RESUMO
Controlled release systems are often integrated into polymeric scaffolds to supply essential biofactors to trigger physiological processes in engineered tissues. Here, we report the modification of chondroitin sulfate (CS) electroactive polymer with gold nanorods (AuNRs) to create hybrid macroporous scaffolds for enhanced on-demand release of growth factors and cytokines. The mechanical properties, porosity and degradation of the hybrid scaffolds were evaluated, and the viability and functionality of seeded cardiac cells were assessed. Following, the ability to control the release of the enzyme lysozyme, and the cytokine, stromal cell-derived factor 1 (SDF-1) by applying electrical stimulation, was demonstrated. The AuNRs were able to increase the current through the scaffolds, providing an efficient on-off release profile of SDF-1, which resulted in higher migration of cells expressing CXCR4 receptor. Finally, the engineered scaffolds were transplanted in rats and SDF-1 was released daily by electrical stimulation, promoting blood vessel-forming cell infiltration and vascularization. We envision that gold nanoparticles and other conducting nanomaterials can be incorporated into different electroactive materials to improve their capabilities not only for tissue engineering applications, but for a variety of biomedical applications, where enhanced electrical stimulation is needed.
Assuntos
Nanopartículas Metálicas , Alicerces Teciduais , Animais , Sulfatos de Condroitina , Ouro , Ratos , Engenharia Tecidual/métodosRESUMO
One of the strategies for heart regeneration includes cell delivery to the defected heart. However, most of the injected cells do not form quick cell-cell or cell-matrix interactions, therefore, their ability to engraft at the desired site and improve heart function is poor. Here, the use of a microfluidic system is reported for generating personalized hydrogel-based cellular microdroplets for cardiac cell delivery. To evaluate the system's limitations, a mathematical model of oxygen diffusion and consumption within the droplet is developed. Following, the microfluidic system's parameters are optimized and cardiac cells from neonatal rats or induced pluripotent stem cells are encapsulated. The morphology and cardiac specific markers are assessed and cell function within the droplets is analyzed. Finally, the cellular droplets are injected to mouse gastrocnemius muscle to validate cell retention, survival, and maturation within the host tissue. These results demonstrate the potential of this approach to generate personalized cellular microtissues, which can be injected to distinct regions in the body for treating damaged tissues.
Assuntos
Transplante de Células , Terapia Baseada em Transplante de Células e Tecidos , Coração , Hidrogéis , Miocárdio , Animais , Transplante de Células/métodos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Injeções , Camundongos , Microfluídica , Modelos Biológicos , Miocárdio/citologia , RatosRESUMO
In cardiac tissue engineering cells are seeded within porous biomaterial scaffolds to create functional cardiac patches. Here, we report on a bottom-up approach to assemble a modular tissue consisting of multiple layers with distinct structures and functions. Albumin electrospun fiber scaffolds were laser-patterned to create microgrooves for engineering aligned cardiac tissues exhibiting anisotropic electrical signal propagation. Microchannels were patterned within the scaffolds and seeded with endothelial cells to form closed lumens. Moreover, cage-like structures were patterned within the scaffolds and accommodated poly(lactic-co-glycolic acid) (PLGA) microparticulate systems that controlled the release of VEGF, which promotes vascularization, or dexamethasone, an anti-inflammatory agent. The structure, morphology, and function of each layer were characterized, and the tissue layers were grown separately in their optimal conditions. Before transplantation the tissue and microparticulate layers were integrated by an ECM-based biological glue to form thick 3D cardiac patches. Finally, the patches were transplanted in rats, and their vascularization was assessed. Because of the simple modularity of this approach, we believe that it could be used in the future to assemble other multicellular, thick, 3D, functional tissues.
