Novel Biodegradable 3D-Printed Analgesics-Eluting-Nanofibers Incorporated Nuss Bars for Therapy of Pectus Excavatum.

A novel hybrid biodegradable Nuss bar model was developed to surgically correct the pectus excavatum and reduce the associated pain during treatment. The scheme consisted of a three-dimensional (3D) printed biodegradable polylactide (PLA) Nuss bar as the surgical implant and electrospun polylactide-polyglycolide (PLGA) nanofibers loaded with lidocaine and ketorolac as the analgesic agents. The degradation rate and mechanical properties of the PLA Nuss bars were characterized after submersion in a buffered mixture for different time periods. In addition, the in vivo biocompatibility of the integrated PLA Nuss bars/analgesic-loaded PLGA nanofibers was assessed using a rabbit chest wall model. The outcomes of this work suggest that integration of PLA Nuss bar and PLGA/analgesic nanofibers could successfully enhance the results of pectus excavatum treatment in the animal model. The histological analysis also demonstrated good biocompatibility of the PLA Nuss bars with animal tissues. Eventually, the 3D printed biodegradable Nuss bars may have a potential role in pectus excavatum treatment in humans.

Effect of Pre-Induced Mesenchymal Stem Cell-Coated Cellulose/Collagen Nanofibrous Nerve Conduit on Regeneration of Transected Facial Nerve.

(1) Objective: In order to evaluate the effect of a pre-induced mesenchymal stem cell (MSC)-coated cellulose/collagen nanofibrous nerve conduit on facial nerve regeneration in a rat model both in vitro and in vivo. (2) Methods: After fabrication of the cellulose/collagen nanofibrous conduit, its lumen was coated with either MSCs or pre-induced MSCs. The nerve conduit was then applied to the defective main trunk of the facial nerve. Rats were randomly divided into three treatment groups (n = 10 in each): cellulose/collagen nanofiber (control group), cellulose/collagen nanofiber/MSCs (group I), and cellulose/collagen nanofiber/pre-induced MSCs (group II). (3) Results Fibrillation of the vibrissae of each group was observed, and action potential threshold was compared 8 weeks post-surgery. Histopathological changes were also observed. Groups I and II showed better recovery of vibrissa fibrillation than the control group. (4) Conclusions: Group II, treated with the pre-induced MSC-coated cellulose/collagen nanofibrous nerve conduit, showed the highest degree of recovery based on functional and histological evaluations.

Polyvinyl Alcohol/Chitosan Single-Layered and Polyvinyl Alcohol/Chitosan/Eudragit RL100 Multi-layered Electrospun Nanofibers as an Ocular Matrix for the Controlled Release of Ofloxacin: an In Vitro and In Vivo Evaluation.

A novel nanofiber insert was prepared with a modified electrospinning method to enhance the ocular residence time of ofloxacin (OFX) and to provide a sustained release pattern by covering hydrophilic polymers, chitosan/polyvinyl alcohol (CS/PVA) nanofibers, with a hydrophobic polymer, Eudragit RL100 in layers, and by glutaraldehyde (GA) cross-linking of CS-PVA nanofibers for the treatment of infectious conjunctivitis. The morphology of the prepared nanofibers was studied using scanning electron microscopy (SEM). The average fiber diameter was found to be 123 ± 23 nm for the single electrospun nanofiber with no cross-linking (OFX-O). The single nanofibers, cross-linked for 10 h with GA (OFX-OG), had an average fiber diameter of 159 ± 30 nm. The amount of OFX released from the nanofibers was measured in vitro and in vivo using UV spectroscopy and microbial assay methods against Staphylococcus aureus, respectively. The antimicrobial efficiency of OFX formulated in cross-linked and non-cross-linked nanofibers was affirmed by observing the inhibition zones of Staphylococcus aureus and Escherichia coli. In vivo studies using the OFX nanofibrous inserts on a rabbit eye confirmed a sustained release pattern for up to 96 h. It was found that the cross-linking of the nanofibers by GA vapor could reduce the burst release of OFX from OFX-loaded CS/PVA in one layer and multi-layered nanofibers. In vivo results showed that the AUC0-96 for the nanofibers was 9-20-folds higher compared to the OFX solution. This study thus demonstrates the potential of the nanofiber technology is being utilized to sustained drug release in ocular drug delivery systems.

