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A major challenge in chronic wound treatment is maintaining an appropriate wound moisture balance throughout the healing process. Wound dehydration hinders wound healing due to impeded molecule transport and cell migration with associated tissue necrosis. In contrast, wounds that produce excess fluid contain high levels of reactive oxygen species and matrix metalloproteases that impede cell recruitment, extracellular matrix reconstruction, and angiogenesis. Dressings are currently selected based on the relative amount of wound exudate with no universal dressing available that can maintain appropriate wound moisture balance to enhance healing. This work aimed to develop a high porosity poly(ethylene glycol) diacrylate hydrogel foam that can both rapidly remove exudate and provide self-tuning moisture control to prevent wound dehydration. A custom foaming device was used to vary hydrogel foam porosity from 25% to 75% by adjusting the initial air-to-solution volume ratio. Hydrogel foams demonstrated substantial improvements in water uptake volume and rate as compared to bulk hydrogels while maintaining similar hydration benefits with slow dehydration rates. The hydrogel foam with the highest porosity (~75%) demonstrated the greatest water uptake and rate, which outperformed commercial dressing products, Curafoam® and Silvercel®, in water absorption, moisture retention, and exudate management. Investigation of the water vapor transmission rates of each dressing at varied hydration levels was characterized and demonstrated the dynamic moisture-controlling capability of the hydrogel foam dressing. Overall, the self-tuning moisture control of this hydrogel foam dressing holds great promise to improve healing outcomes for both dry and exudative chronic wounds.

The attachment of bacteria onto a surface, consequent signaling, and the accumulation and growth of the surface-bound bacterial population are key initial steps in the formation of pathogenic biofilms. While recent reports have hinted that the stiffness of a surface may affect the accumulation of bacteria on that surface, the processes that underlie bacterial perception of and response to surface stiffness are unknown. Furthermore, whether, and how, the surface stiffness impacts biofilm development, after initial accumulation, is not known. We use thin and thick hydrogels to create stiff and soft composite materials, respectively, with the same surface chemistry. Using quantitative microscopy, we find that the accumulation, motility, and growth of the opportunistic human pathogenPseudomonas aeruginosarespond to surface stiffness, and that these are linked through cyclic-di-GMP signaling that depends on surface stiffness. The mechanical cue stemming from surface stiffness is elucidated using finite-element modeling combined with experiments - adhesion to stiffer surfaces results in greater changes in mechanical stress and strain in the bacterial envelope than does adhesion to softer surfaces with identical surface chemistry. The cell-surface-exposed protein PilY1 acts as a mechanosensor, that upon surface engagement, results in higher cyclic-di-GMP levels, lower motility, and greater accumulation on stiffer surfaces. PilY1 impacts the biofilm lag phase, which is extended for bacteria attaching to stiffer surfaces. This study shows clear evidence that bacteria actively respond to different stiffness of surfaces where they adhereviaperceiving varied mechanical stress and strain upon surface engagement.ImportanceBacteria colonize many types of biological and medical surfaces with a large range of stiffnesses. Colonization leads to the formation of biofilms, which cause costly and life-impairing chronic infections. However, whether and how bacteria can sense and respond to the mechanical cue provided by surface stiffness has remained unknown. We find that bacteria do indeed respond to surface stiffness in a way that is both consistent with expectations based on equilibrium continuum mechanics and that quantitatively impacts multiple aspects of early biofilm formation. This is a new understanding for the nascent field of bacterial mechanobiology. Furthermore, this finding suggests the possibility of a new category of approaches to hindering biofilm development by tuning the mechanical properties of biomedical surfaces.

An interpenetrating network design was used to enhance the toughness of PEG-based hydrogel coatings to resist damage during surgical handling.

The high incidence of osteomyelitis associated with critical-sized bone defects raises clinical challenges in fracture healing. Clinical use of antibiotic-loaded bone cement as an adjunct therapy is limited by incompatibility with many antimicrobials, sub-optimal release kinetics, and requirement of surgical removal. Furthermore, overuse of antibiotics can lead to bacterial modifications that increase efflux, decrease binding, or cause inactivation of the antibiotics. Herein, we compared the efficacy of gallium maltolate, a new metal-based antimicrobial, to gentamicin sulfate released from electrospun poly(lactic-co-glycolic) acid (PLGA) wraps in the treatment of osteomyelitis. In vitro evaluation demonstrated sustained release of each antimicrobial up to 14 days. A Kirby Bauer assay indicated that the gentamicin sulfate-loaded wrap inhibited the growth of osteomyelitis-derived isolates, comparable to the gentamicin sulfate powder control. In contrast, the gallium maltolate-loaded wrap did not inhibit bacteria growth. Subsequent microdilution assays indicated a lower than expected sensitivity of the osteomyelitis strain to the gallium maltolate with release concentrations below the threshold for bactericidal activity. A comparison of the selectivity indices indicated that gentamicin sulfate was less toxic and more efficacious than gallium maltolate. A pilot study in a contaminated femoral defect model confirmed that the sustained release of gentamicin sulfate from the electrospun wrap resulted in bacteria density reduction on the surrounding bone, muscle, and hardware below the threshold that impedes healing. Overall, these findings demonstrate the efficacy of a resorbable, antimicrobial wrap that can be used as an adjunct or stand-alone therapy for controlled release of antimicrobials in the treatment of osteomyelitis.

