Staff

Abstract:
Wounds and chronic wounds can be caused by bacterial infections and lead to discomfort in patients. To solve this problem, scientists are working to create modern wound dressings with antibacterial additives, mainly because traditional materials cannot meet the general requirements for complex wounds and cannot promote wound healing. This demand is met by material engineering, through which we can create electrospun wound dressings. Electrospun wound dressings, as well as those based on hydrogels with incorporated antibacterial compounds, can meet these requirements. This manuscript reviews recent materials used as wound dressings, discussing their formation, application, and functionalization. The focus is on presenting dressings based on electrospun materials and hydrogels. In contrast, recent advancements in wound care have highlighted the potential of thermoresponsive hydrogels as dynamic and antibacterial wound dressings. These hydrogels contain adaptable polymers that offer targeted drug delivery and show promise in managing various wound types while addressing bacterial infections. In this way, the article is intended to serve as a compendium of knowledge for researchers, medical practitioners, and biomaterials engineers, providing up-to-date information on the state of the art, possibilities of innovative solutions, and potential challenges in the area of materials used in dressings.

Keywords:
wound dressings, drug delivery systems, thermoresponsive hydrogels

Abstract:
The study explores the in vitro biocompatibility and osteoconductivity of poly(methyl methacrylate)/nano-hydroxyapatite (PMMA/nHA) composite nanofibrous scaffolds for bone tissue engineering (BTE). Electrospun scaffolds, exhibiting both low and high fiber orientation, were investigated. The inclusion of hydroxyapatite nanoparticles enhances the osteoconductivity of the scaffolds while maintaining the ease of fabrication through electrospinning. SEM analysis confirms the high-quality morphology of the scaffolds, with successful incorporation of nHA evidenced by SEM-EDS and FTIR methods. DSC analysis indicates that nHA addition increases the PMMA glass transition temperature (Tg) and reduces stress relaxation during electrospinning. Furthermore, higher fiber orientation affects PMMA Tg and stress relaxation differently. Biological studies demonstrate the composite material’s non-toxicity, excellent osteoblast viability, attachment, spreading, and proliferation. Overall, PMMA/nHA composite scaffolds show promise for BTE applications.

Keywords:
biomaterials, nanofibrous scaffolds, bone tissue engineering

Abstract:
Drugs against bacteria and abnormal cells, such as antibiotics and anticancer drugs, may save human lives. However, drug resistance is becoming more common in the clinical world. Nowadays, a synergistic action of multiple bioactive compounds and their combination with smart nanoplatforms has been considered an alternative therapeutic strategy to fight drug resistance in multidrug-resistant cancers and microorganisms. The present study reports a one-step fabrication of innovative pH-responsive Janus nanofibers loaded with two active compounds, each in separate polymer compartments for synergistic combination therapy. By dissolving one of the compartments from the nanofibers, we could clearly demonstrate a highly yielded anisotropic Janus structure with two faces by scanning electron microscopy (SEM) analysis. To better understand the distinctive attributes of Janus nanofibers, several analytical methods, such as X-ray diffraction (XRD), FTIR spectroscopy, and contact angle goniometry, were utilized to examine and compare them to those of monolithic nanofibers. Furthermore, a drug release test was conducted in pH 7.4 and 6.0 media since the properties of Janus nanofibers correlate significantly with different environmental pH levels. This resulted in the on-demand sequential codelivery of octenidine (OCT) and curcumin (CUR) to the corresponding pH stimulus. Accordingly, the antibacterial properties of Janus fibers against Escherichia coli and Staphylococcus aureus, tested in a suspension test, were pH-dependent, i.e., greater in pH 6 due to the synergistic action of two active compounds, and Eudragit E100 (EE), and highly satisfactory. The biocompatibility of the Janus fibers was confirmed in selected tests.

Keywords:
Janus nanofiber, on-demand release, dual-drug, antibacterial activity, side-by-side electrospinning

Abstract:
Skin wound healing is one of the most challenging processes for skin reconstruction, especially after severe injuries. In our study, nanofiber membranes were prepared for wound healing using an electrospinning process, where the prepared nanofibers were made of different weight ratios of polycaprolactone and bioactive glass that can induce the growth of new tissue. The membranes showed smooth and uniform nanofibers with an average diameter of 118 nm. FTIR and XRD results indicated no chemical interactions of polycaprolactone and bioactive glass and an increase in polycaprolactone crystallinity by the incorporation of bioactive glass nanoparticles. Nanofibers containing 5% w/w of bioactive glass were selected to be loaded with atorvastatin, considering their best mechanical properties compared to the other prepared nanofibers (3, 10, and 20% w/w bioactive glass). Atorvastatin can speed up the tissue healing process, and it was loaded into the selected nanofibers using a dip-coating technique with ethyl cellulose as a coating polymer. The study of the in vitro drug release found that atorvastatin-loaded nanofibers with a 10% coating polymer revealed gradual drug release compared to the non-coated nanofibers and nanofibers coated with 5% ethyl cellulose. Integration of atorvastatin and bioactive glass with polycaprolactone nanofibers showed superior wound closure results in the human skin fibroblast cell line. The results from this study highlight the ability of polycaprolactone-bioactive glass-based fibers loaded with atorvastatin to stimulate skin wound healing.

Keywords:
nanofibers, polycaprolactone, bioactive glass, coating, wound healing

Abstract:
Local bacterial infections lead to delayed wound healing and in extreme cases, such as diabetic foot ulcers, to non-healing due to the impaired cellular function in such wounds. Thus, many scientists have focused on developing advanced therapeutic platforms to treat infections and promote cellular proliferation and angiogenesis. This study presents a facile approach for designing nanofibrous scaffolds in three dimensions (3D) with enhanced antibacterial activity to meet the need of treating chronic diabetic wounds. Being a cationic surfactant as well as an antimicrobial agent, octenidine (OCT) makes a 2D membrane hydrophilic, enabling it to be modified into a 3D scaffold in a “one stone, two birds” manner. Aqueous sodium borohydride (NaBH4) solution plays a dual role in the fabrication process, functioning as both a reducing agent for the in situ synthesis of silver nanoparticles (Ag NPs) anchored on the nanofiber surface and a hydrogen gas producer for expanding the 2D membranes into fully formed 3D nanofiber scaffolds, as demonstrated by morphological analyses. Various techniques were used to characterize the developed scaffold (e.g., SEM, XRD, DSC, FTIR, and surface wettability), demonstrating a multilayered porous structure and superhydrophilic properties besides showing sustained and prolonged release of OCT (61% ± 1.97 in 144 h). Thanks to the synergistic effect of OCT and Ag NPs, the antibacterial performance of the 3D scaffold was significantly higher than that of the 2D membrane. Moreover, cell viability was studied in vitro on mouse fibroblasts L929, and the noncytotoxic character of the 3D scaffold was confirmed. Overall, it is shown that the obtained multifunctional 3D scaffold is an excellent candidate for diabetic wound healing and skin repair.

Abstract:
Aiming at biomedical applications, the present work developed a new bio-nanocomposite fibrous mat based on natural rubber (NR) reinforced with 45S5 bioglass particles (BG) and cellulose nanowhiskers (CNW), which reveals excellent mechanical properties, good biocompatibility and bioactivity properties.
Analyses of the specimens were conducted by means of morphological observa-tions (SEM) and thermal analysis (TG/DTG), as well as mechanical tests used to verify the effect of the incorporation of BG particles and CNW on the ultimate properties of these flexible NR-CWN/BG fibrous membranes. An SEM analysis revealed that all filaments possessed a ribbon-like morphology, with increasing diameters as the BG concentration increased. This likely results from an increased viscosity of the solution used for fiber blowing. In comparison with neat NR fibrous mats, the ultimate mechanical properties of bio-nanocomposites were sig-nificantly improved due to the presence of CNW and BG particles dispersed in the
NR matrix. According to the TG/DTG analysis, the specimens' thermal stability was unaffected by the high BG content, and the thermal profiles were similar,
with isoprene chains decomposition of the NR occurring between 350 and 450C. In-vitro analysis on fibroblasts confirmed that the bio-nanocomposite fibrous mats
are noncytotoxic. It was found that fibrous mats enhanced cellular growth and hold great promise for tissue engineering applications.

Keywords:
bioactive particles,cellulose nanowhiskers,fibrous mat bio-nanocomposite,natural rubber

Abstract:
This research aimed at designing and fabricating a smart thermosensitive injectable methylcellulose/agarose hydrogel system loaded with short electrospun bioactive PLLA/laminin fibers as a scaffold for tissue engineering applications or 3D cell culture models. Considering ECM-mimicking morphology and chemical composition, such a scaffold is capable of ensuring a hospitable environment for cell adhesion, proliferation, and differentiation. Its viscoelastic properties are beneficial from the practical perspective of minimally invasive materials that are introduced to the body via injection. Viscosity studies showed the shear-thinning character of MC/AGR hydrogels enabling the potential injection ability of highly viscous materials. Injectability tests showed that by tuning the injection rate, even a high amount of short fibers loaded inside of hydrogel could be efficiently injected into the tissue. Biological studies showed the non-toxic character of composite material with excellent viability, attachment, spreading, and proliferation of fibroblasts and glioma cells. These findings indicate that MC/AGR hydrogel loaded with short PLLA/laminin fibers is a promising biomaterial for both tissue engineering applications and 3D tumor culture models.

Abstract:
The solution blow spinning technique was used to fabricate a new biocomposite fibrous mat consisting of natural rubber (NR) and polyhydroxybutyrate (PHB) bioblend, with various loads of 45S5 bioglass (BG) particles. According to SEM analysis, NR fibers exhibited ribbon-like morphologies, whereas the addition of PHB resulted in improved fiber formation and a reduction in their diameter. In NR-PHB/BG biocomposites with varying BG loadings, typical thermal degradation events of PHB (stage i) and NR (stage ii) were observed. In comparison with pure PHB, the TG/DTG curves of NR-PHB/BG specimens revealed a lower stage i degradation peak. Such an outcome is possibly due to the fact that PHB in the NR-PHB fibers is located predominantly at the surface, that is, PHB is more susceptible to thermal degradation. The NR-PHB/BG biocomposite possessed an increased stiffness due to the addition of PHB and BG, resulting in an increased stress and a decreased strain at rupture compared to the pure NR and NR-PHB mats. DMA analysis revealed two well-defined regions, above and below the glass transition temperature (Tg), for the storage modulus (E') of the NR-PHB/BG specimens. The values of E' were in both regions for NR-PHB/BG specimens increased at higher BG content. The measured tanδ = E″/E' was used to determine the Tg value for all specimens, with Tg found to be in the −49 to −46°C range. Finally, NR-PHB/BG biocomposite fibrous were proven noncytotoxic by in-vitro testing on fibroblasts. These biocomposites enhanced cell growth, holding great promise for tissue engineering applications.

Keywords:
45S5 bioglass, biocomposite fibrous mat, biomedical applications, natural rubber, polyhydroxybutyrate, solution blow spinning

Abstract:
The electrospinning method has been widely used to produce nano/micro fibers for various applications. As a drug delivery system, electrospun fibers display many advantages such as controlled drug delivery kinetics and the ability to deliver drugs locally. A drug delivery system improves delivery efficiency and reduces possible toxic effects. In particular, multiaxial fibers consisting of two or more fluid components have drawn attention for the simultaneous administration of multiple therapeutic agents for sustained delivery and effective treatment. This review discusses recently studied multi-compartment electrospun fibers, including side-by-side (Janus) and axially symmetric fibers – coaxial and triaxial – from the perspective of multi-drug incorporation. It begins with an overview of conventional uniaxial single-fluid electrospinning methods for drug delivery applications, then highlights the advantages of multi-compartment fibers for multi-substance loading/delivery and the advances in triaxial fibers that seem to be promising from the perspective of challenges for dressings and tissue regeneration. Furthermore, drug release mechanisms and kinetics are discussed in the controlled delivery of multiple therapeutics in fibers. In the conclusion, current biomedical applications of multi-drug delivery systems in selected applications and future perspectives are presented.