Natural Nanofiber Shuttles for Transporting Hydrophobic Cargo into Aqueous Solutions.

Hydrophobic biomolecules realize their functions in vivo in aqueous environments, often through a delicate balance of amphiphilicity and chaperones. Introducing exogenous hydrophobic biomolecules into in vivo aqueous systems is a challenge in drug delivery and regenerative medicine, where labile linkers, carriers, and fusions or chimeric molecules are often designed to facilitate such aqueous interfaces. Here, we utilize naturally derived silk nanofiber shuttles with the capacity to transport hydrophobic cargos directly into aqueous solutions. These nanofibers disperse in organic solvents and in aqueous solutions because of their inherent amphiphilicity, with enriched hydrophobicity and strategically interspersed negatively charged groups. Hydrophobic molecules loaded on these shuttles in organic solvent-water systems separated from the solvent after centrifugation. These concentrated hydrophobic molecule-loaded nanofibers could then be dispersed into aqueous solution directly without modification. These shuttle systems were effective for different hydrophobic molecules such as drugs, vitamins, and dyes. Improved biological stability and functions of hydrophobic cargos after loading on these nanofibers suggest potential applications in drug delivery, cosmetology, medical diagnosis, and related health fields, with a relatively facile process.

Emerging frontiers in drug release control by core-shell nanofibers: a review.

In recent years, core-shell (CS) nanofiber has widely been used as a carrier for controlled drug release. This outstanding attention toward CS nanofiber is mainly due to its tremendous significance in controllable drug release in specific locations. The major advantage of CS nanofibers is forming a highly porous mesh, boosting its performance for many applications, due to its large surface-to-volume ratio. This inherently high ratio has prompted electrospun fibers to be considered one of the best drug-delivery-systems available, with the capacity to enhance properties such as cell attachment, drug loading, and mass transfer. Using electrospun fibers as CS nanofibers to incorporate different cargos such as antibiotics, anticancer agents, proteins, DNA, RNA, living cells, and diverse growth factors would considerably satisfy the need for a universal carrier in the field of nanotechnology. In addition to their high surface area, other benefit included in these nanofibers is the ability to trap drugs, easily controlled morphology, and their biomimetic characteristics. In this review, by taking the best advantages of the preparation and uses of CS nanofibers, a novel work in the domain of the controlled drug delivery by nanofiber-based scaffolds is presented.

Nanofiber Dressings Topically Delivering Molecularly Engineered Human Cathelicidin Peptides for the Treatment of Biofilms in Chronic Wounds.

Biofilms of multidrug-resistant bacteria in chronic wounds pose a great challenge in wound care. Herein, we report the topical delivery of molecularly engineered antimicrobial peptides using electrospun nanofiber dressings as a carrier for the treatment of biofilms of multidrug-resistant bacteria in diabetic wounds. Molecularly engineered human cathelicidin peptide 17BIPHE2 was successfully encapsulated in the core of pluronic F127/17BIPHE2-PCL core-shell nanofibers. The in vitro release profiles of 17BIPHE2 showed an in initial burst followed by a sustained release over 4 weeks. The peptide nanofiber formulations effectively killed methicillin-resistant Staphylococcus aureus (MRSA) USA300. Similarly, the 17BIPHE2 peptide containing nanofibers could also effectively kill other bacteria including Klebsiella pneumoniae (104 to 106 CFU) and Acinetobacter baumannii (104 to 107 CFU) clinical strains in vitro without showing evident cytotoxicity to skin cells and monocytes. Importantly, 17BIPHE2-containing nanofiber dressings without debridement caused five-magnitude decreases of the MRSA USA300 CFU in a biofilm-containing chronic wound model based on type II diabetic mice. In combination with debridement, 17BIPHE2-containing nanofiber dressings could completely eliminate the biofilms, providing one possible solution to chronic wound treatment. Taken together, the biodegradable nanofiber-based wound dressings developed in this study can be utilized to effectively deliver molecularly engineered peptides to treat biofilm-containing chronic wounds.

