In order to make bioplastics accessible for a wider spectrum of applications, ready-to-use plastic material formulations should be available with tailored properties. Ideally, these kinds of materials should also be "home-compostable" to simplify their organic recycling. Therefore, materials based on PLA (polylactid acid) and PHB (polyhydroxybutyrate) blends are presented which contain suitable additives, and some of them contain also thermoplastic starch as a filler, which decreases the price of the final compound. They are intended for various applications, as documented by products made out of them. The produced materials are fully biodegradable under industrial composting conditions. Surprisingly, some of the materials, even those which contain more PLA than PHB, are also fully biodegradable under home-composting conditions within a period of about six months. Experiments made under laboratory conditions were supported with data obtained from a kitchen waste pilot composter and from municipal composting plant experiments. Material properties, environmental conditions, and microbiology data were recorded during some of these experiments to document the biodegradation process and changes on the surface and inside the materials on a molecular level.

• Klíčová slova
• biodegradation, blend polymeric material, home-compost, industrial compost, polyhydroxybutyrate (PHB), polylactic acid (PLA),
• Publikační typ
• časopisecké články MeSH

In this study preparation and characterization of new UV-protecting systems based on liposomes/polyhydroxybutyrate (PHB) with encapsulated coffee extracts are presented. Green and roasted coffee extracts with high phenolics content, high antioxidant activity and sun protection factor (SPF) value 40-50 were used as model organic UV filters and encapsulated into liposomes and PHB-liposomes. Particle size and colloid stability was observed by dynamic light scattering and zeta-potential. Toxicity of particles was tested by MTT and LDH assay on HaCaT cell line. All prepared samples showed moderate or high encapsulation efficiency. Addition of PHB up to 50 % of lecithin led to increased size and stability. As optimal addition of 20 % PHB into liposome particles was found leading to optimum size and processing of particles, to high UV-protective effect as well as to increased colloid stability and SPF value during long-term storage. Significant differences in cell viability were found in cytotoxicity studies after exposure of keratinocytes to liposomes with different PHB content. Newly fabricated PHB-liposome particles with coffee extract were not found as toxic for HaCaT cells and in LDH test up to 12 %. These particles can act as active carriers for organic sunscreen components in combination with UV-protective effect of PHB.

• Klíčová slova
• Coffea arabica, Cytotoxicity, Nanoparticles, PHB-liposomes, Stability, UV protection,
• MeSH
• antioxidancia farmakologie   MeSH
• fenoly * toxicita   MeSH
• liposomy * MeSH
• přípravky chránící proti slunci toxicita   MeSH
• rostlinné extrakty MeSH
• velikost částic MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• antioxidancia MeSH
• fenoly * MeSH
• liposomy * MeSH
• přípravky chránící proti slunci MeSH
• rostlinné extrakty MeSH

The chorioallantoic membrane (CAM) is a highly vascularized avian extraembryonic membrane widely used as an in vivo model to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the use of CAM has become an integral part of the biocompatibility testing process for developing biomaterials intended for regenerative strategies and tissue engineering applications. In this study, we used the chicken ex ovo CAM assay to investigate the angiogenic potential of innovative acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold, which is intended for the treatment of hard tissue defects, depending on treatment with pro- and anti-angiogenic substances. On embryonic day (ED) 7, the experimental biomaterials were placed on the CAM alone or soaked in vascular endothelial growth factor (VEGF-A), saline solution (PHY), or tyrosine kinase inhibitor (SU5402). After 72 h, the formation of vessels was analyzed in the surrounding area of the scaffold and inside the pores of the implants, using markers of embryonic endothelium (WGA, SNA), myofibroblasts (α-SMA), and macrophages (KUL-01). The morphological and histochemical analysis showed strong angiogenic potential of untreated scaffolds without additional effect of the angiogenic factor, VEGF-A. The lowest angiogenic potential was observed in scaffolds soaked with SU5402. Gene expression of pro-angiogenic growth factors, i.e., VEGF-A, ANG-2, and VE-CAD, was upregulated in untreated scaffolds after 72 h, indicating a pro-angiogenic environment. We concluded that the PHB/CHIT has a strong endogenous angiogenic potential and could be promising biomaterial for the treatment of hard tissue defects.

