SBF) from the underlying MAO coating at 37 °C. When the Ag release rate is taken into consideration, it is quantified as 0.0089 and 0.0586 ppm day−1 for PCL‐covered MAO and PCL‐free MAO coatings, respectively. It is finally concluded that the preliminary result of this study can be promising for minimizing the in vivo adverse effects of Ag+ ions arising from rapid release as well as maintaining antibacterial efficacy for prolonged periods, which is ideal for preventing the risk of postimplantation infections.

Towards bone regeneration: Understanding the nucleating ability of proline-rich peptides in biomineralisation

Obtaining rapid mineralisation is a challenge in current bone graft materials, which has been attributed to the difficulty of guiding the biological processes towards osteogenesis. Amelogenin, a key protein in enamel formation, inspired the design of two intrinsically disordered peptides (P2 and P6) that enhance in vivo bone formation, but the process is not fully understood. In this study, we have elucidated the mechanism by which these peptides induce improved mineralisation. Our molecular dynamics analysis demonstrated that in an aqueous environment, P2 and P6 fold to interact with the surrounding Ca2+, PO43− and OH− ions, which can lead to apatite nucleation. Although P2 has a less stable backbone, it folds to a stable structure that allows for the nucleation of larger calcium phosphate aggregates than P6. These results were validated experimentally in a concentrated simulated body fluid solution, where the peptide solutions accelerated the mineralisation process compared to the control and yielded mineral structures mimicking the amorphous calcium phosphate crystals that can be found in lamella bone. A pH drop for the peptide groups suggests depletion of calcium and phosphate, a prerequisite for intrinsic osteoinduction, while S/TEM and SEM suggested that the peptide regulated the mineral nucleation into lamella flakes. Evidently, the peptides accelerate and guide mineral formation, elucidating the mechanism for how these peptides can improve the efficacy of P2 or P6 containing devices for bone regeneration. The work also demonstrates how experimental mineralisation study coupled with molecular dynamics is a valid method for understanding and predicting in vivo performance prior to animal trials.

Calcification alters the viscoelastic properties of tendon fascicle bundles depending on matrix content.

Tendon fascicle bundles are often used as biological grafts and thus must meet certain quality requirements, such as excluding calcification, which alters the biomechanical properties of soft tissues. In this work, we investigate the influence of early-stage calcification on the mechanical and structural properties of tendon fascicle bundles with varying matrix content. The calcification process was modeled using sample incubation in concentrated simulated body fluid. Mechanical and structural properties were investigated using uniaxial tests with relaxation periods, dynamic mechanical analysis, as well as magnetic resonance imaging and atomic force microscopy. Mechanical tests showed that the initial phase of calcification causes an increase in the elasticity, storage, and loss modulus, as well as a drop in the normalized value of hysteresis. Further calcification of the samples results in decreased modulus of elasticity and a slight increase in the normalized value of hysteresis. Analysis via MRI and scanning electron microscopy showed that incubation alters fibrillar relationships within the tendon structure and the flow of body fluids. In the initial stage of calcification, calcium phosphate crystals are barely visible; however, extending the incubation time for the next 14 days results in the appearance of calcium phosphate crystals within the tendon structure and leads to damage in its structure. Our results show that the calcification process modifies the collagen-matrix relationships and leads to a change in their mechanical properties. These findings will help to understand the pathogenesis of clinical conditions caused by calcification process, leading to the development of effective treatments for these conditions. STATEMENT OF SIGNIFICANCE: This study investigates how calcium mineral deposition in tendons affects their mechanical response and which processes are responsible for this phenomenon. By analyzing the elastic and viscoelastic properties of animal fascicle bundles affected by calcification induced via incubation in concentrated simulated body fluid, the study sheds light on the relationship between structural and biochemical changes in tendons and their altered mechanical response. This understanding is crucial for optimizing tendinopathy treatment and preventing tendon injury. The findings provide insights into the calcification pathway and its resulting changes in the biomechanical behaviors of affected tendons, which have been previously unclear.