Fabrication of anisotropic materials is highly desirable in designing biomaterials and tissue engineered constructs. Electrospinning has been broadly adopted due to its versatility in producing non-woven fibrous meshes with tunable fiber diameters (from 10 nanometers to 10 microns), microarchitectures, and construct geometries. A myriad of approaches have been utilized to control fiber alignment of electrospun materials to achieve complex microarchitectures, improve mechanical properties, and provide topographical cellular cues. This review provides a comparative analysis of the techniques developed to generate fiber alignment in electrospun materials. A description of the underlying mechanisms that drive fiber alignment, setup variations for each technique, and the resulting impact on the aligned microarchitecture is provided. A critical analysis of the advantages and limitations of each approach is provided to guide researchers in method selection. Finally, future perspectives of advanced electrospinning methodologies are discussed in terms of developing a scalable method with precise control of microarchitecture.

Hiring practices in academia have critically limited the entry of individuals from historically excluded groups (e.g., people who identify as Black/African American, Hispanic/Latino, American Indian/Alaskan Native, LGBTQ+, people with disabilities) into our biomedical engineering faculty. Excluding such individuals has hindered our profession’s research impact and ability to equitably educate our students. With many departments committed to “do better,” we now need to embrace a new mindset and gain the skills necessary to rectify injustices in our faculty hiring processes. Here, we offer a faculty hiring roadmap that departments can leverage to eliminate many barriers of entry to members of historically excluded groups as faculty in biomedical engineering. We call on our colleagues to recognize the failings of our current hiring practices and adopt the guidelines provided here to diversify biomedical engineering departments so that we might collectively accelerate the impact, innovation, and power of our profession.

Hiring practices in academia have critically limited the entry of individuals from historically excluded groups into our biomedical engineering faculty. Excluding diverse individuals has hindered our profession’s research impact and ability to equitably educate our students. With many departments committed to “do better,” we now need to embrace a new mindset and gain the skills necessary to rectify injustices in our faculty hiring processes. Here, we offer a faculty hiring roadmap that departments can leverage to eliminate many barriers of entry to members of historically excluded groups as faculty in biomedical engineering. We call on our colleagues to recognize the failings of our current hiring practices and adopt the guidelines provided here to diversify biomedical engineering departments so that we might collectively accelerate the impact, innovation, and power of our profession.

There is a high prevalence of intra-abdominal adhesions following bowel resection, which can result in chronic pain, bowel obstruction, and morbidity. Although commercial adhesion barriers have been widely utilized for colonic resections, these barriers do not prevent anastomotic leakage resulting from reduced healing of the anastomosis, which can result in long-term health problems. To address this limitation, we have developed an adhesive bilayer wrap with selective bioactivity to simultaneously prevent intra-abdominal adhesion formation and promote anastomotic healing. Reactive electrospinning was used to generate a crosslinked gelatin mesh to serve as a cell-instructive substrate to improve anastomotic healing. A coating of poly(ethylene glycol) (PEG) foam was applied to the bioactive mesh to generate an antifouling layer and prevent intra-abdominal adhesions. After in vitro confirmation of selective bioactivity, the composite wrap was compared after 2 weeks to a commercial product (InterceedⓇ) in an in vivo rat colonic abrasion model for prevention of intra-abdominal adhesions. The composite bilayer wrap was able to prevent intra-abdominal adhesions when clinical placement was maintained. The composite bilayer wrap was further modified to include tissue adhesive properties for improved efficacy. Preliminary studies indicated that the adhesive composite bilayer wrap maintained a maximum shear strength comparable to InterceedⓇ and greater than fibrin glue. Overall, this work resulted in an initial proof-of-concept device that was shown to effectively prevent intra-abdominal adhesion formation in vivo. The composite bilayer wrap studied here could lead to an improved technology for improved healing of intestinal anastomoses.

Gelatin is used in a broad range of tissue engineering applications because of its bioactivity, mild processing conditions, and ease of modification, which have increased interest in its use as a growth factor delivery vehicle. Traditional methods to control growth factor sequestration and delivery have relied on controlling hydrogel mesh size via chemical crosslinking with corollary changes to the physical properties of the hydrogel. To decouple growth factor release from scaffold properties, affinity sequestration modalities have been developed to preserve the bioactivity of the growth factor through interactions with the modified gelatin. This review provides a summary of these mechanisms, highlights current gelatin growth factor delivery systems, and addresses the future perspective of gelatin matrices for growth factor delivery in tissue engineering.

Scaffold-based tissue engineering requires a resorbable scaffold that can restore function and guide regeneration. Recent advances in material fabrication have expanded our control of compositional and architectural features to approach the complexity of native tissue. However, iterative scaffold design to balance multiple design targets toward optimizing regenerative performance remains both challenging and time-consuming. The number of design parameter combinations for scaffold manufacturing to achieve target properties is nearly limitless. Although a trial-and-error experimental approach may lead to a favorable scaffold design, the time and costs associated with such an approach are enormous. Computational optimization approaches are well suited to sample such a multidimensional design space and streamline the identification of target scaffold parameters for experimental evaluation. In this computational biomechanics approach, target fabrication parameters are identified by using a computational model to iterate across design parameter combinations (input) and predict the resulting graft properties (output). Herein, we describe the three key stages of this model-directed scaffold design: (1) model development, verification, and validation; (2) in silico optimization; and (3) model-directed scaffold fabrication and testing. Although there are several notable examples that have demonstrated the potential of this approach, additional accurate and physiologically appropriate computational simulations are needed to expand its utility across the field. In addition, continued advances in material fabrication are needed to provide the requisite control and resolution to produce the model-directed design. Finally, and most importantly, active collaboration between experts in materials science and computational modeling are critical to realize the full potential of this approach to accelerate scaffold development.