Abstract:
The fundamental aspect of the fabrication of microporous, fibrous biomaterials in form of scaffolds is the optimization of their surface properties to enhance cellular response. In this work, a novel approach to physico-chemical modification and bioactivity enhancement of electrospun fibrous polycaprolactone (PCL) nonwovens using soft X-ray/extreme ultraviolet (SXR/EUV) irradiation and exposure to a low-temperature, SXR/EUV induced, nitrogen and oxygen plasmas is presented for the first time. Chemical alterations and morphology of the fibrous structure of irradiated PCL mats were examined using X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM), respectively. The impact of introduced changes on viability, morphology, and adhesion of L929 mouse fibroblasts was examined. It was found that simultaneous interaction of SXR/EUV radiation and N2 or O2 photoionized plasmas led to strong chemical decomposition of the surface of fibrous PCL mats. Also, mats’ spatial porous structure was not damaged and the fibers were not broken or fused. All modified samples demonstrated cyto-compatible and non-cytotoxic properties. Enhancement of L929 cell adhesion and increased proliferation were also observed.

Keywords:
Soft X-ray/extreme ultraviolet (SXR/EUV) radiation, Low-temperature plasma treatment, Electrospun polycaprolactone (PCL) nonwovens, XPS analysis, L929 mouse fibroblasts, Cytocompatibility enhancement

Abstract:
Anticancer therapies and regenerative medicine are being developed to destroy tumor cells, as well as remodel, replace, and support injured organs and tissues. Nowadays, a suitable three-dimensional structure of the scaffold and the type of cells used are crucial for creating bio-inspired organs and tissues. The materials used in medicine are made of non-degradable and degradable biomaterials and can serve as drug carriers. Developing flexible and properly targeted drug carrier systems is crucial for tissue engineering, regenerative medicine, and novel cancer treatment strategies. This review is focused on presenting innovative biomaterials, i.e., electrospun nanofibers, 3D-printed scaffolds, and hydrogels as a novel approach for anticancer treatments which are still under development and awaiting thorough optimization.

Keywords:
scaffolds, hydrogels, tissue engineering, polymers, anticancer treatments, cancer therapy, regenerative medicine

Abstract:
Polyester-based scaffolds are of research interest for the regeneration of a wide spectrum of tissues. However, there is a need to improve scaffold wettability and introduce bioactivity. Surface modification is a widely studied approach for improving scaffold performance and maintaining appropriate bulk properties. In this study, three methods to functionalize the surface of the poly(lactide-co-ε-caprolactone) PLCL fibres using gelatin immobilisation were compared. Hydrolysis, oxygen plasma treatment, and aminolysis were chosen as activation methods to introduce carboxyl (-COOH) and amino (-NH2) functional groups on the surface before gelatin immobilisation. To covalently attach the gelatin, carbodiimide coupling was chosen for hydrolysed and plasma-treated materials, and glutaraldehyde crosslinking was used in the case of the aminolysed samples. Materials after physical entrapment of gelatin and immobilisation using carbodiimide coupling without previous activation were prepared as controls. The difference in gelatin amount on the surface, impact on the fibres morphology, molecular weight, and mechanical properties were observed depending on the type of modification and applied parameters of activation. It was shown that hydrolysis influences the surface of the material the most, whereas plasma treatment and aminolysis have an effect on the whole volume of the material. Despite this difference, bulk mechanical properties were affected for all the approaches. All materials were completely hydrophilic after functionalization. Cytotoxicity was not recognized for any of the samples. Gelatin immobilisation resulted in improved L929 cell morphology with the best effect for samples activated with hydrolysis and plasma treatment. Our study indicates that the use of any surface activation method should be limited to the lowest concentration/reaction time that enables subsequent satisfactory functionalization and the decision should be based on a specific function that the final scaffold material has to perform.

Keywords:
surface activation,functionalization,electrospun fibres,hydrolysis,plasma,aminolysis

Abstract:
Immobilization of cell adhesive proteins on the scaffold surface has become a widely reported method that can improve the interaction between scaffold and cells. In this study, three nanofibrous scaffolds obtained by electrospinning of poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70:30, or poly(L-lactide) (PLLA) were subjected to chemical immobilization of gelatin based on aminolysis and glutaraldehyde cross-linking, as well as physisorption of gelatin. Two sets of aminolysis conditions were applied to evaluate the impact of amine group content. Based on the results of the colorimetric bicinchoninic acid (BCA) assay, it was shown that the concentration of gelatin on the surface is higher for the chemical modification and increases with the concentration of free NH2 groups. XPS (X-ray photoelectron spectroscopy) analysis confirmed this outcome. On the basis of XPS results, the thickness of the gelatin layer was estimated to be less than 10 nm. Initially, hydrophobic scaffolds are completely wettable after coating with gelatin, and the time of waterdrop absorption was correlated with the surface concentration of gelatin. In the case of all physically and mildly chemically modified samples, the decrease in stress and strain at break was relatively low, contrary to strongly aminolyzed PLCL and PLLA samples. Incubation testing performed on the PCL samples showed that a chemically immobilized gelatin layer is more stable than a physisorbed one; however, even after 90 days, more than 60% of the initial gelatin concentration was still present on the surface of physically modified samples. Mouse fibroblast L929 cell culture on modified samples indicates a positive effect of both physical and chemical modification on cell morphology. In the case of PCL and PLCL, the best morphology, characterized by stretched filopodia, was observed after stronger chemical modification, while for PLLA, there was no significant difference between modified samples. Results of metabolic activity indicate the better effect of chemical immobilization than of physisorption of gelatin.

Keywords:
gelatin, aminolysis, surface modification, electrospinning

Abstract:
Despite the advances in the field of pharmaceutical materials and technology, topical administration remains a method of choice for the treatment of eye diseases such as glaucoma, with eye drops being a leading dosage form. Their main disadvantage is a very short drug residence time and thus poor drug bioavailability, leading to the necessity of continuous repeated dosing. Mucoadhesive electrospun nanofibers are promising candidates for overcoming these challenges, while still benefiting from topical ocular administration. As an alternative for eye drops, a nanofibrous drug delivery system (DDS) for the delivery of brinzolamide (BRZ), based on β-cyclodextrin (β-CD), hydroxypropyl cellulose (HPC) and polycaprolactone (PCL), was designed. The results showed β-CD/BRZ guest–host interactions, successful drug incorporation into the nanofibers, and the possibility of more accurate dosing in comparison with the control eye drops. Drug permeation through sheep corneas was almost linear in time, achieving therapeutic concentrations in the receptor medium, and mucoadhesion to sheep eye mucosa was relatively high in case of formulations with high HPC content. All formulations were biocompatible, their mechanical properties were sufficient to handle them without caution and UV irradiation was suitable to reduce bioburden of the fibers matrix, yet no antibacterial properties of BRZ were observed.

Keywords:
Drug delivery system, Nanofibers, Electrospinning, Cyclodextrins, Glaucoma, Mucoadhesion, Topical administration

Abstract:
In situ crosslinked materials are the main interests of both scientific and industrial research. Methylcellulose (MC) aqueous solution is one of the representatives that belongs to this family of thermosensitive materials. At room temperature, MC is a liquid whereupon during temperature increase up to 37 °C, it crosslinks physically and turns into a hydrogel. This feature makes it unique, especially for tissue engineering applications. However, the crosslinking rate of MC alone is relatively slow considering tissue engineering expectations. According to these expectations, the crosslinking should take place slowly enough to allow for complete injection and fill the injury avoiding clogging in the needle, and simultanously, it should be sufficiently fast to prevent it from relocation from the lesion. One of the methods to overcome this problem is MC blending with another substance that increases the crosslinking rate of MC. In these studies, we used agarose (AGR). These studies aim to investigate the effect of different AGR amounts on MC crosslinking kinetics, and thermal, viscoelastic, and biological properties. Differential Scanning Calorimetry (DSC) and dynamic mechanical analysis (DMA) measurements proved that AGR addition accelerates the beginning of MC crosslinking. This phenomenon resulted from AGR's greater affinity to water, which is crucial in this particular crosslinking part. In vitro tests, carried out using the L929 fibroblast line and mesenchymal stem cells (MSCs), confirmed that most of the hydrogel samples were non-cytotoxic in contact with extracts and directly with cells. Not only does this type of thermosensitive hydrogel system provide excellent mechanical and biological cues but also its stimuli-responsive character provides more novel functionalities for designing innovative scaffold/cell delivery systems for tissue engineering applications.

Abstract:
A drug delivery system (DDSs) was developed in the present study based on textile substrates as drug carriers and electrospun nanofibers as a controller of release rate. Three types of drugs consisting of ciprofloxacin (CIP), clotrimazole (CLO), and benzalkonium chloride (BEN) were loaded into the cover glass (CG) and cotton fabrics (CF1 and CF2) separately. Then, the drug-loaded substrates were coated with polycaprolactone (PCL) and polycaprolactone/gelatin (PCL/Gel) nanofibers with various thicknesses. The morphology and hydrophilicity of the electrospun nanofibers and the release profile of drug-loaded samples were investigated. FTIR, XRD, and in vitro biodegradability analysis were analyzed to characterize the drug delivery system. A morphological study of electrospun fibers showed the mean diameter of the PCL and PCL/Gel nanofibers 127 ± 25 and 178 ± 38 nm, respectively. The drug delivery assay revealed that various factors affect the rate of drug releases, such as the type of drug, the type of drug carrier, and the thickness of the covered nanofibers. The study highlights the ability of drugs to load substrates with coated nanofibers as controlled drug delivery systems. In conclusion, it is shown that the obtained samples are excellent candidates for future wound dressing applications.

Keywords:
electrospinning, controlled drug release, ciprofloxacin

Abstract:
A methylcellulose (MC) is one of the materials representatives performing unique thermal-responsive properties. While reaching a critical temperature upon heating MC undergoes a physical sol-gel transition and consequently becomes a gel. The MC has been studied for many years and researchers agree that the MC gelation is related to the lower critical solution temperature (LCST). Nevertheless, a precise description of the MC gelation mechanism remains under discussion. In this study, we explained the MC gelation mechanism through examination of a wide range of MC concentrations via differential scanning calorimetry (DSC). The results evidenced that MC gelation is a multistep thermoreversible process, manifested by three and two endotherms depending on MC concentration. The occurrence of the three endotherms for low MC concentrations during heating has not been reported in the literature before. We justify this phenomenon by manifestation of three various transitions. The first one manifests water–water interactions, i.e., spanning water network breakdown into small water clusters. It is clearly evidenced by additional normalization to the water content. The second effect corresponds to polymer–water interactions, i.e., breakdown of water cages surrounded methoxy groups of MC. The last one is related to the polymer–polymer interactions, i.e., fibril hydrophobic domain formation. Not only did these results clarify the MC crosslinking mechanism, but also in the future will help to assess MC relevance for various potential application fields.