Bioactive calcium phosphate foam ceramics modified by biomimetic apatite

By combining the method of replication of polyurethane foam matrices at 1200 °C and modification in model SBF (Simulated Body Fluid) solutions of various compositions, open-pore calcium phosphate foam ceramics with a porosity of 53-60 % was obtained. The architecture and morphology of the calcium phosphate foam ceramics surface was formed by using polyurethane foam matrices («Granufoam», «STR») with different porosity and quantity of open pores. Modification of the calcium phosphate foam ceramics in SBF solutions of various compositions leads to a slight decrease in porosity to 3 %, which indicates the formation of an ultrathin apatite layer. The calcium phosphate-modified foam ceramics consisted of β-tricalcium phosphate, β-calcium pyrophosphate, α-tricalcium phosphate, and biomimetic apatite. In the standard SBF solution, the formation of apatite on calcium phosphate foam ceramics occurs slowly (14-56 days) and the strength increases by a factor of 2 as compared to the initial one. Soaking of calcium phosphate foam ceramics in SBF without HCO3- leads to the formation of biomimetic apatite with inclusions of calcium chloride dihydrophosphate in spherulites. Modification in a 5-fold concentrated SBF solution for 3-5 days at 37 °C makes it possible to form 6-10 times more biomimetic apatite compared to standard SBF with a 2.5-fold increase in static strength to 0.05 MPa. It has been established that at 800 °C biomimetic apatite crystallizes into β- tricalcium phosphate.

Surface Functionalization of Three Dimensional-Printed Polycaprolactone-Bioactive Glass Scaffolds by Grafting GelMA Under UV Irradiation

In this study, bioactive glass nanoparticles (BGNPs) with an average diameter of less than 10 nm were synthesized using a sol-gel method and then characterized by transmission electron microscopy (TEM), differential scanning calorimetric (DSC), Fourier transforms infrared spectroscopy (FTIR), and x-ray spectroscopy (XRD). Afterward, three dimensional (3D)-printed polycaprolactone (PCL) scaffolds along with fused deposition modeling (FDM) were incorporated with BGNPs, and the surface of the composite constructs was then functionalized by coating with the gelatin methacryloyl (GelMA) under UV irradiation. Field emission scanning electron microscopy micrographs demonstrated the interconnected porous microstructure with an average pore diameter of 260 µm and homogeneous distribution of BGNPs. Therefore, no noticeable shrinkage was observed in 3D-printed scaffolds compared with the computer-designed file. Besides, the surface was uniformly covered by GelMA, and no effect of surface modification was observed on the microstructure while surface roughness increased. The addition of the BGNPs the to PCL scaffolds showed a slight change in pore size and porosity; however, it increased surface roughness. According to mechanical analysis, the compression strength of the scaffolds was increased by the BGNPs addition and surface modification. Also, a reduction was observed in the absorption capacity and biodegradation of scaffolds in phosphate-buffered saline media after the incorporation of BGNPs, while the presence of the GelMA layer increased the swelling potential and stability of the composite matrixes. Moreover, the capability of inducing bio-mineralization of hydroxyapatite-like layers, as a function of BGNPs content, was proven by FE-SEM micrographs, EDX spectra, and x-ray diffraction spectra (XRD) after soaking the obtained samples in concentrated simulated body fluid. A higher potential of the modified constructs to interact with the aqueous media led to better precipitation of minerals. According to in-vitro assays, the modified scaffolds can provide a suitable surface for the attachment and spreading of the bone marrow mesenchymal stem cells (BMSCs). Furthermore, the number of the proliferated cells confirms the biocompatibility of the scaffolds, especially after a modification process. Cell differentiation was verified by alkaline phosphatase activity as well as the expression of osteogenic genes such as osteocalcin and osteopontin. Accordingly, the scaffolds showed an initial potential for reconstruction of the injured bone.

