Translate this page to:
Choose Language…
In JoVE (1)
Other Publications (11)
- Advanced Drug Delivery Reviews
- Biomedical Materials (Bristol, England)
- Acta Biomaterialia
- Advanced Drug Delivery Reviews
- Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology
- Acta Biomaterialia
- Journal of Biomedical Materials Research. Part A
- Acta Biomaterialia
- The Journal of Spinal Cord Medicine
- Tissue Engineering. Part A
- Biomaterials
Articles by Scott A. Sell in JoVE
JoVE Bioengineering
Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study
1Department of Biomedical Engineering, Virginia Commonwealth University, 2Department of Anatomy and Neurobiology, Virginia Commonwealth University, 3Department of Cardiovascular Surgery, University Hospital of Geneva
The aim of this study was to mimic the native three layered architecture of the arterial wall. To accomplish this, electrospinning was employed with the use of a 3-1 (input-output) nozzle and blends of polycaprolactone, elastin, and collagen.
Other articles by Scott A. Sell on PubMed
Nanofiber Technology: Designing the Next Generation of Tissue Engineering Scaffolds
Tissue engineering is an interdisciplinary field that has attempted to utilize a variety of processing methods with synthetic and natural polymers to fabricate scaffolds for the regeneration of tissues and organs. The study of structure-function relationships in both normal and pathological tissues has been coupled with the development of biologically active substitutes or engineered materials. The fibrillar collagens, types I, II, and III, are the most abundant natural polymers in the body and are found throughout the interstitial spaces where they function to impart overall structural integrity and strength to tissues. The collagen structures, referred to as extracellular matrix (ECM), provide the cells with the appropriate biological environment for embryologic development, organogenesis, cell growth, and wound repair. In the native tissues, the structural ECM proteins range in diameter from 50 to 500 nm. In order to create scaffolds or ECM analogues, which are truly biomimicking at this scale, one must employ nanotechnology. Recent advances in nanotechnology have led to a variety of approaches for the development of engineered ECM analogues. To date, three processing techniques (self-assembly, phase separation, and electrospinning) have evolved to allow the fabrication of nanofibrous scaffolds. With these advances, the long-awaited and much anticipated construction of a truly "biomimicking" or "ideal" tissue engineered environment, or scaffold, for a variety of tissues is now highly feasible. This review will discuss the three primary technologies (with a focus on electrospinning) available to create tissue engineering scaffolds that are capable of mimicking native tissue, as well as explore the wide array of materials investigated for use in scaffolds.
Cross-linking Methods of Electrospun Fibrinogen Scaffolds for Tissue Engineering Applications
The purpose of this study was to enhance the mechanical properties and slow the degradation of an electrospun fibrinogen scaffold, while maintaining the scaffold's high level of bioactivity. Three different cross-linkers were used to achieve this goal: glutaraldehyde vapour, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) in ethanol and genipin in ethanol. Scaffolds with a fibrinogen concentration of 120 mg ml(-1) were electrospun and cross-linked with one of the aforementioned cross-linkers. Mechanical properties were determined through uniaxial tensile testing performed on scaffolds incubated under standard culture conditions for 1 day, 7 days and 14 days. Cross-linked scaffolds were seeded with human foreskin fibroblasts (BJ-GFP-hTERT) and cultured for 7, 14 and 21 days, with histology and scanning electron microscopy performed upon completion of the time course. Mechanical testing revealed significantly increased peak stress and modulus values for the EDC and genipin cross-linked scaffolds, with significantly slowed degradation. However, cross-linking with EDC and genipin was shown to have some negative effect on the bioactivity of the scaffolds as cell migration throughout the thickness of the scaffold was slowed.
Suture-reinforced Electrospun Polydioxanone-elastin Small-diameter Tubes for Use in Vascular Tissue Engineering: a Feasibility Study
This study characterizes the cross-linking of electrospun elastin and the mechanical properties of suture-reinforced 1.5mm internal diameter electrospun tubes composed of blended polydioxanone (PDO) and soluble elastin. Several tube configurations were tested to assess the effects of reinforcement on tube mechanical properties. Between the electrospun layers of each double-layered prosthetic, zero, one or two 6-0 sutures were wound, maintaining 1mm spacing with a pitch of 9 degrees . Single-layered tubes without suture were also examined. Samples were cross-linked and tested for compliance and burst strength. Compliance decreased significantly (p <0.05) and burst strength significantly increased (p <0.01) with reinforcement. Uncross-linked tubes were also tested to determine the effects of cross-linking. Results demonstrated that cross-linking significantly decreases burst strength (p <0.01), while decreases in compliance for cross-linked tubes were not significant. Cross-linked suture-reinforced PDO-elastin tubes had burst pressures more than 10 times greater than normal systolic pressures and exhibited a range of compliance values, including those matching native artery. These tubes display many characteristics of the "ideal" small-diameter graft, having mechanical properties that can be tailored to match those desired in vascular replacement applications.