Keywords:
methylcellulose, thermosensitive hydrogel, crosslinking, DSC

Abstract:
It is reported in the literature that introducing amino groups on the surface improves cellular behaviour due to enhanced wettability and the presence of the positive charge. In this work, electrospun fibers were subjected to aminolysis under various conditions to investigate the impact of reaction parameters on the concentration of free NH2 groups, change of fiber properties, and the response of L929 cells. Three types of electrospun nonwovens obtained from poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70 : 30 and poly(L-lactide) (PLLA) were investigated. For all polymers, the concentration of NH2 groups increased with the diamine concentration and time of reaction. However, it was observed that PCL fibers require much stronger conditions than PLCL and PLLA fibers to reach the same level of introduced amine groups. X-ray photoelectron spectroscopy results clearly demonstrate that an aminolysis reaction is not limited to the surface of the material. Gel permeation chromatography results support this conclusion indicating global molecular weight reduction. However, it is possible to reach a compromise between the concentration of introduced amine groups and the change of mechanical properties. For most of the investigated conditions, aminolysis did not significantly change the water contact angle. Despite this, the change of L929 and MG63 cell shape to being more spread confirmed the positive effect of the presence of the amine groups.

Abstract:
The biomaterial-cells interface is one of the most fundamental issues in tissue regeneration. Despite many years of scientific work, there is no clear answer to what determines the desired adhesion of cells and the synthesis of ECM proteins. Crystallinity is a characteristic of the structure that influences the surface and bulk properties of semicrystalline polymers used in medicine. The crystallinity of polycaprolactone (PCL) was varied by changing the molecular weight of the polymer and the annealing procedure. Measurements of surface free energy showed differences related to substrate crystallinity. Additionally, the water contact angle was determined to characterise surface wettability which was crucial in the analysis of protein absorption. X-ray photoelectron spectroscopy was used to indicate oxygen bonds amount on the surface. Finally, the impact of the crystallinity, and related properties were demonstrated on dermal fibroblasts' response. Cellular proliferation and expression of selected genes: α-SMA, collagen I, TIMP, integrin were analysed.

Abstract:
Injuries of the bone/cartilage and central nervous system are still a serious socio-economic problem. They are an effect of diversified, difficult-to-access tissue structures as well as complex regeneration mechanisms. Currently, commercially available materials partially solve this problem, but they do not fulfill all of the bone/cartilage and neural tissue engineering requirements such as mechanical properties, biochemical cues or adequate biodegradation. There are still many things to do to provide complete restoration of injured tissues. Recent reports in bone/cartilage and neural tissue engineering give high hopes in designing scaffolds for complete tissue regeneration. This review thoroughly discusses the advantages and disadvantages of currently available commercial scaffolds and sheds new light on the designing of novel polymeric scaffolds composed of hydrogels, electrospun nanofibers, or hydrogels loaded with nano-additives.

Keywords:
scaffolds, tissue engineering, polymers, electrospun nanofibers, hydrogels, nanoparticles, composites, injectable materials

Abstract:
Antimicrobial nonwovens for single use applications (e.g., diapers, sanitary napkins, medical gauze, etc.) are of utmost importance as the first line of defense against bacterial infections. However, the utilization of petrochemical nondegradable polymers in such nonwovens creates sustainability-related issues. Here, sustainable poly(hydroxybutyrate) (PHB) and ε-poly-l-lysine (ε-PLL) submicro- and microfiber-based antimicrobial nonwovens produced by a novel industrially scalable process, solution blowing, have been proposed. In such nonwovens, ε-PLL acts as an active material. In particular, it was found that most of ε-PLL is released within the first hour of deployment, as is desirable for the applications of interest. The submicro- and microfiber mat was tested against C. albicans and E. coli, and it was found that ε-PLL-releasing microfibers result in a significant reduction of bacterial colonies. It was also found that ε-PLL-releasing antimicrobial submicro- and microfiber nonwovens are safe for human cells in fibroblast culture. Mechanical characterization of these nonwovens revealed that, even though they are felt as soft and malleable, they possess sufficient strength, which is desirable in the end-user applications.

Keywords:
PHB submicro- and microfibers, antimicrobial nonwovens, ε-PLL release, E. coli, C. albicans

Abstract:
Four chemical crosslinking methods were used in order to prevent gelatin leaching in an aqueous environment, from bicomponent polycaprolactone/gelatin (PCL/Gt) nanofibers electrospun from an alternative solvent system. A range of different concentrations and reaction times were employed to compare genipin, 1-(3-dimethylaminopropyl)-N’-ethylcarbodimide hydrochloride/N-hydroxysuccinimide (EDC/NHS), 1,4-butanediol diglycidyl ether (BDDGE), and transglutaminase. The objective was to optimize and find the most effective method in terms of reaction time and solution concentration, that at the same time provides satisfactory gelatin crosslinking degree and ensures good morphology of the fibers, even after 24 h in aqueous medium in 37 °C. The series of experiments demonstrated that, out of the four compared crosslinking methods, EDC/NHS was able to yield satisfactory results with the lowest concentrations and the shortest reaction times.

Keywords:
crosslinking, gelatin, nanofibers, biodegradable polymers, electrospinning

Abstract:
Tissue engineering (TE) scaffolds have enormous significance for the possibility of regeneration of complex tissue structures or even whole organs. Three-dimensional (3D) printing techniques allow fabricating TE scaffolds, having an extremely complex structure, in a repeatable and precise manner. Moreover, they enable the easy application of computer-assisted methods to TE scaffold design. The latest additive manufacturing techniques open up opportunities not otherwise available. This study aimed to summarize the state-of-art field of 3D printing techniques in applications for tissue engineering with a focus on the latest advancements. The following topics are discussed: systematics of the available 3D printing techniques applied for TE scaffold fabrication; overview of 3D printable biomaterials and advancements in 3D-printing-assisted tissue engineering.

Keywords:
tissue engineering, 3D printing, biomaterials

Abstract:
The growing popularity of solution blow spinning as a method for the production of fibrous tissue engineering scaffolds and the vast range of polymer-solvent systems available for the method raises the need to study the effect of processing conditions on fiber morphology and develop a method for its qualitative assessment. Rheological approaches to determine polymer solution spinnability and image analysis approaches to describe fiber diameter and alignment have been previously proposed, although in a separate manner and mostly for the widely known, well-researched electrospinning method. In this study, a series of methods is presented to determine the processing conditions for the development of submicron fibrous scaffolds. Rheological methods are completed with extensive image analysis to determine the spinnability window for a polymer–solvent system and qualitatively establish the influence of polymer solution concentration and collector rotational speed on fiber morphology, diameter, and alignment. Process parameter selection for a tissue engineering scaffold target application is discussed, considering the varying structural properties of the native extracellular matrix of the tissue of interest.

Keywords:
solution blow spinning, rheology, image analysis, nanofibers, fiber alignment, biodegradable nanofibers

Abstract:
Biodegradability or materials physicochemical stability are the key biomaterials selection parameters for various medical and tissue engineering applications. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a natural copolymer known from its biocompatibility with great support for cells growth and attachment on films and fibers. In our studies, the physicochemical properties of electrospun PHBV fibers and spin-coated films aged for 1, 4 and 8 weeks were analyzed using bulk (FTIR) and surface chemistry (XPS) methods and water contact angle. Further, we characterized the zeta potential changes after aging, by means of electrokinetic measurements, and cell responses to it, using NIH 3T3 murine fibroblasts. Colorimetric MTS cell viability test allowed the assessment of cell proliferation. Additionally, the morphology of fibroblasts and biointerfaces were studied by confocal laser and electron scanning microscopy (CLSM and SEM). These studies indicated that the activity, attachment and proliferation of fibroblasts is independent of aging of PHBV fibers and films. PHBV films show very stable zeta potential over 8 weeks of aging, opposite to PHBV fibers. Importantly, the flat film of PHBV increases cell proliferation, while the fibrous meshes are an excellent support for their stretching. The results of the study revealed clear advantages of PHBV films and fibrous meshes in cell-material interaction.

Keywords:
cell morphology, fibroblast, electrospun fibers, PHBV, Zeta potential

Abstract:
This piece of research explores porous nanocomposite polyurethane (PU) foam synthesis, containing nanolignin (NL), coated with natural antimicrobial propolis for wound dressing. PU foam was synthesized using polyethylene glycol, glycerol, NL, and 1, 6-diisocyanato-hexane (NCO/OH ratio: 1.2) and water as blowing agent. The resultant foam was immersed in ethanolic extract of propolis (EEP). PU, NL-PU, and PU-NL/EEP foams were characterized from mechanical, morphological, and chemical perspectives. NL Incorporation into PU increased mechanical strength, while EEP coating showed lower strength than PU-NL/EEP. Morphological investigations confirmed an open-celled structure with a pore diameter of 150–200 μm, a density of nearly 0.2 g/cm3,, and porosity greater than 85%, which led to significantly high water absorption (267% for PU-NL/EEP). The hydrophilic nature of foams, measured by the contact angle, proved to be increased by NL addition and EEP coating. PU and PU-NL did not show important antibacterial features, while EEP coating resulted in a significant antibacterial efficiency. All foams revealed high biocompatibility toward L929 fibroblasts, with the highest cell viability and cell attachment for PU-NL/EEP. In vivo wound healing using Wistar rats’ full-thickness skin wound model confirmed that PU-NL/EEP exhibited an essentially higher wound healing efficacy compared with other foams. Hence, PU-NL/EEP foam could be a promising wound dressing candidate.

Keywords:
polyurethane foam, nanolignin, propolis, wound dressing

Abstract:
Developing a multifunctional wound dressing that protects, cures and indicates the healing progress, is a new approach of investigation. Red cabbage extract (RCE), consisting of bioactive compounds that have antioxidant, anti-inflammatory, anti-carcinogenic, bactericidal, antifungal, and antiviral activities, was utilized as a natural pH-sensitive indicator. Chitosan-based hydrogel, encapsulating RCE, was developed to obtain a smart therapeutic pH-sensitive wound dressing as antimicrobial bio-matrix provides a comfortable cushion for wound bed and indicates its status. Methacrylated-chitosan was crosslinked by different concentrations of methylenebisacrylamide (MBAA) by which hydrogel mechanical and morphological properties were tuned. The proposed mechanism for hydrogel formation was confirmed by FT-IR. The coloristic properties of the RCE and the changes in color intensity as a function of pH were confirmed by UV–Vis spectroscopy. The effect of MBAA on the mechanical, swelling, release and morphological properties of hydrogel were investigated. MBAA (2.5% wt/v) in 2% wt/v chitosan showed preferable mechanical (20 KPa), swelling (1294% at pH 8 ± 0.2), and release (prolonged up to 5 days) properties. Hydrogel matrices, loaded on cotton gauze submerged in different pH buffer solutions, showed explicit color changes from green to red as pH changed from 9 to 4.

Keywords:
pH-sensitive dye, wound dressing, N-methacylated chitosan

Abstract:
This research work is aimed at studying the effect of ultrasounds on the effectiveness of fiber fragmentation by taking into account the type of sonication medium, processing time, and various PLLA molecular weights. Fragmentation was followed by an appropriate filtration in order to decrease fibers length distribution. It was evidenced by fiber length determination using SEM that the fibers are shortened after ultrasonic treatment, and the effectiveness of shortening depends on the two out of three investigated parameters, mostly on the sonication medium, and processing time. The gel permeation chromatography (GPC) confirmed that such ultrasonic treatment does not change the polymers' molecular weight. Our results allowed to optimize the ultrasonic fragmentation procedure of electrospun fibers while preliminary viscosity measurements of fibers loaded into hydrogel confirmed their potential in further use as fillers for injectable hydrogels for regenerative medicine applications.

