Nerve regeneration following the injury of nerve tissue remains a major issue in the therapeutic medical field. Various bio-mimetic strategies are employed to direct the nerve growth in vitro, among which the chemical and topographical cues elicited by the scaffolds are crucial parameters that is primarily responsible for the axon growth and neurite extension involved in nerve regeneration. We carried out electrospinning for the first time, to fabricate both random and aligned nanofibers of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate; PHBV) and composite PHBV/collagen nanofibers with fiber diameters in the range of 386-472 nm and 205-266 nm, respectively. To evaluate the potential of electrospun aligned nanofibers of PHBV and composite scaffolds as a substrate for nerve regeneration, we cultured nerve cells (PC12) and studied the biocompatibility effect along with neurite extension by immunostaining studies. Cell proliferation assays showed 40.01% and 5.48% higher proliferation of nerve cells on aligned PHBV/Coll50:50 nanofibers compared to cell proliferation on aligned PHBV and PHBV/Col75:25 nanofibers, respectively. Aligned nanofibers of PHBV/Coll provided contact guidance to direct the orientation of nerve cells along the direction of the fibers, thus endowing elongated cell morphology, with bi-polar neurite extensions required for nerve regeneration. Results showed that aligned PHBV/Col nanofibers are promising substrates than the random PHBV/Col nanofibers for application as bioengineered grafts for nerve tissue regeneration.

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The major component of fibrous extracellular matrix of dermis is composed of a complex combination of proteins and polysaccharides. Electrospun cellulose acetate/gelatin might be an effective simulator of the structure and composition of native skin and during this study, we electrospun cellulose acetate/gelatin membranes in various compositions and their performance as a scaffold for either skin tissue engineering or as a wound dressing was evaluated. Skin treatment products, whether tissue-engineered scaffolds or wound dressings, should be sufficiently hydrophilic to allow for gas and fluid exchange and absorb excess exudates while controlling the fluid loss. However, a wound dressing should be easily removable without causing tissue damage and a tissue-engineered scaffold should be able to adhere to the wound, and support cell proliferation during skin regeneration. We showed that these distinct adherency features are feasible just by changing the composition of cellulose acetate and gelatin in composite cellulose acetate/gelatin scaffolds. High proliferation of human dermal fibroblasts on electrospun cellulose acetate/gelatin 25:75 confirmed the capability of cellulose acetate/gelatin 25:75 nanofibers as a tissue-engineered scaffold, while the electrospun cellulose acetate/gelatin 75:25 can be a potential low-adherent wound dressing.

Tissue engineering techniques particularly using electrospun scaffolds have been intensively used in recent years for the development of small diameter vascular grafts. However, the development of a completely successful scaffold that fulfills multiple requirements to guarantee complete vascular regeneration remains challenging. In this study, a hydrophilic and compliant polyurethane namely Tecophilic (TP) blended with gelatin (gel) at a weight ratio of 70:30 (TP(70)/gel(30)) was electrospun to fabricate a tubular composite scaffold with biomechanical properties closely simulating those of native blood vessels. Hydrophilic properties of the composite scaffold induced non-thrombogenicity while the incorporation of gelatin molecules within the scaffold greatly improved the capacity of the scaffold to serve as an adhesive substrate for vascular smooth muscle cells (SMCs), in comparison to pure TP. Preservation of the contractile phenotype of SMCs seeded on electrospun TP(70)/gel(30) was yet another promising feature of this scaffold. The nanostructured TP(70)/gel(30) demonstrated potential feasibility toward functioning as a vascular graft.

The ability of mature smooth muscle cells (SMCs) to modulate their phenotype in response to environmental cues is a critical issue related to vascular diseases. A tissue engineered vascular graft shall promote the contractile phenotype of vascular SMCs. To this aim, Tecophilic/gelatin (TP/gel) was electrospun at different weight ratios of TP/gelatin (100:0, 70:30, 50:50, 30:70), leading to differences in biochemical and mechanical properties of the nanofibers which in turn influenced the phenotype of SMCs. Results indicated that both the substrate with higher ligand density and lower stiffness could enhance SMC contractility and reduce cell proliferation. However, observing the highest SMCs contractility on electrospun TP(70)/gel(30) among the composite scaffolds demonstrated stiffness as the most critical parameter. Due to conflicting effects of softness versus minor fraction of gelatin (reduced ligand density) within TP(70)/gel(30) fibers, a relatively high proliferation of SMCs was still observed on TP(70)/gel(30) scaffold. The surface of TP(70)/gel(30) scaffold was further modified through physical adsorption of gelatin molecules so as to increase the ligand density on its surface, whereby a functional vascular construct that promotes the contractile behavior of SMCs with low cell proliferation was developed.

Aiming to mimic a blood vessel structurally, morphologically, and mechanically, a sequential electrospinning technique using a small diameter mandrel collector was performed and a three-layered tubular scaffold composed of nanofibers of polycaprolactone, collagen, and poly(l-lactic acid) as inner, intermediate, and outer layers, respectively, was developed. Biological performances of the scaffold in terms of compatibility with blood and endothelial cells were assessed to get some insights into its potential use as a tissue engineered small-diameter vascular replacement compared to an expanded polytetrafluoroethylene vascular graft. Due to direct contact of the blood and endothelial cells with inner surface of the scaffold, polycaprolactone fibers were characterized using SEM, water contact angle measurement, and ATR-FTIR. Despite similar surface wettability of the electrospun scaffold and the expanded polytetrafluoroethylene graft, the three-layered scaffold significantly reduced platelet adhesion and hemolysis ratio compared to expanded polytetrafluoroethylene graft while comparable blood clotting profiles were observed for both electrospun scaffold and expanded polytetrafluoroethylene graft. However, inflammatory response to nanofibrous surface of the scaffold was reduced compared to expanded polytetrafluoroethylene graft. The electrospun scaffold also presented a significantly more supportive substrate for endothelialization than the expanded polytetrafluoroethylene graft. The results described herein suggested that the three-layered scaffold has superior biological properties compared to an expanded polytetrafluoroethylene graft for vascular tissue engineering.

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The main challenge encountered in clinical efficacy of tissue engineered vascular grafts is development of a biomimetic scaffold. To establish a scaffold resembling the architecture of the native blood vessels, a bilayered small-diameter nanofibrous tubular scaffold was fabricated by sequential electrospinning process. The inner layer of the scaffold was electrospun from a blend of fast degrading poly(glycerol sebacate) (PGS), a hydrophilic and elastomeric polymer, and slowly degrading polycaprolactone (PCL), with a weight ratio of 2:1, while the outer layer was electrospun using PCL. Our findings elucidated that nanofibrous PCL outer layer can improve the mechanical integrity and at the same time the presence of PGS within the nanofibers of the inner layer provides a nonthrombogenic interface. Additionally, electrospun PGS/PCL nanofibers with an appropriate balance between hydrophilic and hydrophobic characteristics served a suitable substrate for adhesion and proliferation of mesenchymal stem cells (MSCs); meanwhile, sufficient pore size provided by the inner layer facilitated cell infiltration into the interior of the scaffold. KEYWORDS bilayered tubular scaffold, electrospun nanofibers, poly(glycerol sebacate), polycaprolactone, vascular tissue engineering