Bioprosthetic valves (BPVs) have a limited lifespan in the body necessitating repeated surgeries to replace the failed implant. Early failure of these implants has been linked to various surface properties of the valve. Surface properties of BPVs are significantly different from physiological valves because of the fixation process used when processing the xenograft tissue. To improve the longevity of BPVs, efforts need to be taken to improve the surface properties and shield the implant from the bodily interactions that degrade it. Toward this goal, we evaluated the use of hydrogel coatings to attach to the BPV tissue and impart surface properties that are close to physiological. Hydrogels are well characterized for their biocompatibility and highly tunable surface characteristics. Using a previously published coating method, we deposited hydrogel coatings of poly(ethylene glycol)diacrylate (PEGDA) and poly(ethylene glycol)diacrylamide (PEGDAA) atop BPV samples. Coated samples were evaluated against the physiological tissue and uncoated glutaraldehyde-fixed tissue for deposition of hydrogel, surface adherence, mechanical properties, and fixation properties. Results showed both PEGDA- and PEGDAA-deposited coatings were nearly continuous across the valve leaflet surface. Further, the PEGDA- and PEGDAA-coated samples showed restoration of physiological levels of protein adhesion and mechanical stiffness. Interestingly, the coating process rather than the coating itself altered the material behavior yet did not alter the cross-linking from fixation. These results show that the PEG-based coatings for BPVs can successfully alter surface properties of BPVs and help promote physiological characteristics without interfering with the necessary fixation.

Re-entrant arrhythmias—the leading cause of sudden cardiac death—are caused by diseased myocardial tissue and consequent delayed myocardial conduction. Access to the coronary veins that cross the “culprit” scar regions where re-entry originates can provide improved pacing to these delayed regions, offering a novel opportunity to prevent ventricular arrhythmias and circumvent the need for painful defibrillation, risky cardiac ablation, or toxic and often ineffective antiarrhythmic medications. However, there are no pacing electrodes which are small or focal enough to navigate these tributaries. To address this need, we have developed an injectable conductive hydrogel that can fill the epicardial coronary veins and their mid-myocardial tributaries. When connected to a standard pacing lead, these injected hydrogels can be converted into flexible electrodes that directly pace the previously inaccessible mid-myocardial tissue. In our two-component system, hydrogel precursor solutions can be injected through a dual lumen catheter in a minimally invasive deployment strategy to provide direct access to the diseased regions with precision and ease. Mixing of the two solutions upon injection into the vein activates redox-initiated crosslinking of the gel for rapidin situcure without an external stimulus. Anex vivoporcine model was used to identify the requisite viscosity and cure rate for gel retention and homogeneity. Ionic species added to the hydrogel precursor solutions conferred conductivity above target myocardium values that was retained after implantation. Successfulin vivodeployment demonstrated that the hydrogel electrode filled the anterior interventricular vein with extension into the septal (mid-myocardial) venous tributaries to depths far more distal and refined than any current technologies allow. In addition to successful capture and pacing of the heart, analysis of surface ECG tracings revealed a novel pacing observation highly specific for and suggestive of capture of extensive swaths of septal myocardial tissue. This is the first report of an injectable electrode used to successfully pace the mid-myocardium and mimic physiologic conduction. Furthermore,in vivocardiac electroanatomical mapping studies in an ablation scar model showed uniform capture along the hydrogel in the vessels as well as increased capture area compared to point pacing. Collectively, these findings demonstrate that this injectable hydrogel electrode can be deployed to scarred regions of the heart to provide a reliable pacing modality that most closely resembles native conduction with the potential to eliminate delayed myocardial conduction and associated re-entry.One Sentence SummaryInjectable hydrogel electrodes achieve pacing that mimics physiologic conduction by capturing midmyocardial tissue

Biofilm infections can consist of bacterial aggregates that are an order of magnitude larger than neutrophils, phagocytic immune cells that densely surround aggregates but do not enter them. Because a neutrophil is too small to engulf the entire aggregate, it must be able to detach and engulf a few bacteria at a time if it is to use phagocytosis to clear the infection. Current research techniques do not provide a method for determining how the success of phagocytosis, here defined as the complete engulfment of a piece of foreign material, depends on the mechanical properties of a larger object from which the piece must be removed before being engulfed. This article presents a step toward such a method. By varying polymer concentration or cross-linking density, the elastic moduli of centimeter-sized gels are varied over the range that was previously measured for Pseudomonas aeruginosa biofilms grown from clinical bacterial isolates. Human neutrophils are isolated from blood freshly drawn from healthy adult volunteers, exposed to gel containing embedded beads for 1 h, and removed from the gel. The percentage of collected neutrophils that contain beads that had previously been within the gels is used to measure successful phagocytic engulfment. Both increased polymer concentration in agarose gels and increased cross-linking density in alginate gels are associated with a decreased success of phagocytic engulfment. Upon plotting the percentage of neutrophils showing successful engulfment as a function of the elastic modulus of the gel to which they were applied, it is found that data from both alginate and agarose gels collapse onto the same curve. This suggests that gel mechanics may be impacting the success of phagocytosis and demonstrates that this experiment is a step toward realizing methods for measuring how the mechanics of a large target, or a large structure in which smaller targets are embedded, impact the success of phagocytic engulfment.