Electrospinning of Collagen/biopolymers for Regenerative Medicine and Cardiovascular Tissue Engineering
The process of electrospinning has seen a resurgence of interest in the last few decades which has led to a rapid increase in the amount of research devoted to its use in tissue engineering applications. Of this research, the area of cardiovascular tissue engineering makes up a large percentage, with substantial resources going towards the creation of bioresorbable vascular grafts composed of electrospun nanofibers of collagen and other biopolymers. These bioresorbable grafts have compositions that allow for the in situ remodeling of the structure, with the eventual replacement of the graft with completely autologous tissue. This review will highlight some of the work done in the field of electrospinning for cardiovascular applications, with an emphasis on the use of biopolymers such as collagens, elastin, gelatin, fibrinogen, and silk fibroin, as well as biopolymers used in combination with resorbable synthetic polymers.
Nanotechnology in the Design of Soft Tissue Scaffolds: Innovations in Structure and Function
Engineered scaffolds function to supplement or replace injured, missing, or compromised tissue or organs. The current direction in this research area is to create scaffolds that mimic the structure and function of the native extracellular matrix (ECM). It is believed that the fabrication of a scaffold that has both structural integrity and allows for normal cellular function and interaction will bring scaffolds closer to clinical relevance. Nanotechnology innovations have aided in the development of techniques for the production of nanofiber scaffolds. The three major processing techniques, self-assembly, phase separation, and electrospinning, produce fibers that rival the size of those found in the native ECM. However, the simplicity, versatility, and scalability of electrospinning make it an attractive processing method that can be used to reproduce aspects of the complexity that characterizes the native ECM. Novel electrospinning strategies include alterations of scaffold composition and architecture, along with the addition and encapsulation of cells, pharmaceuticals and growth factors within the scaffold. This article reviews the major nanofiber fabrication technologies as well as delves into recent significant contributions to the conception of a meaningful and practical electrospun scaffold.
A Three-layered Electrospun Matrix to Mimic Native Arterial Architecture Using Polycaprolactone, Elastin, and Collagen: a Preliminary Study
Throughout native artery, collagen, and elastin play an important role, providing a mechanical backbone, preventing vessel rupture, and promoting recovery under pulsatile deformations. The goal of this study was to mimic the structure of native artery by fabricating a multi-layered electrospun conduit composed of poly(caprolactone) (PCL) with the addition of elastin and collagen with blends of 45-45-10, 55-35-10, and 65-25-10 PCL-ELAS-COL to demonstrate mechanical properties indicative of native arterial tissue, while remaining conducive to tissue regeneration. Whole grafts and individual layers were analyzed using uniaxial tensile testing, dynamic compliance, suture retention, and burst strength. Compliance results revealed that changes to the middle/medial layer changed overall graft behavior with whole graft compliance values ranging from 0.8 to 2.8%/100 mm Hg, while uniaxial results demonstrated an average modulus range of 2.0-11.8 MPa. Both modulus and compliance data displayed values within the range of native artery. Mathematical modeling was implemented to show how changes in layer stiffness affect the overall circumferential wall stress, and as a design aid to achieve the best mechanical combination of materials. Overall, the results indicated that a graft can be designed to mimic a tri-layered structure by altering layer properties.
Evaluation of Thrombogenic Potential of Electrospun Bioresorbable Vascular Graft Materials: Acute Monocyte Tissue Factor Expression
The purpose of this study was to quantify the acute expression of tissue factor (TF) by monocytes on interaction with electrospun bioresorbable constructs. A minimal expression of TF will demonstrate the potential for scaffolds to be used as a vascular graft without enhanced risk of failure from acute thrombotic occlusion. Polydioxanone (PDO) (60, 80, 120, and 160 mg/mL) and polycaprolactone (PCL) (80, 10, and 160 mg/mL) dissolved in 1,1,1,3,3,3 hexafluoro-2-propanol (HFP) were electrospun to form fibrous scaffolds. Circular discs (10 mm diameter) of each scaffold were disinfected and seeded with human monocytes (50,000 cells/well). The discs were statically cultured under standard conditions (37 degrees C and 5% CO2), and removed after 24 h for TF analysis with an In-Cell Western assay. Fiber diameter was calculated through ImageTool analysis of scanning electron micrographs. Acute monocyte interaction with scaffolds of PCL (120 mg/mL) resulted in the lowest amount of TF expressed (4 ng/disc), whereas scaffolds of 160 mg/mL PDO elicited the highest amount of TF expressed (51 ng/disc). TF levels expressed on all scaffolds were comparable with the amount expressed on e-PTFE (20 ng/disc). Preliminary data for TF expression on scaffolds of silk (70 mg/mL and 150 mg/mL) and silk:PCL (100 mg/mL, v/v) blends (50:50 and 70:30) resulted in values of TF expression ranging from 0 to 24 ng. Results from this study reveal electrospun grafts composed of PDO and PCL provide no greater risk of failure from an acute thrombotic occlusion due to TF expression when compared with that of the standard e-PTFE graft.
