In this paper, the treatment of poly-ε-caprolactone (PCL) nano/micro-mesh system by cryogenic grinding and subsequent characterization of obtained product is described. The PCL nano/micro-mesh layer submerged in appropriate liquid was cryogenically ground and obtained particles were characterized employing mainly laser diffraction and scanning electron microscopy (SEM). In the ground sample, different types of particles (fibrous particles, fibrous fragments, agglomerates with and without an internal fibrous structure, lamellae and nanoparticles) were identified, described and quantified. Parameters of cryogenic grinding (weight of sample, type of liquid medium, and influence of sample storage) were optimized to maximize the yield of particles with desired features. The potential of the system for cell scaffolding was demonstrated by cultivation of 3T3 fibroblasts on the produced microparticles.

• MeSH
• buněčná adheze účinky léků   MeSH
• buněčné linie MeSH
• buňky 3T3 MeSH
• fibroblasty účinky léků   MeSH
• mikroskopie elektronová rastrovací metody   MeSH
• myši MeSH
• nanočástice chemie   MeSH
• polyestery chemie   farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Poly(ε-caprolactone; PCL) is an attractive biodegradable polymer that has been increasingly used to solve environmental problems caused by plastic wastes. In the present study, 468 bacterial isolates were recovered from soil samples and screened for PCL degradation activity. Of the isolates, 37 (7.9%) showed PCL depolymerase activity on PCL agar medium, with the highest activity being by isolate S22 which was identified using 16S rRNA and rpoB gene sequencing as Acinetobacter seifertii. Scanning electron microscopy and Fourier transform infrared spectroscopy confirmed the degradation of PCL films after treatment with A. seifertii S22. The PCL depolymerase activity of A. seifertii S22 relied on the activity of esterase which occurred at an optimum temperature of 30-40 °C. The highest PCL depolymerase activity (35.5 ± 0.7 U/mL) was achieved after culturing A. seifertii S22 for 6 h in mineral salt medium (MSM) containing 0.1% Tween 20 and 0.02% ammonium sulfate as the carbon and nitrogen sources, respectively, which was approximately 20-fold higher than for cultivation in MSM supplemented with 0.1% PCL as sole carbon source. The results suggested that A. seifertii S22 or its enzymes could be used for PCL bioplastic degradation.

• MeSH
• Acinetobacter MeSH
• biodegradace MeSH
• kapronáty MeSH
• laktony MeSH
• polyestery * metabolismus   MeSH
• RNA ribozomální 16S genetika   MeSH
• uhlík * MeSH
• Publikační typ
• časopisecké články MeSH

In this study, we have developed a combined approach to accelerate the proliferation of mesenchymal stem cells (MSCs) in vitro, using a new nanofibrous scaffold made by needleless electrospinning from a mixture of poly-ε-caprolactone and magnetic particles. The biological characteristics of porcine MSCs were investigated while cultured in vitro on composite scaffold enriched with magnetic nanoparticles. Our data indicate that due to the synergic effect of the poly-ε-caprolactone nanofibers and magnetic particles, cellular adhesion and proliferation of MSCs is enhanced and osteogenic differentiation is supported. The cellular and physical attributes make this new scaffold very promising for the acceleration of efficient MSC proliferation and regeneration of hard tissues.

