RESUMEN
Skin identity is controlled by intrinsic features of the epidermis and dermis and their interactions. Modifying skin identity has clinical potential, such as the conversion of residual limb and stump (nonvolar) skin of amputees to pressure-responsive palmoplantar (volar) skin to enhance prosthesis use and minimize skin breakdown. Greater keratin 9 (KRT9) expression, higher epidermal thickness, keratinocyte cytoplasmic size, collagen length, and elastin are markers of volar skin and likely contribute to volar skin resiliency. Given fibroblasts' capacity to modify keratinocyte differentiation, we hypothesized that volar fibroblasts influence these features. Bioprinted skin constructs confirmed the capacity of volar fibroblasts to induce volar keratinocyte features. A clinical trial of healthy volunteers demonstrated that injecting volar fibroblasts into nonvolar skin increased volar features that lasted up to 5 months, highlighting a potential cellular therapy.
Asunto(s)
Refuerzo Biomédico , Bioimpresión , Dermis , Epidermis , Fibroblastos , Queratinocitos , Adulto , Femenino , Humanos , Masculino , Amputados , Diferenciación Celular , Colágeno/metabolismo , Dermis/citología , Dermis/metabolismo , Elastina/metabolismo , Epidermis/metabolismo , Fibroblastos/citología , Fibroblastos/trasplante , Mano , Queratina-9/metabolismo , Queratinocitos/citología , Queratinocitos/metabolismo , Refuerzo Biomédico/métodosRESUMEN
3D bioprinting has become a valuable tool for studying the biology of solid tumors, including glioblastoma multiforme (GBM). Our analysis of publicly available bulk RNA and single-cell sequencing data has allowed us to define the chemotactic profile of GBM tumors and identify the cell types that secrete particular chemokines in the GBM tumor microenvironment (TME). Our findings indicate that primary GBM tissues express multiple chemokines, whereas spherical monocultures of GBM cells significantly lose this diversity. Subsequently, the comparative analysis of GBM spherical monocultures vs. 3D-bioprinted multicultures of cells showed a restoration of chemokine profile diversity in 3D-bioprinted cultures. Furthermore, single-cell RNA-Seq analysis showed that cells of the perivascular niche (pericytes and endocytes) express multiple chemokines in the GBM TME. Next, we 3D-bioprinted cells from two glioblastoma cell lines, U-251 and DK-MG, alone and as co-cultures with mesenchymal stromal cells (representing cells of the perivascular niche) and assessed the chemokine secretome. The results clearly demonstrated that the interaction of tumors and mesenchymal cells leads to in a significant increase in the repertoire and levels of secreted chemokines under culture in 21% O2 and 1% O2. Our study indicates that cells of the perivascular niche may perform a substantial role in shaping the chemokine microenvironment in GBM tumors.
Asunto(s)
Quimiocinas , Técnicas de Cocultivo , Glioblastoma , Células Madre Mesenquimatosas , Microambiente Tumoral , Humanos , Glioblastoma/patología , Glioblastoma/metabolismo , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/patología , Quimiocinas/metabolismo , Línea Celular Tumoral , Impresión Tridimensional , Bioimpresión , Neoplasias Encefálicas/patología , Neoplasias Encefálicas/metabolismoRESUMEN
Spinal cord injury (SCI) is a catastrophic condition that disrupts neurons within the spinal cord, leading to severe motor and sensory deficits. While current treatments can alleviate pain, they do not promote neural regeneration or functional recovery. Three-dimensional (3D) bioprinting offers promising solutions for SCI repair by enabling the creation of complex neural tissue constructs. This review provides a comprehensive overview of 3D bioprinting techniques, bioinks, and stem cell applications in SCI repair. Additionally, it highlights recent advancements in 3D bioprinted scaffolds, including the integration of conductive materials, the incorporation of bioactive molecules like neurotrophic factors, drugs, and exosomes, and the design of innovative structures such as multi-channel and axial scaffolds. These innovative strategies in 3D bioprinting can offer a comprehensive approach to optimizing the spinal cord microenvironment, advancing SCI repair. This review highlights a comprehensive understanding of the current state of 3D bioprinting in SCI repair, offering insights into future directions in the field of regenerative medicine.