Delivery of osteoinductive factors such as bone morphogenetic protein 2 (BMP-2) has emerged as a prominent strategy to improve regeneration in bone grafting procedures. However, it remains challenging to identify a carrier that provides the requisite loading efficiency and release kinetics without compromising the mechanical properties of the bone graft. Previously, we reported on porous, polymerized high internal phase emulsion (polyHIPE) microspheres fabricated using controlled fluidics. Uniquely, this solvent-free method provides advantages over current microsphere fabrication strategies including in-line loading of growth factors to improve loading efficiency. In the current study, we utilized this platform to fabricate protein-loaded microspheres and investigated the effect of particle size (∼400 vs ∼800 μm) and pore size (∼15 vs 30 μm) on release profiles. Although there was no significant effect of these variables on the substantial burst release profile of the microspheres, the incorporation of the protein-loaded microspheres within the injectable polyHIPE resulted in a sustained release of protein from the bulk scaffold over a two-week period with minimal burst release. Bioactivity retention of encapsulated BMP-2 was confirmed first using a genetically-modified osteoblast reporter cell line. A functional assay with human mesenchymal stem cells established that the BMP-2 release from microspheres induced osteogenic differentiation. Finally, microsphere incorporation had minimal effect on the cure and compressive properties of an injectable polyHIPE bone graft. Overall, this work demonstrates that these microsphere-polyHIPE composites have strong potential to enhance bone regeneration through controlled release of BMP-2 and other growth factors. Statement of significance This manuscript describes a method for solvent-free fabrication of porous microspheres from high internal phase emulsions using a controlled fluids setup. The principles of emulsion templating and fluid dynamics provide exceptional control of particle size and pore architecture. In addition to the advantage of solvent-free fabrication, this method provides in-line loading of protein directly into the pores of the microspheres with high loading efficiencies. The incorporation of the protein-loaded microspheres within an injectable polyHIPE scaffold resulted in a sustained release of protein over a two-week period with minimal burst release. Retention of BMP-2 bioactivity and incorporation of microspheres with minimal effect on scaffold compressive properties highlights the potential of these new bone grafts.

Resorbable hydrogels have numerous potential applications in tissue engineering and drug delivery due to their highly tunable properties and soft tissue-like mechanical properties. The incorporation of esters into the backbone of poly(ethylene glycol) hydrogels has been used to develop libraries of hydrogels with tunable degradation rates. However, these synthetic strategies used to increase degradation rate often result in undesired changes in the hydrogel physical properties such as matrix modulus or swelling. In an effort to decouple degradation rate from other hydrogel properties, we inserted thio-β esters into the poly(ethylene glycol)-diacrylate backbone to introduce labile bonds without changing macromer molecular weight. This allowed the number of hydrolytically labile thio-β esters to be controlled through changing the ratios of this modified macromer to the original macromer without affecting network properties. The retention of hydrogel properties at different macromer ratios was confirmed by measuring gel fraction, swelling ratio, and compressive modulus. The tunable degradation profiles were characterized both in vitro and in vivo. Following confirmation of cytocompatibility after exposure to the hydrogel degradation products, the in vivo host response was evaluated in comparison to medical grade silicone. Collectively, this work demonstrates the utility and tunability of these hydrolytically degradable hydrogels for a wide variety of tissue engineering applications.

The scaffold pore size influences many critical physical aspects of tissue engineering, including tissue infiltration, biodegradation rate, and mechanical properties. Manual measurements of pore sizes from scanning electron micrographs using ImageJ/FIJI are commonly used to characterize scaffolds, but these methods are both time-consuming and subject to user bias. Current semi-automated analysis tools are limited by a lack of accessibility or limited sample size in their verification process. The work here describes the development of a new MATLAB algorithm, PoreScript, to address these limitations. The algorithm was verified using three common scaffold fabrication methods (e.g., salt leaching, gas foaming, emulsion templating) with varying pore sizes and shapes to demonstrate the versatility of this new tool. Our results demonstrate that the pore size characterization using PoreScript is comparable to manual pore size measurements. The PoreScript algorithm was further evaluated to determine the effect of user-input and image parameters (relative image magnification, pixel intensity threshold, and pore structure). Overall, this work validates the accuracy of the PoreScript algorithm across several fabrication methods and provides user-guidance for semi-automated image analysis and increased throughput of scaffold characterization.

Our nationwide network of BME women faculty collectively argue that racial funding disparity by the National Institutes of Health (NIH) remains the most insidious barrier to success of Black faculty in our profession. We thus refocus attention on this critical barrier and suggest solutions on how it can be dismantled.

