Biomimetic crimped/aligned microstructure to optimize the mechanics of fibrous hybrid materials for compliant vascular grafts.

The precise mechanical properties of many tissues are highly dependent on both the composition and arrangement of the nanofibrous extracellular matrix. It is well established that collagen nanofibers exhibit a crimped microstructure in several tissues such as blood vessel, tendon, and heart valve. This collagen fiber arrangement results in the classic non-linear 'J-shaped' stress strain curve characteristic of these tissues. Synthetic biomimetic fibrous materials with a crimped microstructure similar to natural collagen demonstrate similar mechanical properties to natural tissues. The following work describes a nanofabrication method based on electrospinning used to fabricate two component hybrid electrospun fibrous materials that mimic the microstructure and mechanical properties of vascular tissue. The properties of these samples can be precisely and predictably optimized by modifying fabrication parameters. Tubular grafts with biomimetic microstructure were constructed to demonstrate the potential of this fabrication method in vascular graft replacement applications. It was possible to closely match both the overall geometry and the compliance of specific blood vessels by optimizing graft microstructure.

Bioinspired Silk Fiber Spinning System via Automated Track-Drawing.

Regenerated silk fibroin (SF) fiber is a multifaceted protein matrix suitable for engineering a wide variety of biological materials. Numerous artificial spinning systems have been developed to mimic the molecular structure and hierarchical properties found in native silks. Here, we show a bioinspired technique that can readily form nanofibers and induce both orientation and structure formation of crystalline ß-sheet assemblies seen in natural silk. In this study, electrospun postdrawn SF nanofibers were fabricated using an automated track-drawing (TD) approach for the continuous production of highly aligned protein nanofibers. This one-step postdrawing process simulates the dominant pulling force seen in natural spinning. The mechanical performance of the postdrawn SF nanofibers with a draw ratio of 2 (DR2) via TD exhibited a 115% increase in Young's modulus and an 80% increase in ultimate tensile strength, compared with the undrawn SF fibers after water treatment. It was also determined that the intermolecular ß-sheet content in DR2 nanofibers increased by 75%. This contribution led to higher glass-transition and degradation temperatures. These biomimetic fibers with structural hierarchy and mechanical properties may be used to build high-performance load-bearing and directionally propagating structures relevant in biomaterial and sustainable material applications.

Effects of Fiber Density and Strain Rate on the Mechanical Properties of Electrospun Polycaprolactone Nanofiber Mats.

This study examines the effects of electrospun polycaprolactone (PCL) fiber density and strain rate on nanofiber mat mechanical properties. An automated track collection system was employed to control fiber number per mat and promote uniform individual fiber properties regardless of the duration of collection. Fiber density is correlated to the mechanical properties of the nanofiber mats. Young's modulus was reduced as fiber density increased, from 14,901 MPa for samples electrospun for 30 s (717 fibers +/- 345) to 3,615 MPa for samples electrospun for 40 min (8,310 fibers +/- 1,904). Ultimate tensile strength (UTS) increased with increasing fiber density, where samples electrospun for 30 s resulted in a UTS of 594 MPa while samples electrospun for 40 min demonstrated a UTS of 1,250 MPa. An average toughness of 0.239 GJ/m3 was seen in the 30 s group, whereas a toughness of 0.515 GJ/m3 was observed at 40 min. The ultimate tensile strain for samples electrospun for 30 s was observed to be 0.39 and 0.48 for samples electrospun for 40 min. The relationships between UTS, Young's modulus, toughness, and ultimate tensile strain with increasing fiber density are the result of fiber-fiber interactions which leads to network mesh interactions.

Microarray Embedding/Sectioning for Parallel Analysis of 3D Cell Spheroids.

Three-dimensional cell spheroid models can be used to predict the effect of drugs and therapeutics and to model tissue development and regeneration. The utility of these models is enhanced by high throughput 3D spheroid culture technologies allowing researchers to efficiently culture numerous spheroids under varied experimental conditions. Detailed analysis of high throughput spheroid culture is much less efficient and generally limited to narrow outputs, such as metabolic viability. We describe a microarray approach that makes traditional histological embedding/sectioning/staining feasible for large 3D cell spheroid sample sets. Detailed methodology to apply this technology is provided. Analysis of the technique validates the potential for efficient histological analysis of up to 96 spheroids in parallel. By integrating high throughput 3D spheroid culture technologies with advanced immunohistochemical techniques, this approach will allow researchers to efficiently probe expression of multiple biomarkers with spatial localization within 3D structures. Quantitative comparison of staining will have improved inter- and intra-experimental reproducibility as multiple samples are collectively processed, stained, and imaged on a single slide.

