'Cookies on a tray': Superselective hierarchical microstructured poly(l-lactide) surface as a decoy for cells.

In this research we developed a micro-sized hierarchical structures on a poly(l-lactide) (PLLA) surface. The obtained structures consist of round-shaped protrusions with a diameter of ~20 µm, a height of ~3 µm, and the distance between them ~30 µm. We explored the effect of structuring PLLA to design a non-cytotoxic material with increased roughness to encourage cells to settle on the surface. The PLLA films were prepared using the casting melt extrusion technique and were modified using ultra-short pulse irradiation - a femtosecond laser operating at λ = 1030 nm. A hierarchical microstructure was obtained resembling 'cookies on a tray'. The cellular response of fibro- and osteoblasts cell lines was investigated. The conducted research has shown that the laser-modified surface is more conducive to cell adhesion and growth (compared to unmodified surface) to such an extent that allows the formation of highly-selectively patterns consisting of living cells. In contrast to eukaryotic cells, the pathogenic bacteria Staphylococcus aureus covered modified and unmodified structures in an even, non-preferential manner. In turn, adhesion pattern of eukaryotic fungus Saccharomyces boulardii resembled that of fibro- and osteoblast cells rather than that of Staphylococcus. The discovered effect can be used for fabrication of personalized and smart implants in regenerative medicine.

Influence of cross-linking time on physico-chemical and mechanical properties of bulk poly(glycerol sebacate).

In the presented study, a PGS prepolymer (pPGS) was synthesized utilizing polycondensation technique (equimolar sebacic acid:glycerol ratio, 130 °C, 24 h). Subsequently, the pPGS was thermally cross-linked in vacuum oven in 130 °C for 84 and 168 h. The cylindrical and dumbbell-shaped samples were subjected for physico-chemical and thorough mechanical analysis including tensile and compressive strength evaluation as well as dynamic mechanical thermal analysis (DMTA). The study allowed for the investigation of alteration of PGS properties during cross-linking and decay of elastomeric properties over prolonged cross-linking time. Moreover, a deconvolution in FTIR analysis allowed to glimpse into the hydrogen bonding structure of the materials which weakens during the cross-linking. The obtained results can be utilized during designing PGS-based bulk materials for biomedical application where bearing mechanical loads and tuned chemical character is of vital importance.

Cytotoxicity Study of UV-Laser-Irradiated PLLA Surfaces Subjected to Bio-Ceramisation: A New Way towards Implant Surface Modification.

In this research we subjected samples of poly(L-lactide) (PLLA) extruded film to ultraviolet (193 nm ArF excimer laser) radiation below the ablation threshold. The modified film was immersed in Simulated Body Fluid (SBF) at 37 °C for 1 day or 7 days to obtain a layer of apatite ceramic (CaP) coating on the modified PLLA surface. The samples were characterized by means of optical profilometry, which indicated an increase in average roughness (Ra) from 25 nm for the unmodified PLLA to over 580 nm for irradiated PLLA incubated in SBF for 1 day. At the same time, the water contact angle decreased from 78° for neat PLLA to 35° for irradiated PLLA incubated in SBF, which suggests its higher hydrophilicity. The obtained materials were investigated by means of cell response fibroblasts (3T3) and macrophage-like cells (RAW 264.7). Properties of the obtained composites were compared to the unmodified PLLA film as well as to the UV-laser irradiated PLLA. The activation of the PLLA surface by laser irradiation led to a distinct increase in cytotoxicity, while the treatment with SBF and the deposition of apatite ceramic had only a limited preventive effect on this harmful impact and depended on the cell type. Fibroblasts were found to have good tolerance for the irradiated and ceramic-covered PLLA, but macrophages seem to interact with the substrate leading to the release of cytotoxic products.

Influence of Thermal Annealing on the Sinterability of Different Grades of Polylactide Microspheres Dedicated for Laser Sintering.

We present a comparison of the influence of the conditioning temperature of microspheres made of medical grade poly(L-lactide) (PLLA) and polylactide with 4 wt % of D-lactide content (PLA) on the thermal and structural properties. The microspheres were fabricated using the solid-in-oil-in-water method for applications in additive manufacturing. The microspheres were annealed below the glass transition temperature (Tg), above Tg but below the onset of cold crystallization, and at two temperatures selected from the range of cold crystallization corresponding to the crystallization of the α' and α form of poly(L-lactide), i.e., at 40, 70, 90, and 120 °C, in order to verify the influence of the conditioning temperature on the sinterability of the microspheres set as the sintering window (SW). Based on differential scanning calorimetry measurements, the SWs of the microspheres were evaluated with consideration of the existence of cold crystallization and reorganization of crystal polymorphs. The results indicated that the conditioning temperature influenced the availability and range of the SWs depending on the D-lactide presence. We postulate the need for an individual approach for polylactide powders in determining the SW as a temperature range free of any thermal events. We also characterized other core powder characteristics, such as the residual solvent content, morphology, particle size distribution, powder flowability, and thermal conductivity, as key properties for successful laser sintering. The microspheres were close to spheres, and the size of the microspheres was below 100 µm. The residual solvent content decreased with the increase of the annealing temperature. The thermal conductivities were 0.073 and 0.064 W/mK for PLA and PLLA microspheres, respectively, and this depended on the spherical shape of the microspheres. The wide angle X-ray diffraction (WAXD) studies proved that an increase in the conditioning temperature caused a slight increase in the crystallinity degree for PLLA microspheres and a clear increase in crystallization for the PLA microspheres.