Assuntos
Materiais Biocompatíveis/química , Doenças Cardiovasculares/cirurgia , Transplante de Coração/métodos , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Adesivos/química , Albuminas/química , Animais , Células Endoteliais , Humanos , Ácido Láctico/química , Masculino , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Porosidade , Ratos , Ratos Sprague-DawleyRESUMO
The development of scaffolding materials that recapitulate the cellular microenvironment and provide cells with physicochemical cues is crucial for successfully engineering functional tissues that can aid in repairing damaged organs. The use of gold nanoparticles for tissue engineering and regenerative medicine has raised great interest in recent years. In this mini review, we describe the shape-dependent properties of gold nanoparticles, and their versatile use in creating tunable nanocomposite scaffolds with improved mechanical and electrical properties for tissue engineering. We further describe using gold nanoparticle-integrated scaffolds to achieve improved stem cells proliferation and differentiation. Finally, we discuss the main challenges and prospects for clinical translation of gold nanoparticles-hybrid scaffolds.
Assuntos
Materiais Biocompatíveis/uso terapêutico , Nanopartículas Metálicas/uso terapêutico , Medicina Regenerativa/tendências , Engenharia Tecidual/tendências , Materiais Biocompatíveis/química , Ouro/química , Humanos , Nanopartículas Metálicas/química , Alicerces TeciduaisRESUMO
The supply of growth factors to engineered tissues is essential for many physiological processes. These processes include the proper organization of the cells into functioning tissues, maintenance of their viability, vasculogenesis, proliferation, and differentiation. Systems to efficiently control the release of growth factors were previously incorporated into tissue engineering scaffolds to affect cells. However, because the initial concentration of the factors in these systems is finite, their ability to provide a long-term physiological effect is limited. Here, we report on a new reloadable system in which 3D fibrous scaffolds conjugated with an anti His-tag antibody enable the retention and controlled release of any His-tag-modified proteinaceous growth factor. The scaffolds can be reloaded in vitro or in vivo with any His-tagged biomolecule at any time according to the physiological need. We show the ability of the scaffolds to release angiogenic factors in a static cell culture or under flow in a microfluidics device and effect on endothelial cells. We also demonstrate the potential of the system to be sequentially reloaded in vivo with various factors, and as a proof of concept, we provide evidence for the efficient in vivo vascularization of scaffolds after reloading with tagged VEGF.
Assuntos
Imunoconjugados/química , Engenharia Tecidual , Alicerces Teciduais/química , Fator A de Crescimento do Endotélio Vascular/química , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Humanos , Imunoconjugados/imunologia , Fator A de Crescimento do Endotélio Vascular/imunologiaRESUMO
Replacement of the damaged scar tissue created by a myocardial infarction is the goal of cardiac tissue engineering. However, once the implanted tissue is in place, monitoring its function is difficult and involves indirect methods, while intervention necessarily requires an invasive procedure and available medical attention. To overcome this, methods of integrating electronic components into engineered tissues have been recently presented. These allow for remote monitoring of tissue function as well as intervention through stimulation and controlled drug release. Here, an improved hybrid microelectronic tissue construct capable of withstanding the dynamic environment of the beating heart without compromising electronic or mechanical functionality is reported. While the reported system is enabled to sense the function of the engineered tissue and provide stimulation for pacing, an electroactive polymer on the electronics enables it to release multiple drugs in parallel. It is envisioned that the integration of microelectronic devices into engineered tissues will provide a better way to monitor patient health from afar, as well as provide facile, more exact methods to control the healing process.
