Improvement of sciatic nerve regeneration by multichannel nanofibrous membrane-embedded electro-conductive conduits functionalized with laminin.

Multichannel structures in the design of nerve conduits offer potential advantages for regeneration of damaged nerves. However, lack of biochemical cues and electrical stimulation could hamper satisfactory nerve regeneration. The aim of this study was to simultaneously evaluate the effects of topographical, biological, and electrical cues on sciatic nerve regeneration. Accordingly, a series of multichannel nerve conduit was made using longitudinally-aligned laminin-coated poly (lactic-co-glycolic acid) (PLGA)/carbon nanotubes (CNT) nanofibers (NF, mean diameter: 455 ± 362 nm) in the lumen and randomly-oriented polycaprolactone (PCL) NF (mean diameter: 340 ± 200 nm) on the outer surface. In vitro studies revealed that the materials were nontoxic and able to promote cell attachment and proliferation on nanofibers and on fibrin gel. To determine the influence of laminin as biological and CNT as electrical cues on nerve regeneration, either of hollow PCL conduits, PLGA NF-embedded, PLGA/CNT NF-embedded or laminin-coated PLGA/CNT NF-embedded PCL conduits were implanted in rats. A new surgery method was utilized and results were compared with an autograft. The results of motor and sensory tests in addition to histopathological examination of the regenerated nerves demonstrated the formation of nerve fibers in laminin-coated PLGA/CNT NF-embedded PCL conduits. Results suggested that these conduits have the potential to improve sciatic nerve regeneration. Graphical abstract.

PCL/gelatin nanofibrous scaffolds with human endometrial stem cells/Schwann cells facilitate axon regeneration in spinal cord injury.

The significant consequences of spinal cord injury (SCI) include sensory and motor disability resulting from the death of neuronal cells and axon degeneration. In this respect, overcoming the consequences of SCI including the recovery of sensory and motor functions is considered to be a difficult tasks that requires attention to multiple aspects of treatment. The breakthrough in tissue engineering through the integration of biomaterial scaffolds and stem cells has brought a new hope for the treatment of SCI. In the present study, human endometrial stem cells (hEnSCs) were cultured with human Schwann cells (hSC) in transwells, their differentiation into nerve-like cells was confirmed by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and immunocytochemistry techniques. The differentiated cells (co-hEnSC) were then seeded on the poly ε-caprolactone (PCL)/gelatin scaffolds. The SEM images displayed the favorable seeding and survival of the cells on the scaffolds. The seeded scaffolds were then transplanted into hemisected SCI rats. The growth of neuronal cells was confirmed with immunohistochemical study using NF-H as a neuronal marker. Finally, the Basso, Beattie, and Bresnahan (BBB) test confirmed the recovery of sensory and motor functions. The results suggested that combination therapy using the differentiated hEnSC seeded on PCL/gelatin scaffolds has the potential to heal the injured spinal cord and to limit the secondary damage.

Novel electro-conductive nanocomposites based on electrospun PLGA/CNT for biomedical applications.

Electro-conductive nanocomposites have several applications in biomedical field. Development of a biocompatible electro-conductive polymeric materials is therefore of prime importance. In this study, electro-conductive nanofibrous mats of PLGA/CNT were fabricated through different methods including blend electrospinning, simultaneous PLGA electrospinning and CNT electrospraying and ultrasound-induced adsorption of CNTs on the electrospun PLGA nanofibers. The morphology and diameter of fibers were characterized by SEM and TEM, showing the lowest average diameters of 477 ± 136 nm for PLGA/MWCNT blend nanocomposites. MWCNT-sprayed PLGA specimens showed significant lower water contact angle (83°), electrical resistance (3.0 × 104 Ω) and higher mechanical properties (UTS: 5.50 ± 0.46 MPa) compared to the untreated PLGA scaffolds. Also, results of PC12 cell study demonstrated highest viability percentage on the MWCNT-sprayed PLGA nanofibers. We propose that the conductive nanocomposites have capability to use as tool for the neural regeneration and biosensors.

A thermoresponsive chitosan-based in situ gel formulation incorporated with 5-FU loaded nanoerythrosomes for fibrosarcoma local chemotherapy.