Electrospun nanofibers of polyelectrolyte-surfactant complexes for antibacterial wound dressing application.

The development of polyelectrolyte-surfactant complexes (PESCs) has attracted extensive research interest in different fields of applications. However, the liquid state of PESCs has limited their utility in applications where solid materials are required. In this study, novel antibacterial fibers were fabricated via electrospinning PESCs in the solid state without any additives. The PESCs were prepared in aqueous mixtures of pre-hydrolyzed polyacrylonitrile (HPAN), a polyelectrolyte, and cetyltrimethyl ammonium chloride (CTAC), an antibacterial cationic surfactant, by taking advantage of the self-aggregation behavior of the polyelectrolyte and surfactant, which increased the antibacterial agent loading ability and, thus, the antibacterial activity of polymers. By release-killing and contact-killing mechanisms, the as-spun PESC nanofibrous membranes exhibited strong antibacterial ability against both Gram-positive and Gram-negative bacteria, killing 5 log CFU of E. coli and S. aureus within a contact time as short as 30 min. Furthermore, PESCs were blended with polycaprolactone (PCL) to prepare composite nanofibrous membranes as a novel wound dressing, which showed excellent antibacterial activity and favorable cytocompatibility, with the mechanical strength high enough to satisfy the clinical application requirements. The PESC fibers with durable antibacterial activity presented in the current work would be promising for medical applications.

Preparation and in vivo evaluation of sodium alginate - poly (vinyl alcohol) electrospun nanofibers of forskolin for glaucoma treatment.

The present investigation is aiming to prepare Sodium Alginate (SA) - Poly (vinyl alcohol) (PVA) nanofibrous mats of Forskolin (FSK) for ocular delivery to treat the glaucoma. Nanofibers of SA: PVA (1:0.25) load with ß- cyclodextrin- FSK solid dispersion were successfully prepared by an electrospinning technique. Eight formulations were Prepared and evaluated for drug content, scanning electron microscopy, degree of swelling, drug release and In Vivo Intra ocular pressure (IOP) reduction studies. The morphological studies revealed that average diameter of prepare nano fibers were decreased for formulations with low polymer concentration. Less diameter and uniform surface was observed for formulations F4 and F8 which are prepared under applied voltage 20kV, Capillary tip-to-Collector distance 15cm conditions. From the degree of swelling studies, it was observed that thinner the nanofiber mats, the greater the degree of swelling. The burst release within one hour was seen for F1 to F4 formulations whereas up to 90 min for F5 to F8 formulations. Release kinetic studies revealed that release of drug from the Nanofibrous mats have followed zero order kinetics. The results of in vivo IOP reduction studies suggested that FSK loaded Nanofibrous mats formulation (F4) produced a significant and controlled reduction in IOP throughout 45h.

Pectin and Mucin Enhance the Bioadhesion of Drug Loaded Nanofibrillated Cellulose Films.

PURPOSE: Bioadhesion is an important property of biological membranes, that can be utilized in pharmaceutical and biomedical applications. In this study, we have fabricated mucoadhesive drug releasing films with bio-based, non-toxic and biodegradable polymers that do not require chemical modifications. METHODS: Nanofibrillar cellulose and anionic type nanofibrillar cellulose were used as film forming materials with known mucoadhesive components mucin, pectin and chitosan as functional bioadhesion enhancers. Different polymer combinations were investigated to study the adhesiveness, solid state characteristics, film morphology, swelling, mechanical properties, drug release with the model compound metronidazole and in vitro cytotoxicity using TR146 cells to model buccal epithelium. RESULTS: SEM revealed lamellar structures within the films, which had a thickness ranging 40-240 µm depending on the film polymer composition. All bioadhesive components were non-toxic and showed high adhesiveness. Rapid drug release was observed, as 60-80% of the total amount of metronidazole was released in 30 min depending on the film formulation. CONCLUSIONS: The liquid molding used was a straightforward and simple method to produce drug releasing highly mucoadhesive films, which could be utilized in treating local oral diseases, such as periodontitis. All materials used were natural biodegradable polymers from renewable sources, which are generally regarded as safe.

Nanofiber technology in the ex vivo expansion of cord blood-derived hematopoietic stem cells.

