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The present study sought to demonstrate the swelling behavior of hydrogel-microcarrier composite constructs to inform their use in controlled release and tissue engineering applications. In this study, gelatin methacrylate (GelMA) and GelMA-gelatin microparticle (GMP) composite constructs were three-dimensionally printed, and their swelling and degradation behavior was evaluated over time and as a function of the degree of crosslinking of included GMPs. GelMA-only constructs and composite constructs loaded with GMPs crosslinked with 10 mM (GMP-10) or 40 mM (GMP-40) glutaraldehyde were swollen in phosphate-buffered saline for up to 28 days to evaluate changes in swelling and polymer loss. In addition, scaffold reswelling capacity was evaluated under five successive drying–rehydration cycles. All printed materials demonstrated shear thinning behavior, with microparticle additives significantly increasing viscosity relative to the GelMA-only solution. Swelling results demonstrated that for GelMA/GMP-10 and GelMA/GMP-40 scaffolds, fold and volumetric swelling were statistically higher and lower, respectively, than for GelMA-only scaffolds after 28 days, and the volumetric swelling of GelMA and GelMA/GMP-40 scaffolds decreased over time. After 5 drying–rehydration cycles, GelMA scaffolds demonstrated higher fold swelling than both GMP groups while also showing lower volumetric swelling than GMP groups. Although statistical differences were not observed in the swelling of GMP-10 and GMP-40 particles alone, the interaction of GelMA/GMP demonstrated a significant effect on the swelling behaviors of composite scaffolds. These results demonstrate an example hydrogel-microcarrier composite system's swelling behavior and can inform the future use of such a composite system for controlled delivery of bioactive molecules in vitro and in vivo in tissue engineering applications. IMPACT STATEMENT: In this study, porous three-dimensional printed (3DP) hydrogel constructs with and without natural polymer microcarriers were fabricated to observe swelling and degradation behavior under continuous swelling and drying–rehydration cycle conditions. Inclusion of microcarriers with different crosslinking densities led to distinct swelling behaviors for each biomaterial ink tested. 3DP hydrogel and hydrogel-microcarrier composite scaffolds have been commonly used in tissue engineering for the delivery of biomolecules. This study demonstrates the swelling behavior of porous hydrogel and hydrogel-microcarrier scaffolds that may inform later use of such materials for controlled release applications in a variety of fields including materials development and tissue regeneration.

In this study of three-dimensional (3D) printed composite β-tricalcium phosphate (β-TCP)-/hydroxyapatite/poly(ɛ-caprolactone)-based constructs, the effects of vertical compositional ceramic gradients and architectural porosity gradients on the osteogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells (MSCs) were investigated. Specifically, three different concentrations of β-TCP (0, 10, and 20 wt%) and three different porosities (33% ± 4%, 50% ± 4%, and 65% ± 3%) were examined to elucidate the contributions of chemical and physical gradients on the biochemical behavior of MSCs and the mineralized matrix production within a 3D culture system. By delaminating the constructs at the gradient transition point, the spatial separation of cellular phenotypes could be specifically evaluated for each construct section. Results indicated that increased concentrations of β-TCP resulted in upregulation of osteogenic markers, including alkaline phosphatase activity and mineralized matrix development. Furthermore, MSCs located within regions of higher porosity displayed a more mature osteogenic phenotype compared to MSCs in lower porosity regions. These results demonstrate that 3D printing can be leveraged to create multiphasic gradient constructs to precisely direct the development and function of MSCs, leading to a phenotypic gradient. IMPACT STATEMENT: In this study, three-dimensional (3D) printed ceramic/polymeric constructs containing discrete vertical gradients of both composition and porosity were fabricated to precisely control the osteogenic differentiation of mesenchymal stem cells. By making simple alterations in construct architecture and composition, constructs containing heterogenous populations of cells were generated, where gradients in scaffold design led to corresponding gradients in cellular phenotype. The study demonstrates that 3D printed multiphasic composite constructs can be leveraged to create complex heterogeneous tissues and interfaces.

