3D Bioprinting Technology Research Articles

Unveiling the impact of hypodermis on gene expression for advancing bioprinted full-thickness 3D skin models.

3D skin models have been explored as an alternative method to the use of animals in research and development. Usually, human skin equivalents comprise only epidermis or epidermis/dermis layers. Herein, we leverage 3D bioprinting technology to fabricate a full-thickness human skin equivalent with hypodermis (HSEH). The collagen hydrogel-based structure provides a mimetic environment for skin cells to adhere, proliferate and differentiate. The effective incorporation of the hypodermis layer is evidenced by scanning electron microscopy, immunofluorescence, and hematoxylin and eosin staining. The transcriptome results underscore the pivotal role of the hypodermis in orchestrating the genetic expression of a multitude of genes vital for skin functionality, including hydration, development and differentiation. Accordingly, we evidence the paramount significance of full-thickness human skin equivalents with hypodermis layer to provide an accurate in vitro platform for disease modeling and toxicology studies.

3D bioprinted in vitro epilepsy models for pharmacological evaluation in temporal lobe epilepsy

This study introduces a novelin vitromodel for intractable temporal lobe epilepsy (TLE) utilizing 3D bioprinting technology, aiming to replicate the complex neurobiological characteristics of TLE more accurately. Primary neural cell constructs were fabricated and subjected to epileptiform-inducing conditions, fostering synaptic proliferation and neuronal loss. Systematically electrophysiological and immunofluorescent analyses indicated that significant synaptic connectivity and sustained epileptiform activities within the constructs akin to those observed in human epilepsy models. Notably, the model responded to treatments with phenytoin and tetrodotoxin, illustrating its potential utility in drug response kinetics studies. Furthermore, we performed drug permeability simulations using COMSOL Multiphysics to analyze the diffusion characteristics of these drugs within the constructs. These results confirm that our 3D bioprinted neural model provides a physiologically relevant and ethically sustainable platform, which is beneficial for studying TLE mechanisms and developing therapeutic strategies with high accuracy and clinical relevance.

Application of 3D printing in bacterial detection

Diseases caused by bacterial infections, especially pathogen variations and drug-resistant bacteria, are a serious threat to human health globally. Accurate and rapid identification of bacterial pathogens is essential for early diagnosis of infections and timely treatment of disease. At present, the routine diagnostic methods for identifying bacterial pathogens in clinic are bacterial culture, immunological detection, genetic analysis, etc. These methods occupy foundational position for bacterial detection that favors for clinical diagnosis of pathogen infection in daily life. Notably, 3D printing technology, together with 3D bioprinting technology, provides more options for bacterial detection due to its personalized design and manufacturing. Compared with traditional methods, 3D printing provides a more convenient, rapid and accurate bacterial detection platform, thus meeting the urgent needs of clinical and scientific research. Here, we have summarized the research progress and given a comprehensive review of 3D printing technology in bacterial detection, including bacterial detection sensors, microfluidic chips, bioprinting microarray technology, AI and spectroscopy technology, providing a scientific reference and filling in gaps in 3D printing-assistance bacterial detection. At the same time, technical advantages, challenges and future development trends will also be discussed in related healthcare fields.

3D Bioprinting in Cancer Modeling and Biomedicine: From Print Categories to Biological Applications.

The continuous interaction between tumor cells and the local microenvironment plays a decisive role in tumor development. Selecting effective models to simulate the tumor microenvironment to study the physiological processes of tumorigenesis and progression is extremely important and challenging. Currently, three-dimensional (3D) bioprinting technology makes it possible to replicate a physiologically relevant tumor microenvironment and induce genomic and proteomic expression to better mimic tumors in vivo. Meanwhile, it plays a crucial role in the prevention and treatment of human diseases, contributing to drug delivery and drug screening, tissue development and regenerative medicine. This paper provides an overview of the categories of 3D bioprinting technology, and the recent advances in the bioinks required for printing. In addition, we summarize the current tumor models based on 3D bioprinting and provide an assessment of possible future biological applications.