Biomimetic in situ precipitation of calcium phosphate containing silver nanoparticles on zirconia ceramic materials for surface functionalization in terms of antimicrobial and osteoconductive properties

ObjectiveZirconia is commonly used for manufacturing of dental implants thanks to its excellent mechanical, biological and aesthetic properties. However, its bioinertness inhibits bonding with the surrounding hard tissue and other surface interactions. In our study, we present a method for multifunctionalization of zirconia surface to improve its osseointegration and to minimize the infection risks. MethodsFor this reason, we introduced antibacterial and bioactive properties to zirconia surfaces by calcium phosphate biomimetic coating. The samples were incubated in vials in horizontal and vertical position in concentrated simulated body fluid (SBF) containing 0.1, 0.5, and 3 g/L of silver nanoparticles (Ag-NPs) and then were tested for their structure, surface properties, cytocompatibility and antibacterial properties. Results and SignificanceThe results demonstrated that our method is suitable to introduce Ag-NPs at different concentrations into the calcium phosphate layer, i.e. from 0.05–26.6 atom% as shown by EDX. According to the results of CFU-assay these coatings exhibited antibacterial properties against S. aureus and E. coli in correlation with the concentration of Ag-NP. The potential cytotoxicity of the coated samples was determined by AlamarBlue® assay and live/dead staining of MG63 osteoblast-like cells in direct contact and by testing the extracts from the materials. Only samples containing 0.05 atom% Ag-NPs, i.e. incubated in vertical position at SBF with 0.01 g/L Ag-NPs, were found cytocompatible in direct contact with MG63 cells. On the contrary in the indirect tests, the extracts from all the materials were found cytocompatible. This method could allow developing the completely new material group, exhibiting not only one but several biological properties, which can improve osseointegration and minimize infection risks.

The effect of biomimetic coating and cuttlebone microparticle reinforcement on the osteoconductive properties of cellulose-based scaffolds

Polymer-based scaffolds have already gained popularity in many biomedical applications due to convenient routes for fabrication and favourable structural, physicochemical and functional characteristics. However, polymeric scaffolds lack osteoconductivity and some synthetic polymers carry the risk of inflammatory response caused by degradation by-products. Those facts limit their practical use in bone tissue engineering. In this study, three-dimensional (3D) porous scaffolds from naturally derived polymer, namely regenerated cellulose, were prepared using a non-hydrolytic sol-gel and lyophilization techniques. To induce osteoconductive properties of the polymeric scaffolds, cuttlebone microparticles were immobilized and the surface coating was achieved via in vitro mineralization using 10-fold concentrated simulated body fluid (10x SBF). Biogenic activity of cuttlebone is explained by its chemical composition, which includes polysaccharide β-chitin and macro-, micro- and trace elements favourable for mineralization. Parallel the scaffolds were examined during long-term (24 weeks) in vitro mineralization in 1x SBF for the purpose to investigate apatite-forming ability of the scaffolds. A nice cauliflower-like structures and needle-like dents of the spherical aggregates, which are characteristic to hydroxyapatite precursors, were observed on the surface of cellulose/cuttlebone scaffolds by SEM. 10x SBF coating enhanced cell attachment to the scaffolds because SBF elements are known to increase bioactivity by inducing re-deposition of carbonate apatite crystallites on scaffold surface. Additionally, calcium and phosphate depositions were clearly observed on the developed scaffolds using von Kossa and Alizarin Red S staining. Proliferative and osteoconductive effects on the osteoblast-like MG-63 cells demonstrate the cellulose/cuttlebone scaffolds soaked in 10x SBF as a favourable material for bone tissue engineering.

In vitro and in vivo assessment of glucose cross-linked gelatin/zein nanofibrous scaffolds for cranial bone defects regeneration.