Assuntos
Liberação Controlada de Fármacos , Eletrônica , Coração/fisiologia , Animais , Animais Recém-Nascidos , Materiais Biocompatíveis/química , Preparações de Ação Retardada/farmacologia , Eletricidade , Nanofibras/química , Nanofibras/ultraestrutura , Ratos Sprague-Dawley , Suínos , Alicerces Teciduais/químicaRESUMO
Peptide-based supramolecular hydrogels are utilized as functional materials in tissue engineering, axonal regeneration, and controlled drug delivery. The Arg-Gly-Asp (RGD) ligand based supramolecular gels have immense potential in this respect, as this tripeptide is known to promote cell adhesion. Although several RGD-based supramolecular hydrogels have been reported, most of them are devoid of adequate resilience and long-range stability for in vitro cell culture. In a quest to improve the mechanical properties of these tripeptide-based gels and their durability in cell culture media, the Fmoc-RGD hydrogelator is non-covalently functionalized with a biocompatible and biodegradable polymer, chitosan, resulting in a composite hydrogel with enhanced gelation rate, mechanical properties and cell media durability. Interestingly, both Fmoc-RGD and Fmoc-RGD/chitosan composite hydrogels exhibit thixotropic properties. The utilization of the Fmoc-RGD/chitosan composite hydrogel as a scaffold for 2D and 3D cell cultures is demonstrated. The composite hydrogel is found to have notable antibacterial activity, which stems from the inherent antibacterial properties of chitosan. Furthermore, the composite hydrogels are able to produce ultra-small, mono-dispersed, silver nanoparticles (AgNPs) arranged on the fiber axis. Therefore, the authors' approach harnesses the attributes of both the supramolecular-polymer (Fmoc-RGD) and the covalent-polymer (chitosan) component, resulting in a composite hydrogel with excellent potential.
Assuntos
Arginina/análogos & derivados , Ácido Aspártico/análogos & derivados , Técnicas de Cultura de Células , Quitosana/química , Fluorenos/química , Glicina/análogos & derivados , Hidrogéis/química , Células 3T3 , Animais , Arginina/química , Ácido Aspártico/química , Células CHO , Cricetulus , Glicina/química , Hidrogéis/síntese química , Nanopartículas Metálicas/química , Camundongos , Microscopia Eletrônica de Varredura , Prata/química , Engenharia TecidualRESUMO
Although cardiac patches hold a promise for repairing the infarcted heart, their integration with the myocardium by sutures may cause further damage to the diseased organ. To address this issue, we developed facile and safe, suture-free technology for the attachment of engineered tissues to organs. Here, nanocomposite scaffolds comprised of albumin electrospun fibers and gold nanorods (AuNRs) were developed. Cardiac cells were seeded within the scaffolds and assembled into a functioning patch. The engineered tissue was then positioned on the myocardium and irradiated with a near IR laser (808 nm). The AuNRs were able to absorb the light and convert it to thermal energy, which locally changed the molecular structure of the fibrous scaffold, and strongly, but safely, attached it to the wall of the heart. Such hybrid biomaterials can be used in the future to integrate any engineered tissue with any defected organs, while minimizing the risk of additional injury for the patient, caused by the conventional stitching methods.
Assuntos
Coração/fisiopatologia , Infarto do Miocárdio/cirurgia , Nanocompostos/uso terapêutico , Nanotubos/química , Albuminas/química , Albuminas/uso terapêutico , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/uso terapêutico , Procedimentos Cirúrgicos Cardíacos , Modelos Animais de Doenças , Ouro/química , Ouro/uso terapêutico , Humanos , Infarto do Miocárdio/patologia , Nanocompostos/química , Ratos , Suturas/efeitos adversos , Engenharia Tecidual , Alicerces Teciduais/químicaRESUMO
In microfluidics-based lab-on-a-chip systems, which are used for investigating the effect of drugs and growth factors on cells, the latter are usually cultured within the device's channels in two-dimensional, and not in their optimal three-dimensional (3D) microenvironment. Herein, we address this shortfall by designing a microfluidic system, comprised of two layers. The upper layer of the system consists of multiple channels generating a gradient of soluble factors. The lower layer is comprised of multiple wells, each deposited with 3D, nanofibrous scaffold. We first used a mathematical model to characterize the fluid flow within the system. We then show that induced pluripotent stem cells can be seeded within the 3D scaffolds and be exposed to a well-mixed gradient of soluble factors. We believe that utilizing such system may enable in the future to identify new differentiation factors, investigate drug toxicity, and eventually allow to perform analyses on patient-specific tissues, in order to fit the appropriate combination and concentration of drugs.