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Bioimpresión , Impresión Tridimensional , Traumatismos de la Médula Espinal , Ingeniería de Tejidos , Andamios del Tejido , Traumatismos de la Médula Espinal/terapia , Humanos , Bioimpresión/métodos , Andamios del Tejido/química , Animales , Ingeniería de Tejidos/métodos , Medicina Regenerativa/métodos , Regeneración NerviosaRESUMEN
The field of bone tissue engineering (BTE) has witnessed a revolutionary breakthrough with the advent of three-dimensional (3D) bioprinting technology, which is considered an ideal choice for constructing scaffolds for bone regeneration. The key to realizing scaffold biofunctions is the selection and design of an appropriate bioink, and existing bioinks have significant limitations. In this study, a composite bioink based on natural polymers (gelatin and alginate) and liver decellularized extracellular matrix (LdECM) was developed and used to fabricate scaffolds for BTE using 3D bioprinting. Through in vitro studies, the concentration of LdECM incorporated into the bioink was optimized to achieve printability and stability and to improve the proliferation and osteogenic differentiation of loaded rat bone mesenchymal stem cells (rBMSCs). Furthermore, in vivo experiments were conducted using a Sprague Dawley rat model of critical-sized calvarial defects. The proposed rBMSC-laden LdECM-gelatin-alginate scaffold, bioprinted layer-by-layer, was implanted in the rat calvarial defect and the development of new bone growth was studied for four weeks. The findings showed that the proposed bioactive scaffolds facilitated angiogenesis and osteogenesis at the defect site. The findings of this study suggest that the developed rBMSC-laden LdECM-gelatin-alginate bioink has great potential for clinical translation and application in solving bone regeneration problems.
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Bioimpresión , Hígado , Células Madre Mesenquimatosas , Osteogénesis , Ratas Sprague-Dawley , Ingeniería de Tejidos , Andamios del Tejido , Animales , Andamios del Tejido/química , Ingeniería de Tejidos/métodos , Bioimpresión/métodos , Ratas , Células Madre Mesenquimatosas/citología , Osteogénesis/fisiología , Hígado/citología , Impresión Tridimensional , Matriz Extracelular Descelularizada/química , Regeneración Ósea/fisiología , Gelatina/química , Diferenciación Celular , Alginatos/química , Proliferación Celular , Matriz Extracelular/química , Huesos/fisiología , TintaRESUMEN
Alginates are salts of alginic acid derived mainly from sea algae of the genus brown algae. They are also synthesized by some bacteria. They belong to negatively charged polysaccharides exhibiting some rheological properties. High plasticity and the ability to modify the structure are the reasons for their application in numerous industries. Moreover, when in contact with the living tissue, they do not trigger an immune response, and for this reason they are the most often tested materials for medical applications. The paper discusses the latest applications, including 3D bioprinting, drug delivery systems, and sorptive properties. Recognizing alginates as biomaterials, it emphasizes the necessity for precise processing and modification to industrialize them for specific uses. This review aims to provide a thorough understanding of the advancements in alginate research, underscoring their potential for innovative applications.
