Poly Ε-caprolactone Nanofibers Research Articles

Synergistic anticancer effect of Pistacia lentiscus essential oils and 5-Fluorouracil co-loaded onto biodegradable nanofibers against melanoma and breast cancer

Chemoresistance and severe toxicities represent major drawbacks of chemotherapy. Natural extracts, including the essential oils of Pistacia lentiscus (PLEO), exhibit substantial anticancer and anti-inflammatory activities where different cancers are reported to dramatically recess following targeting with PLEO. PLEO has promising antimicrobial, anticancer, and anti-inflammatory properties. However, the therapeutic properties of PLEO are restricted by limited stability, bioavailability, and targeting ability. PLEO nanoformulation can maximize their physicochemical and therapeutic properties, overcoming their shortcomings. Hence, PLEO was extracted and its chemical composition was determined by GC–MS. PLEO and 5-Fluorouracil (5FU) were electrospun into poly-ε-caprolactone nanofibers (PCL-NFs), of 290.71 nm to 680.95 nm diameter, to investigate their anticancer and potential synergistic activities against triple-negative breast cancer cells (MDA-MB-231), human adenocarcinoma breast cancer cells (MCF-7), and human skin melanoma cell line (A375). The prepared nanofibers (NFs) showed enhanced thermal stability and remarkable physical integrity and tensile strength. Biodegradability studies showed prolonged stability over 42 days, supporting the NFs use as a localized therapy of breast tissues (postmastectomy) or melanoma. Release studies revealed sustainable release behaviors over 168 h, with higher released amounts of 5FU and PLEO at pH 5.4, indicating higher targeting abilities towards cancer tissues. NFs loaded with PLEO showed strong antioxidant properties. Finally, NFs loaded with either PLEO or 5FU depicted greater anticancer activities compared to free compounds. The highest anticancer activities were observed with NFs co-loaded with PLEO and 5FU. The developed 5FU-PLEO-PCL-NFs hold potential as a local treatment of breast cancer tissues (post-mastectomy) and melanoma to minimize their possible recurrence.Graphical

Biodegradable Poly-ε-Caprolactone Scaffolds with ECFCs and iMSCs for Tissue-Engineered Heart Valves.

Clinically used heart valve prostheses, despite their progress, are still associated with limitations. Biodegradable poly-ε-caprolactone (PCL) nanofiber scaffolds, as a matrix, were seeded with human endothelial colony-forming cells (ECFCs) and human induced-pluripotent stem cells-derived MSCs (iMSCs) for the generation of tissue-engineered heart valves. Cell adhesion, proliferation, and distribution, as well as the effects of coating PCL nanofibers, were analyzed by fluorescence microscopy and SEM. Mechanical properties of seeded PCL scaffolds were investigated under uniaxial loading. iPSCs were used to differentiate into iMSCs via mesoderm. The obtained iMSCs exhibited a comparable phenotype and surface marker expression to adult human MSCs and were capable of multilineage differentiation. EFCFs and MSCs showed good adhesion and distribution on PCL fibers, forming a closed cell cover. Coating of the fibers resulted in an increased cell number only at an early time point; from day 7 of colonization, there was no difference between cell numbers on coated and uncoated PCL fibers. The mechanical properties of PCL scaffolds under uniaxial loading were compared with native porcine pulmonary valve leaflets. The Young’s modulus and mean elongation at Fmax of unseeded PCL scaffolds were comparable to those of native leaflets (p = ns.). Colonization of PCL scaffolds with human ECFCs or iMSCs did not alter these properties (p = ns.). However, the native heart valves exhibited a maximum tensile stress at a force of 1.2 ± 0.5 N, whereas it was lower in the unseeded PCL scaffolds (0.6 ± 0.0 N, p < 0.05). A closed cell layer on PCL tissues did not change the values of Fmax (ECFCs: 0.6 ± 0.1 N; iMSCs: 0.7 ± 0.1 N). Here, a successful two-phase protocol, based on the timed use of differentiation factors for efficient differentiation of human iPSCs into iMSCs, was developed. Furthermore, we demonstrated the successful colonization of a biodegradable PCL nanofiber matrix with human ECFCs and iMSCs suitable for the generation of tissue-engineered heart valves. A closed cell cover was already evident after 14 days for ECFCs and 21 days for MSCs. The PCL tissue did not show major mechanical differences compared to native heart valves, which was not altered by short-term surface colonization with human cells in the absence of an extracellular matrix.

