Plasma-treated Electrospun Research Articles

Plasma-Treated Electrospun PLGA Nanofiber Scaffold Supports Limbal Stem Cells

The corneal epithelial layer is continuously replaced by limbal stem cells. Reconstructing this layer in vitro using synthetic scaffolds is highly needed. Poly-lactic-co-glycolic acid (PLGA) is approved for human use due to its biocompatibility and biodegradability. However, PLGA is hydrophobic, preventing cell adherence to PLGA membranes. PLGA scaffolds were prepared by electrospinning on a custom-made target drum spinning at a rate of 1000 rpm with a flow rate of 0.5 mL/h and voltage at 20 kV, then treated with oxygen plasma at 30 mA using a vacuum coater. Scaffolds were characterized by SEM, mechanically by tensile testing, and thermally by DSC and TGA. In vitro degradation was measured by weight loss and pH drop. Wettability was assessed through water uptake and contact angles measurements. Human limbal stem cells (hLSCs) were isolated and seeded on the scaffolds. Cell attachment and cytotoxicity assay were evaluated on day 1 and 5 after cell seeding. SEM showed regular fiber morphology with diameters ranging between 150 nm and 950 nm. Tensile strength demonstrated similar average stress values for both plasma- and non-plasma-treated samples. Scaffolds also showed gradual degradability over a period of 7–8 weeks. Water contact angle and water absorption were significantly enhanced for plasma-treated scaffolds, indicating a favorable increase in their hydrophilicity. Scaffolds have also supported hLSCs growth and attachment with no signs of cytotoxicity. We have characterized a nanofiber electrospun plasma-treated PLGA scaffold to investigate the mechanical and biological properties and the ability to support the attachment and maintenance of hLSCs.

Investigation of the Cytotoxicity of Electrospun Polysuccinimide-Based Fiber Mats.

This study investigated cell viability in the presence of allylamine-modified and plasma-treated electrospun polysuccinimide fiber mats (PSI-AAmp). Low pressure non-equilibrium plasma was used for crosslinking the PSI-AAm. Comparison of FTIR and XPS analyses demonstrated that crosslinking occurred on the surface of the samples. Cell viability was investigated using the MG-63 osteosarcoma cell line and WST-1 viability reagent. Since PSI hydrolyzes to poly(aspartic acid) (PASP), PASP was used in addition to the regular controls (cells only). Phase contrast showed normal morphology in all cases at 24 h; however, in the presence of PSI-AAmp at 72 h, some rounded, dead cells could also be seen, and proliferation was inhibited. Since proliferation in the presence of PASP alone was not inhibited, the cause of inhibition was not the final product of the hydrolysis. Further investigations will be carried out to pinpoint the cause.

Structure and Properties of Polyamide Fabrics with Insect-Repellent Functionality by Electrospinning and Oxygen Plasma-Treated Surface Coating.

The need for light-weight and high-strength insect-repellant fabrics is of critical importance to the cessation of viral diseases. The goal of the study is to investigate the structure and properties of insect-repellent polyamide fabrics for use in protective garments to guard against mosquitos. Permethrin was applied to the polyamide fabrics through incorporation into the nylon 6 polymer solution during electrospinning and dip coating onto the control untreated and oxygen plasma-treated polyamide fabrics: electropun nylon 6 nanofiber nonwovens, commercially available nylon 6 warp knit tricot, and nylon 66 double weft, knit interlock fabrics. The incorporation of permethrin into the polymer solution before the formation of fibers demonstrated the most efficient way to apply permethrin to the fiber/fabric systems. The plasma treatment significantly increased the amount of permethrin on the surface of the fabrics. All permethrin-containing polyamide fabrics showed excellent fastness of the insecticide to light. The electrospun nylon 6 nonwovens demonstrated the best fastness to washing among the plasma-treated electrospun nylon 6, nylon 66 double weft knit, and nylon 6 warp-knit tricot. All permethrin-treated fabrics were repellent and caused higher percentage of mosquito escape compared to the control untreated fabrics.