• MeSH
• biokompatibilní materiály chemie   farmakologie   MeSH
• buněčná adheze účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• kapronáty chemie   farmakologie   MeSH
• laktony chemie   farmakologie   MeSH
• magnetické nanočástice chemie   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   MeSH
• nanovlákna chemie   MeSH
• polyestery farmakologie   MeSH
• prasata MeSH
• proliferace buněk účinky léků   MeSH
• tkáňové inženýrství MeSH
• tkáňové podpůrné struktury chemie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Polymeric drug carriers exhibit excellent properties that advance drug delivery systems. In particular, carriers based on poly(ethylene oxide)-block-poly(ε-caprolactone) are very useful in pharmacokinetics. In addition to their proven biocompatibility, there are several requirements for the efficacy of the polymeric drug carriers after internalization, e.g., nanoparticle behavior, cellular uptake, the rate of degradation, and cellular localization. The introduction of γ-butyrolactone units into the hydrophobic block enables the tuning of the abovementioned properties over a wide range. In this study, a relatively high content of γ-butyrolactone units with a reasonable yield of ≈60% is achieved by anionic ring-opening copolymerization using 1,5,7-triazabicyclo[4.4.0]dec-5-ene as a very efficient catalyst in the nonpolar environment of toluene with an incorporated γ-butyrolactone content of ≈30%. The content of γ-butyrolactone units can be easily modulated according to the feed ratio of the monomers. This method enables control over the rate of degradation so that when the content of γ-butyrolactone increases, the rate of degradation increases. These findings broaden the application possibilities of polyester-polyether-based nanoparticles for biomedical applications, such as drug delivery systems.

• MeSH
• buněčná smrt MeSH
• buněčné linie MeSH
• gama-butyrolakton chemická syntéza   chemie   MeSH
• intracelulární prostor metabolismus   MeSH
• lidé MeSH
• myši MeSH
• nanočástice chemie   ultrastruktura   MeSH
• nosiče léků chemie   MeSH
• polyestery chemická syntéza   chemie   MeSH
• polyethylenglykoly chemická syntéza   chemie   MeSH
• polymerizace * MeSH
• protonová magnetická rezonanční spektroskopie MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Purpose: Incisional hernia repair is an unsuccessful field of surgery, with long-term recurrence rates reaching up to 50% regardless of technique or mesh material used. Various implants and their positioning within the abdominal wall pose numerous long-term complications that are difficult to treat due to their permanent nature and the chronic foreign body reaction they trigger. Materials mimicking the 3D structure of the extracellular matrix promote cell adhesion, proliferation, migration, and differentiation. Some electrospun nanofibrous scaffolds provide a topography of a natural extracellular matrix and are cost effective to manufacture. Materials and methods: A composite scaffold that was assembled out of a standard polypropylene hernia mesh and poly-ε-caprolactone (PCL) nanofibers was tested in a large animal model (minipig), and the final scar tissue was subjected to histological and biomechanical testing to verify our in vitro results published previously. Results: We have demonstrated that a layer of PCL nanofibers leads to tissue overgrowth and the formation of a thick fibrous plate around the implant. Collagen maturation is accelerated, and the final scar is more flexible and elastic than under a standard polypropylene mesh with less pronounced shrinkage observed. However, the samples with the composite scaffold were less resistant to distracting forces than when a standard mesh was used. We believe that the adverse effects could be caused due to the material assembly, as they do not comply with our previous results. Conclusion: We believe that PCL nanofibers on their own can cause enough fibroplasia to be used as a separate material without the polypropylene base, thus avoiding potential adverse effects caused by any added substances.

• MeSH
• břišní stěna chirurgie   MeSH
• chirurgické síťky * MeSH
• hernie * MeSH
• kolagen metabolismus   MeSH
• miniaturní prasata MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• nanovlákna chemie   MeSH
• operace kýly přístrojové vybavení   metody   MeSH
• polyestery MeSH
• polypropyleny chemie   MeSH
• prasata MeSH
• testování materiálů MeSH
• tkáňové podpůrné struktury chemie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The objective of this work was to characterize the toxicological profile of a newly developed sunscreen formulation based on polymeric nanocapsules (NCs) loading benzophenone-3 (BZP3). NCs composed of poly(ε-caprolactone) carrot oil and Pluronic® F68 were produced by emulsification-diffusion method. Their mean particle size (Z-Ave) ranged from 280 to 420 nm, polydispersity index (PDI) was below 0.37, while zeta potential (ZP) reached about |+11 mV|. No cytotoxic effects were observed in L929 fibroblast cell line for the blank (i.e., non-loaded) NCs and BZP3-loaded NCs (BZP3-NCs). The semi-solid sunscreen formulation was stable over time (centrifugation testing) and exhibited non-Newtonian pseudoplastic behavior, which is typical of products for topical application onto the skin. The sun protection factor (SPF) value reached 8.84, when incorporating BZP3-NCs (SPF of 8.64) into the semi-solid formulation. A synergistic effect was also observed when combining the formulation ingredients of nanocapsules, i.e., SPF of carrot oil was 6.82, blank NCs was 6.84, and BZP3-loaded NCs was 8.64. From the hen's egg-chorioallantoic membrane test (HET-CAM) test, the non-irritation profile of the developed formulations could also be confirmed. The obtained results show a promising use of poly(ε-caprolactone) nanocapsules to be loaded with lipophilic sunscreens as benzophenone-3.