Synthesis and evaluation of electrospun PCL-plasmid DNA nanofibers for post cancer treatments

Poly-ε-caprolactone (PCL) incorporated with plasmid DNA was electrospun, and the resultant nanofibers were used to observe DNA release from the nanofibers. The plasmid DNA enhanced green fluorescent protein (EGFP) with cytomegalovirus (CMV) promoter (PCMVb-GFP) was amplified with E. coli. PCL was chosen because it is biodegradable aliphatic polyester, which plays a critical role in tissue engineering, such as scaffolding, drug, DNA, gene and protein delivery vehicles. Some of the physical and biological properties of the nanofibers were determined using different methods. Scanning electron microscopy (SEM) micrographs showed that nanofibers have an average diameter of about 100 nm. Cytotoxicity tests showed that cell viability for 1 day, 4 days, and 7 days of the tests were above 80 %. These data demonstrated that PCL-plasmid DNA nanofibers have no cytotoxicity and showed benign biocompatibility for biomedical applications. PCMVb-GFP plasmid-linked electrospun nanofibers continuously released double-stranded DNA for at least seven days. For the first 15 min, there was a burst release of about 1.8 ng/ml. For the following hours and days, the release was about to be the same (release of 0.575 ng/ml). Therefore, PCL nanofibers may be an ideal candidate for various biomedical applications such as cancer treatment, scaffolding, tissue engineering, and protein delivery vehicles.

Macrophage membrane-functionalized nanofibrous mats and their immunomodulatory effects on macrophage polarization

Immunomodulation is an important phenomenon in the normal mammalian host response toward an injury, and plays a critical role in tissue regeneration and regenerative medicine. Different phenotypes of macrophages show an array of activation states compassing pro-inflammatory to pro-alleviating cells, which are the critical players to modulate immune response and tissue regeneration. In this study, macrophage membranes of different phenotypes (macrophages (M0), classically activated macrophages (M1) and alternatively activated macrophages (M2)) were coated onto poly-ε-caprolactone (PCL) nanofibers to acquire exterior surface proteins and similar functions of the natural membranes. In vitro results unveiled that these nanofibers, especially the M2-PCL nanofibers, can suppress the activities of inflammatory markers such as TNF-α and IL-1β, and stimulate anti-inflammatory markers such as Arg-1, IL-10 and TGF-β. In a C57BL/6 mouse model, the macrophage membrane-coated nanofibers, especially the M2-PCL nanofibers, displayed minimal cellular infiltration and low collagen deposition, increased anti-inflammatory CD206 and decreased inflammatory CD86 levels. The M2-PCL nanofibers most effectively neutralized inflammatory chemokines, regulated the expression of inflammation-associated genes as well as anti-inflammatory genes, and showed strong immunomodulatory effects than the PCL, M0-PCL and M1-PCL nanofibers. Statement of SignificanceDifferent types of macrophage membrane-functionalized PCL nanofibers were successfully prepared and well characterized. They inherited the surface proteins imitating the source macrophages, and played an important role in limiting cellular infiltration and collagen deposition. These different macrophages and their membrane-coated nanofibers (M0-PCL, M1-PCL and M2-PCL) behaved like their respective source cells. The M2 mimicking M2-PCL nanofibers effectively polarized macrophages to M2 phenotype and decreased the expression of inflammation-associated chemokines and promoted the anti-inflammation in vitro and in vivo, which is critical for tissue regeneration. The mice implanted with the bio-mimicking M2-PCL nanofibers effectively inhibited toll like receptors signaling induced NF-kB and IRF-5 and their target genes such as Edn-1, IL-6, iNOS, TNF-α, etc. compared to the PCL, and M0-PCL and M1-PCL macrophage membrane-coated nanofibers.