The investigation of novel hydrogel systems allows for the study of relationships between biomaterials, cells, and other factors within osteochondral tissue engineering. Three-dimensional (3D) printing is a popular research method that can allow for further interrogation of these questions via the fabrication of 3D hydrogel environments that mimic tissue-specific, complex architectures. However, the adaptation of promising hydrogel biomaterial systems into 3D-printable bioinks remains a challenge. Here, we delineated an approach to that process. First, we characterized a novel methacryloylated gelatin composite hydrogel system and assessed how calcium phosphate and glycosaminoglycan additives upregulated bone- and cartilage-like matrix deposition and certain genetic markers of differentiation within human mesenchymal stem cells (hMSCs), such as RUNX2 and SOX9. Then, new assays were developed and utilized to study the effects of xanthan gum and nanofibrillated cellulose, which allowed for cohesive fiber deposition, reliable droplet formation, and non-fracturing digital light processing (DLP)-printed constructs within extrusion, inkjet, and DLP techniques, respectively. Finally, these bioinks were used to 3D print constructs containing viable encapsulated hMSCs over a 7 d period, where DLP printed constructs facilitated the highest observed increase in cell number over 7 d (∼2.4×). The results presented here describe the promotion of osteochondral phenotypes via these novel composite hydrogel formulations, establish their ability to bioprint viable, cell-encapsulating constructs using three different 3D printing methods on multiple bioprinters, and document how a library of modular bioink additives affected those physicochemical properties important to printability.

Significance Metastatic, castration-resistant prostate cancer (mCRPC) is an advanced prostate cancer with limited therapeutic options and poor patient outcomes. To investigate whether multitargeted kinase inhibitors (KIs) represent an opportunity for mCRPC drug development, we applied machine learning–based functional screening and identified two KIs, PP121 and SC-1, which demonstrated strong suppression of CRPC growth in vitro and in vivo. Furthermore, we show the marked ability of these KIs to improve on standard-of-care chemotherapy in both tumor response and survival, suggesting that combining multitargeted KIs with chemotherapy represents a promising avenue for mCRPC treatment. Overall, our findings demonstrate the application of a multidisciplinary strategy that blends bench science with machine-learning approaches for rapidly identifying KIs that result in desired phenotypic effects., Castration-resistant prostate cancer (CRPC) is an advanced subtype of prostate cancer with limited therapeutic options. Here, we applied a systems-based modeling approach called kinome regularization (KiR) to identify multitargeted kinase inhibitors (KIs) that abrogate CRPC growth. Two predicted KIs, PP121 and SC-1, suppressed CRPC growth in two-dimensional in vitro experiments and in vivo subcutaneous xenografts. An ex vivo bone mimetic environment and in vivo tibia xenografts revealed resistance to these KIs in bone. Combining PP121 or SC-1 with docetaxel, standard-of-care chemotherapy for late-stage CRPC, significantly reduced tibia tumor growth in vivo, decreased growth factor signaling, and vastly extended overall survival, compared to either docetaxel monotherapy. These results highlight the utility of computational modeling in forming physiologically relevant predictions and provide evidence for the role of multitargeted KIs as chemosensitizers for late-stage, metastatic CRPC.

In this study, we describe the additive manufacturing of porous three-dimensionally (3D) printed ceramic scaffolds prepared with hydroxyapatite (HA), β-tricalcium phosphate (β-TCP), or the combination of both with an extrusion-based process. The scaffolds were printed using a novel ceramic-based ink with reproducible printability and storability properties. After sintering at 1200°C, the scaffolds were characterized in terms of structure, mechanical properties, and dissolution in aqueous medium. Microcomputed tomography and scanning electron microscopy analyses revealed that the structure of the scaffolds, and more specifically, pore size, porosity, and isotropic dimensions were not significantly affected by the sintering process, resulting in scaffolds that closely replicate the original dimensions of the 3D model design. The mechanical properties of the sintered scaffolds were in the range of human trabecular bone for all compositions. All ceramic bioinks showed consistent printability over a span of 14 days, demonstrating the short-term storability of the formulations. Finally, the mass loss did not vary among the evaluated compositions over a period of 28 days except in the case of β-TCP scaffolds, in which the structural integrity was significantly affected after 28 days of incubation in phosphate-buffered saline. In conclusion, this study demonstrates the development of storable ceramic inks for the 3D printing of scaffolds of HA, β-TCP, and mixtures thereof with high fidelity and low shrinkage following sintering that could potentially be used for bone tissue engineering in load-bearing applications. IMPACT STATEMENT: In this study, we developed a novel ceramic ink for the preparation of porous ceramic scaffolds useful in bone tissue engineering. The ready-to-use ink can be stored for at least 14 days and printed using printing parameters compatible with any commercially available bioprinter. The composition of the ink can be adapted to incorporate the most relevant ceramics for bone tissue engineering, including hydroxyapatite and β-tricalcium phosphate, without affecting the printability or storability properties. The high ceramic content of the ink and the low shrinkage after sintering enable the custom fabrication of patient-specific tissue-engineered scaffolds.