In this study, a composite scaffold consisting of an electrospun polyurethane and poly(ethylene glycol) hydrogel was investigated for aortic valve tissue engineering. This multilayered approach permitted the fabrication of a scaffold that met the desired mechanical requirements while enabling the 3D culture of cells. The scaffold was tuned to mimic the tensile strength, anisotropy, and extensibility of the natural aortic valve through design of the electrospun polyurethane mesh layer. Valve interstitial cells were encapsulated inside the hydrogel portion of the scaffold around the electrospun mesh, creating a composite scaffold approximately 200 μm thick. The stiffness of the electrospun fibers caused the encapsulated cells to exhibit an activated phenotype that resulted in fibrotic remodeling of the scaffold in a heterogeneous manner. Remodeling was further explored by culturing the scaffolds in both a mechanically constrained state and in a bent state. The constrained scaffolds demonstrated strong fibrotic remodeling with cells aligning in the direction of the mechanical constraint. Bent scaffolds demonstrated that applied mechanical forces could influence cell behavior. Cells seeded on the outside curve of the bend exhibited an activated, fibrotic response, while cells seeded on the inside curve of the bend were a quiescent phenotype, demonstrating potential control over the fibrotic behavior of cells. Overall, these results indicate that this polyurethane/hydrogel scaffold mimics the structural and functional heterogeneity of native valves and warrants further investigation to be used as a model for understanding fibrotic valve disease.

While developments in gynecologic health research continue advancing, relatively few groups specifically focus on vaginal tissue research for areas like wound healing, device development, and/or drug toxicity. Currently, there is no standardized animal or tissue model that mimics the full complexity of the human vagina. Certain practical factors such as appropriate size and anatomy, costs, and tissue environment vary across species and moreover fail to emulate all aspects of the human vagina. Thus, investigators are tasked with compromising specific properties of the vaginal environment as it relates to human physiology to suit their particular scientific question. Our review aims to facilitate the appropriate selection of a model aptly addressing a particular study by discussing pertinent vaginal characteristics of conventional animal and tissue models. In this review, we first cover common laboratory animals studied in vaginal research – mouse, rat, rabbit, mini-pig, and sheep – as well as human, with respect to the estrus cycle and related hormones, basic reproductive anatomy, the composition of vaginal layers, developmental epithelial origin, and microflora. In light of these relevant comparative metrics, we discuss potential selection criteria for choosing an appropriate animal vaginal model. Finally, we allude to the exciting prospects of increasing biomimicry for in vitro applications to provide a framework for investigators to model, interpret, and predict human vaginal health.

Structural hydrogel design precisely controls swelling, challenges fundamental equations, and predicts physical properties., Hydrogel physical properties are tuned by altering synthesis conditions such as initial polymer concentration and polymer–cross-linker stoichiometric ratios. Traditionally, differences in hydrogel synthesis schemes, such as end-linked poly(ethylene glycol) diacrylate hydrogels and cross-linked poly(vinyl alcohol) hydrogels, limit structural comparison between hydrogels. In this study, we use generalized synthesis variables for hydrogels that emphasize how changes in formulation affect the resulting network structure. We identify two independent linear correlations between these synthesis variables and swelling behavior. Analysis through recently updated swollen polymer network models suggests that synthesis-swelling correlations can be used to make a priori predictions of the stiffness and solute diffusivity characteristics of synthetic hydrogels. The same experiments and analyses performed on methacrylamide-modified gelatin hydrogels demonstrate that complex biopolymer structures disrupt the linear synthesis-swelling correlations. These studies provide insight into the control of hydrogel physical properties through structural design and can be used to implement and optimize biomedically relevant hydrogels.

The ability to direct cell behavior has been central to the success of numerous therapeutics to regenerate tissue or facilitate device integration. Biomaterial scientists are challenged to understand and modulate the interactions of biomaterials with biological systems in order to achieve effective tissue repair. One key area of research investigates the use of extracellular matrix-derived ligands to target specific integrin interactions and induce cellular responses such as increased cell migration, proliferation, and differentiation of mesenchymal stem cells (MSCs). These integrin-targeting proteins and peptides have been implemented in a variety of different polymeric scaffolds and devices to enhance tissue regeneration and integration. This review will first present an overview of integrin-mediated cellular processes that have been identified in angiogenesis, wound healing, and bone regeneration. We will then highlight research utilizing biomaterials with integrin-targeting motifs as a means to direct these cellular processes to enhance tissue regeneration. In addition to providing improved materials for tissue repair and device integration, these innovative biomaterials provide new tools to probe the complex processes of tissue remodeling in order to enhance the rational design of biomaterial scaffolds and guide tissue regeneration strategies.

Hydrogels have long been established as materials with tunable stiffness and chemistry that enable controlled cellular interactions. When applied as coatings, hydrogels can be used to introduce biofunctionality to medical devices with minimal effect on bulk properties. However, it remains challenging to uniformly apply hydrogel coatings to three dimensional geometries without substantially changing the manufacturing process and potentially affecting device function. Herein, we report a new redox-based crosslinking method for applying conformable hydrogel coatings with tunable thickness and chemistry. This new diffusion-mediated strategy of redox initiation and hydrogel crosslinking enabled coating of a variety of three dimensional substrates without changing the primary fabrication process. Following adsorption of the reducing agent to the construct, hydrogel coating thickness was readily controlled by immersion time with desorption and diffusion of the reducing agent initiating hydrogel crosslinking from the surface. The process was used to generate a range of hydrogel properties by varying the macromer molecular weight and concentration. In addition, we demonstrated that these coatings can be applied sequentially to generate multilayered constructs with distinct features. Finally, incorporation of proteins into the bulk of the hydrogel coating or as a final surface layer permitted the controlled introduction of bioactivity that supported cell attachment. This work provides a versatile method for assembling bioactive coatings with a simple post-fabrication process that is amenable to diverse geometric substrates and chemistries.

