The Effect of Chitosan on Physicochemical Properties of Whey Protein Isolate Scaffolds for Tissue Engineering Applications.

New scaffolds, based on whey protein isolate (WPI) and chitosan (CS), have been proposed and investigated as possible materials for use in osteochondral tissue repair. Two types of WPI-based hydrogels modified by CS were prepared: CS powder was incorporated into WPI in either dissolved or suspended powder form. The optimal chemical composition of the resulting WPI/CS hydrogels was chosen based on the morphology, structural properties, chemical stability, swelling ratio, wettability, mechanical properties, bioactivity, and cytotoxicity evaluation. The hydrogels with CS incorporated in powder form exhibited superior mechanical properties and higher porosity, whereas those with CS incorporated after dissolution showed enhanced wettability, which decreased with increasing CS content. The introduction of CS powder into the WPI matrix promoted apatite formation, as confirmed by energy dispersive spectroscopy (EDS) and Fourier transform infrared spectroscopy (FTIR) analyses. In vitro cytotoxicity results confirmed the cytocompatibility of CS powder modified WPI hydrogels, suggesting their suitability as cell scaffolds. These findings demonstrate the promising potential of WPI/CS scaffolds for osteochondral tissue repair.

Preliminary Results on Heparin-Modified Double-Layered PCL and PLA-Based Scaffolds for Tissue Engineering of Small Blood Vessels.

Low-diameter blood vessels are challenging to replace with more traditional synthetic vascular grafts. Therefore, the obvious choice is to try to regenerate small veins and arteries through tissue-engineering approaches. However, the layered structure of native vessels and blood compatibility issues make this a very challenging task. The aim of this study is to create double-layered tubular scaffolds with enhanced anticoagulant properties for the tissue engineering of small blood vessels. The scaffolds were made of a polycaprolactone-based porous outer layer and a polylactide-based electrospun inner layer modified with heparin. The combination of thermally induced phase separation and electrospinning resulted in asymmetric scaffolds with improved mechanical properties. The release assay confirmed that heparin is released from the scaffolds. Additionally, anticoagulant activity was shown through APTT (activated partial thromboplastin time) assay. Interestingly, the endothelial cell culture test revealed that after 14 days of culture, HAECs (human aortic endothelial cell lines) tended to organize in chain-like structures, typical for early stages of vascular formation. In the longer culture, HAEC viability was higher for the heparin-modified scaffolds. The proposed scaffold design and composition have great potential for application in tissue engineering of small blood vessels.

Detection of disbonds in adhesively bonded aluminum plates using laser-generated shear acoustic waves.

Adhesively bonded metals are increasingly used in many industries. Inspecting these parts remains challenging for modern non-destructive testing techniques. Laser ultrasound (LU) has shown great potential in high-resolution imaging of carbon-reinforced composites. For metals, excitation of longitudinal waves is inefficient without surface ablation. However, shear waves can be efficiently generated in the thermo-elastic regime and used to image defects in metallic structures. Here we present a compact LU system consisting of a high repetition rate diode-pumped laser to excite shear waves and noncontact detection with a highly sensitive fiber optic Sagnac interferometer to inspect adhesively bonded aluminum plates. Multiphysics finite difference simulations are performed to optimize the measurement configuration. Damage detection is performed for a structure consisting of three aluminum plates bonded with an epoxy film. Defects are simulated by a thin Teflon film. It is shown that the proposed technique can efficiently localize defects in both adhesion layers.

Gradient Chitosan Hydrogels Modified with Graphene Derivatives and Hydroxyapatite: Physiochemical Properties and Initial Cytocompatibility Evaluation.

In this study, we investigated preparation of gradient chitosan-matrix hydrogels through a novel freezing-gelling-thawing method. The influence of three types of graphene family materials (GFM), i.e., graphene oxide (GO), reduced graphene oxide (rGO), and poly(ethylene glycol) grafted graphene oxide (GO-PEG), as well as hydroxyapatite (HAp) on the physicochemical and biological properties of the composite hydrogels was examined in view of their potential applicability as tissue engineering scaffolds. The substrates and the hydrogel samples were thoroughly characterized by X-ray photoelectron spectroscopy, X-ray diffractometry, infrared spectroscopy, digital and scanning electron microscopy, rheological and mechanical analysis, in vitro chemical stability and bioactivity assays, as well as initial cytocompatibility evaluation with human umbilical cord Wharton's jelly mesenchymal stem cells (hUC-MSCs). We followed the green-chemistry approach and avoided toxic cross-linking agents, using instead specific interactions of our polymer matrix with tannic acid, non-toxic physical cross-linker, and graphene derivatives. It was shown that the most promising are the gradient hydrogels modified with GO-PEG and HAp.