Recent developments in 3D printing (3DP) research have led to a variety of scaffold designs and techniques for osteochondral tissue engineering; however, the simultaneous incorporation of multiple types of gradients within the same construct remains a challenge. Herein, we describe the fabrication and mechanical characterization of porous poly(e-caprolactone) (PCL) and PCL-hydroxyapatite (HA) scaffolds with incorporated vertical porosity and ceramic content gradients via a multimaterial extrusion 3DP system. Scaffolds of 0 wt% HA (PCL), 15 wt% HA (HA15), or 30 wt% HA (HA30) were fabricated with uniform composition and porosity (using 0.2 mm, 0.5 mm, or 0.9 mm on-center fiber spacing), uniform composition and gradient porosity, and gradient composition (PCL-HA15-HA30) and porosity. Micro-CT imaging and porosity analysis demonstrated the ability to incorporate both vertical porosity and pore size gradients and a ceramic gradient, which collectively recapitulate gradients found in native osteochondral tissues. Uniaxial compression testing demonstrated an inverse relationship between porosity, ϕ, and compressive modulus, E, and yield stress, σy, for uniform porosity scaffolds, however, no differences were observed as a result of ceramic incorporation. All scaffolds demonstrated compressive moduli within the appropriate range for trabecular bone, with average moduli between 86 ± 14–220 ± 26 MPa. Uniform porosity and pore size scaffolds for all ceramic levels had compressive moduli between 205 ± 37–220 ± 26 MPa, 112 ± 13–118 ± 23 MPa, and 86 ± 14–97 ± 8 MPa respectively for porosities ranging between 14 ± 4–20 ± 6%, 36 ± 3–43 ± 4%, and 54 ± 2–57 ± 2%, with the moduli and yield stresses of low porosity scaffolds being significantly greater (p 0.05) to those of the highest porosity uniform scaffolds (porosity gradient scaffolds 98 ± 23–107 ± 6 MPa, and 102 ± 7 MPa for dual composition/porosity gradient scaffolds), indicating that these properties are more heavily influenced by the weakest section of the gradient. The compression data for uniform scaffolds were also readily modeled, yielding scaling laws of the form E ∼ (1 − ϕ)1.27 and σy ∼ (1 − ϕ)1.37, which demonstrated that the compressive properties evaluated in this study were well-aligned with expectations from previous literature and were readily modeled with good fidelity independent of polymer scaffold geometry and ceramic content. All uniform scaffolds were similarly deformed and recovered despite different porosities, while the large-pore sections of porosity gradient scaffolds were significantly more deformed than all other groups, indicating that porosity may not be an independent factor in determining strain recovery. Moving forward, the technique described here will serve as the template for more complex multimaterial constructs with bioactive cues that better match native tissue physiology and promote tissue regeneration. Statement of significance This manuscript describes the fabrication and mechanical characterization of “dual” porosity/ceramic content gradient scaffolds produced via a multimaterial extrusion 3D printing system for osteochondral tissue engineering. Such scaffolds are designed to better address the simultaneous gradients in architecture and mineralization found in native osteochondral tissue. The results of this study demonstrate that this technique may serve as a template for future advances in 3D printing technology that may better address the inherent complexity in such heterogeneous tissues.

This report seeks to provide an update of the tissue engineering industry from 2011 to 2018. Public tissue engineering companies with a presence in the United States were the focus of this report due to the publicly accessible financial data that they provide on an annual basis. Over the course of this analysis, 49 tissue engineering companies were identified, 21 of which were in the commercial phase of development and had tissue engineering products on the market. These 21 companies made an estimated $9 billion in sales of tissue engineering-related products in 2017. Based on previous reports and market trends, the field of tissue engineering is forecasted to continue to build revenue for the years to come. IMPACT STATEMENT: This report seeks to provide an update of the current landscape of the tissue engineering market in the United States from an unbiased point of view by analyzing the financial reports provided by tissue engineering companies, as well as data from publicly available clinical trials with relevant tissue engineering applications.