Metronidazole- and Amoxicillin-Loaded PLGA and PCL Nanofibers as Potential Drug Delivery Systems for the Treatment of Periodontitis: In Vitro and In Vivo Evaluations.

The purpose of this study was to prepare poly (D-L) lactide-co-glycolide (PLGA) and poly ε-caprolactone (PCL) nanofibers containing metronidazole and amoxicillin using an electrospinning process as intrapocket sustained-release drug delivery systems for the treatment of periodontal diseases. Scanning electron microscopy showed that the drug containing PLGA and PCL nanofibers produced from the electrospinning process was uniform and bead-free in morphology. The obtained nanofibers had a strong structure and resisted external tension according to the tensiometry results. The cytotoxicity results indicated acceptable cell viability (>80%). Quantification by high-performance liquid chromatography showed almost complete in vitro drug release between 7 and 9 days, whereas 14 days were required for complete drug release in vivo. No significant signs of irritation or inflammatory reaction were detected after three weeks of subcutaneous implantation of nanofibers in the animal models, thus indicating suitable compatibility. The results therefore suggest that the designed nanofibers can be used as potential commercial formulations in the treatment of periodontitis as controlled-release intrapocket drug delivery systems that can increase patient compliance. This is due to their ability to reduce the frequency of administration from three times daily in a systemic manner to once weekly as local delivery.

Biocompatible PCL-nanofibers scaffold with immobilized fibronectin and laminin for neuronal tissue regeneration

Recent advances in regenerative medicine have given hope in overcoming and rehabilitating complex medical conditions. In this regard, the biopolymer poly-ε-caprolactone (PCL) may be a promising candidate for tissue regeneration, despite lacking the essential bioactivity. The present study used PCL nanofibers (NFs) scaffold decorated with the extracellular matrix proteins fibronectin and laminin combined for neuronal regeneration. The potential for the dual proteins to support neuronal cells and promote axonal growth was investigated. Two NFs scaffolds were produced with PLC concentrations of 12% or 15%. Under scanning electron microscopy, both scaffolds evidenced uniform diameter distribution in the range of 358 nm and 887 nm, respectively, with >80% porosity. The Brunauer–Emmett–Teller (BET) test confirmed that the fabricated NFs mats had a high surface area, especially for the 12% NFs with 652 m2/g compared to 254 m2/g for the 15% NFs. The proteins of interest were successfully conjugated to the 12% PCL scaffold through chemical carbodiimide reaction as confirmed by Fourier-transform infrared spectroscopy. The addition of fibronectin and laminin together was shown to be the most favorable for cellular attachment and elongation of neuroblastoma SH-SY5Y cells compared to other formulations. Light microscopy revealed longer neurite outgrowth, higher cellular projected area, and lower shape index for the cells cultured on the combined proteins conjugated fibers, indicating enhanced cellular spread on the scaffold. This preliminary study suggests that PCL nanoscaffolding conjugated with matrix proteins can support neuronal cell viability and neurite growth.

Natamycin-loaded electrospun poly(ε-caprolactone) nanofibers as an innovative platform for antifungal applications