Asunto(s)
Alginatos , Alginatos/química , Humanos , Materiales Biocompatibles/química , Animales , Sistemas de Liberación de Medicamentos , Bioimpresión , Ingeniería de Tejidos , Impresión TridimensionalRESUMEN
Vascular permeability is a key factor in developing therapies for disorders associated with compromised endothelium, such as endothelial dysfunction in coronary arteries and impaired function of the blood-brain barrier. Existing fabrication techniques do not adequately replicate the geometrical variation in vascular networks in the human body, which substantially influences disease progression; moreover, these techniques often involve multi-step fabrication procedures that hinder the high-throughput production necessary for pharmacological testing. This paper presents a bioprinting protocol for creating multiple vascular tissues with desired patterns and sizes directly on standard six-well plates, overcoming existing resolution and productivity challenges in bioprinting technology. A simplified fabrication approach was established to construct six hollow, perfusable channels within a hydrogel, which were subsequently lined with human umbilical vein endothelial cells to form a functional and mature endothelium. The computer-controlled nature of 3D bioprinting ensures high reproducibility and requires fewer manual fabrication steps than traditional methods. This highlights VOP's potential as an efficient high-throughput platform for modeling vascular permeability and advancing drug discovery.
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Bioimpresión , Permeabilidad Capilar , Células Endoteliales de la Vena Umbilical Humana , Humanos , Bioimpresión/métodos , Permeabilidad Capilar/fisiología , Hidrogeles/química , Impresión Tridimensional , Ensayos Analíticos de Alto Rendimiento/métodosRESUMEN
Digital light processing (DLP) 3D bioprinting technology has attracted increasing attention in tissue engineering in recent years. However, it still faces significant technical and operational challenges such as cell carcinogenesis caused by prolonged exposure to ultraviolet light and the presence of heavy metal ions in complex photoinitiator systems. In this study, a novel strategy is designed to introduce carbon quantum dots into visible-light-induced silk fibroin bioink as initiators (CDs/SilMA) applied for DLP 3D bioprinting technology. The incorporation of carbon quantum dots facilitates the formation of precise hydrogel structures at 415 nm visible wavelength, enabling the creation of brain, bronchus, spine, and ear models. Replacing heavy metal photoinitiators with carbon quantum dots imparts fluorescence properties to the bioink and enhances its mechanical properties. Meanwhile, the fibroin protein-based hydrogel exhibits favorable properties, such as drug loading, slow release, degradability, and biocompatibility. This is the first study to propose the application of carbon quantum dots in silk fibroin-based bioink. Moreover, the resulting product demonstrates excellent compatibility with the DLP printing process, making it promising for practical applications in various tissue engineering scenarios with specific requirements.
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Bioimpresión , Carbono , Fibroínas , Hidrogeles , Luz , Impresión Tridimensional , Puntos Cuánticos , Puntos Cuánticos/química , Fibroínas/química , Hidrogeles/química , Carbono/química , Animales , Ingeniería de Tejidos/métodos , Ratones , HumanosRESUMEN
The viscosity of gelatin methacryloyl (GelMA)-based bioinks generates shear stresses throughout the printing process that can affect cell integrity, reduce cell viability, cause morphological changes, and alter cell functionality. This study systematically investigated the impact of the viscosity of GelMA-gelatin bioinks on osteoblast-like cells in 2D and 3D culture conditions. Three bioinks with low, medium, and high viscosity prepared by supplementing a 5% GelMA solution with different concentrations of gelatin were evaluated. Cell responses were studied in a 2D environment after printing and incubation in non-cross-linked bioinks that caused the gelatin and GelMA to dissolve and release cells for attachment to tissue culture plates. The increased viscosity of the bioinks significantly affected cell area and aspect ratio. Cells printed using the bioink with medium viscosity exhibited greater metabolic activity and proliferation rate than those printed using the high viscosity bioink and even the unprinted control cells. Additionally, cells printed using the bioink with high viscosity demonstrated notably elevated expression levels of alkaline phosphatase and bone morphogenetic protein-2 genes. In the 3D condition, the printed cell-laden hydrogels were photo-cross-linked prior to incubation. The medium viscosity bioink supported greater cell proliferation compared to the high viscosity bioink. However, there were no significant differences in the expression of osteogenic markers between the medium and high viscosity bioinks. Therefore, the choice between medium and high viscosity bioinks should be based on the desired outcomes and objectives of the bone tissue engineering application. Furthermore, the bioprinting procedure with the medium viscosity bioink was used as an automated technique for efficiently seeding cells onto 3D printed porous titanium scaffolds for bone tissue engineering purposes.