Umbilical cord blood (CB) can be used as an alternative source of hematopoietic stem cells (HSCs) for transplantation in hematological and non-hematological disorders. Despite several recognized advantages the limited cell number in CB one unit still restricts its clinical use. The success of transplantation greatly depends on the levels of total nucleated cell and CD34+ cell counts. Thus, many ex vivo strategies have been developed within the last decade in order to solve this obstacle, with more or less success, mainly determined by the degree of difficulty related with maintaining HSCs self-renewal and stemness properties after long-term expansion. Different research groups have developed very promising and diverse CB-derived HSC expansion strategies using nanofiber scaffolds. Here we review the state-of-the-art of nanofiber technology-based CB-derived HSC expansion.

Fabrication of hybrid scaffold based on hydroxyapatite-biodegradable nanofibers incorporated with liposomal formulation of BMP-2 peptide for bone tissue engineering.

In the present study, we fabricated an efficient, simple biomimetic scaffold to stimulate osteogenic differentiation of mesenchymal stem cells (MSCs). Electrospun poly L-lactic acid nanofibers were employed to mimic the nanofibrillar structure of bone proteins and coated with hydroxyapatite nanoparticles to simulate bone minerals. Thereafter, we regulated the release pattern of BMP-2 peptide through covalent attachment of an optimized liposomal formulation to the scaffold. The fabricated platform provided a sustained release profile of BMP-2 peptide up to 21 days while supporting cellular attachment and proliferation without cytotoxicity. In-vitro results confirmed the superiority of the scaffold containing liposomes through enhancement of growth and differentiation of MSCs. Ectopic bone formation model exhibited significant localized initiation of bone formation of liposome incorporated scaffold. Consequently, these findings demonstrated that our designed platform with modified release properties of BMP-2 peptide considerably promoted osteogenic differentiation of MSCs making it a unique candidate for bone regeneration therapeutics.

Enhanced antibacterial nanocomposite mats by coaxial electrospinning of polycaprolactone fibers loaded with Zn-based nanoparticles.

ZnO and Zn acetate nanoparticles were embedded in polycaprolactone coaxial-fibers and uniaxial-fibers matrices to develop potential antibacterial nanocomposite wound dressings (mats). Morphology, composition, wettability, crystallinity and fiber structure of mats were characterized. Antibacterial properties of mats were tested against E. coli and S. aureus by turbidity and MTT assays. The effect of UVA illumination (prior to bacteria inoculation) on mats' antibacterial activity was also studied. Results showed that a coaxial-fibers design maintained nanoparticles distributed in the outer-shell of fibers and, in general, enhanced the antibacterial effect of the mats, in comparison to conventional uniaxial-fibers mats. Results indicated that mats simultaneously inhibited planktonic and biofilm bacterial growth by, probably, two main antibacterial mechanisms; 1) release of Zn2+ ions (mainly from Zn acetate nanoparticles) and 2) photocatalytic oxidative processes exerted by ZnO nanoparticles. Antibacterial properties of mats were significantly improved by coaxial-fibers design and exposure to UVA-light prior to bacteria inoculation.

Development and Characterization of Novel Medicated Nanofiber for the Treatment of Periodontitis.

Periodontitis is a common microbial infection that involves pocket formation due to the destruction of periodontal ligament. The present work is oriented to provide a holistic approach for the treatment of periodontitis comprising localized delivery of nanometric hydroxyapatite as a reinforcing filler and silver-metronidazole as periodontal pocket disinfectant adjunct to current periodontal therapy because of its broad-spectrum antimicrobial activity and low systemic toxicity. In the present work, electrospinning technique was used to prepare medicated nanofiber enriched with antibacterial-hydroxyapatite layers for dental application. The optimized formulation was characterized by SEM, FTIR, DSC, XRD, etc. Safety assessment and therapeutic potential of optimized formulation was evaluated in both in vitro and in vivo animal models. The newly synthesized complex (silver-metronidazole) exhibited higher antibacterial activity against the selected strain over the referenced silver and metronidazole. Results of in vitro studies suggested good compatibility of the metal complex with the polymer matrix. The drug release behavior from optimized formulation shows constant in vitro release behavior. Both in vitro and in vivo studies show broad-spectrum antimicrobial activity of the metal complex and demonstrate the potential of biomimetic nano-hydroxyapatite for filling periodontal defects. All these observations indicated that the above formulation could play a useful role in the treatment of periodontitis. Graphical Abstract ᅟ.

