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Abstract:
Fused Deposition Modelling (FDM) technique has been widely utilized in fabrication of 3D porous scaffolds for tissue engineering (TE) applications. Surprisingly, although there are many publications devoted to the architectural features of the 3D scaffolds fabricated by the FDM, none of them give us evident information about the impact of the diameter of the fibres on material properties. Therefore, the aim of this study was to investigate, for the first time, the effect of the diameter of 3D-printed PCL fibres on variations in their microstructure and resulting mechanical behaviour. The fibres made of poly(ε-caprolactone) (PCL) were extruded through commonly used types of nozzles (inner diameter ranging from 0.18 mm to 1.07 mm) by means of FDM technique. Static tensile test and atomic force microscopy working in force spectroscopy mode revealed strong decrease in the Young's modulus and yield strength with increasing fibre diameter in the investigated range. To explain this phenomenon, we conducted differential scanning calorimetry, wide-angle X-ray-scattering, Fourier-transform infrared spectroscopy, infrared and polarized light microscopy imaging. The obtained results clearly showed that the most prominent effect on the obtained microstructures and mechanical properties had different cooling and shear rates during fabrication process causing changes in supramolecular interactions of PCL. The observed fibre size-dependent formation of hydrogen bonds affected the crystalline structure and its stability. Summarising, this study clearly demonstrates that the diameter of 3D-printed fibres has a strong effect on obtained microstructure and mechanical properties, therefore should be taken into consideration during design of the 3D TE scaffolds.

Keywords:
fused deposition modelling, polycaprolactone, mechanical properties, hydrogen bonds, microstructure

Abstract:
Development of hybrid scaffolds and their formation methods occupies an important place in tissue engineering. In this paper, a novel method of 3D hybrid scaffold formation is presented as well as an explanation of the differences in scaffold properties, which were a consequence of different crosslinking mechanisms. Scaffolds were formed from 3D freeze-dried gelatin and electrospun poly(lactide-co-glicolide) (PLGA) fibers in a ratio of 1:1 w/w. In order to enhance osteoblast proliferation, the fibers were coated with hydroxyapatite nanoparticles (HAp) using sonochemical processing. All scaffolds were crosslinked using an EDC/NHS solution. The scaffolds' morphology was imaged using scanning electron microscopy (SEM). The chemical composition of the scaffolds was analyzed using several methods. Water absorption and mass loss investigations proved a higher crosslinking degree of the hybrid scaffolds than a pure gelatin scaffold, caused by additional interactions between gelatin, PLGA, and HAp. Additionally, mechanical properties of the 3D hybrid scaffolds were higher than traditional hydrogels. In vitro studies revealed that fibroblasts and osteoblasts proliferated and migrated well on the 3D hybrid scaffolds, and also penetrated their structure during the seven days of the experiment.

Keywords:
hybrid scaffolds, electrospinning, freeze-drying, gelatin, hydroxyapatite, sonochemical covering/grafting

Abstract:
A surface of polymeric nonwovens may be coated with various types of nanoparticles for medical applications, filtration, and so forth. However, quite often methods used for surface modification are difficult to scale up or are not applicable for polymers. In this article, we present one-step process enabling nonwovens functionalization. Poly(l-lactide-co-glicolide) (PLGA) nonwovens were prepared by electrospinning process and coated with hydroxyapatite nanoparticles (HAp) using ultrasonic processing. The effect of the process was evaluated with various techniques. HAp layer was successfully attached without loss of structural properties of HAp or PLGA nonwovens. The analysis confirmed the decrease of hydrophobicity of coated nonwoven, as well as its biocompatibility, making this material valuable from the perspective of medical applications. The sonochemical functionalization of polymeric nonwovens may be considered as an effective and economic method, enhancing surface properties of electrospun nonwovens for various applications.

Keywords:
lectrospinning, fibrous composites, nanoparticles, surface modification, ultrasonic treatment

Abstract:
The degradation in vivo and its effect on the supermolecular structure of poly(caprolactone) was examined. Poly(caprolactone) (PCL) samples were prepared in the form of porous scaffolds implanted into rat calvarial defects. The degradation was investigated by means of gel permeation chromatography, wide angle X-ray scattering (WAXS), scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). The study showed that the observed decrease of PCL crystallinity during degradation is accompanied by reduction of crystal size and/or perfection. The observed phenomenon could be explained by the presence of the high content of the low mobile fraction of investigated polymer, consisting not only almost 50% of crystal fraction but also most probably relatively high fraction of s.c. rigid amorphous fraction (RAF). Considering the type of structure characterized by the dominance of low mobile fraction, it is expected that the degradation will mainly concern these fractions, which in turn will lead to a decrease in the degree of crystallinity as well as crystal size and/or perfection.