This study aimed to develop and characterize poly-e-caprolactone (PCL) nanofibers containing the natural antifungal natamycin by electrospinning technique using four different formulations. Morphological, structural and thermal properties of PCL nanofibers, with or without natamycin, were investigated by scanning electron microscopy, Fourier-transform and near infrared spectroscopy, thermogravimetric analysis and differential scanning calorimetry. Electrospun nanofibers were tested against filamentous fungi and yeasts. SEM images revealed the formation of nanofibers with typical string-like morphology, with mean diameter ranging from 232 to 363 nm. Results from spectroscopy and thermal analyses indicated that no relevant drug-polymer interaction takes place during nanofiber preparation. Natamycin-loaded nanofibers prepared from the formulations NF1 (PLC dissolved in tetrahydrofuran/dimethylformamide) and NF4 (PLC with polyethylene glycol and medium-chain triglycerides dissolved in tetrahydrofuran/chloroform) presented better antifungal activity. A gradual natamycin migration from the PCL nanofibers was observed. The inhibition zones ranged from 4.3 to 25.6 mm, including when fungi were cultivated in skimmed milk agar as a food model system. The antifungal effect of nanofibers was also confirmed in samples of soft cheese. This work provides electrospinning formulations to prepare natamycin-loaded nanofibers as promising platforms to combat filamentous fungi and yeasts.

Aging effect of atmospheric pressure plasma jet treated polycaprolactone polymer solutions on electrospinning properties

ABSTRACTIn previous work, an atmospheric pressure plasma jet was applied to successfully improve the electrospinnability of poly‐ε‐caprolactone (PCL) which enabled the fabrication of beadless nanofibers. In this paper, we report the aging effect of the plasma treatment to evaluate the robustness of the developed process. For this purpose, plasma‐treated PCL polymer solutions with different exposure time were stored for 1 and 8 days and the aged solutions were analyzed in terms of conductivity, viscosity, surface tension, pH, and polymer molecular weight. During storage, the surface tension and acidity of the plasma‐treated solutions were maintained constant. However, the viscosity was found to be significantly lower after 8 days which was attributed to PCL degradation. Electrospinning of all stored PCL solutions resulted in the generation of beadless PCL nanofibers. The plasma treatment effects were thus found to be highly stable over time and capable of producing high‐quality PCL nanofibers up to 8 days. © 2020 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020, 137, 48914.

Comparison of Different Approaches to Surface Functionalization of Biodegradable Polycaprolactone Scaffolds.

Due to their good mechanical stability compared to gelatin, collagen or polyethylene glycol nanofibers and slow degradation rate, biodegradable poly-ε-caprolactone (PCL) nanofibers are promising material as scaffolds for bone and soft-tissue engineering. Here, PCL nanofibers were prepared by the electrospinning method and then subjected to surface functionalization aimed at improving their biocompatibility and bioactivity. For surface modification, two approaches were used: (i) COOH-containing polymer was deposited on the PCL surface using atmospheric pressure plasma copolymerization of CO2 and C2H4, and (ii) PCL nanofibers were coated with multifunctional bioactive nanostructured TiCaPCON film by magnetron sputtering of TiC–CaO–Ti3POx target. To evaluate bone regeneration ability in vitro, the surface-modified PCL nanofibers were immersed in simulated body fluid (SBF, 1×) for 21 days. The results obtained indicate different osteoblastic and epithelial cell response depending on the modification method. The TiCaPCON-coated PCL nanofibers exhibited enhanced adhesion and proliferation of MC3T3-E1 cells, promoted the formation of Ca-based mineralized layer in SBF and, therefore, can be considered as promising material for bone tissue regeneration. The PCL–COOH nanofibers demonstrated improved adhesion and proliferation of IAR-2 cells, which shows their high potential for skin reparation and wound dressing.

Biocompatible Aloe vera and Tetracycline Hydrochloride Loaded Hybrid Nanofibrous Scaffolds for Skin Tissue Engineering.