Heterotopically differentiated PDLSCs-laden 3D bioprinting scaffolds for concurrent oral hard and soft tissue regeneration

Following dental extraction, the alveolar bone and gingival tissues could undergo varying degrees of resorption, affecting subsequent implant integration and aesthetic outcomes. Adequate volume of both hard and soft tissues is essential for optimal results. Three-dimensional (3D) bioprinting technology offers the advantages of biomimicry, personalization, and precise spatial distribution, which are pivotal for enhancing the success and esthetics of dental restorations. In this study, we fabricated a construct with a natural transition and varying material concentrations by 3D bioprinting, comprising an upper layer of Collagen/Alginate/periodontal ligament stem cells (PDLSCs) and a lower layer of Collagen/nano-hydroxyapatite (nHA)/Alginate/PDLSCs. Characterization of the physicochemical properties revealed that the incorporation of nHA significantly enhanced the mechanical properties of both the bioink and the construct. Flow cytometry analysis confirmed the stemness of PDLSCs. Scanning electron microscopy (SEM) revealed that the construct possesses satisfactory pore density and a natural transition at the stratification point. The construct displayed good cell viability and proliferation, with the cellular movement observed at the stratification interface after bioprinting. Differentiation staining and quantitative reverse transcription-polymerase chain reaction (RT-qPCR) results demonstrated that PDLSCs within the 3D construct are capable of both osteogenic and fibroblastic differentiations. Ectopic transplantation in mice confirmed the biocompatibility of the construct. A rat tooth extraction model validated the construct's effectiveness in the integrated regeneration of both hard and soft tissues in alveolar ridge preservation (ARP). In conclusion, this personalized, concentration-varied 3D construct exhibits excellent biocompatibility and tissue preservation effects, holding significant potential for clinical application.  

3D Nanofiber-Assisted Embedded Extrusion Bioprinting for Oriented Cardiac Tissue Fabrication.

Three-dimensional (3D) bioprinting technology stands out as a promising tissue manufacturing process to control the geometry precisely with cell-loaded bioinks. However, the isotropic culture environment within the bioink and the lack of topographical cues impede the formation of oriented cardiac tissue. To overcome this limitation, we present a novel method named 3D nanofiber-assisted embedded bioprinting (3D-NFEP) to fabricate cardiac tissue with an oriented morphology. Aligned 3D nanofiber scaffolds were fabricated by divergence electrospinning, which provided structural support for printing of the low-viscosity bioink and structural induction to cardiomyocytes. Cells adhered to the aligned fibers after hydrogel degradation, and a high degree of cell alignment was observed. This technology was also demonstrated as a feasible solution for multilayer cell printing. Therefore, 3D-NFEP was demonstrated as a promising method for bioprinting oriented cardiac tissue with low-viscosity bioink and is expected to be applied for structured and cardiac tissue engineering.

3D-BIOPRINTING OF BONE TISSUE IN VITRO И IN SITU

Advanced techniques in bone tissue regeneration include innovative achievements in the field of tissue engineering, among which 3D bioprinting technology holds a special place. This method demonstrates significant progress, driven by its numerous advantages, confirmed through extensive scientific research. 3D bioprinting offers the capability of precise printing of bone transplants with specified geometry, incorporating necessary biomaterials such as cells and growth factors, which facilitates effective osteoinduction, osteoconduction, and vascularization. This review article analyzes various modern methods of bone tissue restoration, including the use of autotransplants, allotransplants, and synthetic bone transplants. Particular attention is paid to the latest innovations in 3D bioprinting, opening prospects for creating individualized bone tissues characterized by a high degree of biocompatibility and minimizing the need for invasive procedures. Additionally, the article presents the concept of in situ bioprinting, developed by Weiss and his colleagues in 2007, as a method of direct printing of bio-inks directly in the area of damage in a living organism. This progressive approach provides high precision in the restoration of tissue defects, regardless of their complexity and geometric features. Over the last decade, several successful clinical cases of using printed structures have been registered, demonstrating the significance of 3D bioprinting in treating life-threatening diseases, as well as in accelerating the regeneration process and creating tissue transplants for subsequent implantation.

3D Bioprinted Chondrogenic Gelatin Methacrylate-Poly(ethylene glycol) Diacrylate Composite Scaffolds for Intervertebral Disc Restoration

Abstract Degenerative spine pathologies, including intervertebral disc (IVD) degeneration, present a significant healthcare challenge due to their association with chronic pain and disability. This study explores an innovative approach to IVD regeneration utilizing 3D bioprinting technology, specifically visible light-based digital light processing (VL-DLP), to fabricate tissue scaffolds that closely mimic the native architecture of the IVD. Utilizing a hybrid bioink composed of gelatin methacrylate (GelMA) and poly(ethylene glycol) diacrylate (PEGDA) at a 10% concentration, we achieved enhanced printing fidelity and mechanical properties suitable for load-bearing applications such as the IVD. Preconditioning rat bone marrow-derived mesenchymal stem cell (rBMSC) spheroids with chondrogenic media before incorporating them into the GelMA-PEGDA scaffold further promoted the regenerative capabilities of this system. Our findings demonstrate that this bioprinted scaffold not only supports cell viability and integration but also contributes to the restoration of disc height in a rat caudal disc model without inducing adverse inflammatory responses. The study underscores the potential of combining advanced bioprinting techniques and cell preconditioning strategies to develop effective treatments for IVD degeneration and other musculoskeletal disorders, highlighting the need for further research into the dynamic interplay between cellular migration and the hydrogel matrix.