Regenerative therapies for bone and cartilage injuries are currently unable to replicate the complex microenvironment of native tissue. There are many tissue engineering approaches attempting to address this issue through the use of synthetic materials. Although synthetic materials can be modified to simulate the mechanical and biochemical properties of the cell microenvironment, they do not mimic in full the multitude of interactions that take place within tissue. Decellularized extracellular matrix (dECM) has been established as a biomaterial that preserves a tissue's native environment, promotes cell proliferation, and provides cues for cell differentiation. The potential of dECM as a therapeutic agent is rising, but there are many limitations of dECM restricting its use. This review discusses the recent progress in the utilization of bone and cartilage dECM through applications as scaffolds, particles, and supplementary factors in bone and cartilage tissue engineering.

Here, we demonstrate the in vivo efficacy of glucose microparticles (GMPs) to serve as porogens within calcium phosphate cements (CPCs) to obtain a fast-degrading bone substitute material. Composites were fabricated incorporating 20 wt% GMPs at two different GMP size ranges (100-150 μm (GMP-S) and 150-300 μm (GMP-L)), while CPC containing 20 wt% poly(lactic-co-glycolic acid) microparticles (PLGA) and plain CPC served as controls. After 2 and 8 weeks implantation in a rat femoral condyle defect model, specimens were retrieved and analyzed for material degradation and bone formation. Histologically, no adverse tissue response to any of the CPC-formulations was observed. All CPC-porogen formulations showed faster degradation compared to plain CPC control, but only GMP-containing formulations showed higher amounts of new bone formation compared to plain CPC controls. After 8 weeks, only CPC-porogen formulations with GMP-S or PLGA porogens showed higher degradation compared to plain CPC controls. Overall, the inclusion of GMPs into CPCs resulted in a macroporous structure that initially accelerated the generation of new bone. These findings highlight the efficacy of a novel approach that leverages simple porogen properties to generate porous CPCs with distinct degradation and bone regeneration profiles.

This work investigated a new three-dimensional (3D) printing methodology to prepare porous scaffolds containing horizontal pore and composition gradients. To achieve that, a multimaterial printing technology developed in our laboratory was adapted to incorporate pore gradients. Fibers were printed by welding segments with unique material compositions and fiber diameters. Particularly, we focused on the preparation of model composite poly(ɛ-caprolactone)-based scaffolds with radial gradients of particulate hydroxyapatite (HA) content (higher concentrations in the outer region of the scaffold) and porosity (higher in the inner region). The morphology of the scaffolds revealed that the methodology allowed the fabrication of discrete regions with compressive mechanical properties similar to human trabecular bone while maintaining structural integrity. HA distribution was homogeneous within individual regions and no particle aggregation was detected by microCT analysis and Alizarin Red S staining. Finally, the incubation of the scaffolds in simulated body fluid resulted in the deposition of significantly higher amounts of calcium deposits in the regions of the scaffolds with higher HA content. This work provides a new tool for the preparation of porous scaffolds containing porosity and composition gradients for complex tissue engineering applications. IMPACT STATEMENT: In this study, we report the development of a novel multimaterial segmented three-dimensional printing methodology to fabricate porous scaffolds containing discrete horizontal gradients of composition and porosity. This methodology is particularly beneficial to preparing porous scaffolds with intricate structures and graded compositions for the regeneration of complex tissues. The technique presented is compatible with many commercially available bioprinters commonly used in biofabrication, and can be adapted to better replicate the architectural and compositional requirements of individual tissues compared with traditional scaffold printing methods.

