Instytut Podstawowych Problemów Techniki

Streszczenie:

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.

Słowa kluczowe:

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

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Streszczenie:

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.

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Streszczenie:

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.

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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.

Słowa kluczowe:

gelatin, aminolysis, surface modification, electrospinning

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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.

Słowa kluczowe:

electrospinning, ultrasonication, short fibers, polymers

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This study was conducted as a first step in obtaining eco-friendly fibres for medical applications using a synthesised oligomer poly(glycerol succinate) (PGSu) as an additive for synthetic poly(L-lactic acid) (PLLA) and poly (L-lactide-co-caprolactone) (PLCL). The effects of the oligomer on the structure formation, morphology, crystallisation behaviour, and mechanical properties of electrospun bicomponent fibres were investigated. Nonwovens were investigated by means of scanning electron microscopy (SEM), wide angle X-ray scattering (WAXS), differential scanning calorimetry (DSC), and mechanical testing. The molecular structure of PLLA fibres is influenced by the presence of PGSu mainly acting as an enhancer of molecular orientation. In the case of semicrystalline PLCL, chain mobility was enhanced by the presence of PGSu molecules, and the crystallinity of bicomponent fibres increased in relation to that of pure PLCL. The mechanical properties of bicomponent fibres were influenced by the level of PGSu present and the extent of crystal formation of the main component. An in vitro study conducted using L929 cells confirmed the biocompatible character of all bicomponent fibres.

Słowa kluczowe:

poly(glycerol succinate), plasticiser, eco-friendly polymer, electrospinning, hyperbranched polyester

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Streszczenie:

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.

Słowa kluczowe:

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

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Sodium aluminophosphate glasses were evaluated for their bone repair ability. The glasses belonging to the system 45Na2O–xAl2O3-(55-x)P2O5, with x = (3, 5, 7, 10 mol%) were prepared by a melt-quenching method. We assessed the effect of Al2O3 content on the properties of Na2O–Al2O3–P2O5 (NAP) glasses, which were characterized by density measurements, DSC analyses, solubility, bioactivity in simulated body fluid and cytocompatibility with MG-63 cells. To the best of our knowledge, this is the first investigation of calcium-free Na2O–Al2O3–P2O5 system glasses as bioactive materials for bone tissue engineering.

Słowa kluczowe:

alumina, bioactive glass, bone tissue engineering, calcium-free bioactive glass, phosphate glass, sodium aluminophosphate glass

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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.

Słowa kluczowe:

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

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Two types of phosphate glasses 50Na2O-20B2O3-30P2O5 (NBP) and 30CaO-20Na2O-50P2O5 (CNP) with different content of TiO2 (0, 3 and 5 mol%) have been prepared by melt-quenching process. TiO2 was added to increase glass network stability. Physical properties of glasses were investigated by density measurements, differential scanning calorimetry and degradation in phosphate buffered saline (PBS). Biological performance of glasses in a direct contact with osteoblast-like MG-63 cells was analysed with the use of resazurin test and live-dead staining. The results show that TiO2 addition increased density, glass transition temperature (Tg) and melting temperature (Tm) of both types of glasses. In the case of NBP glasses presence of TiO2 resulted in their fast degradation in PBS and acidification of cell culture medium. As a consequence such glasses did not support cell adhesion and growth, but they can be considered for e.g. drug delivery systems. On the other hand addition of TiO2 to CNP glasses resulted in enhanced cell adhesion and viability. Particularly positive results were found for CNP glass containing 5% TiO2, so it can be a good candidate as a scaffold material for bone tissue engineering.

Słowa kluczowe:

phosphate glasses, bioactive glasses, titanium dioxide, tissue engineering

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Słowa kluczowe:

electrospinning, ultrasonication, short fibers, polymers, scaffold

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Słowa kluczowe:

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

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Słowa kluczowe:

Nanofibers, polyesters, surface functionalization, aminolysis, structure

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Słowa kluczowe:

polymers, aminolysis, surface modification, tissue engineering

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