The purpose of this study was to evaluate the glucose cross-linked gelatin/zein scaffolds for bone regeneration in vitro and in vivo. The nanofibrous scaffolds exhibited fast mineralization in the concentrated simulated body fluid with the deposited octacalcium phosphate and dicalcium phosphate dehydrate. The nanofibrous scaffolds exhibited no cytotoxic effect on MC3T3e1 cells in a CCK-8 test. Additionally, scanning electron microscope and confocal laser scanning microscopy images revealed that all the scaffolds were biocompatible and showed excellent support for MC3T3e1 cells. In the osteogenesis characterizations, Alizarin Red staining experiments indicated the improved calcium deposits on the cross-linked scaffolds, while the alkaline phosphatase activity showed no difference. Furthermore, the in vivo cranial bone regeneration results suggested that the cross-linked gelatin/zein scaffolds presented a strong positive effect on the cranial bone regeneration with the increased new bone volume and connective tissue formation, but the incorporation of zein in the gelatin scaffolds did not favor the bone regeneration. Moreover, the cross-linked gelatin scaffold retarded the bone resorption as indicated by the higher levels of IFN-γ and lower levels of IL-6, which restricted the differentiation of osteoclasts.

Anionic Polymer Brushes for Biomimetic Calcium Phosphate Mineralization-A Surface with Application Potential in Biomaterials.

This article describes the synthesis of anionic polymer brushes and their mineralization with calcium phosphate. The brushes are based on poly(3-sulfopropyl methacrylate potassium salt) providing a highly charged polymer brush surface. Homogeneous brushes with reproducible thicknesses are obtained via surface-initiated atom transfer radical polymerization. Mineralization with doubly concentrated simulated body fluid yields polymer/inorganic hybrid films containing AB-Type carbonated hydroxyapatite (CHAP), a material resembling the inorganic component of bone. Moreover, growth experiments using Dictyostelium discoideum amoebae demonstrate that the mineral-free and the mineral-containing polymer brushes have a good biocompatibility suggesting their use as biocompatible surfaces in implantology or related fields.

Studying Biomimetic Coated Niobium as an Alternative Dental Implant Material to Titanium (in vitro and in vivo study)

Commercially pure titanium (cpTi) is widely used as dental implant material although it was found that titanium exhibited high modulus of elasticity and the lower corrosion tendency in oral environment. Niobium(Nb) was chosen for this study as an alternative to cpTi implant material due to its bioinert behavior and good elastic modulus and moderate cost in addition to corrosion resistance. This study was done to evaluate the effect of biomimetic coating on the surface properties of the commercially pure titanium and niobium implants by in vitro and in vivo experiments. The in vitro study was involved etching the samples of each material in HCl then soaking in 10M NaOH aqueous solution. These samples were then immersed in a 5 times concentrated simulated body fluid for 14 days. Scanning Electron Microscope, Energy Dispersive X-ray, and X-Ray Diffraction tests were done to analyze surface changes. The in vivo study was done by the implantation of screw-shaped implants (two from each material, uncoated and the other was biomimetically coated) in the tibias of New Zealand rabbits. After 2 and 4 weeks of healing period, 20 rabbits were sacrificed for each period. A removal torque was done for ten animals in each group, whereas the other ten were used for histological testing and histomorphometric analysis with optical microscope.The in vitro experiments showed that the use of 14 days immersion in a concentrated simulated body fluid produced a layer of calcium phosphate on metal surfaces. The removal torque values and new bone formation were increased significantly in Nb than Ti, in coated than uncoated screws, and in 4 weeks than 2 weeks healing periods. The Nb implants had better biomechanical and biological properties than the commercially pure titanium implants and can be used as an alternative dental implant.

Studying Biomimetic Coated Niobium as an Alternative Dental Implant Material to Titanium (in vitro and in vivo study)

Commercially pure titanium (cpTi) is widely used as dental implant material although it was found that titanium exhibited high modulus of elasticity and the lower corrosion tendency in oral environment. Niobium(Nb) was chosen for this study as an alternative to cpTi implant material due to its bioinert behavior and good elastic modulus and moderate cost in addition to corrosion resistance. This study was done to evaluate the effect of biomimetic coating on the surface properties of the commercially pure titanium and niobium implants by in vitro and in vivo experiments. The in vitro study was involved etching the samples of each material in HCl then soaking in 10M NaOH aqueous solution. These samples were then immersed in a 5 times concentrated simulated body fluid for 14 days. Scanning Electron Microscope, Energy Dispersive X-ray, and X-Ray Diffraction tests were done to analyze surface changes. The in vivo study was done by the implantation of screw-shaped implants (two from each material, uncoated and the other was biomimetically coated) in the tibias of New Zealand rabbits. After 2 and 4 weeks of healing period, 20 rabbits were sacrificed for each period. A removal torque was done for ten animals in each group, whereas the other ten were used for histological testing and histomorphometric analysis with optical microscope.The in vitro experiments showed that the use of 14 days immersion in a concentrated simulated body fluid produced a layer of calcium phosphate on metal surfaces. The removal torque values and new bone formation were increased significantly in Nb than Ti, in coated than uncoated screws, and in 4 weeks than 2 weeks healing periods. The Nb implants had better biomechanical and biological properties than the commercially pure titanium implants and can be used as an alternative dental implant.