Continuous Dual-Track Fabrication of Polymer Micro-/Nanofibers Based on Direct Drawing.

This manuscript proposes a continuous and straightforward method for fabricating suspended micro- and nanodiameter polymer fibers using an automated single-step drawing system. Termed track spinning, the system is based on a simple manual fiber drawing process that is automated by using two oppositely rotating tracks. Fibers are continuously spun by direct contact of polymer solution coated tracks followed by mechanical drawing as the distance between the tracks increases. The device can draw single or multifilament arrays of micro- and nanofibers from many kinds of polymers and solvent combinations. To demonstrate, fibers were pulled from polymer solutions containing polyvinyl acetate (PVAc) and polyurethane (PU). Fiber morphology was smooth and uniform, and the diameter was sensitive to draw length and polymer solution/melt properties. Polymer nanofibers with diameters as small as 450 nm and length of 255 mm were produced. The track spinning method is able to form fibers from high viscosity solutions and melts that are not compatible with some other nanofiber fabrication methods. Further, the setup is simple and inexpensive to implement and nozzleless and does not require an electric field or high-velocity jets, and the tracks can be widened and patterned/textured to enhance fiber yield and manufacturing precision.

Cardiomyogenic differentiation of human bone marrow-derived mesenchymal stem cell spheroids within electrospun collagen nanofiber mats.

Collagen is the major structural protein in myocardium and contributes to tissue strength and integrity, cellular orientation, and cell-cell and cell-matrix interactions. Significant post-myocardial infarction related loss of cardiomyocytes and cardiac tissue, and their subsequent replacement with fibrous scar tissue, negatively impacts endogenous tissue repair and regeneration capabilities. To overcome such limitations, tissue engineers are working toward developing a 3D cardiac patch which not only mimics the structural, functional, and biological hierarchy of the native cardiac tissue, but also could deliver autologous stem cells and encourage their homing and differentiation. In this study, we examined the utility of electrospun, randomly-oriented, type-I collagen nanofiber (dia = 789 ± 162 nm) mats on the cardiomyogenic differentiation of human bone marrow-derived mesenchymal stem cells (BM-MSC) spheroids, in the presence or absence of 10 µM 5-azacytidine (aza). Results showed that these scaffolds are biocompatible and enable time-dependent evolution of early (GATA binding protein 4: GATA4), late (cardiac troponin I: cTnI), and mature (myosin heavy chain: MHC) cardiomyogenic markers, with a simultaneous reduction in CD90 (stemness) expression, independent of aza-treatment. Aza-exposure improved connexin-4 expression and sustained sarcomeric α-actin expression, but provided only transient improvement in cardiac troponin T (cTnT) expression. Cell orientation and alignment significantly improved in these nanofiber scaffolds over time and with aza-exposure. Although further quantitative in vitro and in vivo studies are needed to establish the clinical applicability of such stem-cell laden collagen nanofiber mats as cardiac patches for cardiac tissue regeneration, our results underscore the benefits of 3D milieu provided by electrospun collagen nanofiber mats, aza, and spheroids on the survival, cardiac differentiation and maturation of human BM-MSCs. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 3303-3312, 2018.

Protein-Based Fiber Materials in Medicine: A Review.

Fibrous materials have garnered much interest in the field of biomedical engineering due to their high surface-area-to-volume ratio, porosity, and tunability. Specifically, in the field of tissue engineering, fiber meshes have been used to create biomimetic nanostructures that allow for cell attachment, migration, and proliferation, to promote tissue regeneration and wound healing, as well as controllable drug delivery. In addition to the properties of conventional, synthetic polymer fibers, fibers made from natural polymers, such as proteins, can exhibit enhanced biocompatibility, bioactivity, and biodegradability. Of these proteins, keratin, collagen, silk, elastin, zein, and soy are some the most common used in fiber fabrication. The specific capabilities of these materials have been shown to vary based on their physical properties, as well as their fabrication method. To date, such fabrication methods include electrospinning, wet/dry jet spinning, dry spinning, centrifugal spinning, solution blowing, self-assembly, phase separation, and drawing. This review serves to provide a basic knowledge of these commonly utilized proteins and methods, as well as the fabricated fibers’ applications in biomedical research.