Aloe vera (AV) and tetracycline hydrochloride (TCH) exhibit significant properties such as anti-inflammatory, antioxidant and anti-bacterial activities to facilitate skin tissue engineering. The present study aims to develop poly-ε-caprolactone (PCL)/ AV containing curcumin (CUR), and TCH loaded hybrid nanofibrous scaffolds to validate the synergistic effect on the fibroblast proliferation and antimicrobial activity against Gram-positive and Gram-negative bacteria for wound healing. PCL/AV, PCL/CUR, PCL/AV/CUR and PCL/AV/TCH hybrid nanofibrous mats were fabricated using an electrospinning technique and were characterized for surface morphology, the successful incorporation of active compounds, hydrophilicity and the mechanical property of nanofibers. SEM revealed that there was a decrease in the fiber diameter (ranging from 360 to 770 nm) upon the addition of AV, CUR and TCH in PCL nanofibers, which were randomly oriented with bead free morphology. FTIR spectra of various electrospun samples confirmed the successful incorporation of AV, CUR and TCH into the PCL nanofibers. The fabricated nanofibrous scaffolds possessed mechanical properties within the range of human skin. The biocompatibility of electrospun nanofibrous scaffolds were evaluated on primary human dermal fibroblasts (hDF) by MTS assay, CMFDA, Sirius red and F-actin stainings. The results showed that the fabricated PCL/AV/CUR and PCL/AV/TCH nanofibrous scaffolds were non-toxic and had the potential for wound healing applications. The disc diffusion assay confirmed that the electrospun nanofibrous scaffolds possessed antibacterial activity and provided an effective wound dressing for skin tissue engineering.

A polypropylene mesh modified with poly-ε-caprolactone nanofibers in hernia repair: large animal experiment.

PurposeIncisional 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 methodsA 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.ResultsWe 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.ConclusionWe 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.

Poly-ε-caprolactone electrospun nanofiber mesh as a gene delivery tool

Poly-e-caprolactone (PCL) is a biodegradable aliphatic polyester which plays critical roles in tissue engineering, such as scaffolds, drug and protein delivery vehicles. PCL nanofiber meshes fabricated by electrospinning technology have been widely used in recent decade. The objective of this study intends to develop a gene-tethering PCL-nanofiber mesh that can be used as a wrapping material during surgical removal of primary bone tumors, and as a gene delivery tool to provide therapeutic means for tumor recurrence. Non-viral plasmid vector encoding green fluorescent protein (eGFP) was incorporated into PCL nanofibers by electron-spinning technique to form multilayer nano-meshes. Our data demonstrated that PCL nanofiber mesh possessed benign biocompatibility in vitro. More importantly, pCMVb-GFP plasmid-linked electrospun nanofiber mesh successfully released the GFP marker gene and incorporated into the co-cultured fibroblast cells, and consequently expressed the transgene product at transcriptional and translational levels. Further investigation is warranted to characterize the therapeutic influence and long-term safety issue of the PCL nanofiber mesh as a gene delivery tool and therapeutic device in orthopedic oncology.

Determining the mechanical properties of electrospun poly-ε-caprolactone (PCL) nanofibers using AFM and a novel fiber anchoring technique

Due to its low cost, biocompatibility and slow bioresorption, poly-ε-caprolactone (PCL) continues to be a suitable material for select biomedical engineering applications. We used a combined atomic force microscopy (AFM)/optical microscopy technique to determine key mechanical properties of individual electrospun PCL nanofibers with diameters between 440–1040nm. Compared to protein nanofibers, PCL nanofibers showed much lower adhesion, as they slipped on the substrate when mechanically manipulated. We, therefore, first developed a novel technique to anchor individual PCL nanofibers to micrometer-sized ridges on a substrate, and then mechanically tested anchored nanofibers. When held at constant strain, tensile stress relaxed with fast and slow relaxation times of 1.0±0.3s and 8.8±3.1s, respectively. The total tensile modulus was 62±26MPa, the elastic (non-relaxing) component of the tensile modulus was 53±36MPa. Individual PCL fibers could be stretched elastically (without permanent deformation) to strains of 19–23%. PCL nanofibers are rather extensible; they could be stretched to a strain of at least 98%, and a tensile strength of at least 12MPa, before they slipped off the AFM tip. PCL nanofibers that had aged for over a month at ambient conditions became stiffer and less elastic. Our technique provides accurate nanofiber mechanical data, which are needed to guide construction of scaffolds for cells and other biomedical devices.