Keywords:
electrospinning, ultrasonication, short fibers, polymers

Abstract:
Electrospinning is a widely investigated process for forming nanofibers. Nanofibers in drug delivery systems are extensively tested due to its remarkable properties e.g. small pore size or large surface area. Recent articles have informed about formation of fibers using triaxial electrospinning in drug delivery systems. This paper summarizes the process of triaxial electrospinning and its application in drug delivery. Triaxial electrospinning has advantages in forming complex nanostructures for specific drug delivery applications. This paper summarizes the possibility to use triaxial electrospinning to resolve the problem of limited drug solubility, to protect biomolecules from hostile environment, and to control drug release kinetics, with the possibility of loading of various drugs. There are literature data evidencing the possibility to achieve sustained release with a border case of zero rate order kinetics. There is no doubt that triaxial electrospinning opens a new way to develop sophisticated nanomaterials for achieving the desired functional performances and to expand the applications in the drug delivery area. Triaxial electrospinning method is interdisciplinary area with great potential in nanotechnology.

Keywords:
triaxial electrospinning, complex nanostructures, drug delivery, desired functional performance, sustained/controlled release

Abstract:
Piezoelectric polymers are promising energy materials for wearable and implantable applications for replacing bulky batteries in small and flexible electronics. Therefore, many research studies are focused on understanding the behavior of polymers at a molecular level and designing new polymer-based generators using polyvinylidene fluoride (PVDF). In this work, we investigated the influence of voltage polarity and ambient relative humidity in electrospinning of PVDF for energy-harvesting applications. A multitechnique approach combining microscopy and spectroscopy was used to study the content of the β-phase and piezoelectric properties of PVDF fibers. We shed new light on β-phase crystallization in electrospun PVDF and showed the enhanced piezoelectric response of the PVDF fiber-based generator produced with the negative voltage polarity at a relative humidity of 60%. Above all, we proved that not only crystallinity but also surface chemistry is crucial for improving piezoelectric performance in PVDF fibers. Controlling relative humidity and voltage polarity increased the d33 piezoelectric coefficient for PVDF fibers by more than three times and allowed us to generate a power density of 0.6 μW·cm^–2 from PVDF membranes. This study showed that the electrospinning technique can be used as a single-step process for obtaining a vast spectrum of PVDF fibers exhibiting different physicochemical properties with β-phase crystallinity reaching up to 74%. The humidity and voltage polarity are critical factors in respect of chemistry of the material on piezoelectricity of PVDF fibers, which establishes a novel route to engineer materials for energy-harvesting and sensing applications.

Keywords:
PVDF, polymer crystallinity, electrospinning, piezoelectricity, voltage polarity

Abstract:
The reactivity of the biobased monomer β-farnesene in the statistical anionic copolymerization with styrene and the effect of the bottlebrush-like polyfarnesene structure on the phase separation behavior were investigated. Furthermore, thermal and material properties of β-farnesene-based thermoplastic elastomers, based on tri- and pentablock copolymers with styrene, and their processing behavior were investigated. As shown by H NMR online kinetics, in analogy to both isoprene and β-myrcene, the direct (i.e., statistical) anionic copolymerization of β-farnesene and styrene in cyclohexane affords block-like, tapered copolymers because of the highly diverging reactivity ratios (rFar = 27; rS = 0.037). Algebraic expressions for both the molar and volume composition profiles were derived, which provide a mathematically accurate picture of the tapered copolymer structure. The one-pot, tapered copolymer approach was used to synthesize series of tri- (ABA) and pentablock (ABABA) copolymers of styrene (A) and β-farnesene (B), varying the polydiene volume fraction between 0.50 and 0.68, respectively. Depending on the polydiene volume fraction, the tapered multiblock copolymers showed phase separation in lamellar or hexagonally packed cylindrical structures, as determined by small-angle X-ray scattering. Well-defined tapered tri- and pentablock copolymers with molecular weights of 120 kg mol^–1 and low dispersity (Đ = 1.05–1.16) were obtained. The order of the tapered poly(farnesene-co-styrene) copolymers bears many similarities (same morphology, practically the same domain spacing, and a similar degree of segregation) to the corresponding polyisoprene copolymers with the same polydiene volume fraction. The similar domain spacing is suggestive of looped configurations mainly in the polyisoprene copolymers that are reduced in the polyterpene copolymers. The influence of the long alkenyl side chains of the polyfarnesene middle blocks on the mechanical properties of the multiblock copolymers was investigated by tensile testing. For this purpose, the respective tri- and pentablock copolymers of isoprene (C5 unit) and β-myrcene (C10) with styrene were synthesized as well, containing equal polydiene volume fractions as their β-farnesene-based (C15) analogs. The mechanical toughness of the polymers increased with decreasing length of the alkenyl side chains (from β-farnesene to isoprene). Furthermore, tapered polyfarnesene tri- and pentablock copolymers with styrene exhibit reduced solution viscosity in comparison to, for example, polyisoprene-based tapered PS-b-P(I-co-S) triblock copolymers, resulting in improved processability by electrospinning. These properties are discussed in terms of the different configurations of the polyterpene blocks and the pronounced differences of the entanglement molecular weights.

Abstract:
Electric field strength and polarity in electrospinning processes and their effect on process dynamics and the physical properties of as-spun fibers is studied. Using a solution of the neutral polymer such as poly(methyl methacrylate) (PMMA) we explored the electrospun jet motion issued from a Taylor cone. We focused on the straight jet section up to the incipient stage of the bending instability and on the radius of the disk of the fibers deposited on the collecting electrode. A new correlation formula using dimensionless parameters was found, characterizing the effect of the electric field on the length of the straight jet, L˜E~E˜0.55. This correlation was found to be valid when the spinneret was either negatively or positively charged and the electrode grounded. The fiber deposition radius was found to be independent of the electric field strength and polarity. When the spinneret was negatively charged, L˜E was longer, the as-spun fibers were wider. The positively charged setup resulted in fibers with enhanced mechanical properties and higher crystallinity. This work demonstrates that often-overlooked electrical polarity and field strength parameters influence the dynamics of fiber electrospinning, which is crucial for designing polymer fiber properties and optimizing their collection.

Keywords:
fibers, electrical polarity, charges, electrospinning, PMMA, mechanical properties

Abstract:
Smart piezoelectric materials are of great interest due to their unique properties. Piezoelectric materials can transform mechanical energy into electricity and vice versa. There are mono and polycrystals (piezoceramics), polymers, and composites in the group of piezoelectric materials. Recent years show progress in the applications of piezoelectric materials in biomedical devices due to their biocompatibility and biodegradability. Medical devices such as actuators and sensors, energy harvesting devices, and active scaffolds for neural tissue engineering are continually explored. Sensors and actuators from piezoelectric materials can convert flow rate, pressure, etc., to generate energy or consume it. This paper consists of using smart materials to design medical devices and provide a greater understanding of the piezoelectric effect in the medical industry presently. A greater understanding of piezoelectricity is necessary regarding the future development and industry challenges.

Keywords:
polymers, smart materials, piezoelectric materials, inorganic materials, organic materials, biomedical devices

Abstract:
Electrospun polymer nanofibers have received much attention in tissue engineering due to their valuable properties such as biocompatibility, biodegradation ability, appropriate mechanical properties, and, most importantly, fibrous structure, which resembles the morphology of extracellular matrix (ECM) proteins. However, they are usually hydrophobic and suffer from a lack of bioactive molecules, which provide good cell adhesion to the scaffold surface. Post-electrospinning surface functionalization allows overcoming these limitations through polar groups covalent incorporation to the fibers surface, with subsequent functionalization with biologically active molecules or direct deposition of the biomolecule solution. Hydrophilic surface functionalization methods are classified into chemical approaches, including wet chemical functionalization and covalent grafting, a physiochemical approach with the use of a plasma treatment, and a physical approach that might be divided into physical adsorption and layer-by-layer assembly. This review discusses the state-of-the-art of hydrophilic surface functionalization strategies of electrospun nanofibers for tissue engineering applications. We highlighted the major advantages and drawbacks of each method, at the same time, pointing out future perspectives and solutions in the hydrophilic functionalization strategies.

Keywords:
surface functionalization, electrospinning, polymers, nanofiber, immobilization, tissue engineering

Abstract:
Polyvinylidene fluoride (PVDF) is one of the most important piezoelectric polymers. Piezoelectricity in PVDF appears in polar β and ɣ phases. Piezoelectric fibers obtained by means of electrospinning may be used in tissue engineering (TE) as a smart analogue of the natural extracellular matrix (ECM). We present results showing the effect of rotational speed of the collecting drum on morphology, phase content and in vitro biological properties of PVDF nonwovens. Morphology and phase composition were analyzed using scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR), respectively. It was shown that increasing rotational speed of the collector leads to an increase in fiber orientation, reduction in fiber diameter and considerable increase of polar phase content, both b and g. In vitro cell culture experiments, carried out with the use of ultrasounds in order to generate electrical potential via piezoelectricity, indicate a positive effect of polar phases on fibroblasts. Our preliminary results demonstrate that piezoelectric PVDF scaffolds are promising materials for tissue engineering applications, particularly for neural tissue regeneration, where the electric potential is crucial.

Keywords:
scaffolds, electrospinning, polyvinylidene fluoride, tissue engineering

Abstract:
Injury to the central or peripheral nervous systems leads to the loss of cognitive and/or sensorimotor capabilities, which still lacks an effective treatment. Tissue engineering in the post-injury brain represents a promising option for cellular replacement and rescue, providing a cell scaffold for either transplanted or resident cells. Tissue engineering relies on scaffolds for supporting cell differentiation and growth with recent emphasis on stimuli responsive scaffolds, sometimes called smart scaffolds. One of the representatives of this material group is piezoelectric scaffolds, being able to generate electrical charges under mechanical stimulation, which creates a real prospect for using such scaffolds in non-invasive therapy of neural tissue. This paper summarizes the recent knowledge on piezoelectric materials used for tissue engineering, especially neural tissue engineering. The most used materials for tissue engineering strategies are reported together with the main achievements, challenges, and future needs for research and actual therapies. This review provides thus a compilation of the most relevant results and strategies and serves as a starting point for novel research pathways in the most relevant and challenging open questions.

Keywords:
neural tissue engineering, piezoelectric scaffolds, smart materials, polymers

Abstract:
Results of the preparation of biodegradable porous scaffolds using an original modification of a wet phase inversion method were presented. Influence of gelatin non‐woven as a non‐classic pore precursor and polyvinylpyrrolidone, Pluronic as classic pore precursors on the structure of obtained scaffolds was analyzed. It was shown that the addition of gelatin non‐wovens enables the preparation of scaffolds, which allow for the growth of cells (size, distribution, and shape of pores). Mechanical properties of the obtained cell scaffolds were determined. The influence of pore precursors on mass absorption of scaffolds against isopropanol and plasma was investigated. Interaction of scaffolds with a T‐lymphocyte line (Jurkat) and with fibroblasts (L929) was investigated. Obtained scaffolds are not cytotoxic and can be used as implants, for example, the regeneration of cartilage tissue.

Keywords:
cell cultures, cytotoxic, fibroblasts, imprinted scaffolds

Abstract:
Since the introduction of tissue engineering as an encouraging method for the repair and regeneration of injured tissue, there have been many attempts by researchers to construct bio-mimetic scaffolds which mimic the native extracellular matrix, with the aim of promoting cell growth, cell proliferation, and restoration of the tissue's native functionality. Among the different materials and methods of scaffold fabrication, one particularly promising class of materials, hydrogels, has been extensively studied, with the inclusion of nano-scaled materials into hydrogels leading to the creation of an exciting new generation of nanocomposites, known as nanocomposite hydrogels. To closely mimic the native tissue behavior, scientists have recently focused on the functionalization of incorporated nanomaterials via chiral biomolecules, with reported results showing great potential. The current article aims to introduce a perspective of nano-scaled cellulose as a promising nanomaterial which can be multi-functionalized for the fabrication of nanocomposite hydrogels with applications in tissue engineering and drug delivery systems. This article also briefly reviews the recently reported literature on nanocomposite hydrogels incorporated with chiral functionalized nanomaterials. Such knowledge paves the path for the development of tailored hydrogels toward practical applications.