Keywords:
PCL degradation, In-vivo conditions, Crystallinity, Rigid amorphous fraction

Abstract:
In this paper, the effect of the presence of L929 fibroblast cells and a cyclic load application on the kinetics of the degradation of amorphous PLGA films was examined. Complex micro and nano morphological, mechanical and physico-chemical studies were performed to assess the degradation of the tested material. For this purpose, molecular weight, glass transition temperature, specimen morphology (SEM, μCT) and topography (AFM) as well as the stiffness of the material were measured. The study showed that the presence of living cells along with a mechanical load accelerates the PLGA degradation in comparison to the degradation occurring in acellular media: PBS and DMEM. The drop in molecular weight observed was accompanied by a distinct increase in the tensile modulus and surface roughness, especially in the case of the film degradation in the presence of cells. The suspected cause of the rise in stiffness during the degradation of PLGA films is a reduction in the molecular mobility of the distinctive superficial layer resulting from severe structural changes caused by the surface degradation. In conclusion, all the micro and nanoscale properties of amorphous PLGA considered in the study are sensitive to the presence of L929 cells, as well as to a cyclic load applied during the degradation process.

Keywords:
L929, aliphatic polyester, stiffness rise

Abstract:
Physical and chemical factors resulting from the sterilization methods may affect the structure and properties of the materials which undergo this procedure. Poly(l-lactide-co-glicolide) (PLGA) is commonly used for medical applications, but, due to its inadequate mechanical properties, it is not recommended for load-bearing applications. One of the methods for improving PLGA mechanical properties is addition of hydroxyapatite nanoparticles (nHAp). The aim of this study was to evaluate the effect of nanoparticles addition on PLGA structure and properties after gamma radiation. According to our results, reduction of the molecular mass caused by gamma radiation was lower for PLGA with nHAp addition. Differential scanning calorimetry (DSC) analysis indicates an increase of crystallinity caused both by nHAp and gamma radiation. The first phenomenon can be explained by heteronucleation, while the second one is most probably related to higher molecular mobility of degrading polymer. Moreover, addition of nanoparticles increases thermal stability and affects the Young's modulus changes after gamma radiation.

Abstract:
Tissue engineering holds great potential in the production of functional substitutes to restore, maintain or improve the functionality in defective or lost tissues. So far, a great variety of techniques and approaches for fabrication of scaffolds have been developed and evaluated, allowing researchers to tailor precisely the morphological, chemical and mechanical features of the final constructs. Electrospinning of biocompatible and biodegradable polymers is a popular method for producing homogeneous nanofibrous structures, which might reproduce the nanosized organization of the tendons. Moreover, composite scaffolds obtained by incorporating nanoparticles within electrospun fibers have been lately explored in order to enhance the properties and the functionalities of the pristine polymeric constructs. The present study is focused on the design and fabrication of biocompatible electrospun nanocomposite fibrous scaffolds for tendon regeneration. A mixture of poly(amide 6) and poly(caprolactone) is electrospun to generate constructs with mechanical properties comparable to that of native tendons. To improve the biological activity of the constructs and modify their topography, wettability, stiffness and degradation rate, we incorporated silica particles into the electrospun substrates. The use of nanosize silica particles enables us to form bead-on-fiber topography, allowing the better exposure of ceramic particles to better profit their beneficial characteristics. In vitro biocompatibility studies using L929 fibroblasts demonstrated that the presence of 20 wt% of silica nanoparticles in the engineered scaffolds enhanced cell spreading and proliferation as well as extracellular matrix deposition. The results reveal that the electrospun nanocomposite scaffold represents an interesting candidate for tendon tissue engineering.