Asunto(s)
Bioimpresión , Gelatina , Tinta , Metacrilatos , Gelatina/química , Viscosidad , Metacrilatos/química , Bioimpresión/métodos , Impresión Tridimensional , Osteoblastos/citología , Osteoblastos/metabolismo , Osteoblastos/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ingeniería de Tejidos , Línea Celular , Animales , Andamios del Tejido/química , Humanos , Supervivencia Celular/efectos de los fármacos , Huesos/citologíaRESUMEN
The incidence of liver diseases is high worldwide. Many factors can cause liver fibrosis, which in turn can lead to liver cirrhosis and even liver cancer. Due to the shortage of donor organs, immunosuppression, and other factors, only a few patients are able to undergo liver transplantation. Therefore, how to construct a bioartificial liver that can be transplanted has become a global research hotspot. With the rapid development of three-dimensional (3D) bioprinting in the field of tissue engineering and regenerative medicine, researchers have tried to use various 3D bioprinting technologies to construct bioartificial livers in vitro. In terms of the choice of bioinks, liver decellularized extracellular matrix (dECM) has many advantages over other materials for cell-laden hydrogel in 3D bioprinting. This review mainly summarizes the acquisition of liver dECM and its application in liver 3D bioprinting as a bioink with respect to availability, printability, and biocompatibility in many aspects and puts forward the current challenges and prospects.
Asunto(s)
Bioimpresión , Matriz Extracelular Descelularizada , Hígado , Impresión Tridimensional , Ingeniería de Tejidos , Humanos , Bioimpresión/métodos , Hígado/metabolismo , Hígado/citología , Ingeniería de Tejidos/métodos , Animales , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/metabolismo , Andamios del Tejido/química , Hidrogeles/química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Materiales Biocompatibles/químicaRESUMEN
Microtumor models, combining cancer and stromal cells within 3D hydrogels, are vital for testing anticancer therapies. Bioprinting hydrogel scaffolds allows tailored in vitro models. We created a 3D microtumor model using a bioprinter, with varying ratios of ovarian stromal cells and leukemia cells (HL-60). PEGylated fibrinogen and alginate hydrogel were used. Cell dynamics and proliferation were assessed via immunofluorescence staining. Microtumors with different HL-60 ratios (1:1, 1:10, 1:100) were cultured for 5 days. Results showed tumor development modulation by cell ratios and culture time. A significant cell density increase occurred in 1:1 ratio microtumors, indicating rapid cancer cell proliferation. No HL-60 cells were found in 1:100 ratio microtumors by day 5. The 1:10 ratio closely mimicked leukemia invasion in ovarian tissue, showing detectable cancer cells by days 3 and 5 without altering total cell density dynamics significantly. This bioprinted leukemia microtumor model offers better physiological relevance than 2D assays, promising applications in cellular analysis and drug screening.
