Vzhledem k obsahu řady růstových faktorů představují trombocyty významný potenciál pro využití v oblasti tkáňového inženýrství a regenerativní medicíny. Růstové faktory uvolněné z krevních destiček ovlivňují buněčnou diferenciaci, proliferaci, transkripci specifických proteinů, chemotaxi a další procesy účastnící se regenerace tkání. In vitro se trombocyty ve formě plazmy bohaté na destičky využívají pro stimulaci buněčné proliferace různých typů tkáňových kultur. Pro účely tkáňového inženýrství buňky porůstají nanovlákenné nosiče (scaffoldy) s cílem vytvořit systém vhodný pro následnou implantaci do organismu. Nanovlákenné scaffoldy se připravují z biokompatibilních a biodegradabilních polymerů technikou elektrostatického zvlákňování. Suspenze trombocytů byla použita pro modifikaci nanovlákenných tkáňových nosičů, které byly připraveny elektrostatickým zvlákňováním biodegradabilního polymeru polykaprolaktonu, který je v tkáňovém inženýrství hojně využíván. Scaffoldy byly modifikovány dvěma metodami: a) smáčením v suspenzi trombocytů a b) sprejováním mezi vznikající nanovlákna během elektrostatického zvlákňování. Tyto modifikované nanovlákenné scaffoldy pak byly testovány in vitro s využitím myších 3T3 fibroblastů a lidských dermálních fibroblastů. Výsledky ukazují, že inkorporace trombocytů do nanovlákenných vrstev napomáhá buněčné proliferaci obou testovaných buněčných typů. Inkorporace trombocytů dovnitř nanovlákenné struktury pomocí sprejování navíc podporuje prorůstání buněk do 3D struktury. Klíčová slova: trombocyty, elektrostatické zvlákňování, tkáňový nosič, nanovlákna, polykaprolakton
Platelets hold a significant promise for the field of regenerative medicine and tissue engineering given their large content of growth factors. Released growth factors promote cell differentiation, proliferation, transcription of specific proteins, chemotaxis and other processes involved in tissue regeneration. Platelets in the form of platelet rich plasma are commonly used in vitro to stimulate cell proliferation of different types of tissue cultures. In the tissue engineering approach, cells are grown on nanofibrous scaffolds in order to create a system suitable for subsequent implantation into the human body. Nanofibrous scaffolds can be prepared from biodegradable and biocompatible polymers using electrospinning. A thrombocyte rich solution was used for modification of nanofibrous electrospun scaffolds made from the biodegradable polymer polycaprolactone, which is widely used in tissue engineering applications. The resulting scaffolds were modified in two ways: a) bathing in thrombocyte rich solution and b) spraying of thrombocyte rich solution in between forming nanofibers during the electrospinning process. Nanofibrous scaffolds were tested in vitro using mouse 3T3 fibroblasts and human dermal fibroblasts. Incorporation of thrombocytes into the nanofibrous layers increased proliferation of both cell types. The use of the spraying technique promotes cell ingrowth into 3D structures. Key words: platelets, electrospinning, scaffold, nanofibers, polycaprolactone
- MeSH
- Fibroblasts MeSH
- Hydrogels chemistry MeSH
- Cells, Cultured * MeSH
- Humans MeSH
- Microscopy, Electron, Scanning MeSH
- Mice MeSH
- Nanofibers * utilization MeSH
- Polyesters chemistry MeSH
- Surface Properties MeSH
- Cell Proliferation physiology MeSH
- In Vitro Techniques MeSH
- Tissue Engineering * methods MeSH
- Tissue Scaffolds * utilization MeSH
- Blood Platelets * cytology metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
