Influence of Addition of Hyaluronic Acid (HA) on the Properties of Collagen/HA Composite Scaffolds

In order to develop a scaffolding material for tissue regeneration, porous matrices containing varying composites of collagen and hyaluronic acid (HA) (from 1:0 to 0:1) were fabricated using a freeze-drying method. The effect of the composition on the morphology, hydrophilicity, swelling behavior, mechanical properties, and in vitro cytotoxicity was investigated. The results showed that all the scaffolds had an interconnected pore structure with sufficient pore size for use as a support for the growth of fibroblasts. The addition of HA improved the swelling property, but reduced the compressive strength. The contact angle decreased with increasing HA content. In in vitro cytotoxicity tests using fibroblastic cells, the collagen/HA scaffolds showed no toxicity. All these results suggest that collagen/HA composite scaffolds are a potential candidate for tissue engineering scaffolds.     

Biopolymer-Hydroxyapatite Scaffolds for Advanced Prosthetics

Biocompatibility in research and development of advanced prosthetics is a current problem faced by medical researchers. A major challenge in tissue engineering is to find materials and processing techniques that allow them to produce extracellular matrices (ECM) mimicking scaffolds that promote cell growth and organization into a specific architecture, inducing cell differentiation and subsequent cell function. The ideal tissue repair material thus should consist of synthetic biomaterials, such as natural polymers mimicking the mechanical and biological functionality of the ECM. Cellulose acetate membranes were used as scaffolds for microvascular cell growth. Hydroxyapatite (HA) is a natural ceramic (responsible for strength and stability in the human skeletal system) operable as a biocomposite coating to improve the biocompatibility of implant substrates. In this work, HA was prepared from low cost natural calcium source — eggshells. Its structural properties were investigated by scanning (SEM), transmission (TEM) electron microscopies and Fourier Transformed Infrared spectroscopy (FT-IR). The composition analyses of HA were measured by the total reflection X-ray fluorescence spectrometer (TXRF) and by prompt gamma activation analysis (PGAA). Hydroxyapatite added biodegradable scaffolds have been prepared by electrospinning method to enhance biological functionality.     

Fabrication of polymer scaffolds for tissue engineering using surface selective laser sintering

A new approach to the fabrication of individual implants and scaffolds for tissue engineering—surface selective laser sintering (SSLS)—is proposed and realized. In contrast to the conventional selective laser sintering, the SSLS method makes it possible to sinter polymer microparticles and melt the near-surface layer rather than the microparticle as a whole. The effect of the laser radiation parameters and the structure and composition of the raw products on the structure and properties of the biomaterials sintered by the laser radiation is analyzed. This approach makes possible both the application of thermally unstable polymers (e.g., polylactides or polylactoglycolides) and the fabrication of scaffolds with incorporated bioactive proteins. The results obtained yield a regular physical basis for a new technology of the fabrication of various polymer scaffolds that represent important materials and elements of modern tissue engineering. The flexibility of the SSLS method is especially important at the stage of investigation of the cell and tissue responses needed for the optimization of the material for a specific application in tissue engineering.     

Designing the mechanical properties of peptide-based supramolecular hydrogels for biomedical applications

Hydrogels are a class of special materials that contain a large amount of water and behave like rubber.These materials have found broad applications in tissue engineering,cell culturing,regenerative medicine etc.Recently,the exploration of peptide-based supramolecular hydrogels has greatly expanded the repertoire of hydrogels suitable for biomedical applications.However,the mechanical properties of peptide-based hydrogels are intrinsically weak.Therefore,it is crucial to develop methods that can improve the mechanical stability of such peptide-based hydrogels.In this review,we explore the factors that determine or influence the mechanical stability of peptide-based hydrogels and summarize several key elements that may guide scientists to achieve mechanically improved hydrogels.In addition,we exemplified several methods that have been successfully developed to prepare hydrogels with enhanced mechanical stability.These mechanically strong peptide-based hydrogels may find broad applications as novel biomaterials.It is still challenging to engineer hydrogels in order to mimic the mechanical properties of biological tissues.More hydrogel materials with optimal mechanical properties suitable for various types of biological applications will be available in the near future.     