In this work, we describe a new 3D printing methodology for the fabrication of multimaterial scaffolds involving the combination of thermoplastic extrusion and low temperature extrusion of bioinks. A fiber engraving technique was used to create a groove on the surface of a thermoplastic printed fiber using a commercial 3D printer and a low viscosity bioink was deposited into this groove. In contrast to traditional extrusion bioinks that rely on increased viscosity to prevent lateral spreading, this groove creates a defined space for bioink deposition. By physically constraining bioink spreading, a broader range of viscosities can be used. As proof-of-concept, we fabricated and characterized a multimaterial scaffold containing poly(e-caprolactone) (PCL) as the thermoplastic polymer and a gelatin-based bioink. A 7.5 w/v% gelatin methacryloyl (GelMA) bioink loaded with either 5 w/v% poly(lactic-co-glycolic acid) (PLGA) microparticles containing fluorescent albumin or mouse fibroblasts (1 ​× ​106 ​cell/mL) was printed at 24 ​°C. The structure of the composite scaffolds had no significant decrease in porosity or mechanical properties as compared to the PCL control scaffolds, demonstrating the engraving technique did not significantly compromise the mechanical or structural integrity of the scaffold. The encapsulated PLGA microparticles were homogeneously distributed in the GelMA and remained in the scaffolds after incubation in PBS for 24 ​h ​at 37 ​°C. In addition, the viability of the fibroblasts encapsulated in the GelMA bioink and printed in the grooves of the PCL scaffolds was confirmed after 24 ​h of incubation. Overall, this work provides a new methodology for the preparation of 3D printed scaffolds containing a robust thermoplastic structure in combination with low viscosity bioinks.

Contains fulltext : 216102.pdf (Publisher’s version ) (Closed access)

In this work, we present a printing method to fabricate scaffolds consisting of multimaterial segmented fibers. Particularly, we developed a reproducible printing process to create single fibers with multiple discrete compositions and control over the distribution of particulate ceramics—namely hydroxyapatite (HA) and β-tricalcium phosphate (TCP)—within poly(ɛ-caprolactone)-based composite scaffolds. Tensile testing revealed that the mechanical integrity of individual segmented fibers was preserved compared with nonsegmented fibers, and microcomputed tomography and thermal analysis confirmed the homogeneous distribution of ceramics incorporated in the fiber compositions. Moreover, we printed and characterized composite scaffolds containing model inverse radial gradients of HA and TCP that could serve as a tunable platform to control the degradation rate of the scaffolds and match bone tissue ingrowth. The morphology of the gradient scaffolds was assessed, and their bulk compressive mechanical properties were found to be in the same range as human trabecular bone. Finally, scaffold degradation was monitored for up to 10 weeks in phosphate-buffered saline pH 7.4 and 0.1 M HCl solution, and scaffolds containing TCP in their composition showed increased degradation compared with those containing HA. This work provides a new methodology for the fabrication and characterization of porous scaffolds containing designer composition gradients that could serve as a platform for the preparation of complex scaffolds for tissue engineering applications. IMPACT STATEMENT: This study introduces a segmented three-dimensional printing methodology to create multimaterial porous scaffolds with discrete gradients and controlled distribution of compositions. This methodology can be adapted for the preparation of complex, multimaterial scaffolds with hierarchical structures and mechanical integrity useful in tissue engineering.

The field of tissue engineering has produced new therapies for the repair of damaged tissues and organs, utilizing biomimetic scaffolds that mirror the mechanical and biological properties of host tissue. The emergence of three-dimensional printing (3DP) technologies has enabled the fabrication of highly complex scaffolds which offer a more accurate replication of native tissue properties and architecture than previously possible. Of strong interest to tissue engineers is the construction of multilayered scaffolds that target distinct regions of complex tissues. Musculoskeletal and dental tissues in particular, such as the osteochondral unit and periodontal complex, are composed of multiple interfacing tissue types, and thus benefit from the usage of multilayered scaffold fabrication. Traditional 3DP technologies such as extrusion printing and selective laser sintering have been used for the construction of scaffolds with gradient architectures and mixed material compositions. Additionally, emerging bioprinting strategies have been used for the direct printing and spatial patterning of cells and chemical factors, capturing the complex organization found in the body. To better replicate the varied and gradated properties of larger tissues, researchers have created scaffolds composed of multiple materials spanning natural polymers, synthetic polymers, and ceramics. By utilizing high precision 3DP techniques and judicious material selection, scaffolds can thus be designed to address the regeneration of previously challenging musculoskeletal, dental, and other heterogeneous target tissues. These multilayered 3DP strategies show great promise in the future of tissue engineering.