Abstract:
Bicomponent polycaprolactone/gelatin and polycaprolactone/collagen nanofibers formed by electrospinning using various solvents were subjected to biodegradation and compared. Hexafluoroisopropanol (HFIP) was used as a reference solvent, while the second, alternative solvent system was the mixture of acetic acid (AA) with formic acid (FA). Biodegradation of investigated materials was manifested mainly by the gelatin leaching, including collagen which is indeed denaturated to gelatin during electrospinning, leading to nanofibers erosion. There was no molecular degradation of PCL during 90 days of biodegradation procedure as deduced from no change in the elongation stress at break. The rate of biopolymer leaching was very fast from all materials during the first 24 h of biodegradation, being related to surface leaching, followed by a slower rate leaching from deeper material layers. Mass measurements showed much faster biopolymer leaching from nanofibers electrospun from AA/FA than from HFIP because of strongly emulsive nature of the solution in the former case. Irrespective of the solvent used, the leaching rate increased with initial content of gelatin. The analysis of Young modulus during biodegradation indicated complex mechanism of changes, including biopolymer mass loss, increase of PCL crystallinity and partial gelatin renaturation.

Keywords:
Bicomponent nanofibers, Biodegradation, Biopolymer

Abstract:
The aim of the present study was preparation, optimization, and systematic characterization of electrospun bionanocomposite fibers based on polyvinyl alcohol (PVA) as matrix and nanohydroxy apatite (nHAp) and cellulose nanofibers (CNF) as nanoreinforcements. The presence of nHAp and nHAp-CNF affected the morphology of electrospun mats and reduced fiber diameter, particularly at a higher content of nanofillers. The obtained results of FTIR, DSC, and WAXS proved the crystallinity reduction of electrospun nancomposites. Both nHAp and nHAp-CNF addition led to a significant increase of Young modulus with the highest stiffness for nanocomposite fibers at 10 wt% of nHAp and 3 wt% of CNF.

Keywords:
Bionanocomposite, cellulose nanofibers, electrospinning, nanohydroxy apatite

Abstract:
Synthetic and natural polymers blends represent a new brand of materials with application in wound healing, scaffolds or drug delivery systems. Polycaprolactone/gelatin (PCL/Gt) blends were analyzed in terms of their miscibility. The PCL structure was investigated as a function of Gt content. Changes in the PCL spherulitic structure with Gt content were investigated by a polarizing-interference microscope. The analysis of the glass transition temperature (Tg) of both components as a function of PCL/Gt ratio by differential scanning calorimetry indicates that the system of polycaprolactone/gelatin belongs to a type of s.c. compatible system, being intermediate between miscible and immiscible systems. There is possibility of very limited miscibility of both components. Supplementary wide angle X-ray scattering results are presented.

Keywords:
lends, compatibility, miscibility, polycaprolactone, gelatin

Abstract:
Introduction In the case of semicrystalline polymers, crystallinity is the parameter determining their physical properties. Some research groups indicate influence of crystallinity on cells response during in- vitro study. Commonly used methods of three-dimensional scaffolds formation do not take into account crystallinity optimisation. The aim of proposed presentation is to evaluate the effect of molecular weight and solvent on crystallinity and crystal size in case of polycaprolactone (PCL) films. Methodology Material: PCL with Mn:10, 45 and 80k g/mol (Sigma Aldrich) was used. As a solvents: Hexafluoroisopropanole, HFIP (Iris Biotech GmbH.), Acetic Acid, AA and Dichloromethane, DCM (Avantor and Chempol respectively) were used. Methods: Films were prepared from the PCL with different molecular weight using various solvents differing in evaporation rate. Characterization: Films were analysed using polarizing-interference microscopy (MPI) allowing characterization ofspherulities morphology. Degree of crystallinity was analysed by differential scanning calorimetry (DSC) and comparatively bywide angle X-ray scattering (WAXS). Results and Discussion It is evident from MPI observations that conditions of PCL films preparation affect the morphology of spherulites. All samples were birefringent, indicating in general crystallinity, being different for particular samples. Sphorulities size depends on Mw and solvent type; sharp Maltese cross was observed on few samples. Crystallinity of PCL films determined from DSC measurements was in the range 0,45-0,68 depending on solvent and polymer Mn used. Generally crystallinity of films formed from DCM is lower than from AA as a result of lower boiling point of DCM. Additional annealing enables increase in crystallinity to 0,8. WAXS crystallinity correlates with values determined by DSC. Changes of full width of half maximum(FWHM) of crystal peaks indicate variations of crystal size and/or defects depending on molecular weight and solvent what correlates with MPI observations also. Conclusions spherulites shape and crystallinity are strongly dependent on Mn and solvent type. Structural parameters of films decide on Young modulus and elasticity in terms of applications

Keywords:
crystallinity, PCL, solvents WAXS, molecular structure