Ultrasound-assisted air-jet spinning of silk fibroin-soy protein nanofiber composite biomaterials

Ultrasound utilizes a non-radiation technology that can meet modern standards to gain access to cheap, reliable and sustainable modern energy. Ultrasound technology can be implemented in the field of biomaterials for its exceptional potential in controlling the shape of nanomaterials. This study presents the first example of the production of soy and silk fibroin protein composite nanofibers in various ratios via combining ultrasonic technology with air-spray spinning. Characterization of ultrasonic spun nanofibers was performed by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric (TG) analysis, water contact angle, water retention, enzymatic degradation, and cytotoxicity assays. The effects that adjustments on the ultrasonic time have on the surface morphology, structures, thermal properties, hydrophilicity, water-uptake, bio-enzyme degradability, mechanical properties, and cytocompatibility of the material were examined. It was discovered that as the sonication time increased from 0 to 180 min, the beading phenomenon disappeared, forming nanofibers with uniform diameter and porosity; while the content of β-sheet crystals in the composites and their thermal stability gradually increased, the materials glass transition temperature decreased, and preferred mechanical properties were obtained. Additional studies show that the hydrophilicity, water retention capacity and enzymatic degradation rate were also enhanced by ultrasound, providing a favorable environment for cell attachment and proliferation. This study highlights the experimental and theoretical methods for ultrasound assisted air-jet spinning of biopolymer nanofibrous materials with tunable properties and high biocompatibility, which provide a wide range of applications in wound dressings and drug-carrying systems. This work shows great potential for a direct road to sustainable development of protein based fibers in the industry, thus promoting economic growth, and improving the health of the general population and well-being of wounded patients worldwide.     

Preparation and Characterization of Poly(L-lactide-co-glycolide-co-ε-caprolactone)/Nano-biaoactive Glass-Nano-β-tricalcium Phosphate Composite Scaffolds

Abstract

Polymeric/ceramic composite scaffolds that are biocompatible and biodegradable are widely used for tissue engineering applications. In this work a series of poly(L-lactide-co-glycolide-co-ε-caprolactone)/nano-biaoactive glass-nano-β-tricalcium phosphate composite scaffolds were successfully fabricated and the influences of the inorganic content and freezing temperature on the physical properties were studied. The composite scaffolds with various inorganic contents showed an interconnected pore structure with irregular shapes. The composite scaffolds had a porosity that was reduced with increasing inorganic content and decreasing freezing temperature. The incorporation of inorganic fillers and decreasing freezing temperature improved the mechanical properties of the hybrid scaffolds. By appropriate control of these two factors (10.0?wt% content of NBAG and β-TCP with freezing at ?30?°C) a suitable composite scaffold was prepared as a potential bone tissue engineering implant.     

Ultrasound elastomicroscopy using water jet and osmosis loading: potentials for assessment for articular cartilage

Research in elasticity imaging typically relies on 1-10 MHz ultrasound. Elasticity imaging at these frequencies can provide strain maps with a resolution in the order of millimeters, but this is not sufficient for applications to skin, articular cartilage, or other fine structures. In this paper, we introduced two methods of ultrasound elastomicroscopy using water jet and osmosis loading for imaging the elasticity of biological soft tissues with high resolutions. In the first system, the specimens were compressed using water jet compression. A water jet was used to couple a focused 20 MHz ultrasound beam into the specimen and meanwhile served as a "soft" indenter. Because there was no additional attenuation when propagating from the ultrasound transducer to the specimen, the ultrasound signal with high signal-to-noise ratio could be collected from the specimens simultaneously with compressing process. The compression was achieved by adjusting the water flow. The pressure measured inside the water pipe and that on the specimen surface was calibrated. This system was easily to apply C-scan over sample surfaces. Experiments on the phantoms showed that this water jet indentation method was reliable to map the tissue stiffness distribution. Results of 1D and 2D scanning on phantoms with different stiffness are reported. In the second system, we used osmotic pressure caused by the ion concentration change in the bathing solutions for the articular cartilage to deform them. When bovine articular cartilage specimens were immerged in solutions with different salt concentration, a 50 MHz focused ultrasound beam was used to monitor the dynamic swelling or shrinkage process. Results showed that the system could reliably map the strain distribution induced by the osmotic loading. We extract intrinsic layered material parameters of the articular cartilage using a triphasic model. In addition to biological tissues, these systems have potential applications for the assessment of bioengineered tissues, biomaterials with fine structures, or some engineering materials. Further studies are necessary to fully realize the potentials of these two new methods.     