Biomimetic surface functionalization of clinically relevant metals used as orthopaedic and dental implants

Titanium and its alloys or tantalum (Ta) are materials used in orthopaedic and dental implants due to their excellent mechanical properties and biocompatibility. However, their bioactivity and osteoconductivity is low. With a view to improving the bioactivity of these materials we hypothesised that the surface of Ta and TiAl6V4 can be functionalised with biomimetic, amorphous nano-sized calcium phosphate (CaP) apatite-like deposits, instead of creating uniform coatings, which can lead to flaking, delamination and poor adherence. We used Ta and TiAl6V4 metal discs with smooth and rough surfaces. Amorphous CaP apatite-like particles were deposited on the different surfaces by a biomimetic rapid two-step soaking method using concentrated simulated body fluid (SBF) solutions without a pre-treatment of the metal surfaces to induce CaP deposition. Immersion times in the second SBF solution of 48 and 18 h for Ta and TiAl6V4 respectively produced CaP deposits composed of amorphous globular nano-sized particles that also contained Mg, C and O. Longer immersion times produced more uniform coatings as well as an undesired calcite mineral phase. Prediction of in vivo behaviour by immersion in regular SBF showed that the obtained CaP deposits would act as a catalyst to rapidly form a Ca deficient CaP layer that also incorporates Mg. The amorphous CaP apatite-like deposits promoted initial attachment, proliferation and osteogenic differentiation of bone marrow derived mesenchymal stem cells. Finally, we used our method to functionalise 3D porous structures of titanium alloy made by selective laser sintering. Our study uses a novel and cost-effective approach to functionalise clinically relevant metal surfaces in order to increase the bioactivity of these materials, which could improve their clinical performance.

Biomimetic Coating of Hydroxyapatite on Glycerol Phosphate-Conjugated Polyurethane via Mineralization.

In this study, glycerol phosphate was introduced into polyurethane (PU) to promote the coating stability of hydroxyapatite (HA) during its mineralization on the PU surface. Glycerol phosphate was successfully conjugated with the PU chain during polymerization. Phosphate groups in glycerol phosphate accelerated the nucleation of HA under calcium phosphate ion-rich conditions (concentrated simulated body fluid), resulting in the enhancement of structural stability. The robust interface between HA and PU also improved mechanical properties. Hydrophilic phosphate groups and bioactive HA improved in vitro cellular responses in terms of the attachment and proliferation of L929 fibroblasts and MC3T3-E1 preosteoblasts. Thus, the highly elastic and bioactive PU-gp-HA could be a promising candidate for tissue engineering applications that experience frequent deformation, including diverse cartilage replacements.

Histological evaluation of bone-implant interface associated with biomimetic coated commercially pure titanium and Ti-6Al-7Nb alloy implants