Mechanical Considerations for Electrospun Nanofibers in Tendon and Ligament Repair.

Electrospun nanofibers possess unique qualities such as nanodiameter, high surface area to volume ratio, biomimetic architecture, and tunable chemical and electrical properties. Numerous studies have demonstrated the potential of nanofibrous architecture to direct cell morphology, migration, and more complex biological processes such as differentiation and extracellular matrix (ECM) deposition through topographical guidance cues. These advantages have created great interest in electrospun fibers for biomedical applications, including tendon and ligament repair. Electrospun nanofibers, despite their nanoscale size, generally exhibit poor mechanical properties compared to larger conventionally manufactured polymer fiber materials. This invites the question of what role electrospun polymer nanofibers can play in tendon and ligament repair applications that have both biological and mechanical requirements. At first glance, the strength and stiffness of electrospun nanofiber grafts appear to be too low to fill the rigorous loading conditions of these tissues. However, there are a number of strategies to enhance and tune the mechanical properties of electrospun nanofiber grafts. As researchers design the next-generation electrospun tendon and ligament grafts, it is critical to consider numerous physiologically relevant mechanical criteria and to evaluate graft mechanical performance in conditions and loading environments that reflect in vivo conditions and surgical fixation methods.

Extracellular matrix particle-glycosaminoglycan composite hydrogels for regenerative medicine applications.

Tissue extracellular matrix (ECM) is a complex material made up of fibrous proteins and ground substance (glycosaminoglycans, GAGs) that are secreted by cells. ECM contains important biological cues that modulate cell behaviors, and it also serves as a structural scaffold to which cells can adhere. For clinical applications, where immune rejection is a constraint, ECM can be processed using decellularization methods intended to remove cells and donor antigens from tissue or organs, while preserving native biological cues essential for cell growth and differentiation. In this study, a decellularized ECM-based composite hydrogel was formulated by using modified GAGs that covalently bind tissue particles. These GAG-ECM composite hydrogels combine the advantages of solid decellularized ECM scaffolds and pepsin-digested ECM hydrogels by facilitating ECM hydrogel formation without a disruptive enzymatic digestion process. Additionally, engineered hydrogels can contain more than one type of ECM (from bone, fat, liver, lung, spleen, cartilage, or brain), at various concentrations. These hydrogels demonstrated tunable gelation kinetics and mechanical properties, offering the possibility of numerous in vivo and in vitro applications with different property requirements. Retained bioactivity of ECM particles crosslinked into this hydrogel platform was confirmed by the variable response of stem cells to different types of ECM particles with respect to osteogenic differentiation in vitro, and bone regeneration in vivo. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 147-159, 2018.

Biocompatible Silk/Polymer Energy Harvesters Using Stretched Poly (vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) Nanofibers.

Energy harvested from human body movement can produce continuous, stable energy to portable electronics and implanted medical devices. The energy harvesters need to be light, small, inexpensive, and highly portable. Here we report a novel biocompatible device made of poly (vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) nanofibers on flexible substrates. The nanofibers are prepared with electrospinning followed by a stretching process. This results in aligned nanofibers with diameter control. The assembled device demonstrates high mechanical-to-electrical conversion performance, with stretched PVDF-HFP nanofibers outperforming regular electrospun samples by more than 10 times. Fourier transform infrared spectroscopy (FTIR) reveals that the stretched nanofibers have a higher ß phase content, which is the critical polymorph that enables piezoelectricity in polyvinylidene fluoride (PVDF). Polydimethylsiloxane (PDMS) is initially selected as the substrate material for its low cost, high flexibility, and rapid prototyping capability. Bombyx Mori silkworm silk fibroin (SF) and its composites are investigated as promising alternatives due to their high strength, toughness, and biocompatibility. A composite of silk with 20% glycerol demonstrates higher strength and larger ultimate strain than PDMS. With the integration of stretched electrospun PVDF-HFP nanofibers and flexible substrates, this pilot study shows a new pathway for the fabrication of biocompatible, skin-mountable energy devices.