3D bioprinting for drug development and screening: Recent trends towards personalized medicine

Personalized medicine, leveraging patient-specific genomic data, is revolutionizing treatment paradigms by enabling tailored therapies to individual needs, thus mitigating adverse effects and enhancing clinical outcomes. One innovative application in this field is the use of 3D bioprinting technology to design, develop and screen patient-customized medicines. This technology meticulously designs and develops therapeutic drugs by depositing active pharmaceutical ingredients layer by layer, culminating in a drug delivery structure optimized for the patient's unique dosage requirements. 3D bioprinting represents an emerging confluence of biological sciences and additive manufacturing, meeting the clinical demand for functional biological tissues through the integration of printing technologies, regenerative medicine principles, and advanced material science. This review provides an overview of the current state of 3D bioprinting applications and explores the transformative potential of 3D bioprinting in personalizing medicine. Moreover, the review elucidates the capabilities of 3D bioprinting in drug design, facilitating the production of complex therapeutic regimens in specific dosage forms, such as drugs with tailored dose concentrations, controlled release kinetics, and the capacity to combine multiple therapeutic regimens into a single, easy-to-administer format. This paper also addresses the challenges and obstacles faced in integrating 3D bioprinting techniques into pharmaceutical practice, ranging from technical and regulatory hurdles to scalability. Further, the efficacy of 3D bioprinting as a tool for advancing personalized medicine helps to utilize the full potential of this technology to enhance patient healthcare regimes.

Effect of Hydroxyapatite Nanowires on Formation and Bioactivity of Osteoblastic Cell Spheroid.

Compared with traditional high-density cell spheroids, which are more prone to core necrosis, nanowires effectively improve the biological activity of core cells in spheroids, emanating more innovations for optimizing the internal cell survival environment and providing differentiation signals. In this study, hydroxyapatite nanowires (HAW), which provide numerous material exchange channels for internal cells by interpenetrating into cell spheroids, were added to osteoblast precursor (MC3T3-E1) cell spheroids. HAW, synthesized using the hydrothermal method, was used as a regulatory material to prepare uniformly sized 3D composite spheroids with good biological activity. Subsequently, material characterization and biocompatibility tests were performed on HAW, and the biological activity and osteogenic differentiation ability of the cell spheroids were tested. Notably, in 2D coculture, HAW displayed a certain attraction to MC3T3-E1 cells and promoted cell aggregation toward it. The content of HAW determined whether composite cell spheroids can form aggregated spherical structures, and incorporation of HAW alleviated core necrosis and enhanced the osteogenic phenotype. In summary, these findings indicate that the prepared HAW-bone cell composite spheroids can potentially be used as building blocks for the construction of large high-density biomimetic tissues and organoids using 3D bioprinting technology.

Stem cell therapy for osteoarthritis: Functional cartilage regeneration using 3d bioprinting technology

Stem cell therapy for osteoarthritis: Functional cartilage regeneration using 3d bioprinting technology Osteoarthritis presents a significant societal and economic burden. Stina Simonsson from the University of Gothenburg explains how EU-funded projects are using 3D bioprinting to create functional cartilage for OA treatment. Osteoarthritis (OA) is a widespread condition that affects millions of people globally. As the population ages and the obesity epidemic continues, the incidence of OA is expected to increase. 3D bioprinting and stem cell therapies are heralded as the future of medical treatments for such conditions. OA affects a significant portion of the Swedish population. Estimates suggest that about 7-8% of the population is diagnosed with OA, which translates to roughly 700,000 to 800,000 individuals. The condition is more common among older adults, with a higher prevalence observed in those over the age of 65.

Advanced roll porous scaffold 3D bioprinting technology.