Assuntos
Técnicas de Cultura de Células/instrumentação , Células-Tronco Pluripotentes Induzidas/citologia , Dispositivos Lab-On-A-Chip , Modelos Estatísticos , Engenharia Tecidual/métodos , Desenho de Equipamento , Humanos , Hidrogéis/química , Células-Tronco Pluripotentes Induzidas/fisiologia , Nanofibras/ultraestrutura , Omento/citologia , Omento/fisiologia , Cultura Primária de Células , Reologia , Engenharia Tecidual/instrumentação , Alicerces TeciduaisRESUMO
In cardiac tissue engineering approaches to treat myocardial infarction, cardiac cells are seeded within three-dimensional porous scaffolds to create functional cardiac patches. However, current cardiac patches do not allow for online monitoring and reporting of engineered-tissue performance, and do not interfere to deliver signals for patch activation or to enable its integration with the host. Here, we report an engineered cardiac patch that integrates cardiac cells with flexible, freestanding electronics and a 3D nanocomposite scaffold. The patch exhibited robust electronic properties, enabling the recording of cellular electrical activities and the on-demand provision of electrical stimulation for synchronizing cell contraction. We also show that electroactive polymers containing biological factors can be deposited on designated electrodes to release drugs in the patch microenvironment on demand. We expect that the integration of complex electronics within cardiac patches will eventually provide therapeutic control and regulation of cardiac function.
Assuntos
Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Nanocompostos/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/cirurgia , Miocárdio/patologia , Miócitos Cardíacos/patologia , Ratos , Ratos Sprague-DawleyRESUMO
Engineered 3D neuronal networks are considered a promising approach for repairing the damaged spinal cord. However, the lack of a technological platform encouraging axonal elongation over branching may jeopardize the success of such treatment. To address this issue we have decorated gold nanoparticles on the surface of electrospun nanofiber scaffolds, characterized the composite material, and investigated their effect on the differentiation, maturation, and morphogenesis of primary neurons and on an immature neuronal cell line. We have shown that the nanocomposite scaffolds have encouraged a longer outgrowth of the neurites, as judged by the total length of the branching trees and the length and total distance of neurites. Moreover, neurons grown on the nanocomposite scaffolds had less neurites originating out of the soma and lower number of branches. Taken together, these results indicate that neurons cultivated on the gold nanoparticle scaffolds prefer axonal elongation over forming complex branching trees. We envision that such cellular constructs may be useful in the future as implantable cellular devices for repairing damaged neuronal tissues, such as the spinal cord.
RESUMO
The cardiac ECM has a unique 3D structure responsible for tissue morphogenesis and strong contractions. It is divided into three fiber groups with specific roles and distinct dimensions; nanoscale endomysial fibers, perimysial fibers with a diameter of 1 µm, and epimysial fibers, which have a diameter of several micrometers. We report here on our work, where distinct 3D fibrous scaffolds, each of them recapitulating the dimension scales of a single fiber population in the heart matrix, were fabricated. We have assessed the mechanical properties of these scaffolds and the contribution of each fiber population to cardiomyocyte morphogenesis, tissue assembly and function. Our results show that the nanoscale fiber scaffolds were more elastic than the microscale scaffolds, however, cardiomyocytes cultured on microscale fiber scaffolds exhibited enhanced spreading and elongation, both on the single cell and on the engineered tissue levels. In addition, lower fibroblast proliferation rates were observed on these microscale topographies. Based on the collected data we have fabricated composite scaffolds containing micro and nanoscale fibers, promoting superior tissue morphogenesis without compromising tissue contraction. Cardiac tissues, engineered within these composite scaffolds exhibited superior function, including lower excitation threshold and stronger contraction forces than tissue engineered within the single-population fiber scaffolds.
Assuntos
Engenharia Tecidual , Alicerces Teciduais , Animais , Técnicas de Cultura de Células , Proliferação de Células , Células Cultivadas , Módulo de Elasticidade , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Fibras Musculares Esqueléticas/química , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Decellularized matrices are valuable scaffolds for engineering functional cardiac patches for treating myocardial infarction. However, the lack of quick and efficient electrical coupling between adjacent cells may jeopardize the success of the treatment. To address this issue, we have deposited gold nanoparticles on fibrous decellularized omental matrices and investigated their morphology, conductivity, and degradation. We have shown that cardiac cells engineered within the hybrid scaffolds exhibited elongated and aligned morphology, massive striation, and organized connexin 43 electrical coupling proteins. Finally, we have shown that the hybrid patches demonstrated superior function as compared to pristine patches, including a stronger contraction force, lower excitation threshold, and faster calcium transients.