Formulation, Characterization, and Optimization of Captopril Fast-Dissolving Oral Films.

This work aimed to using optimization study to formulate a patient-friendly captopril fast-dissolving oral film with satisfactory disintegration time. Films were made with pullulan and hydroxypropyl methyl cellulose (HPMC) by using the solvent-casting method. Cellulose nanofiber (CNF) was used as a compatibilizer and glycerine was used as a plasticizer. In order to find an optimum formulation, a response surface methodology and a central composite design were employed. The concentration percentages of pullulan and glycerine were considered to be the design factors. Disintegration time, tensile strength, percent elongation at break, and folding endurance were considered to be the responses. The results showed that CNF improved the compatibility and tensile strength of the pullulan and HPMC blend. Also, the rigid nature of CNF reduced the film elongation but the addition of glycerine improved its flexibility. All formulations showed an acceptable uniformity content and dissolution rate. Complete dissolution for all formulations occurred within 2 min. Films with 26% pullulan, 74% HPMC, 1% CNF, and 5% glycerine were reported to be optimum formulations for captopril fast-dissolving oral films, with 95% confidence levels. The in vivo comparison of optimized formulation with a conventional captopril sublingual tablet exhibited significant increase in AUC (~ 62%) and Cmax (~ 52%) and a major decrease in Tmax (~ 33%). The overall results showed that the captopril FDF is a promising candidate for enhanced in vivo orotransmucosal absorption.

A new embolic liquid agent comprised of amino acid.

Transcatheter arterial or venous embolization has been widely used to address solid tumors by occluding the tumor-feeding vessels. It is also performed to treat portosystemic shunts and to stop bleeding by repair of the site of trauma. Commonly used embolic materials are gelatin sponges, coils, beads, and liquid agents such as absolute ethanol, histoacyryl, and onyx. In the field of interventional radiology, embolotherapy is performed routinely. Liquid embolization agents have different characteristics. Their coagulation time, the inflammatory reaction of the vascular wall or surrounding tissue, and their adhesion to the vascular wall vary. PuraMatrix, a liquid embolic agent not yet available for clinical use, is comprised of amino acid. We introduce and discuss preliminary experimental studies to examine its potential for use in humans.

Self-Assembled Water-Soluble Nanofibers Displaying Collagen Hybridizing Peptides.

Collagen hybridizing peptides (CHP) have been demonstrated as a powerful vehicle for targeting denatured collagen (dColl) produced by disease or injury. Conjugation of ß-sheet peptide motif to the CHP results in self-assembly of nonaggregating ß-sheet nanofibers with precise structure. Due to the molecular architecture of the nanofibers which puts high density of hydrophilic CHPs on the nanofiber surface at fixed distance, the nanofibers exhibit high water solubility, without any signs of intramolecular triple helix formation or fiber-fiber aggregation. Other molecules that are flanked with the triple helical forming GlyProHyp repeats can readily bind to the nanofibers by triple helical folding, allowing facile display of bioactive molecules at high density. In addition, the multivalency of CHPs allows the nanofibers to bind to dColl in vitro and in vivo with extraordinary affinity, particularly without preactivation that unravels the CHP homotrimers. The length of the nanofibers can be tuned from micrometers down to 100 nm by simple heat treatment, and when injected intravenously into mice, the small nanofibers can specifically target dColl in the skeletal tissues with little target-associated signals in the skin and other organs. The CHP nanofibers can be a useful tool for detecting and capturing dColl, understanding how ECM remodelling impacts disease progression, and development of new delivery systems that target such diseases.

Cellulose Nanofiber Alignment Using Evaporation-Induced Droplet-Casting, and Cell Alignment on Aligned Nanocellulose Surfaces.