Polylactide/Hydroxyapatite Nonwovens Incorporated into Chitosan/Graphene Materials Hydrogels to Form Novel Hierarchical Scaffolds.

In this study, hierarchical, cylindrical scaffolds based on polylactide (PLA) microfibers incorporated into chitosan (CS) hydrogel were prepared for potential use in bone tissue engineering. PLA nonwovens modified with hydroxyapatite particles (HAp) were obtained using the electrospinning method. Then, three-dimensional scaffolds were created by rolling up the nonwovens and immersing them in CS-based solutions with graphene oxide (GO) or reduced graphene oxide (rGO) dispersed in the polymer matrix. Hydrogels were cross-linked using a novel freezing-thawing-gelling method. A broad spectrum of research methods was applied in order to thoroughly characterize both the nanofillers and the composite systems: scanning electron microscopy, X-ray photoelectron spectroscopy, X-ray diffractometry, attenuated total reflection Fourier transform infrared spectroscopy, rheological and mechanical testing, as well as the assessment of chemical stability, bioactivity and cytocompatibility.

Synthesis and Characterization of Chitosan/Reduced Graphene Oxide Hybrid Composites.

Graphene family materials (GFM) are currently considered to be one of the most interesting nanomaterials with a wide range of application. They can also be used as modifiers of polymer matrices to develop composite materials with favorable properties. In this study, hybrid nanocomposites based on chitosan (CS) and reduced graphene oxide (rGO) were fabricated for potential use in bone tissue engineering. CS/rGO hydrogels were prepared by simultaneous reduction and composite formation in acetic acid or lactic acid and crosslinked with a natural agent-tannic acid (TAc). A broad spectrum of research methods was applied in order to thoroughly characterize both the components and the composite systems, i.e., X-ray Photoelectron Spectroscopy, X-ray Diffractometry, Attenuated Total Reflection Fourier-Transform Infrared Spectroscopy, Scanning Electron Microscopy, ninhydrin assay, mechanical testing, in vitro degradation and bioactivity study, wettability, and, finally, cytocompatibility. The composites formed through the self-assembly of CS chains and exfoliated rGO sheets. Obtained results allowed also to conclude that the type of solvent used impacts the polymer structure and its ability to interact with rGO sheets and the mechanical properties of the composites. Both rGO and TAc acted as crosslinkers of the polymer chains. This study shows that the developed materials demonstrate the potential for use in bone tissue engineering. The next step should be their detailed biological examinations.

Biodegradable intramedullary nails reinforced with carbon and alginate fibers: In vitro and in vivo biocompatibility.

BACKGROUND: Commonly, intramedullary nails are made of nondegradable materials, and hence they need to be removed once the bone fracture is healed. We propose a novel composite material consisting of poly-L-lactide matrix modified with carbon and alginate fibers to be used for biodegradable intramedullary fixation. The aim of this study was to make in vitro and in vivo biocompatibility assessments. METHODS: In the in vitro conditions, biocompatibility of biomaterials was compared using normal human osteoblasts. After 3 and 7 days, cytotoxicity, viability and proliferation tests were performed, as well as cell morphology and adhesion observations. In the in vivo experiments, Californian rabbits (approx. 9 months old) were used. The composite nails and controls (Kirschner wires) were used for fixation of distal femoral osteotomy. The evaluation was made on the basis of clinical observations, radiographs taken after 2, 4, 6 and 8 weeks post implantation, and macroscopic and histological observations. RESULTS: Cell tests indicated that both modifiers had a positive influence on cell viability. Biodegradable composite nails led to bony union when used for fixation of distal diaphysis osteotomy in rabbits. Histological analysis showed that the initial focal necrosis should be fully compensated for by the osteoblast proliferation and trabeculae formation. CONCLUSIONS: Both in vitro and in vivo tests confirmed biocompatibility and potential applicability of novel biodegradable intramedullary nails modified with long carbon and alginate fibers for osteosynthesis of bone epiphysis.

Polylactide- and polycaprolactone-based substrates enhance angiogenic potential of human umbilical cord-derived mesenchymal stem cells in vitro - implications for cardiovascular repair.