Tissue matrix arrays for high-throughput screening and systems analysis of cell function.

Cell and protein arrays have demonstrated remarkable utility in the high-throughput evaluation of biological responses; however, they lack the complexity of native tissue and organs. Here we spotted tissue extracellular matrix (ECM) particles as two-dimensional (2D) arrays or incorporated them with cells to generate three-dimensional (3D) cell-matrix microtissue arrays. We then investigated the responses of human stem, cancer and immune cells to tissue ECM arrays originating from 11 different tissues. We validated the 2D and 3D arrays as representative of the in vivo microenvironment by means of quantitative analysis of tissue-specific cellular responses, including matrix production, adhesion and proliferation, and morphological changes after culture. The biological outputs correlated with tissue proteomics, and network analysis identified several proteins linked to cell function. Our methodology enables broad screening of ECMs to connect tissue-specific composition with biological activity, providing a new resource for biomaterials research and further understanding of regeneration and disease mechanisms.

A hyaluronic acid-binding contact lens with enhanced water retention.

PURPOSE: As a main component of an artificial tear or eyedrop, hyaluronic acid (HA) prolongs water retention, slows tear removal, improves tear film stability, reduces protein adsorption at the ocular surface and permits uninterrupted blinking. Here, we hypothesized that the contact lens modified with an HA-binding peptide (HABpep) could locally bind and concentrate exogenous HA present in eyedrops to the modified contact lens surface, which exhibited superior water retention. METHODS: To bind HA, a contact lens surface was covalently modified by HABpep with and without a poly(ethylene glycol) (PEG) spacer. Bound HA and its retention over time on the modified surfaces were evaluated by fluorescence measurements. A comparative water evaporation study was performed to determine water retention in an HA-bound contact lens. RESULTS: Fluorescence studies showed that the contact lens was successfully modified by HABpep with or without a PEG spacer, and HA bound to the contact lens surface. Furthermore, the bound HA via HABpep significantly reduced water loss from the modified contact lens. CONCLUSION: HABpep strategies that locally bind and concentrate HA to create a thin coating of a therapeutic molecule on surfaces could provide physical and biological benefits to treat ocular surface dysfunction. The surface bound HA via HABpep enhanced water retention in the modified contact lens.

The fusion of tissue spheroids attached to pre-stretched electrospun polyurethane scaffolds.

Effective cell invasion into thick electrospun biomimetic scaffolds is an unsolved problem. One possible strategy to biofabricate tissue constructs of desirable thickness and material properties without the need for cell invasion is to use thin (<2 µm) porous electrospun meshes and self-assembling (capable of tissue fusion) tissue spheroids as building blocks. Pre-stretched electrospun meshes remained taut in cell culture and were able to support tissue spheroids with minimal deformation. We hypothesize that elastic electrospun scaffolds could be used as temporal support templates for rapid self-assembly of cell spheroids into higher order tissue structures, such as engineered vascular tissue. The aim of this study was to investigate how the attachment of tissue spheroids to pre-stretched polyurethane scaffolds may interfere with the tissue fusion process. Tissue spheroids attached, spread, and fused after being placed on pre-stretched polyurethane electrospun matrices and formed tissue constructs. Efforts to eliminate hole defects with fibrogenic tissue growth factor-ß resulted in the increased synthesis of collagen and periostin and a dramatic reduction in hole size and number. In control experiments, tissue spheroids fuse on a non-adhesive hydrogel and form continuous tissue constructs without holes. Our data demonstrate that tissue spheroids attached to thin stretched elastic electrospun scaffolds have an interrupted tissue fusion process. The resulting tissue-engineered construct phenotype is a direct outcome of the delicate balance of the competing physical forces operating during the tissue fusion process at the interface of the pre-stretched elastic scaffold and the attached tissue spheroids. We have shown that with appropriate treatments, this process can be modulated, and thus, a thin pre-stretched elastic polyurethane electrospun scaffold could serve as a supporting template for rapid biofabrication of thick tissue-engineered constructs without the need for cell invasion.

Tissue extracellular matrix nanoparticle presentation in electrospun nanofibers.