Influence of Nano-Bioactive Glass (NBG) Content on Properties of Gelatin-Hyaluronic Acid/NBG Composite Scaffolds

The development of three-dimensional (3-D) scaffolds with highly open porous structure is one of the most important issues in tissue engineering. A novel nanocomposite scaffold of gelatin (Gel), hyaluronic acid (HA), and nano-bioactive glass (NBG) was prepared by blending NBG with a Gel and HA solution followed by lyophilization. The effects of NBG content on the properties of the Gel-HA/NBG composite scaffolds, including the morphologies, porosity, compressive strength, swelling behavior, cell viability and alkaline phosphatase (ALP) activity, were investigated. Porous composite scaffolds with interconnected pores were obtained and the pores became cylindrical with increasing NBG content. The porosity percent and swelling ability decreased with increasing NBG content; however, the compressive strength, cell viability and ALP activity were enhanced. All the results showed the addition of NBG particles can improve the physicochemical and biological properties and the Gel-HA/NBG composite scaffolds exhibited good potential for tissue engineering applications.     

Piezoelectric Performance of Microcellular Polypropylene Foams Fabricated Using Foam Injection Molding as a Potential Scaffold for Bone Tissue Engineering

Abstract

Piezoelectric properties and adequate porosity play important roles in bone tissue engineering. In this paper we describe the fabrication of piezoelectric polypropylene (PP) foam using injection molding to be utilized as a potential cost-effective scaffold for bone tissue engineering. One-side mechanical skin removal from the foam was used to investigate the effect of the solid skin on the piezoelectric performance. The microcellular structure, relative density, crystalline structure, mechanical properties, piezoelectric properties under repeated impact pressure and biocompatibility of the scaffolds were investigated using scanning electron microscopy (SEM), water displacement method, differential scanning calorimetry (DSC), uniaxial tension tests, piezoelectric tests and MTT assays, respectively. Uniform spherical cells, with an average size of 75?µm and a density of 1.23?×?10⁶ cells/cm^?3, suitable for bone regeneration, were imaged by SEM. The DSC results showed β crystals formation in the PP foam during the foaming process which would be valuable for mechanical properties. The foaming process did not reduce the mechanical properties significantly. The foaming process promoted the piezoelectric responses by 134, 922, and 87%, respectively, for the PP samples with 3, 2 and 1?mm thickness. The biocompatibility test suggested improved cellular biocompatibility by foaming. Overall, the results demonstrated the development of a cost-effective scaffold for tissue engineering.     

The influence of polymer concentrations on the structure and mechanical properties of porous polycaprolactone-coated hydroxyapatite scaffolds

Polycaprolactone (PCL)-coated porous hydroxyapatite (HA) composite scaffolds were prepared by combining polymer impregnating method with dip-coating method. Three different PCL solution concentrations were used in dip-coating process to improve the mechanical properties of porous HA scaffolds. The results indicated that as the concentration of PCL solution increases the compressive strength significantly increased from 0.09 MPa to 0.51 MPa while the porosity decreased from 90% to 75% for the composite scaffolds. An interlaced structure was found inside the pore wall for all composite scaffolds due to the penetration of PCL. The porous HA/PCL composite scaffolds dip-coated with 10% PCL exhibited optimal combination of mechanical properties and pore interconnectivity, and may be a potential bone candidate for the tissue engineering applications.     