• Klíčová slova
• CAM assay, angiogenesis, biomaterial, bone tissue engineering, chitosan, polyhydroxybutyrate, regeneration,
• Publikační typ
• časopisecké články MeSH

This study focused on material recycling of a biodegradable blend based on PLA and PHB for multiple applications of biodegradable polymeric material under real conditions. In this study, we investigated the effect of multiple processing of a biodegradable polymer blend under the trade name NONOILEN®, which was processed under laboratory as well as industrial conditions. In this article, we report on testing the effect of blending and multiple processing on thermomechanical stability, molecular characteristics, as well as thermophysical and mechanical properties of experimental- and industrial-type tested material suitable for FDM 3D technology. The results showed that the studied material degraded during blending and subsequently during multiple processing. Even after partial degradation, which was demonstrated by a decrease in average molecular weight and a decrease in complex viscosity in the process of multiple reprocessing, there was no significant change in the material's thermophysical properties, either in laboratory or industrial conditions. There was also no negative impact on the strength characteristics of multiple processed samples. The results of this work show that a biodegradable polymer blend based on PLA and PHB is a suitable candidate for material recycling even in industrial processing conditions. In addition, the results suggest that the biodegradable polymeric material NONOILEN® 3D 3056-2 is suitable for multiple uses in FDM technology.

• Klíčová slova
• 3D printing, material recycling, polyhydroxybutyrate, polylactic acid,
• Publikační typ
• časopisecké články MeSH

Vinasse, a recalcitrant waste of the ethanol industry was employed for the production of polyhydroxyalkanoate (PHA) by the extremely halophilic archaeon, Haloarcula marismortui in shake flasks. The PHA was recovered by osmotic lysis of the cells and subsequent purification by sodium hypochlorite and organic solvents. Through UV-vis spectroscopy, differential scanning calorimetry, Fourier transform infrared, and nuclear magnetic resonance spectroscopy, the PHA was found to have characteristics very similar to that of the standard polyhydroxybutyrate (PHB) from Sigma. Inhibitory effect of polyphenols contained in vinasse was assessed by a quick and reliable cup-plate agar-diffusion method. Raw vinasse (10%) was utilized leading to accumulation of 23% PHA (of cell dry weight) and following an efficacious pre-treatment process through adsorption on activated carbon, 100% pre-treated vinasse could be utilized leading to 30% accumulation of PHB by H. marismortui. Maximum specific growth rate, specific production rate, and volumetric productivity attained using 10% raw vinasse were comparable to that obtained using a previously reported nutrient deficient medium (NDM), while the values with 100% pre-treated vinasse were higher than that determined using NDM medium. This is the first report of polyhydroxybutyrate production by a halophilic microorganism utilizing vinasse.

• MeSH
• archeální proteiny genetika   metabolismus   MeSH
• Haloarcula marismortui genetika   růst a vývoj   metabolismus   MeSH
• odpadky - odstraňování MeSH
• polyhydroxyalkanoáty metabolismus   MeSH
• průmyslový odpad analýza   MeSH
• regulace genové exprese u archeí MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• archeální proteiny MeSH
• polyhydroxyalkanoáty MeSH
• průmyslový odpad MeSH

Numerous prokaryotes accumulate polyhydroxybutyrate (PHB) intracellularly as a storage material. It has also been proposed that PHB accumulation improves bacterial stress resistance. Cupriavidus necator and its PHB non-accumulating mutant were employed to investigate the protective role of PHB under hypertonic conditions. The presence of PHB granules enhanced survival of the bacteria after exposure to hypertonic conditions. Surprisingly, when coping with such conditions, the bacteria did not utilize PHB to harvest carbon or energy, suggesting that, in the osmotic upshock of C. necator, the protective mechanism of PHB granules is not associated with their hydrolysis. The presence of PHB granules influenced the overall properties of the cells, since challenged PHB-free cells underwent massive plasmolysis accompanied by damage to the cell membrane and the leakage of cytoplasm content, while no such effects were observed in PHB containing bacteria. Moreover, PHB granules demonstrated "liquid-like" properties indicating that they can partially repair and stabilize cell membranes by plugging small gaps formed during plasmolysis. In addition, the level of dehydration and changes in intracellular pH in osmotically challenged cells were less pronounced for PHB-containing cultures, demonstrating the important role of PHB for bacterial survival under hyperosmotic conditions.