Improvements in the roll porous scaffold (RPS) 3D bioproduction technology will increase print density of 10-15µm cells by ~ 20% up to ~ 1.5 × 108 cells/mL and purity of organoid formation by > 17%. The use of 360 and 1200 dpi inkjet printheads immediately enables biomanufacturing with 10-30µm cells in a single organoid with performance > 1.8 L/h for 15µm layer thickness. The spongy bioresorbable ribbon for RPS technology is designed to solve the problems of precise placement, leakage and increasing in the number of instantly useable cell types and superior to all currently dominant 3D bioprinting methods in speed, volume, and print density without the use of expensive equipment and components. The potential of RPS for parallel testing of new substances studied was not on animals, but using generated 3D biomodels "organ on a chip". Solid organoids are more suitable for personalized medicine with simultaneous checking of several treatment methods and drugs, targeted therapy for a specific patient in vitro using the 3D composition of his personal cells, and selection of the most effective ones with the least toxicity. Overcoming the shortage of organs for implantation and personal hormone replacement therapy for everyone was achieved using printed endocrine glands based on their DNA.

3D bioprinting technology and equipment based on microvalve control.

3D bioprinting technology is widely used in biomedical fields such as tissue regeneration and constructing pathological model. The prevailing printing technique is extrusion-based bioprinting. In this printing method, the bioink needs to meet both printability and functionality, which are often conflicting requirements. Therefore, this study has developed an innovative microvalve-based equipment, incorporating components such as pressure control, a three-dimensional motion platform, and microvalve. Here, we present a droplet-based method for constructing complex three-dimensional structures. By leveraging the rapid switching characteristics of the microvalve, this equipment can achieve precise printing of bio-materials with viscosities as low as 10mPa·s, significantly expanding the biofabrication window for bioinks. This technology is of great significance for 3D bioprinting in tissue engineering and lays a solid foundation for the construction of complex artificial organ tissues.

Advances in 3D bioprinting for environmental remediation and hazardous materials treatment.

The high-throughput method based on the micron-level structure that 3D bioprinting technology offers for various environmental microbiological engineering applications is made possible by its several printing paths and precision programming control. This versatility makes it an on-demand manufacturing technology. A novel 3D manufacturing technique called 3D bioprinting may be used to precisely uptake and disperse bacteria to create microbial active substances with a variety of intricate functionalities for environmental applications. The technological challenges that the current 3D bioprinting technology must face include the mechanical properties of materials, the creation of specific bioinks to adapt to different strains, and the exploration of 4D bioprinting for intelligent applications. Therefore, this analysis delves deeply into the core technological ideas of 3D bioprinting, bioink materials, and their environmental applications. It also offers recommendations about the challenges and opportunities associated with 3D bioprinting. Combined with the present advancements in microbe enhancement technology, 3D bioprinting will provide an enabling platform for multifunctional microorganisms and facilitate the management of in situ directional responses in the environmental domain. This review highlights the applications of 3D bioprinting in the environmental monitoring and bioremediation. 3D printing in solid waste management is also discussed in detail.

Integration of Hydrogels and 3D Bioprinting Technologies for Chronic Wound Healing Management.

The integration of hydrogel-based bioinks with 3D bioprinting technologies presents an innovative approach to chronic wound management, which is particularly challenging to treat because of its multifactorial nature and high risk of complications. Using precise deposition techniques, 3D bioprinting significantly alters traditional wound care paradigms by enabling the fabrication of patient-specific wound dressings that imitate natural tissue properties. Hydrogels are notably beneficial for these applications because of their abundant water content and mechanical properties, which promote cell viability and pathophysiological processes of wound healing, such as re-epithelialization and angiogenesis. This article reviews key 3D printing technologies and their significance in enhancing the structural and functional outcomes of wound-care solutions. Challenges in bioink viscosity, cell viability, and printability are addressed, along with discussions on the cross-linking and mechanical stability of the constructs. The potential of 3D bioprinting to revolutionize chronic wound management rests on its capacity to generate remedies that expedite healing and minimize infection risks. Nevertheless, further studies and clinical trials are necessary to advance these therapies from laboratory to clinical use.

3D Bioprinting of Engineered Living Materials with Extracellular Electron Transfer Capability for Water Purification.