Poly (Vinyl Alcohol)/Chitosan/Akermanite Nanofibrous Scaffolds Prepared by Electrospinning

Abstract

Electrospinning, as an effective method for preparation of scaffolds for cell growth templates, has attracted great attention. In this study electrospinning was used to prepare poly (vinyl alcohol) (PVA)/chitosan scaffolds for bone tissue engineering. In order to improve the bioactivity and mechanical properties of the fibrous scaffolds, 0.5, 1 and 2?wt% akermanite, a calcium silicate based bioceramic, was added to the electrospinning solution. The morphology of the electrospun scaffolds was observed by using field emission-scanning electron microscopy and their mechanical strengths were analyzed by tension tests. The results showed that the formed scaffolds consisted of fibers with less than 100?nm diameter. In the case of the composite containing 1?wt% akermanite, the fibers were more homogeneous and no beads were formed during electrospinning, while in the composite containing 2?wt% akermanite a considerable number of beads were formed which we attribute to an improper viscosity of the electrospinning solution. Among the different compositions, the composite containing 1?wt% akermanite showed higher ultimate tensile strength (10.6?MPa) and fracture strain (9%). These values were increased by crosslinking the scaffold by reaction with glutaraldehyde, up to 13?MPa and 9.4%, respectively.     

Directing the phase behavior of polyelectrolyte complexes using chiral patterned peptides

Polyelectrolyte complexes (PECs) have a broad range of promising applications as soft materials due to their self-assembly and diversity of structure and chemical composition. Peptide polymer PECs are highly biocompatible and biodegradable, making them particularly useful for encapsulation of food additives and flavors, micellar drug delivery, medical and underwater adhesives, fetal membrane patches, and scaffolds for cell growth in tissue engineering. While parameters affecting PEC formation and stability in regards to charge effects are well researched, little is known about the effects of van der Waals interactions, hydrogen bonding, and secondary structure in these materials. Peptide chirality provides a unique opportunity to manipulate PEC phase to modulate the amount of solid-like (precipitate) or liquid-like (coacervate) character by influencing hydrogen bonding interactions among peptide chains. In previous work, we showed that chiral peptides form solid complexes, while complexes with even one racemic peptide were fluid. This raised the interesting question of how long a homochiral sequence must be to result in solid phase formation. In this work, we designed chiral patterned peptides of polyglutamic acid and polylysine ranging from 50 to 90% L-chiral residues with increasing numbers of sequential L-chiral residues before a chirality change. These polymers were mixed together to form PECs. We observed that 8 or more sequential L-chiral residues are necessary to achieve both the appearance of a precipitate phase and sustained β-sheets in the complex, as determined by optical imaging and FTIR Spectroscopy. Less homochiral content results in formation of a coacervate phase. Thus, we show that chiral sequence can be used to control the phase transition of PECs. Understanding how to manipulate PEC phase using chiral sequence as presented here may enable tuning of the material properties to achieve the desired mechanical strength for coatings and polymer brushes, or the most effective molecular release kinetics for drug delivery applications, for example.     

Preparation and Characterization of Chitosan-Gelatin/Glutaraldehyde Scaffolds

Chitosan-gelatin (CG) scaffolds were fabricated with glutaraldehyde as a cross-linker by vacuum freeze-drying. Mixtures of different volumes of chitosan and glutaraldehyde were considered. Morphology, porosity, density, and water absorbency of the scaffolds were studied. Both tensile and compressive properties of the scaffolds were tested. In addition, cellular adherence, proliferation, and morphology on the scaffolds were tested to evaluate the compatibility. It was found that porosity, density, water absorbency, and mechanical properties of CG scaffolds changed with the variation of chitosan or GA content. The adequate adherence, proliferation, and morphology of HaCaT type cells on the scaffolds showed that these scaffolds can be used as carriers for culturing HaCaT. The CG scaffolds, particularly those with chitosan-gelatin volume ratios of 1:1 and adding 6% or 8% volume of 0.25 wt% GA solution, were more suitable than the others through comparing the above properties and could be promising candidates for engineering skin tissue.     