Local administration of drugs at tumor sites over an extended period of time shows potential as a promising approach for cancer treatment. In the present study, the temperature-induced phase transition of chitosan and poloxamer 407 is used to construct an injectable hydrogel encapsulating 5-FU-loaded nanoerythrosome (5-FU-NER-gel). The 5-FU-NERs were found to be spherical, measuring approximately 115 ± 20 nm in diameter and having a surface potential of -7.06 ± 0.4. The drug loading efficiency was approximately 40 %. In situ gel formation took place within 15 s when the gel was exposed to body temperature or subcutaneous injection. A sustained release profile was observed at pH 7.4 and 6.8, with a total 5-FU release of 76.57 ± 4.4 and 98.07 ± 6.31 in 24 h, respectively. MTT, Live/dead, and migration assays confirmed the cytocompatibility of the drug carrier and its effectiveness as a chemotherapeutic formulation. After in vivo antitumor assessment in a subcutaneous autograft model, it was demonstrated that tumor growth inhibition in 14 days was 90 %. Therefore, the obtained injectable chitosan-based hydrogel containing 5-FU-loaded nanoerythrosomes illustrated promising potential as a candidate for local and enhanced delivery of chemotherapeutics at the tumor site.

Co-electrospun PCL-collagen nanofibers containing saffron-synthesized gold nanoparticles as an antioxidant wound dressing.

Oxidant environment and inflammation are the leading cause of chronic wounds such as diabetic ulcers. A dressing containing antioxidants would ensure accelerated wound healing. In this study, electrospun gold nanoparticle (GNP)-embedded nanofibers were developed. GNPs (about 7 nm) were synthesized using saffron extract as a reducing and capping agent (GNP-EXT). For comparison, nanoparticles of the same size were also synthesized using citrate (GNP-CIT). Nanoparticle colloids showed a zeta potential of -27 mV. FTIR confirmed the presence of the extract molecules on the nanoparticles. DPPH assay demonstrated the significant radical scavenging properties of the GNP-EXT. The effect of nanoparticles on the viability of NIH3T3 mouse fibroblast cells was evaluated with an MTT assay that showed no significant toxicity of nanoparticles even in the highest concentration of 250 ppm. Then poly (ɛ-caprolactone) (PCL)- Collagen nanofibers containing GNPs were electrospun. By using SEM, TEM, ATR-FTIR, and contact angle measurement, the nanofibers were characterized. Proper cell adhesion and spreading was observed on nanofibers by SEM and Alamar blue assay illustrated appropriate cyto-compatibility on the obtained nanofibers after 5 days of cell seeding. Wound healing assay also confirmed the cell supporting properties and biocompatibility. The results suggest that saffron-synthesized GNP-loaded nanofibers would be considered as potential wound dressings.

Three-layered PCL-collagen nanofibers containing melilotus officinalis extract for diabetic ulcer healing in a rat model.

Active wound dressing with physicochemical and biological characteristics is more effective in healing diabetic foot ulcer (DFU). In this study, a 3-layer electrospun nanofiber wound dressings was fabricated, while its outer, middle and inner layers of the scaffold were made of PCL, PCL/collagen and collagen nanofibers, respectively. Various amounts of Melilotus officinalis extract were also loaded in the collagen nanofibers as a biologically active compound. The diameter and morphology of the obtained nanofibers were investigated by scanning electron microscopy (SEM) and FT-IR spectroscopy to analyse the composition of prepared dressings. The efficacy of the fabricated dressings as wound healing agent was assessed in streptozotocin-induced diabetic rats. The results demonstrated that the mean diameter of nanofibers are 373 ± 179 nm, 266 ± 108 nm, 160 ± 52 nm, and 393 ± 131 nm for PCL, PCL/collagen, pure collagen, and collagen nanofibers containing 0.08 g extract, respectively. The histo-pathology and histomorphometry assessments demonstrate the herbal extract-loaded electrospun dressings (especially containing 0.08 g of the extract) are promising in improving the diabetic ulcer healing. Our results indicated that the combination of drug did not compromise the physicochemical characteristics of wound dressing, while improving its biological activities.

State-of-the-Art of Nanodiagnostics and Nanotherapeutics against SARS-CoV-2.