• Klíčová slova
• Cupriavidus necator, Hyperosmotic conditions, PHB, Plasmolysis, Poly(3-hydroxybutyrate), Stress conditions,
• MeSH
• časové faktory MeSH
• Cupriavidus necator cytologie   účinky léků   metabolismus   ultrastruktura   MeSH
• cytoplazmatická granula účinky léků   metabolismus   ultrastruktura   MeSH
• elektronová kryomikroskopie MeSH
• fluoresceiny metabolismus   MeSH
• fluorescenční mikroskopie MeSH
• hydroxybutyráty metabolismus   MeSH
• hypertonické roztoky farmakologie   MeSH
• krystalizace MeSH
• mikrobiální viabilita účinky léků   MeSH
• osmotický tlak účinky léků   MeSH
• termogravimetrie MeSH
• voda MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 6-carboxyfluorescein MeSH Prohlížeč
• fluoresceiny MeSH
• hydroxybutyráty MeSH
• hypertonické roztoky MeSH
• voda MeSH

AIM: This study aimed at unprecedented physical and chemical evaluation of the 'green plastics' polyhydroxyalkanoates (PHAs), in an extremely halotolerant Halomonas elongata strain 2FF under high-salt concentration. METHODS AND RESULTS: The investigated bacterial strain was isolated from the surface water of the hypersaline Fără Fund Lake. The 16S rRNA gene sequence phylogeny and phenotypic analysis indicated that the isolate belonged to H. elongata. PHA inclusions were observed by Sudan Black B, Nile Red staining, and transmission electron microscopy during growth at high salinity (10%, w/v, NaCl) on 1% (w/v) d-glucose. The produced polymer was quantitatively and qualitatively assessed using crotonic acid assay, elemental analysis, Fourier transform infrared and Raman spectroscopies. Additionally, X-ray powder diffraction, 1 H-NMR spectroscopy, and differential scanning calorimetry were applied. The investigations showed that the intracellular polymer was polyhydroxybutyrate (PHB) of which the strain produced up to 40 wt% of total cell dry weight after 48 h. The analysis of phaC gene from the isolated H. elongata strain indicated that the encoded PHA synthase belongs to Class I PHA synthase family. CONCLUSIONS: Overall, our investigations pointed out that the halotolerant H. elongata strain 2FF was capable to produce significant amounts of PHB from d-glucose, and PHAs from various carbon substrates at high-salt concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: The tested strain showed the ability for significant production of natural, biodegradable polymers under nutrient limitation and hypersaline conditions suggesting its potentiality for further metabolic and molecular investigations towards enhanced biopolymer production. Additionally, this study reports on the unprecedented use of Raman and XPRD techniques to investigate PHAs of an extremely halotolerant bacterium, thus expanding the repertoire of physical methods to study green plastics derived from extremophilic microorganisms.

• Klíčová slova
• Halomonas sp., green plastics, halotolerant bacteria, high salinity, nutrient limitation, polyhydroxybutyrate,
• MeSH
• biopolymery biosyntéza   MeSH
• chlorid sodný metabolismus   MeSH
• fylogeneze MeSH
• Halomonas genetika   izolace a purifikace   metabolismus   MeSH
• jezera mikrobiologie   MeSH
• polyhydroxyalkanoáty biosyntéza   chemie   MeSH
• RNA ribozomální 16S genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Rumunsko MeSH
• Názvy látek
• biopolymery MeSH
• chlorid sodný MeSH
• polyhydroxyalkanoáty MeSH
• RNA ribozomální 16S MeSH

In this experimental research, different types of essential oils (EOs) were blended with polyhydroxybutyrate (PHB) to study the influence of these additives on PHB degradation. The blends were developed by incorporating three terpenoids at two concentrations (1 and 3%). The mineralization rate obtained from CO2 released from each sample was the factor that defined biodegradation. Furthermore, scanning electron microscope (SEM), differential scanning calorimetry (DSC), and dynamic mechanical analysis (DMA) were used in this research. The biodegradation percentages of PHB blended with 3% of eucalyptol, limonene, and thymol after 226 days were reached 66.4%, 73.3%, and 76.9%, respectively, while the rate for pure PHB was 100% after 198 days, and SEM images proved these results. Mechanical analysis of the samples showed that eucalyptol had the highest resistance level, even before the burial test. The other additives showed excellent mechanical properties although they had less mechanical strength than pure PHB after extrusion. The samples' mechanical properties improved due to their crystallinity and decreased glass transition temperature (Tg). DSC results showed that blending terpenoids caused a reduction in Tg, which is evident in the DMA results, and a negligible reduction in melting point (Tm).