Assuntos
Ouro/química , Nanopartículas Metálicas/química , Miocárdio/citologia , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Células Cultivadas , Conexina 43/análise , Nanopartículas Metálicas/ultraestrutura , RatosRESUMO
In recent years attempts to engineer contracting cardiac patches were focused on recapitulation of the myocardium extracellular microenvironment. We report here on our work, where for the first time, a three-dimensional cardiac patch was fabricated from albumin fibers. We hypothesized that since albumin fibers' mechanical properties resemble those of cardiac tissue extracellular matrix (ECM) and their biochemical character enables their use as protein carriers, they can support the assembly of cardiac tissues capable of generating strong contraction forces. Here, we have fabricated aligned and randomly oriented electrospun albumin fibers and investigated their structure, mechanical properties, and chemical nature. Our measurements showed that the scaffolds have improved elasticity as compared to synthetic electrospun PCL fibers, and that they are capable of adsorbing serum proteins, such as laminin leading to strong cell-matrix interactions. Moreover, due to the functional groups on their backbone, the fibers can be chemically modified with essential biomolecules. When seeded with rat neonatal cardiac cells the engineered scaffolds induced the assembly of aligned cardiac tissues with high aspect ratio cardiomyocytes and massive actinin striation. Compared to synthetic fibrous scaffolds, cardiac cells cultured within aligned or randomly oriented scaffolds formed functional tissues, exhibiting significantly improved function already on Day 3, including higher beating rate (P = 0.0002 and P < 0.0001, respectively), and higher contraction amplitude (P = 0.009 and P = 0.003, respectively). Collectively, our results suggest that albumin electrospun scaffolds can play a key role in contributing to the ex vivo formation of a contracting cardiac muscle tissue.
Assuntos
Substâncias Macromoleculares/metabolismo , Células Musculares/fisiologia , Multimerização Proteica , Soroalbumina Bovina/metabolismo , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Fenômenos Químicos , Substâncias Macromoleculares/química , Ratos , Soroalbumina Bovina/químicaRESUMO
Overcoming the oxygen diffusion limit of approximately 200 µm remains one of the most significant and intractable challenges to be overcome in tissue engineering. The fabrication of hydrogel microtissues and their assembly into larger structures may provide a solution, though these constructs are not without their own drawbacks; namely, these hydrogels are rapidly degraded in vivo, and cells delivered via microtissues are quickly expelled from the area of action. Here, we report the development of an easily customized protocol for creating a protective, biocompatible hydrogel barrier around microtissues. We show that calcium carbonate nanoparticles embedded within an ECM-based microtissue diffuse outwards and, when then exposed to a solution of alginate, can be used to generate a coated layer around the tissue. We further show that this technique can be fine-tuned by adjusting numerous parameters, granting us full control over the thickness of the hydrogel coating layer. The microtissues' protective hydrogel functioned as hypothesized in both in vitro and in vivo testing by preventing the cells inside the tissue from escaping and protecting the microdroplets against external degradation. This technology may provide microtissues with customized properties for use as sources of regenerative therapies.
RESUMO
Tissue engineering is considered a promising approach to treating advanced degenerative maculopathies such as nonexudative age-related macular degeneration (AMD), the leading cause of blindness worldwide. The retina consists of several hierarchical tissue layers, each of which is supported by a layer underneath. Each of these layers has a different morphology and requires distinct conditions for proper assembly. In fact, a prerequisite step for the assembly of each of these layers is the organization of the layer underneath. Advanced retinal degeneration includes degeneration of the other retina layers, including the choroid, the retinal pigmented epithelium (RPE), and the photoreceptors. Here, we report a step-by-step fabrication process of a three-layer retina-like structure. The process included the 3D printing of a choroid-like structure in an extracellular matrix (ECM) hydrogel, followed by deposition of the RPE monolayer. After the formation of the blood vessel-RPE interface, the photoreceptor cells were deposited to interact with the RPE layer. At the end of the fabrication process, each layer was characterized for its morphology and expression of specific markers, and the integration of the three-layer retina was evaluated. We envision that such a retina-like structure may be able to attenuate the deterioration of a degenerated retina and improve engraftment and regeneration. This retinal implant may potentially be suitable for a spectrum of macular degenerative diseases for which there are currently no cures and may save millions from complete blindness.