Background: The clinical success of implants has been achieved not only because of the mechanical strength or excellent biocompatibility of the implant material but also because of other characteristics such as surface properties. In am attempt to modify the surface properties of an implant, this study was conducted to evaluate the effect of biomimetic calcium phosphate coating on the bone-implant interface of screw-shaped implants made from the commercially pure titanium and Ti-6Al-7Nb alloy by histological analysis with optical microscope. Materials and methods: The screws were biomimetically coated with calcium phosphate by immersion in a concentrated simulated body fluid (5 times), which simulates the inorganic part of human blood plasma, under static conditions in a biological thermostat at 37oC for 6 days. The uncoated screws were passivated with 28% nitric acid. The tibias of 15 white New Zealand rabbits were chosen as implantation sites for 4 implants (two from each material, one is uncoated and the other is biomimetically coated). After 2, 6, and 18 weeks of healing period, 5 rabbits were sacrificed for each period. The influence of modified surface on the bone-implant interaction was analyzed by histological testing with optical microscope. Results: The results obtained from this experiment revealed that the quality of bone response was improved among the coated screws. Furthermore, the coated Ti-6Al-7Nb alloy implants had better properties than the coated commercially pure titanium implants. Conclusion: Biomimetic coatings play an active role in the bone-remodeling process by creating a friendly surface for cell adhesion and proliferation, which is a key issue for bone regeneration.

Bioactive cellulose grafted soy protein isolate towards biomimetic calcium phosphate mineralization

The current study is a new approach to prepare an environmentally friendly and bioactive material from cellulose grafted soy protein isolate (SPI) for biomimetic calcium phosphate mineralization. Neat cellulose was oxidized using periodate then chemically modified by reacting with SPI followed by soaking in doubly concentrated simulated body fluid (2xSBF) solution. Scanning electron microscopy/energy dispersive X-ray spectroscopy, X-ray diffraction and transmission electron microscopy suggested the formation of uniform hydroxyapatite rod-like nanocrystals with ∼50nm diameter. The chemical composition of the prepared materials was further investigated using infrared spectroscopy and thermogravimetric analysis. The cytotoxicity of cellulose/SPI/calcium phosphate hybrid was evaluated using animal fibroblast baby hamster kidney cells (BHK-21). The cytotoxicity results suggested cellulose/SPI/calcium phosphate hybrid as potentially useful scaffold for regenerative therapies. The current article suggests a green strategy for preparing a new biohybrid material for tissue engineering.

Electrochemical corrosion of Ti6Al4V, Ti and AISI 316L SS after immersed in concentrated simulated body fluid

The biomimetic method is used to obtain hydroxyapatite (HAP) coatings on Ti6Al4V, Ti and AISI 316L SS substrates. These substrates with different pretreatment surface operations (HNO3, anodic polarization, base-acid) were immersed in concentrated simulated body fluids (SBF) for different days at physiologic conditions of 37°C, initial pH of 7.4. Then the corrosion behaviours of substrates after immersion in concentrated SBF were examined by electrochemical methods in Ringer’s and 0.9 wt% NaCl solutions at a temperature of 37°C. Ions concentrations and pH analyses were carried out after incubation in concentrated SBF. After immersion in SBF for different days, the surface morphology remains almost unchanged and no apatite formation is observed. Corrosion currents of substrates increased after immersion. Ions concentrations and pH values were shown variability according to soaking time and pretreatment surface operations.

Hydroxyapatite-hybridized chitosan/chitin whisker bionanocomposite fibers for bone tissue engineering applications

Biomimetic nanofibrous scaffolds derived from natural biopolymers for bone tissue engineering applications require good mechanical and biological performances including biomineralization. The present work proposes the utility of chitin whisker (CTWK) to enhance mechanical properties of chitosan/poly(vinyl alcohol) (CS/PVA) nanofibers and to offer osteoblast cell growth with hydroxyapatite (HA) mineralization. By using diacid as a solvent, electrospun CS/PVA nanofibrous membranes containing CTWK can be easily obtained. The dimension stability of nanofibrous CS/PVA/CTWK bionanocomposite is further controlled by exposing to glutaraldehyde vapor. The nanofibrous membranes obtained allow mineralization of HA in concentrated simulated body fluid resulting in an improvement of Young’s modulus and tensile strength. The CTWK combined with HA in bionanocomposite is a key to promote osteoblast cell adhesion and proliferation. The present work, for the first time, demonstrates the use of CTWKs for bionanocomposite fibers of chitosan and its hydroxyapatite biomineralization with the function in osteoblast cell culture. These hydroxyapatite-hybridized CS/PVA/CTWK bionanocomposite fibers (CS/PVA/CTWK-HA) offer a great potential for bone tissue engineering applications.