Biomaterials derived from the decellularization of mature tissues retain biological and architectural features that profoundly influence cellular activity. However, the clinical utility of such materials remains limited as the shape and physical properties are difficult to control. In contrast, scaffolds based on synthetic polymers can be engineered to exhibit specific physical properties, yet often suffer from limited biological functionality. This study characterizes composite materials that present decellularized extracellular matrix (DECM) particles in combination with synthetic nanofibers and examines the ability of these materials to influence stem cell differentiation. Mechanical processing of decellularized tissues yielded particles with diameters ranging from 71 to 334 nm. Nanofiber scaffolds containing up to 10% DECM particles (wt/wt) derived from six different tissues were engineered and evaluated to confirm DECM particle incorporation and to measure bioactivity. Scaffolds containing bone, cartilage, and fat promoted osteogenesis at 1 and 3 weeks compared to controls. In contrast, spleen and lung DECM significantly reduced osteogenic outcomes compared to controls. These findings highlight the potential to incorporate appropriate source DECM nanoparticles within nanofiber composites to design a scaffold with bioactivity targeted to specific applications.

Precisely Assembled Nanofiber Arrays as a Platform to Engineer Aligned Cell Sheets for Biofabrication.

A hybrid cell sheet engineering approach was developed using ultra-thin nanofiber arrays to host the formation of composite nanofiber/cell sheets. It was found that confluent aligned cell sheets could grow on uniaxially-aligned and crisscrossed nanofiber arrays with extremely low fiber densities. The porosity of the nanofiber sheets was sufficient to allow aligned linear myotube formation from differentiated myoblasts on both sides of the nanofiber sheets, in spite of single-side cell seeding. The nanofiber content of the composite cell sheets is minimized to reduce the hindrance to cell migration, cell-cell contacts, mass transport, as well as the foreign body response or inflammatory response associated with the biomaterial. Even at extremely low densities, the nanofiber component significantly enhanced the stability and mechanical properties of the composite cell sheets. In addition, the aligned nanofiber arrays imparted excellent handling properties to the composite cell sheets, which allowed easy processing into more complex, thick 3D structures of higher hierarchy. Aligned nanofiber array-based composite cell sheet engineering combines several advantages of material-free cell sheet engineering and polymer scaffold-based cell sheet engineering; and it represents a new direction in aligned cell sheet engineering for a multitude of tissue engineering applications.

A novel method to precisely assemble loose nanofiber structures for regenerative medicine applications.

Polymer nanofibers are favorable for tissue engineering scaffolds because of their high surface-to-volume ratio and biomimicry of the extracellular matrix. Random and uniaxially oriented polymer nanofibers are easily fabricated by conventional electrospinning techniques; however, control over fiber organization within nanofiber structures is limited when they are collected directly from an electrospinning jet. The regenerative medicine applications of electrospun scaffolds could be expanded by developing assembly methods that allow better control of fiber organization. Here, a novel technique is presented that utilizes parallel automated tracks to orient and collect nanofibers from an electrospinning jet. The stabilized fibers are then subsequently assembled into desirable structures. It is difficult to assemble complex structures directly from an electrospinning jet because of high electrical charge and velocities, so this technology adds an intermediate step where nanofibers are immobilized on automated tracks. The result is a continuous steady-state delivery of static stabilized nanofibers that provides a unique and promising platform for automated post processing into useful nanofiber structures. This technique also allows for an indefinite amount of time, as determined by design parameters, for fibers to dry or cool before they contact other nanofibers in the collection site, thus eliminating potential for fiber-to-fiber adhesions even with slow evaporating solvents or high-temperature melts. To demonstrate potential in regenerative medicine applications, several nanofiber structures were fabricated, including: 2D structures with well-controlled fiber density; 3D loosely assembled aligned nanofiber structures with good cell penetration properties; and, complex layer-by-layer 3D aligned fiber structures assembled by integration with post-processing techniques.

Strength comparison of mitral annuloplasty ring and suturing combinations: an in-vitro study.