Brief review on poly(glycerol sebacate) as an emerging polyester in biomedical application: Structure, properties and modifications.

Poly(glycerol sebacate) (PGS) is an aliphatic polyester which attracted significant scientific attention in recent years due to its vast potential in biomedical applications with regard to tissue engineering. It has been presented in the literature in the form of 2D films, porous scaffolds or nonwovens, to name just a few. Moreover, various applications have been proposed as a component of composite materials or polymer blends. Its physicochemical properties can be significantly adjusted by means of synthesis and post-synthetic modifications, including cross-linking or chemical modification, such as copolymerization. Many scientists have discussed PGS as a new-generation polymer for biomedical applications. Its regenerative potential has been confirmed, in particular, in tissue engineering of soft tissues (including nerve, cartilage and cardiac tissues). Therefore, we must anticipate a growing importance of PGS in contemporary biomedical applications. This brief review aims to familiarize the readers with this relatively new polymeric material for tissue engineering applications.

Pectin-organophilized ZnO nanoparticles as sustainable fillers for high-density polyethylene composites.

A series of nanocomposites made of high-density polyethylene (HDPE) and 10 wt% zinc oxide nanoparticles (ZnO NPs) were produced by extrusion and injection molding. The nanoparticles were prepared via a green way using the pectin-based banana peel extract as the stabilizer and a proper dispersion-providing agent. The fillers were well-dispersed in the matrix and the composites exhibited improved functional characteristics such as increased thermal stability and mechanical properties. The presence of the pectin-organophilized filler had a significant impact on the crystallization process of HDPE. The kinetics of the degradation process was also altered in comparison to the pure polymer. The fire properties of the composites were enhanced as the amount of the gas products produced during their degradation was reduced, what was confirmed by thermogravimetric analysis coupled with gas products analyses (TGA/FTIR/QMS). The structure and morphology of the materials were characterized by scanning electron microscope (SEM), infrared spectroscopy (FTIR), X-ray diffraction (XRD), and differential scanning calorimetry (DSC). Additionally, the mechanical properties were tested by tensile tests. An in-depth analysis revealed that the HDPE-pectin-ZnO interactions are crucial for the structural and performance properties of the final composite. The used biopolymer reacts with ZnO via ionic interaction and through hydrogen bond in the case of HDPE.

New approach to modification of poly (l-lactic acid) with nano-hydroxyapatite improving functionality of human adipose-derived stromal cells (hASCs) through increased viability and enhanced mitochondrial activity.

The aim of this study was to determine the cytocompatibility of poly (l-lactide) (PLLA) scaffolds fabricated using co-rotating twin screw extrusion technique and functionalized with different concentrations of nano-hydroxyapatite (nHAp). The efforts were aimed on the designing bioactive scaffolds improving the viability and metabolic activity of human adipose-derived multipotent stromal cells (hASCs). The in vitro study was designed to determine the optimal nHAp concentration, based on analysis of hASCs morphology, adhesion rate, as well as metabolic and proliferative potential. Initially, the PLLA filled with three different concentrations of the nHAp were tested i.e. 5%, 10% and 15 wt%. The obtained results indicated that the 10 wt% nHAp in the PLLA (10% nHAp/PLLA) matrices improved the adhesion and proliferation of the hASCs, what was in good agreement with the results of tensile properties of the composites. Further, we performed profound studies regarding the cytotoxicity of 10% nHAp/PLLA. The analysis included the evaluation of the biomaterial influence on viability, apoptosis-related markers expression profile and mitochondrial function. The cytocompatibility of 10% nHAp/PLLA scaffolds toward the hASCs was confirmed. The hASCs propagated on 10% nHAp/PLLA were more viable then those propagated on the plain PLLA. The level of pro-apoptotic markers, i.e. caspase-3 and Bax in cultures on 10% nHAp/PLLA was significantly decreased. Obtained results correlated with higher mitochondrial membrane potential of hASCs in those cultures. The obtained composites may improve therapeutic potential of hASCs via directing their adhesion, enhancing proliferation and viability as well as increasing mitochondrial potential, thus may find potential application in tissue engineering.