Nanofibrous Substrate Research Articles

Mesenchymal stem cell differentiation to neuronal cells on electrospun nanofibrous substrates for nerve tissue engineering

Bone marrow Mesenchymal stem cells capable of differentiating into neuronal cells on engineered nanofibrous scaffolds have great potential for bionanomaterial–cell transplantation therapy of neurodegenerative diseases and injuries of the nervous system. MSCs have been the highlight of many tissue engineering studies mainly because of their multipotential properties. We investigated the potential of human bone marrow derived Mesenchymal stem cells (MSCs) for neuronal differentiation in vitro on poly( l-lactic acid)-co-poly-(3-caprolactone)/Collagen (PLCL/Coll) nanofibrous scaffolds. PLCL and PLCL/Coll nanofibrous scaffolds were fabricated by electrospinning process and their chemical and mechanical characterizations were carried out using SEM, contact angle, FTIR, and tensile instrument. The differentiation of MSCs was carried out using neuronal inducing factors including β-mercaptoethanol, epidermal growth factor, nerve growth factor and brain derived growth factor in DMEM/F12 media. The proliferations of MSCs evaluated by MTS assay showed that the cells grown on PLCL/Coll nanofibrous scaffolds were comparatively higher (80%) than those on PLCL. Scanning electron microscopy results showed that MSCs differentiated on PLCL/Coll nanofibrous scaffolds showed neuronal morphology, with multipolar elongations and expressed neurofilament and nestin protein by immuno-fluorescent microscopy. Our studies on the differentiation of MSCs to neuronal cells on nanofibrous scaffolds suggest their potential application towards nerve regeneration.

Continuous Nanostructures for the Controlled Release of Drugs

The annual world wide market for controlled release of polymer systems which extends beyond drug delivery is now estimated to $60 billion and these systems are used by over 100 million people each year. It was estimated that drug delivery will play a pivotal role in approximately 40% of all pharmaceutical sales in near future. Novel methods of drug delivery will not only result in more effective and efficacious treatments but also generates new niche markets to provide greater intellectual property protection to already existing drug formulations. Recently, biodegradable electrospun polymer nanofibrous substrate as drug carrier seems to be a promising method for delivering anticancer drugs, especially in postoperative local chemotherapy. Alternatively drug release can be triggered by the environment or other external events such as changes in pH, temperature, or the presence of analyte such as glucose. In general, controlled release of polymer systems delivering drugs in the optimum dosage for long periods is to increase the efficacy of drug, reducing patient compliance. Recent research for the use of nanotechnology (nanoparticle and nanofibers) in drug delivery suggests that the technology might solve problems in the areas such as controlled release, various topical administration, gut absorption and targeted systemic delivery. This review article described the applications of polymer nanoparticles and nanofibers for loading potential drugs for the controlled release to target incurable diseases.

Nanofibrous membrane of collagen–polycaprolactone for cell growth and tissue regeneration

Nanofibrous substrates of synthetic polymers including polycaprolactone (PCL) have shown considerable potential in tissue regeneration. This paper reports the use of PCL/collagen nanofibers to improve the in vitro osteoblastic responses for the applications in bone regeneration area. Collagen and PCL were dissolved in a co-solvent, and the resulting solution was electrospun into a nanofibrous web. Nonwoven fibrous matrices were successfully produced at various compositional ratios (PCL/collagen = 1/3, 1 and 3 by weight). Although the PCL nanofiber was hydrophobic, the presence of collagen significantly improved the water affinity, such as the water contact angle and water uptake capacity. Tensile mechanical tests showed that the collagen-PCL nanofiber had a significantly higher extension rate (approximately 2.8-fold) than the PCL while maintaining the maximum tensile load in a similar range. The osteoblastic cells cultured on the collagen-PCL nanofibrous substrate showed better initial adhesion and a higher level of growth than those cultured on the PCL nanofiber. Furthermore, real-time RT-PCR revealed the expression of a series of bone-associated genes, including osteopontin, collagen type I and alkaline phosphatase. The expression of these genes was significantly higher on the collagen-PCL nanofiber than on the PCL nanofiber. When subcutaneously implanted in mouse the collagen-PCL membrane facilitated tissue cells to well penetrate into the nanofibrous structure at day 7, whilst no such cell penetration was noticed in the pure PCL nanofiber. Overall, the presence of collagen within the PCL nanofiber improves the water affinity, tensile extension rate, and the tissue cell responses, such as initial adhesion, growth, penetration and the expression of bone-associated genes. Therefore, the collagen-PCL nanofibrous membrane may have potential applications in the cell growth and bone tissue regeneration.