Quantitative imaging of electrospun fibers by PeakForce Quantitative NanoMechanics atomic force microscopy using etched scanning probes

Electrospun polymeric submicron and nanofibers can be used as tissue engineering scaffolds in regenerative medicine. In physiological conditions fibers are subjected to stresses and strains from the surrounding biological environment. Such stresses can cause permanent deformation or even failure to their structure. Therefore, there is a growing necessity to characterize their mechanical properties, especially at the nanoscale.Atomic force microscopy is a powerful tool for the visualization and probing of selected mechanical properties of materials in biomedical sciences. Image resolution of atomic force microscopy techniques depends on the equipment quality and shape of the scanning probe. The probe radius and aspect ratio has huge impact on the quality of measurement.In the presented work the nanomechanical properties of four different polymer based electrospun fibers were tested using PeakForce Quantitative NanoMechanics atomic force microscopy, with standard and modified scanning probes. Standard, commercially available probes have been modified by etching using focused ion beam (FIB). Results have shown that modified probes can be used for mechanical properties mapping of biomaterial in the nanoscale, and generate nanomechanical information where conventional tips fail.     

Functionally graded PCL/β-TCP biocomposites in a multilayered structure for bone tissue regeneration

Functionally graded (FG) composites consisting of polycaprolactone (PCL) and beta-tricalcium phosphate (β-TCP) particles were fabricated with a multilayered structure using a melt plotter with a two-heating-barrel system. Using this process, the concentration of β-TCP particles varied in each layered strut. Scanning electron microscopy (SEM) and energy dispersive spectroscopy mapping of calcium on the fabricated scaffolds indicated that the β-TCP particles were well distributed in each PCL strut, according to conceptual design. By incorporating β-TCP, the FG-PCL/β-TCP scaffolds had meaningful increases in water absorption (30 % increase) and showed good mechanical properties, although the mechanical properties are slightly low compared to pure PCL/β-TCP composite. We performed biological assessments to evaluate the capability of these FG scaffolds to act as a biomaterial for bone tissue regeneration with osteoblast-like cells (MG63). SEM images of cell-seeded FG scaffolds showed that the concentrated β-TCP struts were affected as good cell attachment/proliferation sites. Additionally, calcium deposition on the FG scaffolds was higher than that of normal scaffolds after 14 days. In particular, we observed high levels of mineralization in the highly concentrated β-TCP struts in the FG scaffolds. Based on these results, we believe that the FG scaffolds having various spatially designed structures with graded properties will be widely applicable for hard tissue engineering applications.     

Ultrasound regulated flexible protein materials: Fabrication,structure and physical-biological properties

Ultrasound can be used in the biomaterial field due to its high efficiency, easy operation, no chemical treatment, repeatability and high level of control. In this work, we demonstrated that ultrasound is able to quickly regulate protein structure at the solution assembly stage to obtain the designed properties of protein-based materials. Silk fibroin proteins dissolved in a formic acid-CaCl₂ solution system were treated in an ultrasound with varying times and powers. By altering these variables, the silks physical properties and structures can be fine-tuned and the results were investigated with Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic mechanical analysis (DMA), gas permeability and water contact angle measurements. Ultrasonic treatment aids the interactions between the calcium ions and silk molecular chains which leads to increased amounts of intermolecular β-sheets and α-helix. This unique structural change caused the silk film to be highly insoluble in water while also inducing a hydrophilic swelling property. The ultrasound-regulated silk materials also showed higher thermal stability, better biocompatibility and breathability, and favorable mechanical strength and flexibility. It was also possible to tune the enzymatic degradation rate and biological response (cell growth and proliferation) of protein materials by changing ultrasound parameters. This study provides a unique physical and non-contact material processing method for the wide applications of protein-based biomaterials.     