This study evaluated the structural, mechanical, and cytocompatibility changes of three-dimensional (3D) printed porous polymer scaffolds during degradation. Three porous scaffold designs were fabricated from a poly(propylene fumarate) (PPF) resin. PPF is a hydrolytically degradable polymer that has been well characterized for applications in bone tissue engineering. Over a 224 day period, scaffolds were hydrolytically degraded and changes in scaffold parameters, such as porosity and pore size, were measured nondestructively using micro-computed tomography. In addition, changes in scaffold mechanical properties were also measured during degradation. Scaffold degradation was verified through decreasing pH and increasing mass loss as well as the formation of micropores and surface channels. Current methods to evaluate polymer cytotoxicity have been well established; however, the ability to evaluate toxicity of an absorbable polymer as it degrades has not been well explored. This study, therefore, also proposes a novel method to evaluate the cytotoxicity of the absorbable scaffolds using a combination of degradation extract, phosphate-buffered saline, and cell culture media. Fibroblasts were incubated with this combination media, and cytotoxicity was evaluated using XTT assay and fluorescence imaging. Cell culture testing demonstrated that the 3D-printed scaffold extracts did not induce significant cell death. In addition, results showed that over a 224 day time period, porous PPF scaffolds provided mechanical stability while degrading. Overall, these results show that degradable, 3D-printed PPF scaffolds are suitable for bone tissue engineering through the use of a novel toxicity during degradation assay.

Contains fulltext : 196228.pdf (Publisher’s version ) (Closed access) This study investigated the effects of incorporating glucose microparticles (GMPs) and poly(lactic-co-glycolic acid) microparticles (PLGA MPs) within a calcium phosphate cement on the cement's handling, physicochemical properties, and the respective pore formation. Composites were fabricated with two different weight fractions of GMPs (10 and 20wt%) and two different weight fractions of PLGA MPs (10 and 20wt%). Samples were assayed for porosity, pore morphology, and compressive mechanical properties. An in vitro degradation study was also conducted. Samples were exposed to a physiological solution for 3days, 4wks, and 8wks in order to understand how the inclusion of GMPs and PLGA MPs affects the composite's porosity and mass loss over time. GMPs and PLGA MPs were both successfully incorporated within the composites and all formulations showed an initial setting time that is appropriate for clinical applications. Through a main effects analysis, we observed that the incorporation of GMPs had a significant effect on the overall porosity, mean pore size, mode pore size, and in vitro degradation rate of PLGA MPs as early as after 3days (p

One of the primary challenges in translating tissue engineering to clinical applicability is adequate, functional vascularization of tissue constructs. Vascularization is necessary for the long-term viability of implanted tissue expanded and differentiated in vitro. Such tissues may be derived from various cell sources, including mesenchymal stem cells (MSCs). MSCs, able to differentiate down several lineages, have been extensively researched for their therapeutic capabilities. In addition, MSCs have a variety of roles in the vascularization of tissue, both through direct contact and indirect signaling. The studied relationships between MSCs and vascularization have been utilized to further the necessary advancement of vascularization in tissue engineering concepts. This review aims to provide a summary of relevant relationships between MSCs, vascularization, and other relevant cell types, along with an overview discussing applications and challenges related to the roles and relationships of MSCs and vascular tissues.

Due to the lack of success in small-diameter (

Adjusting the mechanical properties of polyester-based vascular grafts is crucial to achieving long-term success in vivo. Although previous studies using a fabric-based approach have achieved some success, a central issue with pure poly(lactic acid) (PLA) or poly(glycolic acid) (PGA) grafts sealed with poly(DL-caprolactone-co-lactic acid) (P(CL/LA)) has been stenosis. Intimal hyperplasia, a leading cause of stenosis, can be caused by the mechanical incompatibility of synthetic vascular grafts. Investigating the performance of poly(glycolic-co-lactic acid) (PGLA) grafts could lead to insight into whether graft stenosis stems from mechanical issues such as noncompliance and unfavorable degradation times. This could be achieved by examining grafts with tunable mechanical properties between the ranges of such properties in pure PGA and PLA-based grafts. In this study, we examined PGLA-based grafts sealed with different P(CL/LA) solutions to determine the PGLA-P(CL/LA) grafts' mechanical properties and tissue functionality. Cell attachment and proliferation on graft surfaces were also observed. For in vivo assessment, grafts were implanted in a mouse model. Mechanical properties and degradation times appeared adequate compared to recorded values of vessels used in autograft procedures. Initial neotissue formation was observed in the grafts and patency maintained during the pilot study. This study presents a ∼1-mm diameter degradable graft demonstrating suitable mechanical properties and in vivo pilot study success, enabling further investigation into the tuning of mechanical properties to reduce complications in degradable polyester fabric-based vascular grafts.