Polycaprolactone nanofibers are used as scaffolds in the field of tissue engineering for tissue regeneration or drug delivery. Polycaprolactone (PCL) is a biodegradable hydrophobic polyester used to obtain implantable nanostructures, which are clinically applicable due to their biological safety. Polydatin (PD), a glycosidic precursor of resveratrol, is known for its antioxidant, antitumor, antiosteoporotic, and bone regeneration activities. We aimed to use the osteogenic capacity of polydatin to create a biomimetic innovative and patented scaffold consisting of PCL-PD for bone tissue engineering. Both osteosarcoma cells (Saos-2) and mesenchymal stem cells (MSCs) were used to test the in vitro cytocompatibility of the PD-PCL scaffold. Reverse-phase (RP) HPLC was used to evaluate the timing release of PD from the PCL-PD nanofibers and the MTT assay, scanning electron microscopy, and alkaline phosphatase (ALP) activity were used to evaluate the proliferation, adhesion, and cellular differentiation in both osteosarcoma and human mesenchymal stem cells (MSCs) seeded on PD-PCL nanofibers. The proliferation of osteosarcoma cells (Saos-2) on the PD-PCL scaffold decreased when compared to cells grown on PLC nanofibers, whereas the proliferation of MSCs was comparable in both PCL and PD-PCL nanofibers. Noteworthy, after 14 days, the ALP activity was higher in both Saos-2 cells and MSCs cultivated on PD-PCL than on empty scaffolds. Moreover, the same cells showed a spindle-shaped morphology after 14 days when grown on PD-PCL as shown by SEM. In conclusion, we provide evidence that nanofibers appropriately coated with PD support the adhesion and promote the osteogenic differentiation of both human osteosarcoma cells and MSCs.
- Publication type
- Journal Article MeSH
The alveolar-capillary interface is the key functional element of gas exchange in the human lung, and disruptions to this interface can lead to significant medical complications. However, it is currently challenging to adequately model this interface in vitro, as it requires not only the co-culture of human alveolar epithelial and endothelial cells but mainly the preparation of a biocompatible scaffold that mimics the basement membrane. This scaffold should support cell seeding from both sides, and maintain optimal cell adhesion, growth, and differentiation conditions. Our study investigates the use of polycaprolactone (PCL) nanofibers as a versatile substrate for such cell cultures, aiming to model the alveolar-capillary interface more accurately. We optimized nanofiber production parameters, utilized polyamide mesh UHELON as a mechanical support for scaffold handling, and created 3D-printed inserts for specialized co-cultures. Our findings confirm that PCL nanofibrous scaffolds are manageable and support the co-culture of diverse cell types, effectively enabling cell attachment, proliferation, and differentiation. Our research establishes a proof-of-concept model for the alveolar-capillary interface, offering significant potential for enhancing cell-based testing and advancing tissue-engineering applications that require specific nanofibrous matrices.
BACKGROUND/AIM: This study investigated the therapeutic potential of lipophosphonoxin (LPPO), an antibacterial agent, loaded into polycaprolactone nanofiber dressings (NANO-LPPO) for full-thickness wound healing. Using a porcine model, we aimed to assess the impact of areal weight of the dressing (10, 20 and 30 g/m2) on wound-healing outcomes and validate findings from previous murine studies. MATERIALS AND METHODS: Full-thickness wounds were created on porcine skin and treated with the NANO-LPPO dressings of differing thickness. Positive control (Aquacel Ag+) and standard control (Jelonet) groups were included for comparison. Wound-healing progression was evaluated macroscopically and on the histological level. RESULTS: Macroscopic observations indicated no signs of infection in any group, with wounds covered by scabs by day 14. Thicker dressings (areal weights of 30 and 20 g/m2) demonstrated superior performance in promoting the formation of granulation tissue and healing compared to the thinner version (areal weight of 10 g/m2). LPPO-loading enhanced scaffold wettability and biodegradability without impairing healing outcomes. Both control groups exhibited similar healing characteristics. CONCLUSION: The findings underscore the importance of optimizing dressing thickness for effective wound healing. NANO-LPPO dressings exhibit translational potential as a therapeutic option for full-thickness wounds, warranting further preclinical and regulatory evaluation to support clinical application.