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Bioimpresión , Proliferación Celular , Hidrogeles , Ovario , Humanos , Femenino , Bioimpresión/métodos , Células HL-60 , Ovario/patología , Ovario/metabolismo , Neoplasias Ováricas/patología , Neoplasias Ováricas/metabolismo , Andamios del Tejido/química , Impresión Tridimensional , AlginatosRESUMEN
Current research practice for optimizing bioink involves exhaustive experimentation with multi-material composition for determining the printability, shape fidelity and biocompatibility. Predicting bioink properties can be beneficial to the research community but is a challenging task due to the non-Newtonian behavior in complex composition. Existing models such as Cross model become inadequate for predicting the viscosity for heterogeneous composition of bioinks. In this paper, we utilize a machine learning framework to accurately predict the viscosity of heterogeneous bioink compositions, aiming to enhance extrusion-based bioprinting techniques. Utilizing Bayesian optimization (BO), our strategy leverages a limited dataset to inform our model. This is a technique especially useful of the typically sparse data in this domain. Moreover, we have also developed a mask technique that can handle complex constraints, informed by domain expertise, to define the feasible parameter space for the components of the bioink and their interactions. Our proposed method is focused on predicting the intrinsic factor (e.g. viscosity) of the bioink precursor which is tied to the extrinsic property (e.g. cell viability) through the mask function. Through the optimization of the hyperparameter, we strike a balance between exploration of new possibilities and exploitation of known data, a balance crucial for refining our acquisition function. This function then guides the selection of subsequent sampling points within the defined viable space and the process continues until convergence is achieved, indicating that the model has sufficiently explored the parameter space and identified the optimal or near-optimal solutions. Employing this AI-guided BO framework, we have developed, tested, and validated a surrogate model for determining the viscosity of heterogeneous bioink compositions. This data-driven approach significantly reduces the experimental workload required to identify bioink compositions conducive to functional tissue growth. It not only streamlines the process of finding the optimal bioink compositions from a vast array of heterogeneous options but also offers a promising avenue for accelerating advancements in tissue engineering by minimizing the need for extensive experimental trials.
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Teorema de Bayes , Bioimpresión , Aprendizaje Automático , Bioimpresión/métodos , Viscosidad , Tinta , Animales , RatonesRESUMEN
Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer, a leading cause of cancer-related deaths globally. Initial lesions of PDAC develop within the exocrine pancreas' functional units, with tumor progression driven by interactions between PDAC and stromal cells. Effective therapies require anatomically and functionally relevantin vitrohuman models of the pancreatic cancer microenvironment. We employed tomographic volumetric bioprinting, a novel biofabrication method, to create human fibroblast-laden constructs mimicking the tubuloacinar structures of the exocrine pancreas. Human pancreatic ductal epithelial (HPDE) cells overexpressing the KRAS oncogene (HPDE-KRAS) were seeded in the multiacinar cavity to replicate pathological tissue. HPDE cell growth and organization within the structure were assessed, demonstrating the formation of a thin epithelium covering the acini inner surfaces. Immunofluorescence assays showed significantly higher alpha smooth muscle actin (α-SMA) vs. F-actin expression in fibroblasts co-cultured with cancerous versus wild-type HPDE cells. Additionally,α-SMA expression increased over time and was higher in fibroblasts closer to HPDE cells. Elevated interleukin (IL)-6 levels were quantified in supernatants from co-cultures of stromal and HPDE-KRAS cells. These findings align with inflamed tumor-associated myofibroblast behavior, serving as relevant biomarkers to monitor early disease progression and target drug efficacy. To our knowledge, this is the first demonstration of a 3D bioprinted model of exocrine pancreas that recapitulates its true 3-dimensional microanatomy and shows tumor triggered inflammation.
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Bioimpresión , Fibroblastos , Páncreas Exocrino , Humanos , Páncreas Exocrino/metabolismo , Fibroblastos/metabolismo , Fibroblastos/citología , Impresión Tridimensional , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/metabolismo , Carcinoma Ductal Pancreático/patología , Carcinoma Ductal Pancreático/metabolismo , Línea Celular Tumoral , Tomografía , Actinas/metabolismo , Interleucina-6/metabolismo , Ingeniería de Tejidos , Técnicas de Cocultivo , Proteínas Proto-Oncogénicas p21(ras)/metabolismo , Proteínas Proto-Oncogénicas p21(ras)/genéticaRESUMEN
This study explores the bioprinting of a smooth muscle cell-only bioink into ionically crosslinked oxidized methacrylated alginate (OMA) microgel baths to create self-supporting vascular tissues. The impact of OMA microgel support bath methacrylation degree and cell-only bioink dispensing parameters on tissue formation, remodeling, structure and strength was investigated. We hypothesized that reducing dispensing tip diameter from 27 G (210µm) to 30 G (159µm) for cell-only bioink dispensing would reduce tissue wall thickness and improve the consistency of tissue dimensions while maintaining cell viability. Printing with 30 G tips resulted in decreased mean wall thickness (318.6µm) without compromising mean cell viability (94.8%). Histological analysis of cell-only smooth muscle tissues cultured for 14 d in OMA support baths exhibited decreased wall thickness using 30 G dispensing tips, which correlated with increased collagen deposition and alignment. In addition, a TUNEL assay indicated a decrease in cell death in tissues printed with thinner (30 G) dispensing tips. Mechanical testing demonstrated that tissues printed with a 30 G dispensing tip exhibit an increase in ultimate tensile strength compared to those printed with a 27 G dispensing tip. Overall, these findings highlight the importance of precise control over bioprinting parameters to generate mechanically robust tissues when using cell-only bioinks dispensed and cultured within hydrogel support baths. The ability to control print dimensions using cell-only bioinks may enable bioprinting of more complex soft tissue geometries to generatein vitrotissue models.