• Klíčová slova
• biodegradation, essential oils (EOs), mechanical properties, polyhydroxybutyrate (PHB),
• MeSH
• antibakteriální látky MeSH
• antiinfekční látky * MeSH
• butyráty * MeSH
• eukalyptol MeSH
• polyestery chemie   MeSH
• terpeny MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• antiinfekční látky * MeSH
• butyráty * MeSH
• eukalyptol MeSH
• polyestery MeSH
• terpeny MeSH

Biopolymer composites allow the creation of an optimal environment for the regeneration of chondral and osteochondral defects of articular cartilage, where natural regeneration potential is limited. In this experimental study, we used the sheep animal model for the creation of knee cartilage defects. In the medial part of the trochlea and on the medial condyle of the femur, we created artificial defects (6 × 3 mm2) with microfractures. In four experimental sheep, both defects were subsequently filled with the porous acellular polyhydroxybutyrate/chitosan (PHB/CHIT)-based implant. Two sheep had untreated defects. We evaluated the quality of the newly formed tissue in the femoral trochlea defect site using imaging (X-ray, Computer Tomography (CT), Magnetic Resonance Imaging (MRI)), macroscopic, and histological methods. Macroscopically, the surface of the treated regenerate corresponded to the niveau of the surrounding cartilage. X-ray examination 6 months after the implantation confirmed the restoration of the contour in the subchondral calcified layer and the advanced rate of bone tissue integration. The CT scan revealed a low regenerative potential in the bone zone of the defect compared to the cartilage zone. The percentage change in cartilage density at the defect site was not significantly different to the reference area (0.06-6.4%). MRI examination revealed that the healing osteochondral defect was comparable to the intact cartilage signal on the surface of the defect. Hyaline-like cartilage was observed in most of the treated animals, except for one, where the defect was repaired with fibrocartilage. Thus, the acellular, chitosan-based biomaterial is a promising biopolymer composite for the treatment of chondral and osteochondral defects of traumatic character. It has potential for further clinical testing in the orthopedic field, primarily with the combination of supporting factors.

• Klíčová slova
• biopolymer, cartilage, chitosan, regeneration, sheep model,
• Publikační typ
• časopisecké články MeSH

This work presents a comprehensive analysis of the biodegradation of polyhydroxybutyrate (PHB) and chemically modified PHB with different chemical and crystal structures in a soil environment. A polymer modification reaction was performed during preparation of the chemically modified PHB films, utilizing 2,5-dimethyl-2,5-di(tert-butylperoxy)-hexane as a free-radical initiator and maleic anhydride. Films of neat PHB and chemically modified PHB were prepared by extrusion and thermocompression. The biological agent employed was natural mixed microflora in the form of garden soil. The course and extent of biodegradation of the films was investigated by applying various techniques, as follows: a respirometry test to determine the production of carbon dioxide through microbial degradation; scanning electron microscopy (SEM); optical microscopy; fluorescence microscopy; differential scanning calorimetry (DSC); and X-ray diffraction (XRD). Next-generation sequencing was carried out to study the microbial community involved in biodegradation of the films. Findings from the respirometry test indicated that biodegradation of the extruded and chemically modified PHB followed a multistage (2-3) course, which varied according to the spatial distribution of amorphous and crystalline regions and their spherulitic morphology. SEM and polarized optical microscopy (POM) confirmed that the rate of biodegradation depended on the availability of the amorphous phase in the interspherulitic region and the width of the interlamellar region in the first stage, while dependence on the size of spherulites and thickness of spherulitic lamellae was evident in the second stage. X-ray diffraction revealed that orthorhombic α-form crystals with helical chain conformation degraded concurrently with β-form crystals with planar zigzag conformation. The nucleation of PHB crystals after 90 days of biodegradation was identified by DSC and POM, a phenomenon which impeded biodegradation. Fluorescence microscopy evidenced that the crystal structure of PHB affected the physiological behavior of soil microorganisms in contact with the surfaces of the films.

• MeSH
• hydroxybutyráty * chemie   MeSH
• kyselina 3-hydroxymáselná MeSH
• polyestery * chemie   MeSH
• půda MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hydroxybutyráty * MeSH
• kyselina 3-hydroxymáselná MeSH
• poly-beta-hydroxybutyrate MeSH Prohlížeč
• polyestery * MeSH
• polyhydroxybutyrate MeSH Prohlížeč
• půda MeSH