• Publikační typ
• časopisecké články MeSH

Bone and cartilage are tissues of a three-dimensional (3D) nature. Therefore, scaffolds for their regeneration should support cell infiltration and growth in all 3 dimensions. To fulfill such a requirement, the materials should possess large, open pores. Centrifugal spinning is a simple method for producing 3D fibrous scaffolds with large and interconnected pores. However, the process of bone regeneration is rather complex and requires additional stimulation by active molecules. In the current study, we introduced a simple composite scaffold based on platelet adhesion to poly-ε-caprolactone 3D fibers. Platelets were used as a natural source of growth factors and cytokines active in the tissue repair process. By immobilization in the fibrous scaffolds, their bioavailability was prolonged. The biological evaluation of the proposed system in the MG-63 model showed improved metabolic activity, proliferation and alkaline phosphatase activity in comparison to nonfunctionalized fibrous scaffold. In addition, the response of cells was dose dependent with improved biocompatibility with increasing platelet concentration. The results demonstrated the suitability of the system for bone tissue.

• MeSH
• adhezivita trombocytů účinky léků   MeSH
• alkalická fosfatasa metabolismus   MeSH
• kinetika MeSH
• lékové transportní systémy metody   MeSH
• lidé MeSH
• mezibuněčné signální peptidy a proteiny aplikace a dávkování   farmakologie   MeSH
• nádorové buněčné linie MeSH
• osteoblasty cytologie   účinky léků   ultrastruktura   MeSH
• osteogeneze účinky léků   MeSH
• polyestery chemie   farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury chemie   MeSH
• trombocyty účinky léků   metabolismus   ultrastruktura   MeSH
• tvar buňky účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: We prepared 3D poly (ε-caprolactone) (PCL) nanofibre scaffolds and tested their use for seeding, proliferation, differentiation and migration of mesenchymal stem cell (MSCs). MATERIALS AND METHODS: 3D nanofibres were prepared using a special collector for common electrospinning; simultaneously, a 2D PCL nanofibre layer was prepared using a classic plain collector. Both scaffolds were seeded with MSCs and biologically tested. MSC adhesion, migration, proliferation and osteogenic differentiation were investigated. RESULTS: The 3D PCL scaffold was characterized by having better biomechanical properties, namely greater elasticity and resistance against stress and strain, thus this scaffold will be able to find broad applications in tissue engineering. Clearly, while nanofibre layers of the 2D scaffold prevented MSCs from migrating through the conformation, cells infiltrated freely through the 3D scaffold. MSC adhesion to the 3D nanofibre PCL layer was also statistically more common than to the 2D scaffold (P < 0.05), and proliferation and viability of MSCs 2 or 3 weeks post-seeding, were also greater on the 3D scaffold. In addition, the 3D PCL scaffold was also characterized by displaying enhanced MSC osteogenic differentiation. CONCLUSIONS: We draw the conclusion that all positive effects observed using the 3D PCL nanofibre scaffold are related to the larger fibre surface area available to the cells. Thus, the proposed 3D structure of the nanofibre layer will find a wide array of applications in tissue engineering and regenerative medicine.