Recent approaches in tissue regeneration focus on combining innovative achievements of stem cell biology and biomaterial sciences to develop novel therapeutic strategies for patients. Growing recent evidence indicates that mesenchymal stem cells harvested from human umbilical cord Wharton's jelly (hUC-MSCs) are a new valuable source of cells for autologous as well as allogeneic therapies in humans. hUC-MSCs are multipotent, highly proliferating cells with prominent immunoregulatory activity. In this study, we evaluated the impact of widely used FDA approved poly(α-esters) including polylactide (PLA) and polycaprolactone (PCL) on selected biological properties of hUC-MSCs in vitro. We found that both polymers can be used as non-toxic substrates for ex vivo propagation of hUC-MSCs as shown by no major impact on cell proliferation or viability. Moreover, PCL significantly enhanced the migratory capacity of hUC-MSCs. Importantly, genetic analysis indicated that both polymers promoted the angiogenic differentiation potential of hUC-MSCs with no additional chemical stimulation. These results indicate that PLA and PCL enhance selected biological properties of hUC-MSCs essential for their regenerative capacity including migratory and proangiogenic potential, which are required for effective vascular repair in vivo. Thus, PLA and PCL-based scaffolds combined with hUC-MSCs may be potentially employed as future novel grafts in tissue regeneration such as blood vessel reconstruction.

Influence of the intramedullary nail preparation method on nail's mechanical properties and degradation rate.

When it comes to the treatment of long bone fractures, scientists are still investigating new materials for intramedullary nails and different manufacturing methods. Some of the most promising materials used in the field are resorbable polymers and their composites, especially since there is a wide range of potential manufacturing and processing methods. The aim of this work was to select the best manufacturing method and technological parameters to obtain multiphase, and multifunctional, biodegradable intramedullary nails. All composites were based on a poly(l-lactide) matrix. Either magnesium alloy wires or carbon and alginate fibres were introduced in order to reinforce the nails. The polylactide matrix was also modified with tricalcium phosphate and gentamicin sulfate. The composite nails were manufactured using three different methods: forming from solution, injection moulding and hot pressing. The effect of each method of manufacturing on mechanical properties and degradation rate of the nails was evaluated. The study showed that injection moulding provides higher uniformity and homogeneity of the particle-modified polylactide matrix, whereas hot pressing favours applying higher volume fractions of fibres and their better impregnation with the polymer matrix. Thus, it was concluded that the fabrication method should be individually selected dependently on the nail's desired phase composition.

Gentamicin release from biodegradable poly-l-lactide based composites for novel intramedullary nails.

One of the major problems in orthopedic surgery is infection associated with implantation. The treatment is a very difficult and long-term process. A solution to this issue can be the use of implants which additionally constitute an antibiotic carrier preventing the development of an infection. Prototypes of biodegradable intramedullary nails made of three different composites with a poly(L-lactide) matrix were designed. The nails served as gentamicin sulfate (GS) carrier - an antibiotic commonly used in the treatment of osteomyelitis. The matrix was reinforced with carbon fibers (CF), alginate fibers (Alg) and magnesium alloy wires (Mg), as well as modified with bioactive particles of tricalcium phosphate (TCP) in various systems. In this way, novel, multi-phase and multifunctional degradable intramedullary nails were obtained. The tests demonstrated strong dependence between the type of the modifying phase introduced into the composite, and the rate of drug release. Introduction of gentamicin into the nail structure strengthened and prolonged antibacterial activity of the nails.

Magnesium alloy wires as reinforcement in composite intramedullary nails.

A promising group of biomaterials assigned for the production of intramedullary nails are composites with a polylactide (PLA) matrix, reinforced with wires made of magnesium alloys and carbon fibres. The paper describes the effect of the composition of magnesium alloy wires, their number and orientation in the composite, as well as their connection with differently directed long carbon fibres, on the mechanical properties and the degradation rate of the obtained intramedullary nails. Among the tested implant prototypes, the best mechanical characteristics and a gradual and uniform course of magnesium alloy wires were exhibited by the PLA+CF1D+MgI composite nails (with a unidirectional orientation of carbon fibres and an axially oriented single Mg alloy wire). The strength of these nails became gradually decreased with the incubation time, which should allow for a gradual loading of the bone. In the case of the PLA with only magnesium alloy wires (without carbon fibres), the increase of the number of wires, on the one hand, stimulates the improvement of the nails' strength, yet on the other hand, a higher content of magnesium alloys in the PLA matrix affects the nails' faster resorption.