Aligned and random nanofibrous substrate for the in vitro culture of Schwann cells for neural tissue engineering

The current challenge in peripheral nerve tissue engineering is to produce an implantable scaffold capable of bridging long nerve gaps that will produce results similar to autograft without requiring the harvest of autologous donor tissue. Aligned and random polycaprolactone/gelatin (PCL/gelatin) nanofibrous scaffolds were fabricated for the in vitro culture of Schwann cells that assist in directing the growth of regenerating axons in nerve tissue engineering. The average fiber diameter attained by electrospinning of polymer blend (PCL/gelatin) ranged from 232 ± 194 to 160 ± 86 nm with high porosity (90%). Blending PCL with gelatin resulted in increased hydrophilicity of nanofibrous scaffolds and yielded better mechanical properties, approaching those of PCL nanofibers. The biocompatibility of fabricated nanofibers was assessed for culturing and proliferation of Schwann cells by MTS assay. The results of the MTS assay and scanning electron microscopy confirmed that aligned and random PCL/gelatin nanofibrous scaffolds are suitable substrates for Schwann cell growth as compared to PCL nanofibrous scaffolds for neural tissue engineering.

Electrospun Biocomposite Nanofibrous Scaffolds for Neural Tissue Engineering

Bridging of nerve gaps after injury is a major problem in peripheral nerve regeneration. Considering the potential application of a bio-artificial nerve guide material, polycaprolactone (PCL)/chitosan nanofibrous scaffolds was designed and evaluated in vitro using rat Schwann cells (RT4-D6P2T) for nerve tissue engineering. PCL, chitosan, and PCL/chitosan nanofibers with average fiber diameters of 630, 450, and 190 nm, respectively, were fabricated using an electrospinning process. The surface chemistry of the fabricated nanofibers was determined using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Simple blending of PCL with chitosan proved an easy and efficient method for fabricating PCL/chitosan nanofibrous scaffolds, whose surface characteristics proved more hydrophilic than PCL nanofibers. Evaluation of mechanical properties showed that the Young's modulus and strain at break of the electrospun PCL/chitosan nanofibers were better than those of the chitosan nanofibers. Results of cell proliferation studies on nanofibrous scaffolds using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay showed 48% more cell proliferation on PCL/chitosan scaffolds than on PCL scaffolds after 8 days of culture. PCL/chitosan scaffolds showed better cell proliferation than PCL scaffolds and maintained their characteristic cell morphology, with spreading bipolar elongations to the nanofibrous substrates. This electrospun nanofibrous matrix thus proved of specific interest in tissue engineering for peripheral nerve regeneration.

Surface-mineralized polymeric nanofiber for the population and osteogenic stimulation of rat bone-marrow stromal cells

Nanofibrous biomaterials made of degradable polymers, including poly(ɛ-caprolactone) (PCL), are considered as a potential substrate for populating and differentiating tissue cells. The surface modification of biomaterials is of utmost importance in regulating cell functions. This study examined the effects of an apatite-mineralization of the PCL nanofiber surface on the growth behavior and osteogenic differentiation of rat bone-marrow stromal cells (rBMSCs). BMSCs isolated from adult rats were seeded on the PCL nanofiber with mineralization (PCLnf-M) and without it as a control (PCLnf), and cultured for up to 28 days. Initially (for 1 h), the cells adhered better on the PCLnf-M than on the control, showing favorable cell affinity to the mineralized nanofibrous surface. The secretion of collagen by the cells was shown to increase with culturing time on both types of the nanofiber. The cell viability was similar at day 7 but was higher on the control at a prolonged period of day 14. However, the alkaline phosphatase (AP) activity was noticed to significantly higher level on the PCLnf-M than on the control at days 14 and 28. Overall, the surface-mineralized PCL nanofibrous substrate was shown to support the adhesion and growth of BMSCs and to stimulate the differentiation into an osteogenic lineage. This type of nanofibrous sheet is considered to be useful as a matrix for the regeneration and engineering of skeletal tissues.