BACKGROUND AND AIM OF THE STUDY: While mattress sutures are commonly used to secure annuloplasty rings during mitral valve repair, the use of a flexible ring secured with a running polypropylene suture has recently been advocated. The study aim was to assess the separation tensions of semi-rigid and flexible rings using mattress and running suture techniques in an in-vitro static load model. METHODS: Semi-rigid and flexible annuloplasty rings were sutured with either mattress or running sutures (RM, RR, FM, FR) in four groups, of eight hearts each. Twelve additional sutures were passed through the surfaces of each ring to fix the preparation to a testing machine. In a fifth (control) group the mattress sutures securing a flexible ring (FMS) were connected directly to the machine. Each preparation was subjected to progressive axially directed (base-apex) tension until ring-tissue separation occurred. RESULTS: The first major decrease in tension (defined as > or = 10 N in < or = 1.5 s) typically occurred with the separation of at least three adjacent sutures. These starting tensions (N) were: FMS 117 +/- 32.6, RR 131.7 +/- 30.5, RM 137.4 +/- 35.3, FM 152.1 +/- 32.3, and FR 213.2 +/- 30.5. The magnitudes of tension decrease with separation (and percentage of starting tensions) were: FMS 25.4 (21.2%), RR 26.8 (17.6%), RM 28.9 (21.6%), FM 24.6 (17.6%), and FR 22.5 (10.8%). The FR group required more tension to separate than the other groups (p < 0.001), but had a lower magnitude of force drop at dehiscence. CONCLUSION: Flexible rings secured with a running suture required more force to separate than other ring-suture combinations. The lower magnitude of force drop in this group indicated a better tension distribution than in the other groups. Semi-rigid rings separated with a lower force, and had larger drops in tension, regardless of the suture technique used.

An in vitro model of the glomerular capillary wall using electrospun collagen nanofibres in a bioartificial composite basement membrane.

The filtering unit of the kidney, the glomerulus, contains capillaries whose walls function as a biological sieve, the glomerular filtration barrier. This comprises layers of two specialised cells, glomerular endothelial cells (GEnC) and podocytes, separated by a basement membrane. Glomerular filtration barrier function, and dysfunction in disease, remains incompletely understood, partly due to difficulties in studying the relevant cell types in vitro. We have addressed this by generation of unique conditionally immortalised human GEnC and podocytes. However, because the glomerular filtration barrier functions as a whole, it is necessary to develop three dimensional co-culture models to maximise the benefit of the availability of these cells. Here we have developed the first two tri-layer models of the glomerular capillary wall. The first is based on tissue culture inserts and provides evidence of cell-cell interaction via soluble mediators. In the second model the synthetic support of the tissue culture insert is replaced with a novel composite bioartificial membrane. This consists of a nanofibre membrane containing collagen I, electrospun directly onto a micro-photoelectroformed fine nickel supporting mesh. GEnC and podocytes grew in monolayers on either side of the insert support or the novel membrane to form a tri-layer model recapitulating the human glomerular capillary in vitro. These models will advance the study of both the physiology of normal glomerular filtration and of its disruption in glomerular disease.

Polymer nanofibrous structures: Fabrication, biofunctionalization, and cell interactions.

Extracellular matrix fibers (ECM) such as collagen, elastin, and keratin provide biological and physical support for cell attachment, proliferation, migration, differentiation and ultimately cell fate. Therefore, ECM fibers are an important component in tissue and organ development and regeneration. Meanwhile, polymer nanofibers could play the same critical role in tissue regeneration process. Fibrous structures can be fabricated from a variety of materials and methods with diameters ranging throughout the size scale where cells can sense individual fibers (several nanometers to several microns). Polymer nanofiber scaffolds can be designed in a way that predictably modulates a variety of important cell behaviors towards a desired overall function. The nanofibrous topography itself, independent of the fiber material, has demonstrated the potential to modulate cell behaviors desirable in tissue engineering such as: unidirectional alignment; increased viability, attachment, and ECM production; guided migration; and controlled differentiation. The versatility of polymer nanofibers for functionalization with biomolecules opens the door to vast opportunities for the design of tissue engineering scaffolds with even greater control over cell incorporation and function. Despite the promise of polymer nanofibers as tissue engineering scaffolds there have been few clinically relevant successes because no single fabrication technique currently combines control over structural arrangement, material composition, and biofunctionalization, while maintaining reasonable cost and yield. Promising strategies are currently being investigated to allow for the fabrication of optimal polymer nanofiber tissue engineering scaffolds with the goal of treating damaged and degenerated tissues in a clinical setting.