Keywords:
scaffold, nanocomposite hydrogels, biodegradable hydrogels, chiral biomolecules, self-healing

Abstract:
Highly efficient stoichiometric coupling of sterically hindered polystyryllithium (PS-Li) chain ends was achieved using tetra[3-(chloro-dimethylsilyl)propyl]silane (TCDMSPS) as the linking agent. Based on the disparate reactivities of isoprene (I, rI = 11.0) and styrene (S, rS = 0.049) in the anionic copolymerization in nonpolar media, poly(isoprene0.5-grad-styrene0.5) (P(I0.5-grad-S0.5)) tapered 4-arm star copolymers were synthesized in only two steps. The tapered 4-arm star copolymers (Mwtargeted = 40 to 160 kg mol^-1) were synthesized with high star functionalities f (Mw,star/Mw,arm = 3.68 - 3.98), low dispersity (Ð = 1.06 - 1.15) and minimal residual precursor content (2-8 wt%), avoiding fractionation or other purification methods. Coupling kinetics measurements revealed that for the synthesis of polystyrene (PS) 4-arm stars (12 kg mol^-1) a coupling efficiency of 98% was already achieved within 2 minutes. All star polymers were analyzed by size exclusion chromatography (SEC) viscometry with universal calibration (UC) as well as NMR spectroscopy. Well-defined nanofibers from the tapered copolymer stars were obtained via electrospinning.

Abstract:
Two types of poly(glycerol sebacate) (PGS) prepolymers were synthesized and electrospun with poly(l-lactic acid) (PLA), resulting in bicomponent nonwovens. The obtained materials were pre-heated in a vacuum, at different times, to crosslink PGS and investigate morphological and structural dependencies in that polymeric, electrospun system. As both PGS and PLA are sensitive to pre-heating (crosslinking) conditions, research concerns both components. More interest is focused on the properties of PGS, considering further research for mechanical properties and subsequent experiments with PGS synthesis. Electrospinning of PGS blended with PLA does not bring difficulties, but obtaining elastomeric properties of nonwovens is problematic. Even though PGS has many potential advantages over other polyesters when soft tissue engineering is considered, its full utilization via the electrospinning process is much harder in practice. Further investigations are ongoing, especially with the promising PGS prepolymer with a higher esterification degree and its variations.

Keywords:
electrospinning, degradable polymers, synthesis, structure, crosslinking

Abstract:
Thermosensitive, physically crosslinked injectable hydrogels are in the area of interests of various scientific fields. One of the representatives of this materials group is an aqueous solution of methylcellulose. At ambient conditions, methylcellulose (MC) is a sol while on heating up to 37 °C, MC undergoes physical crosslinking and transforms into a gel. Injectability at room temperature, and crosslinkability during subsequent heating to physiological temperature raises hopes, especially for tissue engineering applications. This research work aimed at studying crosslinking kinetics, thermal, viscoelastic, and biological properties of MC aqueous solution in a broad range of MC concentrations. It was evidenced by Differential Scanning Calorimetry (DSC) that crosslinking of MC is a reversible two-stage process, manifested by the appearance of two endothermic effects, related to the destruction of water cages around methoxy groups, followed by crosslinking via the formation of hydrophobic interactions between methoxy groups in the polymeric chains. The DSC results also allowed the determination of MC crosslinking kinetics. Complementary measurements of MC crosslinking kinetics performed by dynamic mechanical analysis (DMA) provided information on the final storage modulus, which was important from the perspective of tissue engineering applications. Cytotoxicity tests were performed using mouse fibroblasts and showed that MC at low concentration did not cause cytotoxicity. All these efforts allowed to assess MC hydrogel relevance for tissue engineering applications.

Keywords:
methylcellulose, thermosensitive hydrogel, crosslinking kinetics, DSC, DMA, cellular tests

Abstract:
Each year, new glaucoma drug delivery systems are developed. Due to the chronic nature of the disease, it requires the inconvenient daily administration of medications. As a result of their elution from the eye surface and penetration to the bloodstream through undesired permeation routes, the bioavailability of active compounds is low, and systemic side effects occur. Despite numerous publications on glaucoma drug carriers of controlled drug release kinetics, only part of them consider drug permeation routes and, thus, carriers' location, as an important factor affecting drug delivery. In this paper, we try to demonstrate the importance of the delivery proximal to glaucoma drug targets. The targeted delivery can significantly improve drug bioavailability, reduce side effects, and increase patients' compliance compared to both commercial and scientifically developed formulations that can spread over the eye surface or stay in contact with conjunctival sac. We present a selection of glaucoma drug carriers intended to be placed on cornea or injected into the aqueous humor and that have been made by advanced materials using hi-tech forming methods, allowing for effective and convenient sustained antiglaucoma drug delivery.

Keywords:
hydrogels, nanofibers, electrospinning, glaucoma, ophthalmology

Abstract:
Surface functionalization of polymer scaffolds is a method used to improve interactions of materials with cells. A frequently used method for polyesters is aminolysis reaction, which introduces free amine groups on the surface. In this study, nanofibrous scaffolds and films of three different polyesters–polycaprolactone (PCL), poly(lactide-co-caprolactone) (PLCL), and poly(l-lactide) (PLLA) were subjected to this type of surface modification under the same conditions. Efficiency of aminolysis was evaluated on the basis of ninhydrin tests and ATR–FTIR spectroscopy. Also, impact of this treatment on the mechanical properties, crystallinity, and wettability of polyesters was compared and discussed from the perspective of aminolysis efficiency. It was shown that aminolysis is less efficient in the case of nanofibers, particularly for PCL nanofibers. Our hypothesis based on the fundamentals of classical high speed spinning process is that the lower efficiency of aminolysis in the case of nanofibers is associated with the radial distribution of crystallinity of electrospun fiber with more crystalline skin, strongly inhibiting the reaction. Moreover, the water contact angle results demonstrate that the effect of free amino groups on wettability is very different depending on the type and the form of polymer. The results of this study can help to understand fundamentals of aminolysis-based surface modification.

Keywords:
aminolysis, polyester, electrospinning, nanofibers, film, surface chemical modification

Abstract:
The current work studies the electrospun poly (vinyl alcohol) (PVA) nanofibers and its nanocomposites including nanohydroxy apatite (nHAp) and nHAp/cellulose nanofibers (CNFs), emphasizing the impact of nanofillers on the toughness of nanofibers. PVA nanofibers were incorporated with 10 wt% of nHAp and then various amounts of CNF were added to subsequent PVA/nHAp fibrous nanocomposites. The morphology of nonwoven mats was studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). While neat PVA nanofibers were smooth and uniform in thickness, the nanofiller loading resulted in thinner fibers with less uniformity. Furthermore, the thermal properties of the nonwoven network of fibers were characterized employing thermogravimetric analysis (TGA). Although the maximum loss mass temperature of PVA was partially reduced upon addition of nanofillers, the onset of decomposition was not altered. The mechanical characterizations were performed using static tensile and dynamic mechanical analysis (DMA). Compared to neat PVA mats, the tensile test of nanocomposites mats demonstrated the significant increase in Young's modulus; however, strain at break was dramatically reduced. In addition, the fracture work was assessed from the area under the stress-strain curve, which showed brittleness of fibrous nanocomposites due to the nanofiller incorporation. Field emission SEM (FE-SEM) was employed to scan the fracture surface of stretched fibers. The increase in modulus of electrospun mats was also shown by DMA in frequency mode. In parallel, both tensile test and DMA confirmed the change in fracture of PVA fibers from a tough to brittle mode, due to the nanofiller addition.

Keywords:
electrospun nanocomposites, nanofillers, toughness, mechanical properties

Abstract:
The present study aims to theoretically model and verify the mechanical behavior of electrospun fibers of poly(vinyl alcohol) (PVA) reinforced by nanohydroxy apatite (nHAp) and cellulose nanofibers (CNF), the three composites designated as PVA/nHAp, PVA/CNF, and PVA/nHAp/CNF. Tensile tests and AFM nanoindentation studies were used to measure tensile modulus of electrospun scaffolds and single fibers respectively. Halpin–Tsai and Ouali models were applied to predict the stiffness of electrospun mats. Theoretical analysis according to the Halpin–Tsai model showed that CNF have no preferred orientation in the electrospun fibers, particularly at higher filler content. Additionally, this model provided a better prediction than Ouali model, especially at lower filler content. Theoretical models based on the geometry of an unit cell in open-cell structure such as honeycomb, tetrakaidecahedron and cube models simulate electrospun scaffolds. Among the structural models for analysis of porous scaffolds, the honeycomb model showed the best prediction, tetrakaidecahedron model—a moderate one, and cube model was the worst. In general, it was proved by both experiment and theory that the porous structure of electrospun mat caused significant modulus reduction of nanocomposites.

Keywords:
Nanocomposites, Cellulose nanofibers, Electrospinning, Modulus

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:
Approach. Injuries of the central nervous system (CNS) can cause serious and permanent disability due to limited regeneration ability of the CNS. Presently available therapies are focused on lesion spreading inhibition rather than on tissue regeneration. Recent investigations in the field of neural tissue engineering indicate extremely promising properties of novel injectable and non-injectable hydrogels which are tailored to serve as biodegradable scaffolds for CNS regeneration. Objective. This review discusses the state-of-the-art and barriers in application of novel polymer-based hydrogels without and with nanoparticles for CNS regeneration. Main results. Pure hydrogels suffer from lack of similarities to natural neural tissue. Many of the biological studies indicated nano-additives in hydrogels may improve their topography, mechanical properties, electroconductivity and biological functions. The most promising biomaterials which meet the requirements of CNS tissue engineering seem to be injectable thermosensitive hydrogels loaded with specific micro-and nanoparticles. Significance. We highlight injectable hydrogels with various micro-and nanoparticles, because of novelty and attractiveness of this type of materials for CNS regeneration and future development perspectives.

Keywords:
hydrogels, nanoparticles, injectable, microparticles, nanofibers, central nervous system

Abstract:
Cartilage loss due to age‐related degeneration and mechanical trauma is a significant and challenging problem in the field of surgical medicine. Unfortunately, cartilage tissue can be characterized by the lack of regenerative ability. Limitations of conventional treatment strategies, such as auto‐, allo‐ and xenografts or implants stimulate an increasing interest in the tissue engineering approach to cartilage repair. This review discusses the application of polymer‐based scaffolds, with an emphasis on hydrogels in cartilage tissue engineering. We highlight injectable hydrogels with various micro‐ and nanoparticles, as they constitute a novel and attractive type of scaffolds. We discuss advantages, limitations and future perspectives of injectable nanocomposite hydrogels for cartilage tissue regeneration.