Asunto(s)
Alginatos , Bioimpresión , Vasos Coronarios , Miocitos del Músculo Liso , Ingeniería de Tejidos , Miocitos del Músculo Liso/citología , Vasos Coronarios/fisiología , Vasos Coronarios/citología , Animales , Alginatos/química , Supervivencia Celular , Andamios del Tejido/química , Tinta , Resistencia a la TracciónRESUMEN
In situ additive biomanufacturing of structures may boost regenerative medicine.
Asunto(s)
Bioimpresión , Tratamiento Basado en Trasplante de Células y Tejidos , Medicina Regenerativa , Animales , Humanos , Bioimpresión/métodos , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Medicina Regenerativa/métodos , Andamios del TejidoRESUMEN
BACKGROUND: Tissue engineering seeks to improve, maintain, or replace the biological functions of damaged organs or tissues with biological substitutes such as the development of scaffolds. In the case of bone tissue, they must have excellent mechanical properties like native bone. OBJECTIVE: In this work, three geometric models were designed for scaffolds with different structure lattices and porosity that could be biomechanically suitable and support cell growth for trabecular bone replacement applications in tissue engineering and regenerative medicine to the proximal femur area. METHODS: Geometries were designed using computer-aided design (CAD) software and evaluated using finite element analysis in compression tests. Three loads were considered according to the daily activity: 1177 N for slow walking, 2060 N for fast walking, and 245.25 N for a person in a bipedal position. All these loads for an adult weight of 75 kg. For each of them, three biomaterials were assigned: two polymers (poly-glycolic acid (PGA) and poly-lactic acid (PLA)) and one mineral (hydroxyapatite (HA)). 54 tests were performed: 27 for each of the tests. RESULTS: The results showed Young's modulus (E) between 1 and 4 GPa. CONCLUSION: If the resultant E is in the range of 0.1 to 5 GPa, the biomaterial is considered an appropriate alternative for the trabecular bone which is the main component of the proximal bone. However, for the models applied in this study, the best option is the poly-lactic acid which will allow absorbing the acting loads.
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Diseño Asistido por Computadora , Análisis de Elementos Finitos , Ingeniería de Tejidos , Andamios del Tejido , Andamios del Tejido/química , Humanos , Ingeniería de Tejidos/métodos , Durapatita/química , Módulo de Elasticidad , Bioimpresión/métodos , Poliésteres/química , Porosidad , Simulación por Computador , Materiales Biocompatibles/química , Sustitutos de Huesos/química , Ácido Poliglicólico/química , Impresión Tridimensional , Ensayo de Materiales , HuesosRESUMEN
Digital light processing (DLP) bioprinting, known for its high resolution and speed, enables the precise spatial arrangement of biomaterials and has become integral to advancing tissue engineering and regenerative medicine. Nevertheless, inherent light scattering presents significant challenges to the fidelity of the manufactured structures. Herein, we introduce a photoinhibition strategy based on Rutin nanoparticles (Rnps), attenuating the scattering effect through concurrent photoabsorption and free radical reaction. Compared to the widely utilized biocompatible photoabsorber tartrazine (Tar), Rnps-infused bioink enhanced printing speed (1.9×), interlayer homogeneity (58% less overexposure), resolution (38.3% improvement), and print tolerance (3× high-precision range) to minimize trial-and-error. The biocompatible and antioxidative Rnps significantly improved cytocompatibility and exhibited resistance to oxidative stress-induced damage in printed constructs, as demonstrated with human induced pluripotent stem cell-derived endothelial cells (hiPSC-ECs). The related properties of Rnps facilitate the facile fabrication of multimaterial, heterogeneous, and cell-laden biomimetic constructs with intricate structures. The developed photoinhibitor, with its profound adaptability, promises wide biomedical applications tailored to specific biological requirements.