Modified chitosan scaffolds: Proliferative, cytotoxic, apoptotic, and necrotic effects on Saos-2 cells and antimicrobial effect on Escherichia coli

Scaffolds used in tissue engineering applications should have high biocompatibility with minimum allergic, toxic, apoptotic, or necrotic effects on the growing cells and newly forming tissue and, if possible, have antimicrobial property to prevent infection at the host site. In this study, novel micro-fibrous chitosan scaffolds, having mineralized bioactive surface to enhance cell adhesion and a model antibiotic (gentamicin) to prevent bacterial attack, were prepared. The effects of the scaffolds on proliferation, viability, apoptosis, and necrosis of Saos-2 cells are reported for the first time. Wet spinning technique was used in the scaffold preparation and biomineralization was achieved by incubating them in five-time concentrated simulated body fluid for 2, 7, or 14 days (coded as CH-BM/2, CH-BM/7, and CH-BM/14, respectively). Gentamicin, an effectively used antibiotic in bone treatments, was loaded by vacuum-pressure cycle. Energy-dispersive X-ray results demonstrated that Ca/P ratio of the mineral phase varies depending on the incubation period. When the scaffolds were cultured with Saos-2 cells, cell adhesion and extracellular matrix formation occurred on all types of scaffolds. Alamar Blue cytotoxicity tests showed correlation among mineral concentration and cytotoxicity where CH-BM/2 had significantly more favorable properties. For all types of scaffolds, apoptosis and necrosis were less than 10%, meaning the samples are biocompatible. Gentamicin-loaded scaffolds showed high antimicrobial efficacy against Escherichia coli. The presence of mineral phase enhanced the adhesive capacity of cells and entrapment efficiency of antibiotic. These results suggest that the bioactive and antimicrobial scaffolds prepared in this study can act as promising matrices in bone tissue engineering applications.

Biomimetic Hydroxyapatite Growth on Functionalized Surfaces of Ti-6Al-4V and Ti-Zr-Nb Alloys.

A biomimetic approach for coating titanium-containing alloys with hydroxyapatite (HA) is reported in the article. Two types of Ti-containing alloys were chosen as an object for coating: Ti-6Al-4V (recommended for orthopedic application) and a novel highly biocompatible Ti-Zr-Nb alloy, with good mechanical compatibility due to a modulus that is more close to that of human bones (E ≈ 50 GPa instead of 110 GPa in Ti-6Al-4V). Coating process was carried out in a 10×-concentrated simulated body fluid (SBF)—synthetic analog of human body plasma. The effect of oxidized and carboxylated alloy surface on formation of biomimetic hydroxyapatite has been studied. By XRD, we found influence of thermal conditions on HA crystal formation and size. SEM images and Fourier transform infrared confirmed that hydroxyapatite with different morphology, crystallinity, and Ca/P ratio formed on metallic surfaces. X-ray photoelectron spectroscopy showed that in the Ti-6AL-4V sample the observed Ca/P ratio reach 0.97, whereas in the Ti-Zr-Nb sample the observed Ca/P ratio reach 1.15.

Fabrication of biomimetic PCL scaffold using rapid prototyping for bone tissue engineering

We report the fabrication of a porous three-dimensional (3D) scaffold using a 3D plotting system, with applications in bone tissue engineering. Biomimetic surface coatings of hydroxyapatite were formed using concentrated simulated body fluid (SBF). The mineralized scaffold had a uniform interconnected porous structure. The apatite that was formed on the surface of the scaffold was characterized by using energy dispersive spectroscopy (EDS), X-ray powder diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The biological properties of the scaffold were evaluated using WST-1 assays of alkaline phosphatase (ALP) activity and gene expression of D1 mouse mesenchymal stem cells. The results show that the biomimetic scaffolds exhibited good apatite-forming properties, with favorable cell proliferation and differentiation for bone formation. Therefore, these biomimetic scaffolds with hydroxyapatite coatings have potential applications in bone regeneration.