Acrylic acid-grafted pre-plasma nanofibers for efficient removal of oil pollution from aquatic environment

Oily wastewater is a worldwide problem threatening the environment and humans. High flux and low-energy consumption separation of oil and water is urgently required but still faces great challenges. In this study, nanofibrous membranes with superhydrophilic and underwater superoleophobic surfaces were fabricated by grafting acrylic acid onto plasma-treated electrospun polystyrene/polyacrylonitrile (PS/PAN) membranes. The morphologies, chemical compositions, mechanical and surface properties of the membranes were examined in detail. The water contact angles of the PS/PAN membranes were 137.4°, 130.1°, 119.5°, 88.1° and 80.2°, respectively, which decreased to 76.5°, 47.9°, 34.4°, 0° and 0° after grafting treatment, proving that the modification improved the surface hydrophilicity of the membranes due to the introduction of hydrophilic groups. In addition, a gravity-driven filtration device was utilized to investigate the oil/water separation potential of the membranes. The results indicated that the grafted PS/PAN membranes separated the layered oil/water mixtures with permeate flux up to 57509 L m-2 h-1, while high fluxes of 1390-6460 L m-2 h-1 for the separation of different oil-in-water emulsions. Importantly, the membranes still maintained high flux and efficiency even after several cycles of separation. Therefore, the reusable membranes can be expected to be potential cost-effective materials for oil/water treatment.

Osteoconduction of Unrestricted Somatic Stem Cells on an Electrospun Polylactic-Co-Glycolic Acid Scaffold Coated with Nanohydroxyapatite

The limitation of traditional bone grafts could be overcome by applying engineered bone constructs, which are mainly produced by seeding suitable stem cells on appropriate scaffolds. So far, bone marrow-derived stromal cells have been the most applied cells in bone tissue engineering, but current data show that unrestricted somatic stem cells (USSCs) from human cord blood might actually be a better stem cell source due to the accessibility and noninvasive procedure of collection. In this study, we cultured USSCs on a plasma-treated electrospun polylactic-co-glycolic acid (PLGA) scaffold coated with nanohydroxyapatite (nHA). Adhesion and proliferation of USSCs on PLGA/nHA were assessed by scanning electron microscopy and MTT assay. Osteogenic differentiation of USSCs into osteoblast lineage cells was evaluated via alkaline phosphatase (ALP) activity and real-time polymerase chain reaction. Our observation showed that USSCs attached and proliferated on PLGA/nHA. Osteogenic differentiation was confirmed by increased ALP activity and OSTEONECTIN expression in USSCs on PLGA/nHA after the 1st week of the osteogenic period. Therefore, using USSCs on electrospun PLGA/nHA is a promising approach in bone tissue engineering.

Plasma-treated electrospun nanofibers as a template for the electrostatic assembly of silver nanoparticles

Silver nanoparticles assembled on a plasma treated electrospun nanofiber membrane could show excellent SERS effect.

Ammonia plasma-treated electrospun polyacrylonitryle nanofibrous membrane: the robust substrate for protein immobilization through glutaraldhyde coupling chemistry for biosensor application

The surface of polyacrylonitrile electrospun nanofibrous membrane (PAN NFM) was aminated by the ammonia plasma treatment. The content of amine groups has been estimated for different time of plasma treatment. The newly generated amine groups were successfully activated by glutaraldehyde (Ga) for the covalent attachment of the protein molecules on the NFM surface. Bio-functionalization of ammonia plasma treated PAN NFM was carried out by the primary antibodies (Ab) immobilization as a protein model through Ga coupling chemistry. For comparison, the immobilization of Ab was also performed through physical interactions. Attenuated total reflection-fourier transform infrared spectroscopy (ATR-FTIR) was used for the characterization of surface functional groups of PAN NFM after different modifications. The surface morphology of the NFM after immobilization was characterized using scanning electron microscope (SEM). The efficacy of Ab immobilization was estimated by enzyme-linked immuno sorbent assay (ELISA) method. X- Ray photoelectron spectroscopy (XPS) was performed to confirm the covalent immobilization of Ab on the modified PAN NFM. Results show that ammonia plasma treatment effectively increased the amount of Ab immobilization through Ga coupling chemistry. Our findings suggest that this is a versatile model for the preparation of stable bio-functionalized NFM which is applicable in different field of biomedical science.