Two Pole Air Gap Electrospinning: Fabrication of Highly Aligned, Three-dimensional Scaffolds for Nerve Reconstruction
We describe the structural and functional properties of three-dimensional (3D) nerve guides fabricated from poly-ε-caprolactone (PCL) using the air gap electrospinning process. This process makes it possible to deposit nano-to-micron diameter fibers into linear bundles that are aligned in parallel with the long axis of a cylindrical construct. By varying starting electrospinning conditions it is possible to modulate scaffold material properties and void space volume. The architecture of these constructs provides thousands of potential channels to direct axon growth. In cell culture functional assays, scaffolds composed of individual PCL fibers ranging from 400 to 1500 nm supported the penetration and growth of axons from rat dorsal root ganglion. To test the efficacy of our guide design we reconstructed 10mm lesions in the rodent sciatic nerve with scaffolds that had fibers 1 μm in average diameter and void volumes >90%. Seven weeks post implantation, microscopic examination of the regenerating tissue revealed dense, parallel arrays of myelinated and non-myelinated axons. Functional blood vessels were scattered throughout the implant. We speculate that end organ targeting might be improved in nerve injuries if axons can be directed to regenerate along specific tissue planes by a guide composed of 3D fiber arrays.
A Case Report on the Use of Sustained Release Platelet-rich Plasma for the Treatment of Chronic Pressure Ulcers
Chronic pressure ulcers affect patient health, emotional state, and quality of life, causing considerable morbidity and mortality in addition to contributing to significant health care costs from lengthy hospitalizations to advanced home care and surgical care costs. The conventional treatment of these wounds can be slow due to their chronic inflammatory state and the senescence of local reparative cells. Platelet-rich plasma (PRP) therapy has been growing as a viable treatment alternative for a number of clinical applications and has potential benefit for use in chronic wounds. The sustained release of large quantities of autologous growth factors, cytokines, and other mediators found in PRP plus the favorable mononuclear cell profile of PRP may help us to stimulate wound healing and resolve chronic inflammation.
Incorporating Platelet-rich Plasma into Electrospun Scaffolds for Tissue Engineering Applications
Platelet-rich plasma (PRP) therapy has seen a recent spike in clinical interest due to the potential that the highly concentrated platelet solutions hold for stimulating tissue repair and regeneration. The aim of this study was to incorporate PRP into a number of electrospun materials to determine how growth factors are eluted from the structures, and what effect the presence of these factors has on enhancing electrospun scaffold bioactivity. PRP underwent a freeze-thaw-freeze process to lyse platelets, followed by lyophilization to create a powdered preparation rich in growth factors (PRGF), which was subsequently added to the electrospinning process. Release of protein from scaffolds over time was quantified, along with the quantification of human macrophage and adipose-derived stem cell (ADSC) chemotaxis and proliferation. Protein assays demonstrated a sustained release of protein from PRGF-containing scaffolds at up to 35 days in culture. Scaffold bioactivity was enhanced as ADSCs demonstrated increased proliferation in the presence of PRGF, whereas macrophages demonstrated increased chemotaxis to PRGF. In conclusion, the work performed in this study demonstrated that the incorporation of PRGF into electrospun structures has a significant positive influence on the bioactivity of the scaffolds, and may prove beneficial in a number of tissue engineering applications.
The Use of Air-flow Impedance to Control Fiber Deposition Patterns During Electrospinning
Electrospun non-woven structures have the potential to form bioresorbable vascular grafts that promote tissue regeneration in situ as they degrade and are replaced by autologous tissue. Current bioresorbable grafts lack appropriate regeneration potential since they do not have optimal architecture, and their fabrication must be altered by the manipulation of process parameters, especially enhancing porosity. We describe here an air-impedance process where the solid mandrel is replaced with a porous mandrel that has pressurized air exiting the pores to impede fiber deposition. The mandrel design, in terms of air-flow rate, pore size, and pore distribution, allows for control over fiber deposition and scaffold porosity, giving greater cell penetration without a detrimental loss of mechanical properties or structural integrity.