A critical challenge to the success of biodegradable vascular grafts is the establishment of a healthy endothelium. To establish this monolayer of endothelial cells (ECs), a variety of techniques have been developed, including cell seeding. Vascular grafts may be seeded with relevant cell types and allowed to mature before implantation. Due to the low proliferative ability of adult ECs and issues with donor site morbidity, there has been increasing interest in using endothelial progenitor cells (EPCs) for vascular healing procedures. In this work, we combined the proliferative and differentiation capabilities of a commercial cell line of early EPCs with an established bioreactor system to support the maturation of cell-seeded vascular grafts. All components of the vascular graft and bioreactor setup are commercially available and allow for complete customization of the scaffold and culturing system. This bioreactor setup enables the control of flow through the graft, imparting fluid shear stress on EPCs and affecting cellular proliferation and differentiation. Grafts cultured with EPCs in the bioreactor system demonstrated greatly increased cell populations and neotissue formation compared with grafts seeded and cultured in a static system. Increased expression of markers for mature endothelial tissues were also observed in bioreactor-cultured EPC-seeded grafts. These findings suggest the distinct advantages of a customizable bioreactor setup for the proliferation and maturation of EPCs. Such a strategy may be beneficial for utilizing EPCs in vascular tissue engineering applications.

Although several strategies to three-dimensionally print cardiovascular grafts exist, these technologies either have required extensive culturing of cells on scaffolds prior to implantation to impart mechanical stability, necessitate additional fabricated components after 3D printing, or have not been proven in vivo. Here, we demonstrate a material and methodology utilizing digital stereolithography for the fabrication of non-cellular biodegradable polymeric vascular grafts. Using this approach, we show, for the first time, the functionality of a 3D printed vascular graft in vivo as an inferior vena cava conduit interposition graft. Our 3D printing materials and methodology should provide a platform for future efforts to fabricate wholly 3D printed, custom-tailored cardiovascular grafts. Such a platform will also enable the precise control over both macroscale—like curvature and bifurcations—and microscale features—like porosity and surface roughness—to improve the performance and integration of these patient-specific vascular scaffolds.

The emerging technique of three-dimensional (3D) printing provides a revolutionary way to fabricate objects with biologically realistic geometries. Previously we have performed optical and morphological characterization of basic 3D printed tissue-simulating phantoms and found them suitable for use in evaluating biophotonic imaging systems. In this study we assess the potential for printing phantoms with irregular, image-defined vascular networks that can be used to provide clinically-relevant insights into device performance. A previously acquired fundus camera image of the human retina was segmented, embedded into a 3D matrix, edited to incorporate the tubular shape of vessels and converted into a digital format suitable for printing. A polymer with biologically realistic optical properties was identified by spectrophotometer measurements of several commercially available samples. Phantoms were printed with the retinal vascular network reproduced as ~1.0 mm diameter channels at a range of depths up to ~3 mm. The morphology of the printed vessels was verified by volumetric imaging with μ-CT. Channels were filled with hemoglobin solutions at controlled oxygenation levels, and the phantoms were imaged by a near-infrared hyperspectral reflectance imaging system. The effect of vessel depth on hemoglobin saturation estimates was studied. Additionally, a phantom incorporating the vascular network at two depths was printed and filled with hemoglobin solution at two different saturation levels. Overall, results indicated that 3D printed phantoms are useful for assessing biophotonic system performance and have the potential to form the basis of clinically-relevant standardized test methods for assessment of medical imaging modalities.

A major challenge in the development of artificial organs is vascularization. In artificial organ fabrication, it may be advantageous to design custom vasculature for adequate vascularization. One such technique is bioprinting. There is a tremendous demand for the fabrication of macroscale vascular grafts and microscale vessels. We introduce motivations for vessel printing, along with an introduction to vessel biology. Then, we discuss challenges associated with vessel bioprinting, followed by an examination of methods. For direct bioprinting, we will examine inkjet/deposition, laser-based, extrusion-based, stereolithography, and syringe-based printing. We will next review hybrid techniques and alternative printing techniques. We will then provide a discussion of postprinting considerations for vessel bioprinting. Finally, we provide a glimpse of how vessel bioprinting fits into the bioprinting arena, along with future trends.

