Highly Compliant Vascular Grafts with Gelatin-Sheathed Coaxially Structured Nanofibers.

We have developed three types of materials composed of polyurethane-gelatin, polycaprolactone-gelatin, or polylactic acid-gelatin nanofibers by coaxially electrospinning the hydrophobic core and gelatin sheath with a ratio of 1:5 at fixed concentrations. Results from attenuated total reflection-Fourier transformed infrared spectroscopy demonstrated the gelatin coating around nanofibers in all of the materials. Transmission electron microscopy images further displayed the core-sheath structures showing the core-to-sheath thickness ratio varied greatly with the highest ratio found in polyurethane-gelatin nanofibers. Scanning electron microscopy images revealed similar, uniform fibrous structures in all of the materials, which changed with genipin cross-linking due to interfiber interactions. Thermal analyses revealed varied interactions between the hydrophilic sheath and hydrophobic core among the three materials, which likely caused different core-sheath structures, and thus physicomechanical properties. The addition of gelatin around the hydrophobic polymer and their interactions led to the formation of graft scaffolds with tissue-like viscoelasticity, high compliance, excellent swelling capability, and absence of water permeability while maintaining competent tensile modulus, burst pressure, and suture retention. The hydrogel-like characteristics are advantageous for vascular grafting use, because of the capability of bypassing preclotting prior to implantation, retaining vascular fluid volume, and facilitating molecular transport across the graft wall, as shown by coculturing vascular cells sandwiched over a thick-wall scaffold. Varied core-sheath interactions within scaffolding nanofibers led to differences in graft functional properties such as water swelling ratio, compliance, and supporting growth of cocultured vascular cells. The PCL-gelatin scaffold with thick gelatin-sheathed nanofibers demonstrated a more compliant structure, elastic mechanics, and high water swelling property. Our results demonstrate a feasible approach to produce new hybrid, biodegradable nanofibrous scaffold biomaterials with interactive core-sheath structure, good biocompatibility, and tissue-like viscoelasticity, which may reduce potential problems with the use of individual polymers for vascular grafts.

Development of Tripolymeric Triaxial Electrospun Fibrous Matrices for Dual Drug Delivery Applications.

Since the first work by Laurencin and colleagues on the development of polymeric electrospinning for biomedical purposes, the use of electrospinning technology has found broad applications in such areas of tissue regeneration and drug delivery. More recently, coaxial electrospinning has emerged as an important technique to develop scaffolds for regenerative engineering incorporated with drug(s). However, the addition of a softer core layer leads to a reduction in mechanical properties. Here, novel robust tripolymeric triaxially electrospun fibrous scaffolds were developed with a polycaprolactone (PCL) (core layer), a 50:50 poly (lactic-co-glycolic acid) (PLGA) (sheath layer) and a gelatin (intermediate layer) with a dual drug delivery capability was developed through modified electrospinning. A sharp increase in elastic modulus after the incorporation of PCL in the core of the triaxial fibers in comparison with uniaxial PLGA (50:50) and coaxial PLGA (50:50) (sheath)-gelatin (core) fibers was observed. Thermal analysis of the fibrous scaffolds revealed an interaction between the core-intermediate and sheath-intermediate layers of the triaxial fibers contributing to the higher tensile modulus. A simultaneous dual release of model small molecule Rhodamine B (RhB) and model protein Fluorescein isothiocynate (FITC) Bovine Serum Albumin (BSA) conjugate incorporated in the sheath and intermediate layers of triaxial fibers was achieved. The tripolymeric, triaxial electrospun systems were seen to be ideal for the support of mesenchymal stem cell growth, as shrinkage of fibers normally found with conventional electrospun systems was minimized. These tripolymeric triaxial electrospun fibers that are biomechanically competent, biocompatible, and capable of dual drug release are designed for regenerative engineering and drug delivery applications.

Coaxially-structured fibres with tailored material properties for vascular graft implant.