Ultrasound impact on whey protein concentrate-pectin complexes and in the O/W emulsions with low oil soybean content stabilization

Consumers’ preference for products with reduced levels of fat increased in the last years. Proteins and polysaccharides have an important role due to their functional and interaction properties because, when combined in ratios and pH of higher potential for electrostatic interactions they may act as emulsifiers or stabilizers. This study evaluated the ultrasound impact on the electrostatic interaction between pectin (PEC) and whey protein concentrate (WPC) at different WPC:PEC ratios (1:1 to 5:1), and its effect on the emulsification and stability of emulsions formulated with WPC:PEC blends (1:1, 4:1) at low soybean oil contents (5 to 15%). Zeta potential analysis showed greater interactions between biopolymers at pH 3.5, which was proven in FTIR spectra. Rheology and turbidimetry showed that the ultrasound reduced the suspension viscosity and the size of the biopolymer complexes. Suspensions were Newtonian, whereas the emulsions showed shear-thinning behavior with slight increase in apparent viscosity as a function of oil content, and remained stable for seven days, with small droplets (<8 μm) stabilized and entrapped in a pectin network evidenced by confocal laser microscopy. Sonication was successfully applied to emulsion stabilization, improving the functional properties of WPC:PEC blends and enabling their application as low-fat systems, providing healthier products to consumers.     

Graphene and Graphene‐Based Materials in Biomedical Science

Graphene and its composite materials are very important in many disciplines of science and have been used enormously by researchers since their discovery in 2004. These are a new group of compounds, and are also wonderful model systems for quantum behavior studies. Their properties like exceptional conductivity, biocompatibility, surface area, mechanical strength, and thermal properties make them rising stars in the scientific community. Graphene and its composite compounds are utilized widely in different medical applications, for example, biosensing of biological compounds responsible for disease development, bioimaging of various cells, tissues, microorganisms, animal models, etc. In addition, they are used for enhancing and supporting the stem cell differentiation, i.e., regenerative medicine for regeneration studies of various human organs, tissue engineering in biology for the development of carrier materials, as well as in bone reformation. This review focuses on the modification procedure involved in the fabrication of graphene‐based biomaterials for various applications and recent developments in research related to graphene and graphene‐based materials in biosensing, optical sensing, gas sensing, drug, gene, protein delivery, tissue engineering, and bioimaging. In addition, the potential toxicological effects of graphene‐based biomaterials are discussed.     

A facile method to determine pore size distribution in porous scaffold by using image processing

Image processing permits scientists to investigate morphological properties of three-dimensional structures starting from their bi-dimensional gray-scale representation. In many cases porous structure with complex architecture has to be designed in order to attempt specific properties such in the case of scaffold for tissue engineering.Traditional morphological characterization, like scanning electron microscopy, should be coupled with quantitative information such as pore size distribution (PSD) in order to get a deeper understanding of the influence of the porous structure on tissue regeneration processes and on other related applications, it is remarkable to study a quantitative analysis of porosity and of pores dimension.In this work it was developed as a software able to accomplish the segmentation of images containing pores of any geometry in a semi-automatic way with the aim to measure the PSD. Case study constituted by PLA porous scaffolds with different pore size was adopted. Results indicate that image processing methods well fit the pore size features of PLA scaffolds, overcoming the limits of the more invasive porosimetry techniques.     

New bioactive hybrid material of nano-hydroxyapatite based on N-carboxyethylchitosan for bone tissue engineering

N-carboxyethylchitosan/nano-hydroxyapatite (NCECS/HA) composite films were fabricated and their potential applications in guiding bone regeneration were investigated in terms of their in vitro cellular activity. Fourier ransform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and thermogravimetric analysis (TGA) were used to investigate the structure and composition of the composite film. Field Emission scanning electron microscopy (FESEM) revealed that HA nanoparticles were dispersed homogeneously in NCECS matrix. The composite film has sufficient mechanical properties for tissue engineering scaffold. The composite film was found to have better cartilage cell adhesion and growth than pure NCECS film.