The pandemic outbreak of SARS-CoV-2, with millions of infected patients worldwide, has severely challenged all aspects of public health. In this regard, early and rapid detection of infected cases and providing effective therapeutics against the virus are in urgent demand. Along with conventional clinical protocols, nanomaterial-based diagnostics and therapeutics hold a great potential against coronavirus disease 2019 (COVID-19). Indeed, nanoparticles with their outstanding characteristics would render additional advantages to the current approaches for rapid and accurate diagnosis and also developing prophylactic vaccines or antiviral therapeutics. In this review, besides presenting an overview of the coronaviruses and SARS-CoV-2, we discuss the introduced nanomaterial-based detection assays and devices and also antiviral formulations and vaccines for coronaviruses.

Preparation and characterization of polyurethane/chitosan/CNT nanofibrous scaffold for cardiac tissue engineering.

Myocardial infarction of cardiomyocytes is a leading cause of heart failure (HF) worldwide. Since heart has very limited regeneration capacity, cardiac tissue engineering (TE) to produce a bioactive scaffold is considered. In this study, a series of polyurethane solutions (5-7%wt) in aqueous acetic acid were prepared using electrospinning. A variety of Polyurethane (PU)/Chitosan (Cs)/carbon nanotubes (CNT) composite nanofibrous scaffolds with random and aligned orientation were fabricated to structurally mimic the extracellular matrix (ECM). Electrospun nanofibers were then characterized using field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), water contact angle, degradation studies, tensile tests, electrical resistance measurement and cell viability assay. The biocompatibility of electrospun random and aligned nanofibrous scaffolds with H9C2 Cells was confirmed. The results revealed that fabricated PU/Cs/CNT composite nanofibrous scaffolds were electro-conductive and aligned nanofibers could be considered as promising scaffolds with nano-scale features for regeneration of infarcted myocardium.

Biomimetic modification of polyurethane-based nanofibrous vascular grafts: A promising approach towards stable endothelial lining.

The emerging demand for small caliber vascular grafts to replace damaged vessels has attracted research attention. However, there is no perfect replacement in clinical use yet, mainly due to low patency rate of synthetic small caliber grafts. The main pathology behind low patency rate include thrombosis and graft/vessel hemodynamic mismatch, leading to intimal hyperplasia. Rapid in-situ endothelialization of vascular grafts is considered as one of the best strategies to overcome these complications. In the present study, Heparin and VEGF were immobilized via self-polymerization and deposition of polydopamine (PDA) on polyurethane (PU) nanofibrous scaffolds to improve endothelialization. Polyurethane nanofibrous scaffold (PUNF) that mimics vascular extracellular matrix (ECM) was chosen owing to its biocompatibility, biodegradability. Scanning electron microscopy (SEM), water contact angle (CA) measurement and Raman spectroscopy were used to characterize the surface, and tensile test was used to analyze mechanical properties before and after surface modification of the scaffolds. It was found that tensile strength and young's modulus were significantly increased after PDA coating on PUNF membranes. The hemocompatibility tests revealed that surface heparinization significantly inhibited the adhesion of platelet on the scaffolds. Immobilization of VEGF on the scaffolds significantly enhanced the proliferation of human umbilical vein endothelial cells (HUVECs) through enhanced cells adhesion and improved cell-scaffold interactions. The results suggest that dual-factor immobilization resulted in not only confluent monolayer of endothelial cells but also conferred excellent antithrombotic properties to the surface. This method of surface modification (immobilization of Heparin, VEGF by PDA layer) is suggested as a promising modification technique to increase hemocompatibility of small-diameter vascular grafts.

Preparation of collagen/polyurethane/knitted silk as a composite scaffold for tendon tissue engineering.