Apatite‐mineralized polycaprolactone nanofibrous web as a bone tissue regeneration substrate

Degradable synthetic polymers with a nanofibrous structure have shown great promise in populating and recruiting cells for the reconstruction of damaged tissues. However, poor cell affinity and lack of bioactivity have limited their potential usefulness in bone regeneration. We produced polymeric nanofiber poly(epsilon-caprolactone) (PCL) with its surface mineralized with bone-like apatite for use as bone regenerative and tissue engineering matrices. PCL was first electrospun into a nanofibrous web, and the surface was further mineralized with apatite following a series of solution treatments. The surface of the mineralized PCL nanofiber was observed to be almost fully covered with nanocrystalline apatites. Through mineralization, the wettability of the nanofiber matrix was greatly improved. Moreover, the murine-derived osteoblastic cells were shown to attach and grow actively on the apatite-mineralized nanofibrous substrate. In particular, the mineralized PCL nanofibrous substrate significantly stimulated the expression of bone-associated genes, including Runx2, collagen type I, alkaline phosphatase, and osteocalcin, when compared with the pure PCL nanofiber substrate without mineralization. The currently developed polymer nanofibrous web with the bioactive mineralized surface is considered to be potentially useful as bone regenerative and tissue engineering matrices.

Nanofibrous matrices of poly(lactic acid) and gelatin polymeric blends for the improvement of cellular responses

Substrates with a nanofibrous morphology are considered as a prospective matrix to populate and support cells in the tissue regeneration area. Although the nanofibers made of synthetic degradable polymers, including poly(lactic acid) (PLA), have been well studied, their poor cell affinity has restricted wider applications. Herein, we produced blending nanofibers made of PLA and gelatin to improve the cellular responses of PLA. For this, both PLA and gelatin were dissolved in an organic solvent, varying the compositions of PLA:gelatin at 1:3 and 1:1 by weight, and the solutions were electrospun into nanofibers. At all compositions, nanofibers could be successfully generated with diameters of approximately hundreds of nanometers. The addition of gelatin into PLA markedly improved the wettability of the nanofibrous substrate. The osteoblastic cells attached and spread well on all the blending nanofibers and pure PLA. In particular, the cellular growth was significantly higher on the gelatin-blended PLA than on the pure PLA nanofiber. On the basis of this study, the PLA/gelatin blending polymeric nanofibers are considered to be useful as a bone cell supporting matrix in the tissue regeneration area.

Bone regeneration on computer-designed nano-fibrous scaffolds

The ability to control architectural features in tissue engineering scaffolds is critical to the success of neo-tissue regeneration. In this work, reverse solid freeform fabrication and thermal phase separation of poly( l-lactic acid) (PLLA) solutions were used to create three-dimensional nano-fibrous (NF) scaffolds with complex geometries on the macro- and micro-scales. This approach allows for the fabrication of NF matrices while having precise control of internal pore size and structure, as well as external scaffold shape including architectures generated from computed-tomography scans and histological sections. In vitro cell cultivation experiments with MC3T3-E1 preosteoblasts were performed on NF scaffolds and on similarly designed solid-walled (SW) scaffolds that did not have nano-fibers. Proliferation studies showed significantly more cells on NF scaffolds after 7 d. In differentiation studies, the NF scaffolds displayed more uniform matrix and mineral production throughout. Real-time PCR also showed significantly higher expression of osteocalcin and bone sialoprotein mRNAs after 2 and 6 weeks in the NF scaffolds. Expression of type I collagen mRNA was lower in NF scaffolds which possibly indicates quicker differentiation on the NF substrate. In summary, we controlled the geometry of NF PLLA scaffolds at multiple size scales, and the in vitro results showed that these NF scaffolds were advantageous to control scaffolds for bone tissue engineering.

High Flux Filtration Medium Based on Nanofibrous Substrate with Hydrophilic Nanocomposite Coating

A novel high flux filtration medium, consisting of a three-tier composite structure, i.e., a nonporous hydrophilic nanocomposite coating top layer, an electrospun nanofibrous substrate midlayer, and a conventional nonwoven microfibrous support, was demonstrated for oil/water emulsion separations for the first time. The nanofibrous substrate was prepared by electrospinning of poly(vinyl alcohol) (PVA) followed by chemical cross-linking with glutaraldehyde (GA) in acetone. The resulting cross-linked PVA substrates showed excellent water resistance and good mechanical properties. The top coating was based on a nanocomposite layer containing hydrophilic polyether-b-polyamide copolymer or a cross-linked PVA hydrogel incorporated with surface-oxidized multiwalled carbon nanotubes (MWNTs). Scanning electron microscopy (SEM) examinations indicated that the nanocomposite layer was nonporous within the instrumental resolution and MWNTs were well dispersed in the polymer matrix. Oil/ water emulsion tests showed that this unique type of filtration media exhibited a high flux rate (up to 330 L/m2-h at the feed pressure of 100 psi) and an excellent total organic solute rejection rate (99.8%) without appreciable fouling. The increase in the concentration of surface-oxidized MWNT in the coating layer generally improves the flux rate, which can be attributed to the generation of more effective hydrophilic nanochannels for water passage in the composite membranes.