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Bioimpresión , Luz , Nanopartículas , Rutina , Humanos , Rutina/química , Rutina/farmacología , Nanopartículas/química , Ingeniería de Tejidos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Células Madre Pluripotentes Inducidas/citología , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacosRESUMEN
Sodium alginate (SA) has gained widespread acclaim as a carrier medium for three-dimensional (3D) bioprinting of cells and a diverse array of bioactive substances, attributed to its remarkable biocompatibility and affordability. The conventional approach for fabricating alginate-based tissue engineering constructs entails a post-treatment phase employing a calcium ion solution. However, this method proves ineffectual in addressing the predicament of low precision during the 3D printing procedure and is unable to prevent issues such as non-uniform alginate gelation and substantial distortions. In this study, we introduced borate bioactive glass (BBG) into the SA matrix, capitalizing on the calcium ions released from the degradation of BBG to incite the cross-linking reaction within SA, resulting in the formation of BBG-SA hydrogels. Building upon this fundamental concept, it unveiled that BBG-SA hydrogels greatly enhance the precision of SA in extrusion-based 3D printing and significantly reduce volumetric contraction shrinkage post-printing, while also displaying certain adhesive properties and electrical conductivity. Furthermore, in vitro cellular experiments have unequivocally established the excellent biocompatibility of BBG-SA hydrogel and its capacity to actively stimulate osteogenic differentiation. Consequently, BBG-SA hydrogel emerges as a promising platform for 3D bioprinting, laying the foundation for the development of flexible, biocompatible electronic devices.
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Alginatos , Materiales Biocompatibles , Bioimpresión , Boratos , Calcio , Vidrio , Hidrogeles , Impresión Tridimensional , Alginatos/química , Alginatos/farmacología , Bioimpresión/métodos , Boratos/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Calcio/química , Hidrogeles/química , Vidrio/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Humanos , Diferenciación Celular/efectos de los fármacos , Osteogénesis/efectos de los fármacosRESUMEN
Gallbladder carcinoma (GBC) is a malignant hepatobiliary cancer characterized by an intricate tumor microenvironments (TME) and heterogeneity. The traditional GBC 2D culture models cannot faithfully recapitulate the characteristics of the TME. Three-dimensional (3D) bioprinting enables the establishment of high-throughput and high-fidelity multicellular GBC models. In this study, we designed a concentric cylindrical tetra-culture model to reconstitute the spatial distribution of cells in tumor tissue, with the inner portion containing GBC cells, and the outer ring containing a mixture of endothelial cells, fibroblasts, and macrophages. We confirmed the survival, proliferation, biomarker expression and gene expression profiles of GBC 3D tetra-culture models. Hematoxylin-eosin (HE) and immunofluorescence staining verified the morphology and robust expression of GBC/endothelial/fibroblast/macrophage biomarkers in GBC 3D tetra-culture models. Single-cell RNA sequencing revealed two distinct subtypes of GBC cells within the model, glandular epithelial and squamous epithelial cells, suggesting the mimicry of intratumoral heterogeneity. Comparative transcriptome profile analysis among variousin vitromodels revealed that cellular interactions and the TME in 3D tetra-culture models reshaped the biological processes of tumor cells to a more aggressive phenotype. GBC 3D tetra-culture models restored the characteristics of the TME as well as intratumoral heterogeneity. Therefore, this model is expected to have future applications in tumor biology research and antitumor drug development.