Surface modification of biodegradable vascular grafts is an important strategy to improve the in situ endothelialization of tissue engineered vascular grafts (TEVGs) and prevent major complications associated with current synthetic grafts. Important strategies for improving endothelialization include increasing endothelial cell mobilization and increased endothelial cell capture through biofunctionalization of TEVGs. The objective of this study was to assess two biofunctionalization strategies for improving endothelialization of biodegradable polyester vascular grafts. These techniques consisted of cross-linking heparin to graft surfaces to immobilize vascular endothelial growth factor (VEGF) or antibodies against CD34 (anti-CD34Ab). To this end, heparin, VEGF, and anti-CD34Ab attachment and quantification assays confirmed the efficacy of the modification strategy. Cell attachment and proliferation on these groups were compared to unmodified grafts in vitro and in vivo. To assess in vivo graft functionality, the grafts were implanted as inferior vena cava interpositional conduits in mice. Modified vascular grafts displayed increased endothelial cell attachment and activity in vivo, according to microscopy techniques, histological results, and eNOS expression. Inner lumen diameter of the modified grafts was also better maintained than controls. Overall, while both functionalized grafts outperformed the unmodified control, grafts modified with anti-CD34Ab appeared to yield the most improved results compared to VEGF-loaded grafts.

Adjusting the mechanical properties of polyester-based vascular grafts is crucial to achieving long-term success in vivo. Although previous studies using a fabric-based approach have achieved some success, a central issue with pure poly(lactic acid) (PLA) or poly(glycolic acid) (PGA) grafts sealed with poly(DL-caprolactone-co-lactic acid) (P(CL/LA)) has been stenosis. Intimal hyperplasia, a leading cause of stenosis, can be caused by the mechanical incompatibility of synthetic vascular grafts. Investigating the performance of poly(glycolic-co-lactic acid) (PGLA) grafts could lead to insight into whether graft stenosis stems from mechanical issues such as noncompliance and unfavorable degradation times. This could be achieved by examining grafts with tunable mechanical properties between the ranges of such properties in pure PGA and PLA-based grafts. In this study, we examined PGLA-based grafts sealed with different P(CL/LA) solutions to determine the PGLA-P(CL/LA) grafts' mechanical properties and tissue functionality. Cell attachment and proliferation on graft surfaces were also observed. For in vivo assessment, grafts were implanted in a mouse model. Mechanical properties and degradation times appeared adequate compared to recorded values of vessels used in autograft procedures. Initial neotissue formation was observed in the grafts and patency maintained during the pilot study. This study presents a ∼1-mm diameter degradable graft demonstrating suitable mechanical properties and in vivo pilot study success, enabling further investigation into the tuning of mechanical properties to reduce complications in degradable polyester fabric-based vascular grafts.

The emerging technique of rapid prototyping with three-dimensional (3-D) printers provides a simple yet revolutionary method for fabricating objects with arbitrary geometry. The use of 3-D printing for generating morphologically biomimetic tissue phantoms based on medical images represents a potentially major advance over existing phantom approaches. Toward the goal of image-defined phantoms, we converted a segmented fundus image of the human retina into a matrix format and edited it to achieve a geometry suitable for printing. Phantoms with vessel-simulating channels were then printed using a photoreactive resin providing biologically relevant turbidity, as determined by spectrophotometry. The morphology of printed vessels was validated by x-ray microcomputed tomography. Channels were filled with hemoglobin (Hb) solutions undergoing desaturation, and phantoms were imaged with a near-infrared hyperspectral reflectance imaging system. Additionally, a phantom was printed incorporating two disjoint vascular networks at different depths, each filled with Hb solutions at different saturation levels. Light propagation effects noted during these measurements—including the influence of vessel density and depth on Hb concentration and saturation estimates, and the effect of wavelength on vessel visualization depth—were evaluated. Overall, our findings indicated that 3-D-printed biomimetic phantoms hold significant potential as realistic and practical tools for elucidating light–tissue interactions and characterizing biophotonic system performance.