Comparative of fibroblast and osteoblast cells adhesion on surface modified nanofibrous substrates based on polycaprolactone.

One of the determinant factors for successful bioengineering is to achieve appropriate nano-topography and three-dimensional substrate. In this research, polycaprolactone (PCL) nano-fibrous mat with different roughness modified with O2 plasma was fabricated via electrospinning. The purpose of this study was to evaluate the effect of plasma modification along with surface nano-topography of mats on the quality of human fibroblast (HDFs) and osteoblast cells (OSTs)-substrate interaction. Surface properties were studied using scanning electron microscopy (SEM), atomic force microscopy (AFM), contact angle, Fourier-transformation infrared spectroscopy. We evaluated mechanical properties of fabricated mats by tensile test. The viability and proliferation of HDFs and OSTs on the substrates were followed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT). Mineralization of the substrate was determined by alizarin red staining method and calcium content of OSTs was determined by calcium content kit. Cells morphology was studied by SEM analysis. The results revealed that the plasma-treated electrospun nano-fibrous substrate with higher roughness was an excellent designed substrate. A bioactive topography for stimulating proliferation of HDFs and OSTs is to accelerate the latter’s differentiation time. Therefore, the PCL substrate with high density and major nano-topography were considered as a bio-functional and elegant bio-substrate for tissue regeneration applications.

Fabrication of electrospun polycaprolactone coated withchitosan-silver nanoparticles membranes for wound dressing applications

In this study, electrospun polycaprolactone membrane coated with chitosan-silver nanoparticles (CsAg), electrospun polycaprolactone/chitosan/Ag nanoparticles, was fabricated by immersing the plasma-treated electrospun polycaprolactone membrane in the CsAg gel. The plasma modification of electrospun polycaprolactone membrane prior to CsAg coating was tested by methylene blue stain and scanning electron microscope. The presence of silver and chitosan on the plasma-treated electrospun polycaprolactone membrane was confirmed by energy-dispersive X-ray spectroscopy and FT-IR spectrum. Scanning electron microscope observation was employed to observe the morphology of the membranes. The release of Ag ions from electrospun polycaprolactone/chitosan/Ag nanoparticles membrane was tested using atomic absorption spectrometry. Electrospun polycaprolactone/chitosan/Ag nanoparticles membrane inherited advantages from both CsAg gel and electrospun polycaprolactone membrane such as: increasing biocompatibility, mechanical strength, and antibacterial activity against both Gram-negative and Gram-positive bacteria. Thus, this investigation introduces a highly potential membrane that can increase the efficacy of the wound dressing process.

Surface Plasma Treatment of Poly(caprolactone) Micro, Nano, and Multiscale Fibrous Scaffolds for Enhanced Osteoconductivity

In this study, poly(caprolactone) (PCL) was electrospun to nano, micro, and multiscale (micro-nano) fibers, which were then subjected to low pressure argon and nitrogen plasma treatment. The electrospun fibers contain microfibers of diameter 8-10 μm and nanofibers of diameter 200-300 nm. Characterization of the plasma-treated fibers showed that treatment using less oxidizing gas like nitrogen and inert gas like argon functionalize the surface with polar groups that significantly modify the properties of the scaffold. Highly hydrophobic PCL fibrous scaffolds were rendered hydrophilic, with significantly improved biomineralization after the plasma treatment. While plasma treatment on micro and multiscale fibers enhanced their protein adsorption, cell attachment, spreading, elongation, and proliferation, nanofibers showed remarkably improved cell attachment. The applicability of plasma-treated electrospun fibers for differentiation of mesenchymal stem cell toward osteogenic lineage was also studied. Accelerated differentiation toward osteoblast lineage, with maximum alkaline phosphatase (ALP) activity in 14 days was achieved in plasma-treated fibers. Another remarkable outcome was the enhanced ALP activity of the microfibers after plasma treatment, compared with multiscale and nanofibers. Alizarin red staining further confirmed the mineralization of the plasma-treated scaffolds, indicative of maturation of the differentiated cells. This work thus concentrates on harnessing the potential of plasma treatment, for improving the osteoconductivity of fibrous scaffolds, which could be used for bone tissue engineering/regenerative medicine.