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Bioimpresión , Neoplasias de la Vesícula Biliar , Impresión Tridimensional , Microambiente Tumoral , Humanos , Neoplasias de la Vesícula Biliar/patología , Neoplasias de la Vesícula Biliar/metabolismo , Línea Celular Tumoral , Macrófagos/metabolismo , Macrófagos/patología , Macrófagos/citología , Proliferación CelularRESUMEN
Three-dimensional (3D) bioprinting culture models capable of reproducing the pathological architecture of diseases are increasingly advancing. In this study, 3D scaffolds were created using extrusion-based bioprinting method with alginate, gelatin, and hyaluronic acid to investigate the effects of hyaluronic acid on the physical properties of the bioscaffold as well as on the formation of liver cancer spheroids. Conformational analysis, rheological characterization, and swelling-degradation tests were performed to characterize the scaffolds. After generating spheroids from hepatocellular carcinoma cells on the 3D scaffolds, cell viability and proliferation assays were performed. Flow cytometry and immunofluorescence microscopy were used into examine the expression of albumin, CD44, and E-cadherin to demonstrate functional capability and maturation levels of the spheroid-forming cells. The results show that hyaluronic acid in the scaffolds correlates with spheroid formation and provides high survival rates. It is also associated with an increase in CD44 expression and a decrease in E-cadherin, while there is no significant change in the albumin expression in the cells. Overall, the findings demonstrate that hyaluronic acid in a 3D hydrogel scaffold supports spheroid formation and may induce stemness. We present a promising 3D scaffold model for enhancing liver cancer spheroid formation and mimicking solid tumors. This model also has the potential for further studies to examine stem cell properties in 3D models.
Asunto(s)
Receptores de Hialuranos , Ácido Hialurónico , Células Madre Neoplásicas , Esferoides Celulares , Andamios del Tejido , Ácido Hialurónico/farmacología , Ácido Hialurónico/química , Humanos , Esferoides Celulares/efectos de los fármacos , Esferoides Celulares/metabolismo , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/patología , Andamios del Tejido/química , Receptores de Hialuranos/metabolismo , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/metabolismo , Supervivencia Celular/efectos de los fármacos , Cadherinas/metabolismo , Proliferación Celular/efectos de los fármacos , Bioimpresión/métodos , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/metabolismo , Línea Celular Tumoral , Gelatina/química , Hidrogeles/química , Hidrogeles/farmacologíaRESUMEN
Three-dimensional bioprinting leverages computer-aided design to construct tissues and organs with specialized bioinks. A notable biomaterial for this purpose is chitosan, a natural polysaccharide sourced from crustacean exoskeletons. Chitosan's biocompatibility, biodegradability, non-toxicity, and ability to promote cell adhesion and proliferation make it an excellent component for bioinks. Initially, the rheological properties of chitosan presented challenges for its use in bioprinting. Enhancements in its printability and stability were achieved by integrating it with other natural or synthetic polymers, facilitating its successful application in bioprinting. Chitosan-based bioinks are particularly promising for controlled drug delivery. Incorporating pharmaceuticals directly into the bioink enables the printed structures to serve as localized, sustained-release systems. This approach offers multiple advantages, including precise drug delivery to targeted disease sites, increased therapeutic efficiency, and reduced systemic side effects. Moreover, bioprinting allows for the customization of drug delivery mechanisms to meet individual patient requirements. Although there have been considerable advancements, the use of chitosan-based bioinks in drug delivery is still an emerging field. This review highlights chitosan's essential role in both systemic and localized drug delivery, underscoring its significance and discussing ongoing trends in its application for pharmaceutical purposes.