Keywords:
polymers, hydrogels, injectable hydrogels, injectable nanocomposite hydrogels, cartilage repair, cartilage tissue engineering

Abstract:
Bicomponent polycaprolactone/gelatin and polycaprolactone/collagen fibres were formed by electrospinning using two kinds of solvents: a representative of commonly used solvents with this polymer composition, highly toxic hexafluoroisopropanol (HFIP) and alternative, less harmful one, the mixture of acetic (AA) and formic (FA) acids. Both material types were subjected to investigations of structure and in-vitro cellular activity. Viscosity and Fourier transform infrared spectroscopy (FTIR) measurements shown that the type of solvent used influences the structure of solution and conformation of polymer molecules. In-vitro quantitative tests as well as cell culture morphology observations proved that materials electrospun with the use of 'green' solvents can yield similar results to those obtained by made with toxic ones. Slightly better cellular response to materials electrospun from HFIP can be explained by relatively well dispersed components within the fibre and more expanded conformation of molecules, resulting in better exposition of RGD (Arg-Gly-Asp) binding sites to cells' integrin receptors.

Keywords:
Cellular tests, Electrospinning, Biopolymers, Viscosity, Solvents

Abstract:
The article is focused on the role of nanohydroxy apatite (nHAp) and cellulose nanofibers (CNFs) as fillers in the electrospun poly (vinyl alcohol) (ES-PVA) nanofibers for bone tissue engineering (TE). Fibrous scaffolds of PVA, PVA/nHAp (10 wt.%), and PVA/nHAp(10 wt.%)/CNF(3 wt.%) were successfully fabricated and characterized. Tensile test on electrospun PVA/nHAp10 and PVA/nHAp10/CNF3 revealed a three-fold and seven-fold increase in modulus compared with pure ES-PVA (45.45 ± 4.77). Although, nanofiller loading slightly reduced the porosity percentage, all scaffolds had porosity higher than 70%. In addition, contact angle test proved the great hydrophilicity of scaffolds. The presence of fillers reduced in vitro biodegradation rate in PBS while accelerates biomineralization in simulated body fluid (SBF). Furthermore, cell viability, cell attachment, and functional activity of osteoblast MG-63 cells were studied on scaffolds showing higher cellular activity for scaffolds with nanofillers. Generally, the obtained results confirm that the 3-componemnt fibrous scaffold of PVA/nHAp/CNF has promising potential in hard TE.

Keywords:
electrospinning, PVA bionanocomposites, scaffolds, bone tissue engineering, cell culture

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:
The aim of this research was to study the effect of charge polarity applied to the spinning nozzle on the structure and properties of polycaprolactone/chitosan (PCL/CHT) blends, in particular the efficiency of further surface modification by chondroitin sulphate (CS). The observed differences in the morphology and properties of fibres formed at different polarities were interpreted in terms of molecular interactions occurring in the system. FTIR results indicate stronger PCL-chitosan interactions at negative polarity, resulting in lower PCL crystallinity and crystal size distribution determined by DSC, as well as lower wettability. The charge polarity influences PCL/CHT fibre morphology and tailors some of their properties, e.g. wettability, mechanical properties and the efficiency of surface modification. Better efficiency of CS attachment was observed at negative polarity using atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS) is most probably related to higher chitosan content at the fibres' surface being attracted by the negative external potential.

Keywords:
Polycaprolactone/chitosan nanofibres, Charge potential effect in electrospinning, Polycaprolactone-chitosan interactions

Abstract:
Electrospinning of chitosan blends is a reasonable idea to prepare fibre mats for biomedical applications. Synthetic and natural components provide, for example, appropriate mechanical strength and biocompatibility, respectively. However, solvent characteristics and the polyelectrolyte nature of chitosan influence the spinnability of these blends. In order to compare the effect of solvent on polycaprolactone/chitosan fibres, two types of the most commonly used solvent systems were chosen, namely 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) and acetic acid (AA)/formic acid (FA). Results obtained by various experimental methods clearly indicated the effect of the solvent system on the structure and properties of electrospun polycaprolactone/chitosan fibres. Viscosity measurements confirmed different polymer–solvent interactions. Various molecular interactions resulting in different macromolecular conformations of chitosan influenced its spinnability and properties. HFIP enabled fibres to be obtained whose average diameter was less than 250 nm while maintaining the brittle and hydrophilic character of the nonwoven, typical for the chitosan component. Spectroscopy studies revealed the formation of chitosan salts in the case of the AA/FA solvent system. Chitosan salts visibly influenced the structure and properties of the prepared fibre mats. The use of AA/FA caused a reduction of Young's modulus and wettability of the proposed blends. It was confirmed that wettability, mechanical properties and the antibacterial effect of polycaprolactone/chitosan fibres may be tailored by selecting an appropriate solvent system. The MTT cell proliferation assay revealed an increase of cytotoxicity to mouse fibroblasts in the case of 25% w/w of chitosan in electrospun nonwovens.

Keywords:
chitosan, electrospinning, PCL/chitosan fibres, solvent system, chitosan salts

Abstract:
Crystallization from melt of poly(vinylidene fluoride) was studied by thin film chip calorimetry at cooling rates from 500 to 100,000 Ks−1 and isothermally down to 76°C. At ca. 70°C, for cooling rates higher than 2000 Ks−1, there appears a change in crystallization from high temperature α phase to low temperature β phase. The amorphous state is preserved at cooling rate 100,000 Ks−1. Analysis of the crystallization kinetics with Ziabicki model reveals maximum of the steady-state crystallization rate of β phase as 2200 s−1 at 22°C, and the highest crystallization rate of α phase as 200 s−1 at 70°C. Approximation of the temperature dependent steady-state crystallization rate with the Turnbull and Fisher nucleation model results in the equilibrium melting temperatures 227 and 173°C for the α and β phase, respectively, and in the energy barrier for short-distance transport, ED, as 70–80 kJ mol−1 at high supercooling.

Keywords:
Poly(vinylidene fluoride), Ultra-fast calorimetry, Crystallization kinetics

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:
This study aims to explore crystallinity variations of polyvinyl alcohol (PVA) as a result of electrospinning, filler addition, and heat treatment. Pure PVA and PVA nanocomposite fibers containing only nanohydroxy apatite (nHAp) and together with cellulose nanofibers (CNF) were electrospun. Electrospun nanofibers were heat treated at 180°C for 8 h. The morphology of electrospun fibers was evaluated by scanning electron microscopy (SEM) while Fourier transform infrared spectroscopy, differential scanning calorimetry, and wide angle X-ray scattering were used to analyze nanofibers crystallinity. Un-treated electrospun nanofibers were shrank and lost their porous structure in water, while heat treatment of nanofibers caused stabilization of fibrous mats in boiling water. It was concluded that the crystallinity of electrospun PVA were considerably reduced compared to PVA powder due to formation of metastable—small and/or defective crystals. Adding small content (1 wt%) of nHAp led to increase in electrospun nanofibers crystallinity. However, incorporation of higher content of nHAp and CNF caused reduction of crystallinity most probably due to possible interactions among components which interrupt the orientation of macromolecules. All analyzing methods proved the crystallinity enhancement of nanofibers upon heat treatment which can be attributed mostly to water evaporation from electrospun fibers structure.

Keywords:
Polyvinyl alcohol, Crystallinity, Electrospinning, Nanofiber, Nanofiller, Heat treatment

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:
Biological interaction between cells and scaffolds is mediated through events at surfaces. Proteins present in the culture medium adsorb on substrates, generating a protein adlayer that triggers further downstream events governing cell adhesion. Polymer blends often combine the properties of the individual components, for example, can provide mechanical as well as surface properties in one fibre. Therefore, mixtures of synthetic polycaprolactone and gelatin as a denatured form of collagen were electrospun at selected conditions and polymer weight ratios. Fibre morphologies and chemical properties of the surfaces were analysed. These scaffolds were seeded with human mesenchymal stromal cells and their viability was studied. Gelatin addition to polycaprolactone leads to a reduction in fibre diameter. A linear increase in gelatin at the fibre surface was observed in function of the weighed polymers, except for polycaprolactone/gelatin fibres incorporating equal weight ratios. Thereby, a depletion of gelatin at the fibre surface is stated for equally mixed polymers. The depletion of gelatin at the fibre surface is most probably due to hydrophobic interactions between hydrophobic segments of polycaprolactone and gelatin, affecting the spinning mechanism and thus fibre structure. Furthermore, polycaprolactone/gelatin blends show enhanced wettability properties compared to pure gelatin, at least partly due to molecular segregation. Results of in vitro studies reveal an increase in cellular viability and proliferation for cells cultivated on nanofibres containing gelatin, caused by the cell-attractive surface composition as well as the hydrophilic nature of the scaffolds. Contact guidance of cells seeded on parallelised fibres is observed, and DNA tests show evidently enhanced cell numbers on nanofibres containing 20 wt% of gelatin.

Keywords:
Mesenchymal stromal cells, electrospinning, surface, blends, biocompatibility, polymers, bioactivity

Abstract:
Bicomponent polycaprolactone/gelatin (PCL/Gt) nanofibers were successfully formed for the first time by electrospinning using a novel polymer–solvent system with solvents being alternative to the commonly used toxic solvents like fluorinated alcohols. The mixture of acetic acid (AA) with formic acid (FA; 90:10) was applied. Stable electrospinning was possible despite the fact the mixture of PCL and gelatin in AA/FA solvent showed emulsive structure. From the practical perspective, there is no doubt that it is possible to obtain PCL/Gt fibers using AA/FA mixture with morphology similar to that for fibers spun from hexafluoroisopropanol (HFIP) solutions.

Keywords:
Alternative solvents, electrospinning, gelatin, nanofibers, polycaprolactone, structure

Abstract:
The effect of electrospinning parameters on morphology, molecular, and supermolecular structure of polycaprolactone (PCL) fibers was analyzed, with respect to tissue engineering applications. Fibers morphology and structure are mainly determined by solution concentration and collector type. Applied voltage does not significantly influence supermolecular structure (crystallinity) and mechanical stiffness. There is correlation between changes in structure and proliferation of 3T3 cells as evidenced by in vitro study. Processing window of optimal scaffolds is relatively wide, however, variation of electrospinning parameters do not significantly affect their biological functionality.

Keywords:
3T3 cells, crystallinity, electrospinning, molecular orientation, polycaprolactone, porosity, tissue engineering

Abstract:
Gelatin is one of the most promising materials in tissue engineering as a scaffold component. This biopolymer indicates biocompatibility and bioactivity caused by the existence of specific amino acid sequences, being preferred sites for interactions with cells, with high similarity to natural extracellular matrix. The present paper does not aspire to be a full review of electrospinning of gelatin and gelatin containing nanofibers as scaffolds in tissue engineering. It is focused on the still open question of the role of the higher order structures of gelatin in scaffold’s bioactivity/functionality. Gelatin molecules can adopt various conformations depending on temperature, solvent, pH, etc. Our review indicates the potential ways for formation of α-helix conformation during electrospinning and the methods of further structure stabilization. It is intuitively expected that the native α-helix conformation appearing as a result of partial renaturation of gelatin can be beneficial from the viewpoint of bioactivity of scaffolds, providing thus a much cheaper alternative approach as opposed to expensive electrospinning of native collagen.

Keywords:
gelatin, molecular conformation, electrospinning, nanofibers, scaffolds

Abstract:
Novel polyurethanes based on synthetic, atactic poly[(R, S)-3-hydroxybutyrate] (a-PHB) and polycaprolactone (PCL) or polyoxytetramethylene (PTMG) diols were synthesized. It was shown that the presence of a-PHB within soft segments reduces crystallinity of PUR. Because of the low melting temperature for polyurethanes with PCL in soft segments, at this stage of work, electrospinning was limited to polyurethanes containing PTMG and a-PHB. Polyurethane containing 80% of PTMG and 20% of a-PHB was electrospun at various parameters from hexafluoro-2-propanole solution, resulting in formation of fibers with the average diameter ca. 2 μm. The fiber diameter decreased with decreasing polymer concentration in a solution and was practically insensitive to the needle-collector distance in the applied range of distances.