The main objective of this study was to prepare a hybrid three-dimensional scaffold that mimics natural tendon tissues. It has been found that a knitted silk shows good mechanical strength; however, cell growth on the bare silk is not desirable. Hence, electrospun collagen/polyurethane combination was used to cover knitted silk. A series of collagen and polyurethane solutions (4%-7% w/v) in aqueous acetic acid were prepared and electrospun. According to obtained scanning electron microscopy images from pure collagen and polyurethane nanofibers, concentration was set constant at 5% (w/v) for blend solutions of collagen/polyurethane. Afterward, blend solutions with the weight ratios of 75/25, 50/50 and 25/75 were electrospun. Scanning electron microscopy images demonstrated the smooth and uniform morphology for the optimized nanofibers. The least fibers diameter among three weight ratios was found for collagen/polyurethane (25/75) which was 100.86 ± 40 nm and therefore was selected to be electrospun on the knitted silk. Attenuated total reflectance-Fourier transform infrared spectra confirmed the chemical composition of obtained electrospun nanofibers on the knitted silk. Tensile test of the specimens including blend nanofiber, knitted silk and commercial tendon substitute examined and indicated that collagen/polyurethane-coated knitted silk has appropriate mechanical properties as a scaffold for tendon tissue engineering. Then, Alamar Blue assay of the L929 fibroblast cell line seeded on the prepared scaffolds demonstrated appropriate viability of the cells with a significant proliferation on the scaffold containing more collagen content. The results illustrate that the designed structure would be promising for being used as a temporary substitute for tendon repair.

Electrospun PLLA nanofiber scaffolds for bladder smooth muscle reconstruction.

PURPOSE: Urinary bladder may encounter several pathologic conditions that could lead to loss of its function. Tissue engineering using electrospun PLLA scaffolds is a promising approach to reconstructing or replacing the problematic bladder. METHODS: PLLA nanofibrous scaffolds were prepared utilizing single-nozzle electrospinning. The morphology and distribution of fiber diameters were investigated by scanning electron microscopy (SEM). Human bladder smooth muscle cells (hBSMCs) were isolated from biopsies and characterized by immunocytochemistry (ICC). Then, the cells were seeded on the PLLA nanofibers and Alamar Blue assay proved the biocompatibility of prepared scaffolds. Cell attachment on the nanofibers and also cell morphology over fibrous scaffolds were observed by SEM. RESULTS: The results indicated that electrospun PLLA scaffold provides proper conditions for hBSMCs to interact and attach efficiently to the fibers. Alamar Blue assay showed the compatibility of the obtained electrospun scaffolds with hBSMCs. Also, it was observed that the cells could achieve highly elongated morphology and their native aligned direction besides each other on the random electrospun scaffolds and in the absence of supporting aligned nanofibers. CONCLUSION: Electrospun PLLA scaffold efficiently supports the hBSMCs growth and alignment and also has proper cell compatibility. This scaffold would be promising in urinary bladder tissue engineering.

Comparison of Capability of Human Bone Marrow Mesenchymal Stem Cells and Endometrial Stem Cells to Differentiate into Motor Neurons on Electrospun Poly(ε-caprolactone) Scaffold.

Human endometrial and bone marrow-derived mesenchymal stem cells can be differentiated into a number of cell lineages. Mesenchymal stem cells (MSCs) are potential candidates for cellular therapy. The differentiation of human bone marrow MSCs (hBM-MSCs) and endometrial stem cells (hEnSCs) into motor neuron-like cells has been rarely investigated previously; however, the comparison between these stem cells when they are differentiated into motor neuron-like cell is yet to be studied. The aim of this study was therefore to investigate and compare the capability of hBM-MSCs and hEnSCs cultured on tissue culture polystyrene (TCP) and poly ε-caprolactone (PCL) nanofibrous scaffold to differentiate into motor neuron-like cells in the presence of neural inductive molecules. Engineered hBM-MSCs and hEnSCs seeded on PCL nanofibrous scaffold were differentiated into beta-tubulin III, islet-1, Neurofilament-H (NF-H), HB9, Pax6, and choactase-positive motor neurons by immunostaining and real-time PCR, in response to the signaling molecules. The data obtained from PCR and immunostaining showed that the expression of motor neuron markers of both hBM-MSCs and hEnSCs differentiated cells on PCL scaffold are significantly higher than that of the control group. The expression of these markers in hEnSCs differentiated cells was higher than that in hBM-MSCs. However, this difference was not statistically significant. In conclusion, differentiated hBM-MSCs and hEnSCs on PCL can provide a suitable three-dimensional situation for neuronal survival and outgrowth for regeneration of the central nervous system. Both cells may be potential candidates for cellular therapy in motor neuron disorders. However, differentiation of hEnSCs into motor neuron-like cells was better than hBM-MSCs.