Instytut Podstawowych Problemów Techniki

Streszczenie:

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.

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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.

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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.

Słowa kluczowe:

methylcellulose, thermosensitive hydrogel, crosslinking, DSC

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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.

Słowa kluczowe:

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

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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.

Słowa kluczowe:

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

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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.

Słowa kluczowe:

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

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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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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.

Słowa kluczowe:

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

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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.

Słowa kluczowe:

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

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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.

Słowa kluczowe:

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

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Słowa kluczowe:

electrospinning, ultrasonication, short fibers, polymers, scaffold

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Słowa kluczowe:

electrospinning, microfibers, nanofibers, in vivo, animal model

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Słowa kluczowe:

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

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