- MeSH
- Anti-Bacterial Agents pharmacology administration & dosage chemistry MeSH
- Wound Healing * drug effects MeSH
- Skin drug effects pathology MeSH
- Lipoxins * chemistry pharmacology administration & dosage MeSH
- Disease Models, Animal MeSH
- Nanofibers * chemistry MeSH
- Bandages * MeSH
- Polyesters * chemistry MeSH
- Swine MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
Polycaprolactone composite nanofibers coated with a polydopamine layer are introduced as a new type of absorption material for on-line solid phase extraction (SPE) in chromatographic system. A hybrid technology combining the electrospinning and melt blowing was used for the preparation of 3D-structured microfiber/nanofibrous polycaprolactone composite. The dopamine coating was then applied to functionalize the micro/nanofibers. Polydopamine-coated polycaprolactone fibers were tested as an extraction phase in on-line SPE prior to HPLC separation and UV detection. Four groups of biologically active substances including bisphenols (Bisphenol S, Bisphenol AF, Bisphenol A, Bisphenol C, Bisphenol AP, Bisphenol Z, Bisphenol BP, and Bisphenol M), betablockers (Timolol, Metoprolol, Labetalol, and Propranolol), nonsteroidal antiphlogistic drugs (Salicylic acid, Ketoprofen, Naproxen, Indomethacin, Diclofenac, Ibuprophen, and Meclofenamic acid), and phenolic acids (Chlorogenic acid, Caffeic acid, Sinapic acid, m-Coumaric acid, Benzoic acid, and Cinnamic acid) were used as the model analytes. Neat and coated fibers were compared and applied as sorbents for the on-line extraction set-up. Both materials produced good extraction potential for the determination of bisphenols and nonsteroidal drugs in model biological and environmental samples including river water, human urine, and blood serum. However, the polydopamine layer significantly increased the extraction efficiency of polar drugs. Typical repeatability of on-line extraction procedure on polydopamine coated fibers was in the range 0.12-4.11% for bisphenols, 0.55-1.41% for antiphlogistic drugs, 0.59-2.52% for phenolic acids, and 1.01-1.65% for betablockers. Graphical abstract Schematic representation of polycaprolactone composite nanofibers coated with a polydopamine layer as an advanced absorption material for on-line solid phase extraction in chromatography.
- MeSH
- Anti-Inflammatory Agents, Non-Steroidal analysis isolation & purification MeSH
- Adrenergic beta-Antagonists analysis isolation & purification MeSH
- Water Pollutants, Chemical analysis isolation & purification MeSH
- Cinnamates analysis isolation & purification MeSH
- Solid Phase Extraction methods MeSH
- Phenols analysis isolation & purification MeSH
- Indoles chemistry MeSH
- Nanofibers chemistry MeSH
- Polyesters chemistry MeSH
- Polymerization MeSH
- Polymers chemistry MeSH
- Rivers chemistry MeSH
- Reproducibility of Results MeSH
- Chromatography, High Pressure Liquid MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
For biodegradable porous scaffolds to have a potential application in cartilage regeneration, they should enable cell growth and differentiation and should have adequate mechanical properties. In this study, our aim was to prepare biocompatible scaffolds with improved biomechanical properties. To this end, we have developed foam scaffolds from poly-epsilon-caprolactone (PCL) with incorporated chitosan microparticles. The scaffolds were prepared by a salt leaching technique from either 10 or 15 wt% PCL solutions containing 0, 10 and 20 wt% chitosan microparticles, where the same amount and size of NaCl was used as a porogen in all the cases. PCL scaffolds without and with low amounts of chitosan (0 and 10 wt% chitosan) showed higher DNA content than scaffolds with high amounts of chitosan during a 22-day experiment. 10 wt% PCL with 10 and 20 wt% chitosan showed significantly increased viscoelastic properties compared to 15 wt% PCL scaffolds with 0 and 10 wt% chitosan. Thus, 10 wt% PCL scaffolds with 0 wt% and 10 wt% chitosan are potential scaffolds for cartilage regeneration.