Keywords:
polyurethane, polyhydroxybutyrate, electrospinning, scaffolds

Abstract:
Wide Angle X-ray Scattering (WAXS), Differential Scanning Calorimetry (DSC) and Fourier Transform Infrared spectroscopy (FTIR) analyses of phase composition and of thermal properties of PVDF samples, crystallized at temperatures 27 - 155°C by casting from N,N-dimethyl formamide (DMF) solution, are reported. Samples obtained at 27°C contain only β crystal phase and with increase of casting temperature content of β phase decreases in favor of α phase. Evaluation of combined: phase content (WAXS) and melting heat (DSC), leads to two fold higher than for 100 % α phase value of 100% β melting enthalpy, ΔHβ0= 219.7 J.g-1, which may be justified by strong polar interactions in β phase TTT conformation. The relation ΔHβ0 > ΔHα0 leads either to the thermodynamic stability of β phase in whole temperature range (if Tmβ0 ≥ Tmα0) or to the limited temperature range of thermodynamic stability of α phase (if Tmβ0 < Tmα0).

Keywords:
pvdf, WAXS, FTIR, DSC, crystallinity, polymorphism

Abstract:
Analysis of the thermo-mechanical behavior of electrospun thermoplastic polyurethane (TPU) block co-polymer nanofibers (glass transition temperature ∼−50°C) is presented. Upon heating, nanofibers began to massively contract, at ∼70°C, whereas TPU cast films started to expand. Radial wide-angle X-ray scattering (WAXS) profiles of the nanofibers and the films showed no diffraction peaks related to crystals, whereas their amorphous halo had an asymmetric shape, which can be approximated by two components, associated with hard and soft segments. During heating, noticeable changes in the contribution of these components were only observed in nanofibers. These changes, which were accompanied with an endothermic DSC peak, coinciding with the start of the nanofibers contraction, can be attributed to relaxation of an oriented stretched amorphous phase created during electrospinning. Such structure relaxation becomes possible when a portion of the hard segment clusters, forming an effective physical network, is destroyed upon heating.

Keywords:
Block-copolymer, Electrospinning, Nanofibers, Thermo-mechanical properties

Abstract:
Blends of polycaprolactone (PCL) and gelatine (Ge), being effective materials for tissue engineering strategies, were electrospun at various conditions and polymer weight ratios. The morphology, the supermolecular structure as well as the mechanical properties of resulting submicron sized fibres have been analyzed in relation to electrospinning conditions and PCL/Ge weight ratio. Compared to pure PCL, Ge addition leads to large reduction of fibre diameter and finally to changes of fibre morphology. For parallelised fibres collected on a rotating drum, preferred molecular orientation of PCL crystals is found. With increasing Ge content a general reduction of molecular orientation is observed. In addition, there is peculiar dependence of polycaprolactone crystallinity on the content of Ge, showing maximum at low Ge concentration (20%) as determined by differential scanning calorimetry (DSC) and wide angle X-ray scattering (WAXS). Such a trend can be explained by hydrophobic interactions in the system containing PCL, gelatine and water, being additional driving forces for crystallization of nonpolar PCL molecules. The presence of water within investigated blend systems has been evidenced experimentally using thermal gravimetric analysis (TGA). Young’s modulus of nonwovens, as determined by uniaxial tensile testing, indicates the effect of additivity of the stiffness of both polymers as well as the influence of preferred molecular orientation. Additional experiments were performed using collagen (Col) as a biopolymeric alternative to Ge. WAXS results show evidently amorphous structure of Col within the blended fibres, indicating strong tendency for denaturation of collagen into gelatine under the influence of hexafluoroisopropanol as a solvent.

Keywords:
Electrospinning, Nanofibres, Blend, Gelatine polycaprolactone, Molecular structure

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:
The potential of polarizing-interference Pluta microscope for determination of optical birefringence of individual nanofibers formed by electrospinning was shown. This technique can be applied for measurements of fiber birefringence, practically at diameter above 300 nm. The molecular orientation of individual polycaprolactone (PCL) nanofibers was determined from birefringence assuming the same orientation of both phases, crystal and amorphous. The molecular orientation was determined using DSC crystallinity, crystal intrinsic birefringence calculated for the first time for PCL from bond polarizabilities as well as estimated value of amorphous intrinsic birefringence. Our results indicate that the birefringence and thus molecular orientation are strongly inhomogeneous along the nanofibers, reflecting a complex nature of forces acting during electrospinning process. The average molecular orientation is weak if any, being dependent together with fiber thickness and crystallinity on electrospinning parameters, like applied voltage, concentration and type of solvent. The obtained results indicate that the average molecular orientation displays similar dependence on applied voltage as fiber diameter. Relatively low melting temperature of electrospun nanofibers suggests low crystal size and/or high concentration of defects in crystals. This observation corresponds with low crystallinity and molecular orientation, indicating together relatively low degree of crystal ordering due to high rate of cooling and solvent evaporation during electrospinning, limiting thus crystallization process.

Keywords:
Nanofibers, Electrospinning, Birefringence, Polarizing-interference microscopy, Polycaprolactone

Abstract:
The kinetics of isothermal crystallization of various polymers was investigated by light depolarization technique (LDT) using the new setup with direct registration of depolarization ratio. Experimental data were analyzed using new method proposed by Ziabicki who shown that degree of crystallinity is a non-linear function of degree of depolarization, crystal thickness, and its birefringence. Other experimental methods were involved providing supplementary information on crystal thickness (SAXS) and allowing comparison of crystallization kinetics (WAXS, DSC). The advantage of LDT relies on high sampling rate allowing on-line measurements and lack of inertia effects that exist in other methods like calorimetry. The limitations of the applicability of the method are discussed. The method needs supplementary information not only on crystal thickness but also on variable optical birefringence of real crystals. Our results show that LDT can be used in a simple way for investigation of crystallization kinetics at relatively high temperatures, providing large and perfect crystals. In such a case it is sufficient to use crystal intrinsic birefringence and final crystal thickness typical at particular temperature of crystallization. On the other hand, depolarization ratio combined with measurements by other methods (crystallinity and crystal thickness) can be used for estimation of crystal birefringence.

Keywords:
Polymer, Crystallization kinetics, Light depolarization, Crystal birefringence

Abstract:
Phase transitions in isotactic polypropylene were investigated during isothermal crystallization and heating after isothermal crystallization using various experimental techniques. The results obtained by wide angle x-ray scattering (WAXS), light depolarization technique (LDT), differential scanning calorimetry (DSC) and optical microscopy show that crystallization of isotactic polypropylene can result in simultaneous formation of two crystal modifications, alpha and beta. There is clear experimental evidence that beta phase tends to convert into alpha modification during crystallization as well as during subsequent heating. Experimental results are compared with numerical simulation performed according to the model of nucleation-controlled phase transitions in multiphase systems. The results of simulation show that beta phase is not thermodynamically stable in any temperature range. The reason for the appearance of beta phase is related to low interfacial tension of melt vs. beta. It has been also shown that maximum crystallinity reached in experiments does not exceed 40–50% in agreement with the concept of constrained amorphous phase.

Keywords:
polypropylene, polymorphism, metastability, crystallization

Abstract:
A new setup for light depolarization measurements was designed. Two innovative elements have been introduced. The first is an electronic system which enables depolarization ratio to be registered directly. The second is a system of temperature control allowing effective implementation of a temperature–time program according to the particular requirements. Direct registration of depolarization ratio instead of intensity of depolarized light for individual components (parallel and perpendicular), as is performed in the usual apparatus, allows elimination of light scattering effects because of the insensitivity of depolarization ratio to the scattering level. Application of the new setup was shown for crystallization and melting of isotactic polypropylene (i-PP). Comparison of phase transitions in i-PP, as registered by light depolarization and DSC, indicates some differences. Possible sources of the observed differences are discussed.

Keywords:
Light depolarization, Polymers, Crystallization, Melting, Kinetics of phase transitions

Abstract:
The time dependence of nucleation rate in isothermal crystallization of poly(3-hydroxybutyrate) was experimentally shown, both in heterogeneous and homogeneous nucleation. The time dependence of nucleation rate is one of the important limitations for the applicability of the simplified form of Kolmogoroff- Avrami-Evans model with time independent kinetic characteristics. The presented results are interpreted in terms of non-steady-state cluster size distribution underlying transient nature of nucleation. The relaxation time needed for reaching a steady-state cluster size distribution and thus steady-state nucleation rate is relatively long, exceeding the time of exhaustion of heterogeneities. The relaxation time estimated from homogeneous process was tens of seconds in the temperature range between 83 and 120 oC. Application of Arrhenius law allows estimation of relaxation time in broader temperature range, showing an increase of relaxation time with decreasing temperature.

Keywords:
PHB, isothermal crystallization, nucleation rate

Abstract:
Studies of kinetics of polymer crystallization are generally performed by heating the material above the melting point, in order to erase previous thermal and mechanical history, followed by rapid cooling to the desired crystallization temperature or by cooling at a constant rate. For poly(3-hydroxybutyrate) this procedure implies some degradation of the polymer chain, which starts below the onset of melting. In this article the effects of melting conditions on the subsequent crystallization kinetics are discussed. It is shown that in order to sufficiently cancel memories of previous crystalline order of the analyzed PHB, it is necessary to bring the material at a temperature higher than 192 °C. Thermal treatments conducted at lower temperatures are not sufficient to destroy all solid aggregates, and crystallization of PHB has an anticipated onset of crystallization due to nucleation occurring via self-seeding. The chain degradation attained upon exposure at high temperatures has much lesser influence on crystallization kinetics than incomplete melting, with some effects detectable on the spherulitic morphology and on the final degree of crystallinity.

Keywords:
PHB, thermal history, crystallization, degradation

Abstract:
Melt-crystallization of polyvinylidene fluoride (PVDF) was investigated in non-isothermal mode at ultra-high cooling rates ranging between 30–3000 K/s as well as at constant temperatures after quenching at 6000 K/s. An increase of the cooling rate above 150 K/s leads to the formation of betaphase manifested by a low temperature shoulder of crystallization exotherm in addition to the alphamodification. At the cooling rates above 2000 K/s there is only low temperature exothermic peak that is attributed to the crystallization of pure betamodification. Isothermal crystallization was possible to realize at 110 oC as the lowest, resulting in form. Much higher crystallization rate in submicrogram samples, as compared to standard DSC experiments, is also reported.

Keywords:
Polyvinylidene fluoride, Crystallization, Ultra-fast calorimetry, Polymorphism

Abstract:
This article details a study of irregularly shaped DSC exotherms of poly(3-hydroxybutyrate) (PHB) developed during isothermal and non-isothermal crystallization. Due to the extreme purity of the polymer, PHB crystallization is mainly initiated by homogeneous nucleation, with the formation of very large spherulites, especially under slow nucleation conditions. When the number of growing spherulites is low, the evolution of latent heat is very sensitive to every act of nucleation as well as to the space limitations in the process of growth, resulting in non-monotonous development of latent heat, with sudden increases and decreases in crystallization rates. This results in non conventional DSC exotherms, under given crystallization conditions, characterized by spikes or shoulders associated to nucleation of new spherulites.