Significant improvement of biocompatibility of polypropylene mesh for incisional hernia repair by using poly-ε-caprolactone nanofibers functionalized with thrombocyte-rich solution.

Incisional hernia is the most common postoperative complication, affecting up to 20% of patients after abdominal surgery. Insertion of a synthetic surgical mesh has become the standard of care in ventral hernia repair. However, the implementation of a mesh does not reduce the risk of recurrence and the onset of hernia recurrence is only delayed by 2–3 years. Nowadays, more than 100 surgical meshes are available on the market, with polypropylene the most widely used for ventral hernia repair. Nonetheless, the ideal mesh does not exist yet; it still needs to be developed. Polycaprolactone nanofibers appear to be a suitable material for different kinds of cells, including fibroblasts, chondrocytes, and mesenchymal stem cells. The aim of the study reported here was to develop a functionalized scaffold for ventral hernia regeneration. We prepared a novel composite scaffold based on a polypropylene surgical mesh functionalized with poly-ε-caprolactone (PCL) nanofibers and adhered thrombocytes as a natural source of growth factors. In extensive in vitro tests, we proved the biocompatibility of PCL nanofibers with adhered thrombocytes deposited on a polypropylene mesh. Compared with polypropylene mesh alone, this composite scaffold provided better adhesion, growth, metabolic activity, proliferation, and viability of mouse fibroblasts in all tests and was even better than a polypropylene mesh functionalized with PCL nanofibers. The gradual release of growth factors from biocompatible nanofiber-modified scaffolds seems to be a promising approach in tissue engineering and regenerative medicine.

Nanofibers prepared by needleless electrospinning technology as scaffolds for wound healing

Electrospun gelatin and poly-ε-caprolactone (PCL) nanofibers were prepared using needleless technology and their biocompatibility and therapeutic efficacy have been characterized in vitro in cell cultures and in an experimental model of a skin wound. Human dermal fibroblasts, keratinocytes and mesenchymal stem cells seeded on the nanofibers revealed that both nanofibers promoted cell adhesion and proliferation. The effect of nanofibers on wound healing was examined using a full thickness wound model in rats and compared with a standard control treatment with gauze. Significantly faster wound closure was found with gelatin after 5 and 10 days of treatment, but no enhancement with PCL nanofibers was observed. Histological analysis revealed enhanced epithelialisation, increased depth of granulation tissue and increased density of myofibroblasts in the wound area with gelatin nanofibers. The results show that gelatin nanofibers produced by needleless technology accelerate wound healing and may be suitable as a scaffold for cell transfer and skin regeneration.

Controlled release of metronidazole benzoate from poly ε-caprolactone electrospun nanofibers for periodontal diseases

Poly ε-caprolactone (PCL) nanofibers containing metronidazole benzoate (MET) were successfully electrospun and evaluated for periodontal diseases. Solutions of 10.5% w/v PCL and 5–15% w/w MET in mixtures of dichloromethane (DCM)/ N, N-dimethylformamide (DMF) with ratios of 90:10, 80:20 and 70:30 v/v were prepared, and the nanofibers were produced by electrospinning technique. Scanning electron microscopy (SEM) was used to investigate the morphology and average diameter of the electrospun nanofibers. DSC results indicated a molecular dispersion of MET in the PCL nanofibers and showed a decrease in crystallinity of PCL nanofibers by adding MET. Results showed that an increase in the DCM:DMF ratio led to a decrease in the solution conductivity and an increase in the solution viscosity as well as in the nanofibers diameter. Also increasing metronidazole benzoate concentration caused an increase in the solution conductivity and a decrease in the solution viscosity as well as in the nanofibers diameter. In vitro drug release studies in phosphate buffer solution (pH 7.4) showed that the drug release rate was affected by the solvents ratio and the drug concentration. Moreover, the burst release was low, and sustained drug release was prolonged to at least 19 days.