Attention is widely drawn to the extracellular electron transfer (EET) process of electroactive bacteria (EAB) for water purification, but its efficacy is often hindered in complex environmental matrices. In this study, the engineered living materials with EET capability (e-ELMs) were for the first time created with customized geometric configurations for pollutant removal using three-dimensional (3D) bioprinting platform. By combining EAB and tailored viscoelastic matrix, a biocompatible and tunable electroactive bioink for 3D bioprinting was initially developed with tuned rheological properties, enabling meticulous manipulation of microbial spatial arrangement and density. e-ELMs with different spatial microstructures were then designed and constructed by adjusting the filament diameter and orientation during the 3D printing process. Simulations of diffusion and fluid dynamics collectively showcase internal mass transfer rates and EET efficiency of e-ELMs with different spatial microstructures, contributing to the outstanding decontamination performances. Our research propels 3D bioprinting technology into the environmental realm, enabling the creation of intricately designed e-ELMs and providing promising routes to address the emerging water pollution concerns.

A biomimetic skin microtissue biosensor for the detection of fish parvalbumin

In this paper, a biomimetic skin microtissue biosensor was developed based on three-dimensional (3D) bioprinting to precisely and accurately determine fish parvalbumin (FV). Based on the principle that allergens stimulate cells to produce ONOO− (peroxynitrite anion), a screen-printed electrode for the detection nanomolar level ONOO− was innovatively prepared to indirectly detect FV based on the level of ONOO− release. Gelatin methacryloyl (GelMA), RBL-2H3 cells, and MS1 cells were used as bio-ink for 3D bioprinting. The high-throughput and standardized preparation of skin microtissue was achieved using stereolithography 3D bioprinting technology. The printed skin microtissues were put into the self-designed 3D platform that integrated cell culture and electrochemical detection. The experimental results showed that the sensor could effectively detect FV when the optimized ratio of RBL-2H3 to MS1 cells and allergen stimulation time were 2:8 and 2 h, respectively. The linear detection range was 0.125–3.0 μg/mL, and the calculated lowest detection limit was 0.122 μg/mL. In addition, the sensor had excellent selectivity, specificity, stability, and reliability. Thus, this study successfully constructed a biomimetic skin microtissue electrochemical sensor for PV detection.

Advances in 3D Bioprinting for Neuroregeneration: A Literature Review of Methods, Bioinks, and Applications

Recent advancements in 3D-bioprinting technology have sparked a growing interest in its application for brain repair, encompassing tissue regeneration, drug delivery, and disease modeling. This literature review examines studies conducted over the past five years to assess the current state of research in this field. Common bioprinting methods and key parameters influencing their selection are explored, alongside an analysis of the diverse types of bioink utilized and their associated parameters. The extrusion-based 3D-bioprinting method emerged as the most widely studied and popular topic, followed by inkjet-based and laser-based bioprinting and stereolithography. Regarding bioinks, fibrin-based and collagen-based bioinks are predominantly utilized. Furthermore, this review elucidates how 3D bioprinting holds promise for neural tissue repair, regeneration, and drug screening, detailing the steps involved and various approaches employed. Neurovascular 3D printing and bioscaffold 3D printing stand out as the top two preferred methods for brain repair. The recent studies’ shortcomings and potential solutions to address them are also examined and discussed. Overall, by synthesizing recent findings, this review provides valuable insights into the potential of 3D bioprinting for advancing brain repairment strategies.

The Impact of the Methacrylation Process on the Usefulness of Chitosan as a Biomaterial Component for 3D Printing.

Chitosan is a very promising material for tissue model printing. It is also known that the introduction of chemical modifications to the structure of the material in the form of methacrylate groups makes it very attractive for application in the bioprinting of tissue models. The aim of this work is to study the characteristics of biomaterials containing chitosan (BCH) and its methacrylated equivalent (BCM) in order to identify differences in their usefulness in 3D bioprinting technology. It has been shown that the BCM material containing methacrylic chitosan is three times more viscous than its non-methacrylated BCH counterpart. Additionally, the BCM material is characterized by stability in a larger range of stresses, as well as better printability, resolution, and fiber stability. The BCM material has higher mechanical parameters, both mechanical strength and Young's modulus, than the BCH material. Both materials are ideal for bioprinting, but BCM has unique rheological properties and significant mechanical resistance. In addition, biological tests have shown that the addition of chitosan to biomaterials increases cell proliferation, particularly in 3D-printed models. Moreover, modification in the form of methacrylation encourages reduced toxicity of the biomaterial in 3D constructs. Our investigation demonstrates the suitability of a chitosan-enhanced biomaterial, specifically methacrylate-treated, for application in tissue engineering, and particularly for tissues requiring resistance to high stress, i.e., vascular or cartilage models.

Composite bioink incorporating cell-laden liver decellularized extracellular matrix for bioprinting of scaffolds for bone tissue engineering

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.