Keywords:
PHB, crystallization, nucleation, DSC

Abstract:
The analysis of WAXS profiles for various polyethylenes indicates that the proper description of a structure needs the introduction of a kind of ‘third phase’ in addition to the classical crystalline and amorphous phases. The structure of the additional phase is intermediate between that of the amorphous and crystalline phase. With increasing branch content and molecular weight the intermediate phase becomes more similar to the structure of amorphous phase. The experimental evidence for the intermediate phase is derived not only from the crude approximation of WAXS profiles based on the two phase model but also from the unexpected behavior of the parameters of amorphous halo during crystallization. When crystallization is started, an analysis based upon two-phase model results in an apparent increase of the diffraction angle and width of amorphous halo with time above the values anticipated from the range before the start of crystallization. This is caused by the fact that the amorphous fitting function tries to cover a peak of the intermediate component that appears between morphous halo and (110) reflection of crystalline phase. The conventionally applied two-phase model leads to several serious errors in determination of structural parameters of both phases. The analysis of crystallization kinetics using three-phase model provides additional information on the nature of crystallization itself.

Keywords:
Polyethylene, Crystallization, Intermediate phase

Abstract:
Crystallization of polypropylene (PP) can result in formation of different crystal modifications depending on external conditions. The mechanisms of formation of various crystal modifications in polypropylene are still under discussion.We have investigated non-isothermal melt crystallization of isotactic polypropylene at cooling rates ranging from 1 up to 180,000 K/min using two types of differential scanning calorimeters-standard device Perkin–Elmer DSC Pyris-1 and ultra-fast calorimeter. Additional results were obtained by means of wide angle X-ray scattering and optical microscopy. At cooling rates below 6000 K/min there is only one exothermic peak corresponding to simultaneous crystallization ofalpha andbeta modifications. At cooling rates higher than 6000 K/min there is additional low temperature DSC peak corresponding to formation of mesomorphic phase. At the rates higher than 36,000 K/min there is no trace of formation of any ordered phase. In our opinion this complex behavior observed during crystallization of polypropylene can be explained using the concept of metastable phases. An increase ofbeta content in samples with quinacridone pigment has been observed only at very low cooling rates, corresponding to high temperatures of crystallization and low homogeneous nucleation rate.

Keywords:
Polypropylene, Crystallization, Utra-fast calorimetry, Crystallographic modifications, Metastable phases

Abstract:
In the present study, a full bionanocomposite scaffold from poly (vinyl alcohol) (PVA), nanohydroxy apatite (nHAp) and cellulose nanofibers (CNF) was fabricated by electospinning and its potential application for bone tissue engineering was investigated. Morphology of the electrospun scaffolds was seen by field emission scanning electron microscope (FE-SEM). Both nHAp and CNF enhanced the tensile modulus of the scaffolds; however, both tensile strength as well as slongation at break showed reduced behaviour. Porosity measurement showed that scaffolds had porosity more than 70% which is appropriate for tissue engineering scaffolds. Contact angle test proved high hydrophilicty of electrospun mats while nanofiller incorporation promoted hydrophilicity. Biodegradability was investigated in phosphate buffer saline (PBS). In vitro biomineralization in simulated body fluid (SBF) and MTT cytotoxicity analysis showed that addition of nHAp and CNF increased bioactivity and cell viability of the scaffolds. The obtained results offered a 3-component promising scaffold for bone tissue engineering.

Keywords:
Bionanocomposite, Scaffold, Electrospinning, Poly (vinyl alcohol) and Bone tissue engineering

Abstract:
Three types of membrane structures with wide pores were compared in this study. One of the membranes was obtained from polyethersulfone using cellulose fibers as the macropore precursors. Two of the fibers were obtained from poly(L-lactide). As the macropore precursors olyvinylpyrrolidone (1.2 MDa) and pork gelatin non-woven were used, the influence of non-woven fibers on the structure of membranes was shown. Necessity of specific membrane structure application was explained. The hoice of polymers and co-polymers with a range of biodegradation times can determine the scaffold type suitable for the age of a patient.

Keywords:
Polysulfone membrane, Polyester membranes, Membrane structures, Biodegradable membranes, 3D scaffold

Keywords:
electrospinning, ultrasonication, short fibers, polymers, scaffold

Keywords:
Piezo-nerve, scaffold, nanofibers, tissue engineering, stem cells

Keywords:
coatings, fibre-based biomaterials incl. electrospinning, material/tissue interfaces

Keywords:
Nanofibers, polyesters, surface functionalization, aminolysis, structure

Keywords:
scaffolds, electro spinning, tissue engineering

Keywords:
scaffolds, electrospinning, polyvinylidene fluoride, tissue engineering

Keywords:
polymers, aminolysis, surface modification, tissue engineering

Keywords:
methylcellulose, agarose, hydrogel, cross-linking kinetics, DMA, modulus

Abstract:
Electrospun nonwovens from PCL/gelatin and PCL/collagen structurally mimic native extracellular matrix and provide cells with chemical cues affecting them. Electrospinning of bicomponent nanofibres requires the use of a solvent which dissolves both of the polymers.
We have optimized the process of electrospinning of PCL/gelatin and PCL/collagen nanofibers based on the use of non-toxic, alternative solvents: acetic acid and formic acid (AA/FA) as previously described [1].
Bicomponent PCL/gelatin and PCL/collagen nanofibers were formed by electrospinning using the mixture of acetic acid and formic acid (9:1 w/w ratio), while hexafluoroisopropanol (HFIP) was used as a reference solvent. Nonwoven materials were subjected to cellular in vitro and biodegradation tests and compared.
All in vitro tests were performed using L929 mouse fibroblast cells. Cytotoxicity test was carried out on extracts and showed that all type of materials are not cytotoxic. Materials with 10% biopolymer content as well as made from PCL only underwent experiment in direct contact. Cells were cultured on materials for 3, 5 and 7 days and afterwards taken for SEM as well as fluorescent dying of nuclei and cytoskeleton. Obtained results proved that the addition of Arg-Gly-Asp (RGD) amino acid sequences from biopolymer, in comparison to pure PCL materials, facilitates cell adhesion and spreading on the surface of nonwovens regardless of solvent used in electrospinning.
PCL/gelatin and PCL/collagen nonwovens underwent biodegradation in PBS solution at 37°C. After different times, ranging from 1 to 90 days samples were subjected for comparative analysis via various methods.
Despite the fact that bicomponent nanofibers electrospun from alternative solvents have similar morphology to those electrospun from perfluorinated alcohols, they differ in the internal structure which seriously affects biodegradation process. Biodegradation of investigated materials is manifested mainly by the gelatin leaching, which leads to nanofibers erosion, particularly large for nanofibers spun from AA/FA.

Keywords:
electrospinning, bicomponent nanofibers, biodegradation, cellular studies

Keywords:
chitosan, PCL, cellular responce, electrospinning

Keywords:
chitozan, polikaprolakton, electrospinning, L929, cytotoksyczność

Keywords:
electrospinning, fibres, surface, chitosan

Abstract:
Cells sense subtract stiffness, elasticity and transduce that information into morphological changes and lineage specification. Polymer molecular order and mechanical properties, specially stiffness and elasticity indicate influence on cellular response during in- vitro study [e.g. Bershadskye et al 2013]. The aim of proposed presentation is to evaluate the effect of tailored crystallinity and mechanical properties of one- and bicomponent polymer films in terms of cells morphology and proliferation without changing other parameters. Polycaprolactone (PCL) and Gelatin (Ge) were used. As a solvents: Hexafluoroisopropanole (H), Acetic Acid (AA) were chosen. Two methods of foil preparation were analysed: forming from melt (onecomponent), forming from solution (one- and bicomponent).In both methods, the degree of crystallinity was modified mainly by the different type of PCL molecular weight, solvent type and/or annealing. Films were analysed using polarizinginterference microscopy allowing characterization of spherulities morphology. Degree of crystallinity was analysed by differential scanning calorimetry. Foils topography was analysed by atomic force microscopy, selected mechanical properties and hydrophilicity (contact angle) as the significant from the viewpoint of cellular activity were determined as well. L929 cells adhesion and morphology ware analysed by immunohistochemical staining for actin and nuclei. Cell activity and proliferation were analysed also. It is evident that conditions of PCL films preparation affect the morphology of spherulites. All samples were birefringent, indicating in general crystallinity, being different for particular samples. Maltese cross was observed in few samples. Crystallinity of PCL films determined from DSC measurements was in range 0,45-0,70 depending on solvent and polymer molecular weight used. Young Modulus strongly depends on Mw of PCL and Ge additive. L929 cells interact with subtract; morphology and proliferation degree change with crystallinity and elasticity of one- and bicomponet films.

Keywords:
crystallinity, PCL, mechanical properties, casted films

Abstract:
Bicomponent polycaprolactone/gelatin nanofibers were formed by electrospinning as previously described [1] using a novel polymer – solvent system with solvents being alternative to the commonly used toxic solvents like fluorinated alcohols. PCL/Gelatin nanofibres were electrospun from the mixture of acetic acid (AA) with formic acid (FA) (90:10) and from hexafluoroisopropanol (HFIP), that was used as reference solvent. PCL/Gelatin nanofibres with polymers w/w ratios 9:1, 8:2 and 7:3, underwent biodegradation in PBS solution at 37°C. After different times, ranging from 1 to 90 days, they were rinsed in demineralized water and dried. Weight loss and FTIR tests were performed to assess the kinetics of gelatin leaching, while SEM imaging and hydrophobicity tests to show its depletion from the surface. DSC measurements were carried out to examine changes in fibres’ internal structure and uniaxial tensile testing to compare their mechanical properties. Morphology of PCL/Gt fibers obtained from AA/FA is similar to that obtained from HFIP. Despite similar morphology, the internal structure of nanofibers formed from alternative solvents is different, reflecting the emulsive nature of PCL/gelati n mixture in AA/FA solvents contrary to clear, transparent solutions in HFIP. This apparent difference affects strongly the kinetics of leaching of gelatin from bicomponent fibres and thus how their mechanical and bioactive properties are changing in time after placing in living organism. There is substantial difference in kinetics of gelatin leaching depending on solvent used. Mass measurements show much faster gelatin degradation in nanofibres electrospun from AA/FA than from HFIP. For instance, for PCL/Gt 7:3 samples, gelatin content loss is 85% for AA/FA and 68% for HFIP after 90 days. Moreover, irrespective of the solvent used, the degradation rate increases with initial content of gelatin and is the highest in the first 24 hours: 27% for AA/FA 9:1 and 67% for 7:3 and 13% and 32% for HFIP respectively. The observed changes can be explained by nonuniform distribution of gelatin within fibres spun form AA/FA due to emulsive character of solution. Comparison of SEM images reveals linear groove-like sites remaining after gelatin leaching on a surface of fibres spun from AA/FA solvent. Contrary to this, fibres spun from HFIP remain smooth which can be attributed to molecular dispersion of both components.

Keywords:
nanofibers, biodegradation, 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

Abstract:
In this study bicomponent polycaprolactone/gelatin nanofibers were successfully formed by electrospinning using for the first time a novel polymer – solvent system consisting of acetic acid and formic acid. Such solvent system is alternative to the commonly used toxic solvents like fluorinated alcohols, mainly hexafluoroisopropanol. The effect of electrospinning conditions on morphology and structure of nanofibers were investigated.

Keywords:
nanofibers, electrospinning, polycaprolactone, gelatin, alternative solvents, structure

Abstract:
Few research groups have highlighted the unexpected degree of cell proliferation depending on the degree of crystallinity of the substrate. Commonly used methods of forming three-dimensional scaffolds do not take into account crystallinity optimisation.
The aim of proposed presentation is to investigate polycaprolactone (PCL) substrate supermolecular structure effect, mainly crystallinity, on cells spreading, activity and proliferation.

Keywords:
crystalllinity, PCL, solvent, molecular structure, L929