RESUMO
Cardiac monitoring after heart surgeries is crucial for health maintenance and detecting postoperative complications early. However, current methods like rigid implants have limitations, as they require performing second complex surgeries for removal, increasing infection and inflammation risks, thus prompting research for improved sensing monitoring technologies. Herein, we introduce a nanosensor platform that is biodegradable, biocompatible, and integrated with multifunctions, suitable for use as implants for cardiac monitoring. The device has two electrochemical biosensors for sensing lactic acid and pH as well as a pressure sensor and a chemiresistor array for detecting volatile organic compounds. Its biocompatibility with myocytes has been tested in vitro, and its biodegradability and sensing function have been proven with ex vivo experiments using a three-dimensional (3D)-printed heart model and 3D-printed cardiac tissue patches. Moreover, an artificial intelligence-based predictive model was designed to fuse sensor data for more precise health assessment, making it a suitable candidate for clinical use. This sensing platform promises impactful applications in the realm of cardiac patient care, laying the foundation for advanced life-saving developments.
Assuntos
Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Humanos , Inteligência Artificial , Próteses e Implantes , Monitorização FisiológicaRESUMO
The development of three-dimensional (3D) synthetic biomaterials as structural and bioactive scaffolds is central to fields ranging from cellular biophysics to regenerative medicine. As of yet, these scaffolds cannot electrically probe the physicochemical and biological microenvironments throughout their 3D and macroporous interior, although this capability could have a marked impact in both electronics and biomaterials. Here, we address this challenge using macroporous, flexible and free-standing nanowire nanoelectronic scaffolds (nanoES), and their hybrids with synthetic or natural biomaterials. 3D macroporous nanoES mimic the structure of natural tissue scaffolds, and they were formed by self-organization of coplanar reticular networks with built-in strain and by manipulation of 2D mesh matrices. NanoES exhibited robust electronic properties and have been used alone or combined with other biomaterials as biocompatible extracellular scaffolds for 3D culture of neurons, cardiomyocytes and smooth muscle cells. Furthermore, we show the integrated sensory capability of the nanoES by real-time monitoring of the local electrical activity within 3D nanoES/cardiomyocyte constructs, the response of 3D-nanoES-based neural and cardiac tissue models to drugs, and distinct pH changes inside and outside tubular vascular smooth muscle constructs.
Assuntos
Materiais Biocompatíveis/química , Nanotecnologia/instrumentação , Nanofios/química , Alicerces Teciduais/química , Transistores Eletrônicos , Humanos , Músculo Liso Vascular/citologia , Porosidade , Silício/química , Engenharia TecidualRESUMO
Despite advances in biomaterials engineering, a large gap remains between the weak mechanical properties that can be achieved with natural materials and the strength of synthetic materials. Here, a method is presented for reinforcing an engineered cardiac tissue fabricated from differentiated induced pluripotent stem cells (iPSCs) and an extracellular matrix (ECM)-based hydrogel in a manner that is fully biocompatible. The reinforcement occurs as a post-fabrication step, which allows for the use of 3D-printing technology to generate thick, fully cellularized, and vascularized cardiac tissues. After tissue assembly and during the maturation process in a soft hydrogel, a small, tissue-penetrating reinforcer is deployed, leading to a significant increase in the tissue's mechanical properties. The tissue's robustness is demonstrated by injecting the tissue in a simulated minimally invasive procedure and showing that the tissue is functional and undamaged at the nano-, micro-, and macroscales.