- MeSH
- Biocompatible Materials administration & dosage chemistry MeSH
- Cartilage cytology physiology MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Microspheres * MeSH
- Polyesters administration & dosage chemistry MeSH
- Cell Proliferation drug effects physiology MeSH
- Guided Tissue Regeneration methods MeSH
- Tissue Scaffolds * MeSH
- Cell Survival drug effects physiology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
OBJECTIVES: Faulty wound healing is a global healthcare problem. Chronic wounds are generally characterized by a reduction in availability of growth factors. New strategies are being developed to deliver growth factors more effectively. METHODS: In this study, we introduced electrospun scaffolds composed of polycaprolactone (PCL) nanofibers functionalized with adhered platelets, as a source of numerous growth factors. Three concentrations of platelets were immobilized to nanofibrous scaffolds by simple adhesion, and their influence on adhesion, proliferation and metabolic activity of seeded cells (murine fibroblasts, keratinocytes and melanocytes) was investigated. RESULTS: The data obtained indicated that presence of platelets significantly promoted cell spreading, proliferation and metabolic activity in all the skin-associated cell types. There were no significant differences among tested concentrations of platelets, thus even the lowest concentration sufficiently promoted proliferation of the seeded cells. CONCLUSIONS: Such complex stimulation is needed for improved healing of chronic wounds. However, the nanofibrous system can be used not only as a skin cover, but also in broader applications in regenerative medicine.
- MeSH
- Cell Adhesion MeSH
- Cell Line MeSH
- Fibroblasts cytology metabolism MeSH
- Wound Healing MeSH
- Keratinocytes cytology metabolism MeSH
- Melanocytes cytology metabolism MeSH
- Mice MeSH
- Nanofibers chemistry ultrastructure MeSH
- Polyesters chemistry MeSH
- Cell Proliferation * MeSH
- Tissue Engineering MeSH
- Tissue Scaffolds chemistry MeSH
- Blood Platelets cytology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
A novel sorbent for solid phase extraction (SPE) based on hybrid nanofibrous polycaprolactone containing graphene nanoparticles has been prepared. The preparation of hybrid polymer nanofibers with a very high 1:1 polymer/graphene ratio was achieved for the first time using alternating current electrospinning. The final appearance of these nanofibers was a thick porous layer that was cut into the shape of easy-to-handle extraction discs. Based on the preliminary study in which the graphene content varied, 30% graphene-doped nanofibers (w/w) exhibited the highest recoveries and enabled a significant increase in the retention of analytes, 2-25 times in comparison to PCL. The incorporation of graphene resulted in a higher surface area of 12 g/m2 compared to 2 g/m2 determined for the native polycaprolactone (PCL) nanofibers. This unique material was applied for a simple stirred disc sorptive extraction and preconcentration of trace levels of emerging organic environmental contaminants, bisphenols A, AF, AP, C, S, Z, 3-chlorophenol, and pesticides fenoxycarb, deltamethrin, and kadethrin from surface waters prior to HPLC-DAD determination. This was accomplished by stirring the unsupported nanofiber disc in a large-volume sample with RSD of five extractions of 3-15%. Recoveries yielded 87-120%, except 52% for bisphenol S due to its high polarity. Optimization of the extraction procedure included conditioning, sample volume, extraction time, and elution solvent. Our novel desorption procedure carried out in a vial used for the direct injection into the HPLC system significantly reduced sample handling and minimized potential human error.