Enhanced reconstruction of rat calvarial defects achieved by plasma-treated electrospun scaffolds and induced pluripotent stem cells

Tissue engineering with a combination of stem cells and nanofibrous scaffolds has attracted interest with regard to bone regeneration applications. In the present study, human induced pluripotent stem cells (iPSCs) were cultured on polymeric nanofibrous polyethersulfone (PES) with and without plasma treatment. The capacity of PES and plasma-treated PES (Plasma-PES) scaffolds to support the proliferation and osteogenic differentiation of iPSCs was investigated by MTT assay and for common osteogenic markers such as alkaline phosphatase activity, calcium mineral deposition and bone-related genes. Plasma-PES scaffolds with or without iPSCs were subsequently used to evaluate bone regeneration of critical-size defects in the rat by digital mammography, multislice spiral-computed tomography imaging and histological analysis. The results of in vitro analysis showed that plasma treatment significantly enhanced iPSC proliferation and osteogenesis. After 8 weeks of iPSC-loaded Plasma-PES implantation, no mortality or complication was observed in animals or at the site of surgery. Imaging analysis revealed more extensive bone reconstruction in rats receiving nanofibers compared with untreated control groups. Moreover, Plasma-PES seeded with iPSCs induced the highest regeneration of bone defects among all groups. These findings were confirmed by histological staining. Affective osseointegration was observed in implanted scaffolds. Thus, plasma-treated nanofibrous scaffolds are suitable tissue-engineered matrices for supporting the proliferation and osteogenic differentiation of iPSCs and might also be appropriate for the reconstruction of bone defects.

Bladder tissue engineering using biocompatible nanofibrous electrospun constructs: feasibility and safety investigation.

To investigate the feasibility and safety of using biocompatible, nanofibrous electrospun polycaprolactone (PCL) and combination of polylactic acid (PLLA) and PCL mats in a canine model. Plasma-treated electrospun unseeded mats were implanted in three dogs. The first dog was sacrificed after 3 months and the second and third ones after 4 months, and then, the graft was examined macroscopically with subsequent morphological and histochemical evaluation. Both films showed high levels of cell infiltration and tissue formation, but body response to PLLA/PCL mat in comparison to PCL mat was very low. All three implantation models showed the same light microscopic morphology, immunohistochemistry, and scanning electron microscopy results; nevertheless, only the PCL/PLLA model showed favorable clinical results. Based on these data, nanofibrous PLLA/PCL scaffolding could be a suitable material for the bladder tissue engineering; however, it deserves further investigations.

Poly (epsilon-caprolactone) nanofibrous ring surrounding a polyvinyl alcohol hydrogel for the development of a biocompatible two-part artificial cornea.

The study aimed to fabricate and characterize a 2-part artificial cornea as a substitute for penetrating keratoplasty in patients with corneal blindness. The peripheral part of the artificial cornea consisted of plasma-treated electrospun poly (ɛ-caprolactone) (PCL) nanofibers, which were attached to a hydrogel disc of polyvinyl alcohol (PVA) as a central optical part. The physical properties of the prepared artificial cornea, including morphology, mechanical properties, light transmittance, and contact angle, were assessed. Cell attachment and proliferation studies were performed on rabbit limbal stem cells. The SEM image of the polymeric system showed that the peripheral part formed a highly porous scaffold that could facilitate tissue biointegration. Assessment of the mechanical properties of the peripheral nanofibrous part and the hydrogel optical part showed suitable elasticity. Young’s modulus values of the electrospun PCL skirt and PVA hydrogel core were 7.5 and 5.3 MPa, respectively, which is in line with the elasticity range of natural human cornea (0.3–7 MPa). The light transmittance of the central part was >85% when measured in the 400–800 nm wavelength range. The plasma-treated PCL nanofibrous scaffold promoted limbal stem cell adhesion and proliferation within 10 days. These results confirmed that the polymeric artificial cornea showed suitable physical properties and good biocompatibility and epithelialization ability.