The design of tissue engineered scaffolds based on polymerized high internal phase emulsions (polyHIPEs) has emerged as a promising bone grafting strategy. We previously reported the ability to 3D print emulsion inks to better mimic the structure and mechanical properties of native bone while precisely matching defect geometry. In the current study, redox-initiated hydrogel carriers were investigated for in situ delivery of human mesenchymal stem cells (hMSCs) utilizing the biodegradable macromer, poly(ethylene glycol)-dithiothreitol. Hydrogel carrier properties including network formation time, sol-gel fraction, and swelling ratio were modulated to achieve rapid cure without external stimuli and a target cell-release period of 5-7 days. These in situ carriers enabled improved distribution of hMSCs in 3D printed polyHIPE grafts over standard suspension seeding. Additionally, carrier-loaded polyHIPEs supported sustained cell viability and osteogenic differentiation of hMSCs post-release. In summary, these findings demonstrate the potential of this in situ curing hydrogel carrier to enhance the cell distribution and retention of hMSCs in bone grafts. Although initially focused on improving bone regeneration, the ability to encapsulate cells in a hydrogel carrier without relying on external stimuli that can be attenuated in large grafts or tissues is expected to have a wide range of applications in tissue engineering.

Sustained biomaterial thromboresistance has long been a goal and challenge in blood-contacting device design. Endothelialization is one of the most successful strategies to achieve long-term thromboresistance of blood-contacting devices, with the endothelial cell layer providing dynamic hemostatic regulation. It is well established that endothelial cell behavior is influenced by interactions with the underlying extracellular matrix (ECM). Numerous researchers have sought to exploit these interactions to generate improved blood-contacting devices by investigating the expression of hemostatic regulators in endothelial cells on various ECM coatings. The ability to select substrates that promote endothelial cell-mediated thromboresistance is crucial to advancing material design strategies to improve cardiovascular device outcomes. This review provides an overview of endothelial cell regulation of hemostasis, the major components found within the cardiovascular basal lamina, and the interactions of endothelial cells with prominent ECM components of the basement membrane. A summary of ECM-mimetic strategies used in cardiovascular devices is provided with a focus on the effects of key adhesion modalities on endothelial cell regulators of hemostasis.

Small-caliber vascular grafts used in coronary artery bypass procedures typically fail due to the development of intimal hyperplasia or thrombosis. Our laboratory has developed a multilayered vascular graft with an electrospun polyurethane outer layer with improved compliance matching and a hydrogel inner layer that is both thromboresistant and promotes endothelialization. Initial in vivo studies showed that hydrogel particulates were dislodged from the hydrogel layer of the grafts during suturing. To address this problem, we developed and characterized a new hydrogel formulation that resists damage during suturing. Introduction of sacrificial, hydrogen bonds to poly(ethylene glycol)-based hydrogels via co-polymerization with n-vinyl pyrrolidone (NVP) increased the fracture energy as determined by single edge notch testing. This enhanced defect tolerance resulted in a hydrogel layer that was resistant to suture-induced damage with no dislodged particles observed. Importantly, the incorporation of NVP did not affect the thromboresistance, bioactivity, or biostability of the hydrogel layer. In addition to eliminating complications due to hydrogel particle generation in our multilayer graft design, this defect tolerant hydrogel formulation has broad potential use in many cardiovascular and soft tissue applications. Statement of Significance: Small-caliber vascular grafts used in coronary artery bypass procedures typically fail due to development of intimal hyperplasia or thrombosis. Our laboratory has developed a multilayered vascular graft with an electrospun polyurethane outer layer with improved compliance matching and a hydrogel inner layer that is both thromboresistant and promotes endothelialization. However, hydrogel particulates were dislodged from the hydrogel layer during suturing in vivo. This work describes a hydrogel formulation based on poly(ethylene glycol) that is resistant to suture-induced damage. The introduction of sacrificial, hydrogen bonds by co-polymerization with n-vinyl pyrrolidone (NVP) resulted in an increase fracture energy without affecting the thromboresistance, bioactivity, or biostability. This defect-tolerant hydrogel formulation and the methodology to assess hydrogel defect tolerance has broad potential use in cardiovascular and soft tissue applications.