- Publication type
- Journal Article MeSH
PURPOSE: Electrospun meshes may be considered as substitutes to textile polypropylene implants. We compared the host response and biomechanical properties of the rat abdominal wall following reinforcement with either polycaprolactone (PCL) modified with ureidopyrimidinone-motifs (UPy) or polypropylene mesh. METHODS: First we measured the response to cyclic uniaxial load within the physiological range both dry (room temperature) and wet (body temperature). 36 rats underwent primary repair of a full-thickness abdominal wall defect with a polypropylene suture (native tissue repair), or reinforced with either UPy-PCL or ultra-light weight polypropylene mesh (n = 12/group). Sacrifice was at 7 and 42 days. Outcomes were compliance of explants, mesh dimensions, graft related complications and semi-quantitative assessment of inflammatory cell (sub) types, neovascularization and remodeling. RESULTS: Dry UPy-PCL implants are less stiff than polypropylene, both are more compliant in wet conditions. Polypropylene loses stiffness on cyclic loading. Both implant types were well incorporated without clinically obvious degradation or herniation. Exposure rates were similar (n = 2/12) as well as mesh contraction. There was no reinforcement at low loads, while, at higher tension, polypropylene explants were much stiffer than UPy-PCL. The latter was initially weaker yet by 42 days it had a compliance similar to native abdominal wall. There were eventually more foreign body giant cells around UPy-PCL fibers yet the amount of M1 subtype macrophages was higher than in polypropylene explants. There were less neovascularization and collagen deposition. CONCLUSION: Abdominal wall reconstruction with electrospun UPy-PCL mesh does not compromise physiologic tissue biomechanical properties, yet provokes a vivid inflammatory reaction.
- MeSH
- Biomechanical Phenomena MeSH
- Abdominal Wall physiology MeSH
- Surgical Mesh * MeSH
- Fascia physiology MeSH
- Muscle, Skeletal physiology MeSH
- Rats MeSH
- Polyesters MeSH
- Rats, Sprague-Dawley MeSH
- Pyrimidinones MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Polycaprolactone (PCL) was electrospun with the addition of arginine (Arg), an α-amino acid that accelerates the healing process. The efficient needleless electrospinning technique was used for the fabrication of the nanofibrous layers. The materials produced consisted mainly of fibers with diameters of between 200 and 400 nm. Moreover, both microfibers and beads were present within the layers. Higher bead sizes were observed with the increased addition of arginine. The arginine content within the layers as well as the weight of the resultant electrospun materials were enhanced with the increased addition of arginine to the electrospinning solution (1, 5 and 10 wt%). The PCL + 1% Arg nanofibrous layer contained 5.67 ± 0.04% of arginine, the PCL + 5% Arg layer 22.66 ± 0.24% of arginine and the PCL + 10% Arg layer 37.33 ± 0.39% of arginine according to the results of the elemental analysis. A high burst release within 5 h of soaking was recorded for the PCL + 5% and PCL + 10% nanofibrous layers. However, the release rate of arginine from the PCL + 1% Arg was significantly slower, reaching a maximum level after 72 h of soaking. The resulting materials were hydrophobic. Hemocompatibility testing under static conditions revealed no effect on hemolysis following the addition of arginine and the prolongation of the prothrombin time with the increased addition of arginine, thus exerting an influence on the extrinsic and common pathway of coagulation activation. The results of the dynamic hemocompatibility assessment revealed that the numbers of blood cells and platelets were not affected significantly by the various electrospun samples during incubation. The TAT, β-thromboglobulin and SC5-b9 concentrations were characterized by a moderate increase in the PCL group compared to those of the control group. The presence of arginine resulted in a decrease in the investigated hemocompatibility markers. The PMN elastase levels were comparable with respect to all the groups.
- MeSH
- Arginine chemistry MeSH
- Biocompatible Materials chemistry MeSH
- Electricity MeSH
- Hemolysis * MeSH
- Wound Healing * MeSH
- Humans MeSH
- Nanofibers chemistry MeSH
- Polyesters chemistry MeSH
- Prothrombin Time MeSH
- Materials Testing methods MeSH
- Tissue Engineering MeSH
- Tissue Scaffolds chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