This work investigates droplet-evaporated cellulose nanofiber (CNF) alignment and cell responses on CNF surfaces. Surfaces of unmodified (u-), anionic (a-), and cationic (c-) CNFs were fabricated using an evaporation-induced droplet-casting method and characterized in terms of degree of orientation. Circular variance (CV) values obtained using Cytospectre software to analyze the degree of orientation from AFM images showed a significantly higher degree of orientation on c- and u-CNF surfaces (average CV 0.27 and 0.24, respectively) compared to a-CNF surfaces (average CV 0.76). Quantitative analysis of surface roughness plots obtained from AFM images confirmed the difference between the direction of alignment versus the direction perpendicular to alignment. AFM images as well as observations during droplet evaporation indicated c-CNF alignment parallel to a dry-boundary line during droplet evaporation. Fibroblasts were cultured on the u-, a-, and c-CNF surfaces with or without a fibronectin (FN) coating for 48 h, and the cell response was evaluated in terms of cell viability, proliferation, morphology, and degree of orientation. Cell viability and proliferation were comparable to that on a control surface on the a-CNF and c-CNF surfaces. Although an FN coating slightly enhanced cell growth on the studied surfaces, uncoated a-CNF and c-CNF surfaces were able to support cell growth as well. The results showed cell orientation on aligned c-CNF surfaces, a finding that could be further utilized when guiding the growth of cells. We also showed that the alignment direction of c-CNFs and thus the cell orientation direction can be controlled with a contact-dispensing technique.

High-Strength Films Consisted of Oriented Chitosan Nanofibers for Guiding Cell Growth.

Chitosan has biocompatibility and biodegradability; however, the practical use of the bulk chitosan materials is hampered by its poor strength, which can not satisfy the mechanical property requirement of organs. Thus, the construction of highly strong chitosan-based materials has attracted much attention. Herein, the high strength nanofibrous hydrogels and films (CS-E) were fabricated from the chitosan solution in LiOH/KOH/urea aqueous system via a mild regenerating process. Under the mild condition (ethanol at low temperature) without the severe fluctuation in the system, the alkaline-urea shell around the chitosan chains was destroyed, and the naked chitosan molecules had sufficient time for the orderly arrangement in parallel manner to form relatively perfect nanofibers. The nanofibers physically cross-linked to form CS-E hydrogels, which could be easily oriented by drawing to achieve a maximum orientation index of 84%, supported by the scanning electron microscopy and two-dimensional wide-angle X-ray diffraction. The dried CS-E films could be bent and folded arbitrarily to various complex patterns and shapes. The oriented CS-E films displayed even ultrahigh tensile strength (282 MPa), which was 5.6× higher than the chitosan films prepared by the traditional acid dissolving method. The CS-E hydrogels possessed hierarchically porous structure, beneficial to the cell adhesion, transportation of nutrients, and removal of metabolic byproducts. The cell assay results demonstrated that the CS-E hydrogels were no cytotoxicity, and osteoblastic cells could adhere, spread, and proliferate well on their surface. Furthermore, the oriented CS-E hydrogels could regulate the directional growth of osteoblastic cells along the orientation direction, on the basis of the filopodia of the cells to extend and adhere on the nanofibers. This work provided a novel approach to construct the oriented high strength chitosan hydrogels and films.

Gelatin/nanoceria nanocomposite fibers as antioxidant scaffolds for neuronal regeneration.

BACKGROUND: The design of efficient nerve conduits able to sustain the axonal outgrowth and its guidance towards appropriate targets is of paramount importance in nerve tissue engineering. METHODS: In this work, we propose the preparation of highly aligned nanocomposite fibers of gelatin/cerium oxide nanoparticles (nanoceria), prepared by electrospinning. Nanoceria are powerful self-regenerative antioxidant nanomaterials, that behave as strong reactive oxygen species scavengers, and among various beneficial effects, they have been proven to inhibit the cell senescence and to promote the neurite sprouting. RESULTS: After a detailed characterization of the developed substrates, they have been tested on neuron-like SH-SY5Y cells, demonstrating strong antioxidant properties and beneficial multi-cue effects in terms of neurite development and alignment. CONCLUSIONS: Obtained findings suggest efficiency of the proposed substrates in providing combined topographical stimuli and antioxidant effects to cultured cells. GENERAL SIGNIFICANCE: Proposed nanocomposite scaffolds represent a promising approach for nerve tissue engineering and regenerative medicine.