Although xenograft biomaterials have been used for decades in replacement heart valves, they continue to face multiple limitations, including limited durability, mineralization, and restricted design space due to their biological origins. These issues necessitate the need for novel replacement heart valve biomaterials that are durable, non-thrombogenic, and compatible with transcatheter aortic valve replacement devices. In this study, we explored the suitability of an electrospun poly(carbonate urethane) (ES-PCU) mesh coated with a poly(ethylene glycol) diacrylate (PEGDA) hydrogel as a synthetic biomaterial for replacement heart valve leaflets. In this material design, the mesh provides the mechanical support, while the hydrogel provides the required surface hemocompatibility. We conducted a comprehensive study to characterize the structural and mechanical properties of the uncoated mesh as well as the hydrogel-coated mesh (composite biomaterial) over the estimated operational range. We found that the composite biomaterial was functionally robust with reproducible stress–strain behavior within and beyond the functional ranges for replacement heart valves, and was able to withstand the rigors of mechanical evaluation without any observable damage. In addition, the composite biomaterial displayed a wide range of mechanical anisotropic responses, which were governed by fiber orientation of the mesh, which in turn, was controlled with the fabrication process. Finally, we developed a novel constitutive modeling approach to predict the mechanical behavior of the composite biomaterial under in-plane extension and shear deformation modes. This model identified the existence of fiber–fiber mechanical interactions in the mesh that have not previously been reported. Interestingly, there was no evidence of fiber-hydrogel mechanical interactions. This important finding suggests that the hydrogel coating can be optimized for hemocompatibility independent of the structural mechanical responses required by the leaflet. This initial study indicated that the composite biomaterial has mechanical properties well-suited for replacement heart valve applications and that the electrospun mesh microarchitecture and hydrogel biological properties can be optimized independently. It also reveals that the structural mechanisms contributing to the mechanical response are more complicated than what was previously established and paves the pathway for more detailed future studies.

Congenital heart defects (CHDs) affect 1 in 120 newborns in the United States. Surgical repair of structural heart defects often leads to arrhythmia and increased risk of heart failure. Our laboratory previously developed an acellular fibrin patch reinforced with a biodegradable poly(ether ester urethane) urea mesh that resulted in improved heart function when tested in a rat right ventricle wall replacement model compared to fixed pericardium. However, this patch did not drive significant neotissue formation. In this study, we modified the patch materials and prevascularized this patch with human umbilical vein endothelial cells and c-Kit(+) human amniotic fluid stem cells. Rudimentary capillary-like networks formed in the fibrin after culture of cell-encapsulated patches for 3 days in vitro. Prevascularized patches and non-cell loaded patch controls were implanted onto full-thickness heart wall defects created in the right ventricle of athymic nude rats. Two months after surgery, defect repair with prevascularized patches resulted in improved heart function and the patched heart area exhibited greater vascularization and muscularization, less fibrosis, and increased M2 macrophage infiltration compared to acellular patches.

The highly tunable mechanical properties and resilience of polyurethanes make them promising candidates for tissue engineering applications. Biodegradability is conferred by incorporation of hydrolytically or enzymatically cleavable moieties into the polyurethane structure. A common choice for the biodegradable soft segment is a poly(ether ester) triblock copolymer synthesized by ring opening polymerization of the polyester from a polyether macroinitiator. Herein, we describe a new "plug-and-play" approach for triblock synthesis based on urethane block coupling that enables finer control of block lengths and ease of segmental tuning. The inclusion of urethane linkages in the soft segment was also hypothesized to promote hydrogen bonding between the segments with an associated increase in modulus, tensile strength, and ultimate elongation. Hard segment content of the biodegradable polyurethane urea was varied to demonstrate the tunable tensile properties and degradation rate. As expected, increasing hard segment content led to large increases in initial secant modulus and tensile strength. A corollary decrease in ultimate elongation, elastic recovery, and degradation rate was also observed with increasing hard segment content. Finally, cytocompatibility and hydrolytic degradation of electrospun polyurethane meshes were evaluated to establish the potential use of these biodegradable matrixes as tissue engineering scaffolds. All of the polyurethane formulations displayed comparable cytocompatibilty to tissue culture plastic controls and hydrolytic chain scission of the polyester soft segment. Overall, this synthetic approach provides a platform to produce biodegradable polyurethane ureas with enhanced control over segmental chemistry, mechanical properties, and degradation rate.

Although a variety of fabrication methods have been developed to generate electrospun meshes with gradient properties, no platform has yet to achieve fiber alignment in the direction of the gradient that mimics the native tendon-bone interface. In this study, we present a method combining in-line blending and air-gap electrospinning to address this limitation in the field. A custom collector with synced rotation permitted fiber collection with uniform mesh thickness and periodic copper wires were used to induce fiber alignment. Two poly(ester urethane ureas) with different hard segment contents (BPUR 50, BPUR 10) were used to generate compositional gradient meshes with and without fiber alignment. The compositional gradient across the length of the mesh was characterized using a fluorescent dye and the results indicated a continuous transition from the BPUR 50 to the BPUR 10. As expected, the fiber alignment of the gradient meshes induced a corresponding alignment of adherent cells in static culture. Tensile testing of the sectioned meshes confirmed a graded transition in mechanical properties and an increase in anisotropy with fiber alignment. Finite element modeling was utilized to illustrate the gradient mechanical properties across the full length of the mesh and lay the foundation for future computational development work. Overall, these results indicate that this electrospinning method permits the fabrication of macromolecular gradients in the direction of fiber alignment and demonstrate its potential for use in interfacial tissue engineering. Statement of Significance The native tendon-bone interface contains a gradient of properties that ensures stability of the joint. Without this transition, failure can occur due to stress concentration at the bone insertion site. Electrospinning is a method commonly used to produce fibrous grafts with gradient properties; however, no current method allows for gradients in the direction of fiber alignment. This work details a novel electrospinning method to produce gradients in the direction of fiber alignment in order to better mimic transitional zones and improve regeneration of the tendon-bone interface. In addition to the biomechanical gradients demonstrated here, this method may also be used to generate gradients of macromolecular, biochemical, and cellular cues with broad potential utility in tissue engineering.