• MeSH
• buněčná diferenciace * MeSH
• buněčné kultury přístrojové vybavení   metody   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• nanovlákna chemie   ultrastruktura   MeSH
• osteogeneze MeSH
• osteokalcin metabolismus   MeSH
• pohyb buněk MeSH
• polyestery chemie   MeSH
• povrchové vlastnosti MeSH
• proliferace buněk MeSH
• pružnost MeSH
• regenerativní lékařství MeSH
• sialoprotein vázající integrin metabolismus   MeSH
• tkáňové inženýrství MeSH
• tkáňové podpůrné struktury * MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The study involved the electrospinning of the copolymer poly(L-lactide-co-ε-caprolactone) (PLCL) into tubular grafts. The subsequent material characterization, including micro-computed tomography analysis, revealed a level of porosity of around 70%, with pore sizes of 9.34 ± 0.19 μm and fiber diameters of 5.58 ± 0.10 μm. Unlike fibrous polycaprolactone, the electrospun PLCL copolymer promoted fibroblast and endothelial cell adhesion and proliferation in vitro. Moreover, the regeneration of the vessel wall was detected following implantation and, after six months, the endothelialization of the lumen and the infiltration of arranged smooth muscle cells producing collagen was observed. However, the degradation rate was found to be accelerated in the rabbit animal model. The study was conducted under conditions that reflected the clinical requirements-the prostheses were sutured in the end-to-side fashion and the long-term end point of prosthesis healing was assessed. The regeneration of the vessel wall in terms of endothelialization, smooth cell infiltration and the presence of collagen fibers was observed after six months in vivo. A part of the grafts failed due to the rapid degradation rate of the PLCL copolymer.

• MeSH
• aorta patologie   MeSH
• arteriae carotides patologie   MeSH
• buněčná adheze MeSH
• buňky 3T3 MeSH
• cévní protézy * MeSH
• endoteliální buňky pupečníkové žíly (lidské) MeSH
• endoteliální buňky MeSH
• fibroblasty cytologie   MeSH
• kolagen metabolismus   MeSH
• králíci MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• myocyty hladké svaloviny cytologie   MeSH
• myši MeSH
• polyestery chemie   MeSH
• polymery chemie   MeSH
• poréznost MeSH
• prasata MeSH
• psi MeSH
• regenerace MeSH
• rentgenová mikrotomografie MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury MeSH
• transplantace cév * MeSH
• zobrazování trojrozměrné MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In this study, we developed a three-dimensional (3D) in vitro infection model to investigate the crosstalk between phagocytes and microbes in inflammation using a nanofibrous membrane (NM). Poly(ε-caprolactone) (PCL)-NMs (PCL-NMs) were generated via electrospinning of PCL in chloroform. Staphylococcus aureus and phagocytes were able to adhere to the nanofibers and phagocytes engulfed S. aureus in the PCL-NM. The migration of phagocytes to S. aureus was evaluated in a two-layer co-culture system using PCL-NM. Neutrophils, macrophages and dendritic cells (DCs) cultured in the upper PCL-NM layer migrated to the lower PCL-NM layer containing bacteria. DCs migrated to neutrophils that cultured with bacteria and then engulfed neutrophils in two-layer system. In addition, phagocytes in the upper PCL-NM layer migrated to bacteria-infected MLE-12 lung epithelial cells in the lower PCL-NM layer. S. aureus-infected MLE-12 cells stimulated the secretion of tumor necrosis factor-α and IL-1α in 3D culture conditions, but not in 2D culture conditions. Therefore, the PCL-NM-based 3D culture system with phagocytes and bacteria mimics the inflammatory response to microbes in vivo and is applicable to the biomimetic study of various microbe infections.

• Publikační typ
• časopisecké články MeSH