Readily-available small-diameter arterial grafts require a great combination of materials properties, including high strength, compliance, suturability, blood sealing and anti-thrombogenicity, as well as anti-kinking property for those used in challenging anatomical situations. We have constructed grafts composed of coaxially-structured polycaprolactone (PCL)/gelatin nanofibres, and tailored the material structures to achieve high strength, compliance and kink resistance, as well as excellent water sealing and anti-thrombogenicity. Coaxially-structured fibres in the grafts provided mechanical stability through the core, while flexibility and cell adhesion through the sheath. Results showed that graft compliance increased while strength decreased with the concentration ratio between core and sheath polymers. Compared to pure PCL fibrous surfaces, coaxial PCL/gelatin fibrous surfaces potently inhibited platelet adhesion and activation, providing excellent anti-thrombogenicity. To render sufficient burst strength and suturability, an additional layer of pure PCL was necessary to cap the layer of coaxial PCL/gelatin fibres. The two-layered grafts with the wall thickness comparable to native arteries demonstrated artery-like compliance and kink resistance, properties important to arteries under complex mechanical loading. The in vivo evaluation was performed using the interposition carotid artery graft model in rabbits for three months. Interestingly, results from ultrasonic imaging and histological analysis demonstrated that the two-layered grafts with a thinner outer PCL layer, which possessed higher compliance and kink resistance, showed increased blood flow, minimal lumen reduction and fibrosis. All vascular grafts exhibited patency and induced limited cell infiltration. Together, we presented a facile and useful approach to fabricate vascular grafts with superior graft performances, biomechanical properties, and blood compatibility. Grafts with artery-like compliance and flexibility have demonstrated improved implantation outcomes.

Evaluation of electrospun PLLA/PEGDMA polymer coatings for vascular stent material.

The field of percutaneous coronary intervention has seen a plethora of advances over the past few decades, which have allowed for its development into safe and effective treatments for patients suffering from cardiovascular diseases. However, stent thrombosis and in-stent restenosis remain clinically significant problems. Herein, we describe the synthesis and characterization of fibrous polymer coatings on stent material nitinol, in the hopes of developing a more suitable stent surface to enhance re-endothelialization. Electrospinning technique was used to fabricate polyethylene glycol dimethacrylate/poly l-lactide acid (PEGDMA/PLLA) blend fiber substrate with tunable elasticity and hydrophilicity for use as coatings. Attachment of platelets and arterial smooth muscle cells (SMC) onto the coatings as well as the secretory effect of mesenchymal stem cells cultured on the coatings on the proliferation and migration of arterial endothelial cells and SMCs were assessed. It was demonstrated that electrospun PEGDMA/PLLA coating with 1:1 ratio of the components on the nitinol stent-reduced platelet and SMC attachment and increased stem cell secretory factors that enhance endothelial proliferation. We therefore postulate that the fibrous coating surface would possess enhanced biological compatibility of nitinol stents and hold the potential in preventing stent failure through restenosis and thrombosis.

Biomimetic interconnected porous keratin-fibrin-gelatin 3D sponge for tissue engineering application.

The medicated wound dressing material with highly interconnected pores, mimicking the function of the extracellular matrix was fabricated for the promotion of cell growth. In this study, keratin (K), fibrin (F) and gelatin (G) composite scaffold (KFG-SPG) was fabricated by freeze drying technique and the mupirocin (D) drug was successfully incorporated with KFG-SPG (KFG-SPG-D) intended for tissue engineering applications. The fabrication of scaffold was performed without the use of any strong chemical solvents, and the solid sponge scaffold was obtained with well interconnected pores. The porous morphology of the scaffold was confirmed by SEM analysis and exhibited competent mechanical properties. KFG-SPG and KFG-SPG-D possess high level of biocompatibility, cell proliferation and cell adhesion of NIH 3T3 fibroblast and human keratinocytes (HaCaT) cell lines thereby indicating the scaffolds potential as a suitable medicated dressing for wound healing.

Development and characterization of coaxially electrospun gelatin coated poly (3-hydroxybutyric acid) thin films as potential scaffolds for skin regeneration.

The morphology of fibers synthesized through electrospinning has been found to mimic extracellular matrix. Coaxially electrospun fibers of gelatin (sheath) coated poly (3-hydroxybutyric acid) (PHB) (core) was developed using 2,2,2 trifluoroethanol(TFE) and 1,1,1,3,3,3 hexafluoro-2-propanol(HFIP) as solvents respectively. The coaxial structure and coating of gelatin with PHB fibers was confirmed through transmission electron microscopy (TEM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Thermal stability of the coaxially electrospun fibers was analyzed using thermogravimetric analysis(TGA), differential scanning calorimetry(DSC) and differential thermogravimetric analysis(DTA). Complete evaporation of solvent and gelatin grafting over PHB fibers was confirmed through attenuated total reflection-Fourier transformed infrared spectroscopy (ATR-FTIR). The coaxially electrospun fibers exhibited competent tensile properties for skin regeneration with high surface area and porosity. In vitro degradation studies proved the stability of fibers and its potential applications in tissue engineering. The fibers supported the growth of human dermal fibroblasts and keratinocytes with normal morphology indicating its potential as a scaffold for skin regeneration.