Electrospinning, a technique used to fabricate fibrous scaffolds, has gained popularity in recent years as a method to produce tissue engineered grafts with architectural similarities to the extracellular matrix. Beyond its versatility in material selection, electrospinning also provides many tools to tune the fiber morphology and scaffold geometry. Recent efforts have focused on extending the capabilities of electrospinning to produce scaffolds that better recapitulate tissue properties and enhance regeneration. This review highlights these advancements by providing an overview of the processing variables and setups used to modulate scaffold architecture, discussing strategies to improve cellular infiltration and guide cell behavior, and providing a summary of electrospinning applications in tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2892-2905, 2017.

Additive manufacturing, broadly known as 3D printing, is a class of manufacturing processes in which a 3D construct is built through sequential layer fabrication. This rapid prototyping methodology is transforming how medical devices are designed, developed, and manufactured by enabling low-volume and on-demand production without dedicated equipment or tooling. In brief, computer-aided design software is used to generate a 3D model that is then exported into a file format that describes the volume or surface mesh in 3D space. Another program, generally known as a “slicer,” is then used to translate the 3D data into individual layers to be printed using a multiaxis positioning system through a variety of processes, including extrusion deposition, solidification, polymerization, sintering, or binding. A subset of additive manufacturing includes living cells during the printing process and has been termed bioprinting. Specifically, bioprinting enables robotic deposition of cells and materials into custom shapes and patterns to replicate anatomical complexity associated with tissues and organs. Bioprinting has been adopted for tissue engineering with the goal of developing highly customized cell-laden scaffolds to enable healthy human tissue to be regrown from a patient’s own stem cells. This chapter provides an overview of common 3D printing modalities and design considerations for bioprinting. Current applications of 3D printing technologies in different facets of the medical field are highlighted to demonstrate the current and future impact of additive manufacturing in healthcare.

Polymer foams are becoming increasingly important in industry, especially biodegradable polymer foams are in demand. Depending on the application, polymer foams need to have characteristic properties, which include connectivity and polydispersity. We show how polymer foams with tailor-made structures can be synthesized from water-in-monomer emulsions which were generated via microfluidics. As monomer we used 1,4-butanediol dimethacrylate (1,4-BDDMA). Firstly, we synthesised monodisperse open- and closed-cell poly(1,4-BDDMA) foams with either spherical or hexagonal pore shapes by varying the locus of initiation. Secondly, we were able to control the pore diameters and obtained polymer foams of both connectivities and pore shapes with pore sizes from ∼70 μm up to ∼120 μm by means of one microfluidic chip. Finally, we synthesized poly(1,4-BDDMA) foams with controllable polydispersity. Here, the mean droplet diameter was the same as that of the monodisperse counterparts in order to be able to compare the properties of the resulting polymer foams.

Purpose of review The current review highlights the complexity of the pediatric and adolescent gynecology subspecialty as well as the recent and exciting opportunities for innovation within the field. Recent findings The opportunities for concept, treatment, instrument, and knowledge-transfer innovation to better serve the specific needs of pediatric gynecology patients include novel approaches to neovagina creation using magnets, improving postoperative vaginal wound healing through newly designed and degradable vaginal stents, and complex Mullerian reconstructive surgical planning using virtual reality immersive experiential training. Summary There is a significant window of opportunity to address the needs of pediatric, adolescent and adult gynecological patients with new innovative concepts and tools.

Although skeletal muscle has a high potential for self-repair, volumetric muscle loss can result in impairment beyond the endogenous regenerative capacity. There is a clinical need to improve on current clinical treatments that fail to fully restore the structure and function of lost muscle. Decellularized extracellular matrix (dECM) scaffolds have been an attractive platform for regenerating skeletal muscle, as dECM contains many biochemical cues that aid in cell adhesion, proliferation, and differentiation. However, there is limited capacity to tune physicochemical properties in current dECM technologies to improve outcome. In this study, we aim to create a novel, high-throughput technique to fabricate dECM scaffolds with tunable physicochemical properties while retaining proregenerative matrix components. We demonstrate a successful decellularization protocol that effectively removes DNA. We also identified key steps for the successful production of electrospun muscle dECM without the use of a carrier polymer; electrospinning allows for rapid scaffold fabrication with high control over material properties, which can be optimized to mimic native muscle. To this end, fiber orientation and degree of crosslinking of these dECM scaffolds were modulated and the corollary effects on fiber swelling, mechanical properties, and degradation kinetics were investigated. Beyond application in skeletal muscle, the versatility of this technology has the potential to serve as a foundation for dECM scaffold fabrication in a variety of tissue engineering applications. IMPACT STATEMENT: This study develops a method to decellularize skeletal muscle and fabricate electrospun scaffolds from the decellularized tissue without the need for a carrier polymer. In addition, the resulting scaffolds have tunable physicochemical properties, including fiber alignment, while retaining important extracellular matrix components for regeneration of skeletal muscle.