3D Bioprinting Technology Research Articles

Applications of Light-Based 3D Bioprinting and Photoactive Biomaterials for Tissue Engineering

The emergence of additive manufacturing, commonly referred to as 3D printing, has led to a revolution in the field of biofabrication. Numerous types of 3D bioprinting, including extrusion bioprinting, inkjet bioprinting, and lithography-based bioprinting, have been developed and have played pivotal roles in driving a multitude of pioneering breakthroughs in the fields of tissue engineering and regenerative medicine. Among all the 3D bioprinting methods, light-based bioprinting utilizes light to crosslink or solidify photoreactive biomaterials, offering unprecedented spatiotemporal control over biomaterials and enabling the creation of 3D structures with extremely high resolution and precision. However, the lack of suitable photoactive biomaterials has hindered the application of light-based bioprinting in tissue engineering. The development of photoactive biomaterials has only recently been expanded. Therefore, this review summarizes the latest advancements in light-based 3D bioprinting technologies, including the development of light-based bioprinting techniques, photo-initiators (PIs), and photoactive biomaterials and their corresponding applications. Moreover, the challenges facing bioprinting are discussed, and future development directions are proposed.

Natural biopolymer for 3D printing

Three-dimensional (3D) bioprinting is a promising technological approach for various applications in the biomedical field. Natural polymers, which comprise the majority of 3D printable “bioinks”, have played a crucial role in various 3D bioprinting technologies during the layered 3D manufacturing processes in the last decade. However, the polymers must be customized for printing and effector function needs in cancer, dental care, oral medicine and biosensors, cardiovascular disease, and muscle restoration. This review provides an overview of 3D bio-printed natural polymers—commonly employed in various medical fields—and their recent development.

Advances in volumetric bioprinting.

The three-dimensional (3D) bioprinting technologies are suitable for biomedical applications owing to their ability to manufacture complex and high-precision tissue constructs. However, the slow printing speed of current layer-by-layer (bio)printing modality is the major limitation in biofabrication field. To overcome this issue, volumetric bioprinting (VBP) is developed. VBP changes the layer-wise operation of conventional devices, permitting the creation of geometrically complex, centimeter-scale constructs in tens of seconds. VBP is the next step onward from sequential biofabrication methods, opening new avenues for fast additive manufacturing in the fields of tissue engineering, regenerative medicine, personalized drug testing, and soft robotics, etc. Therefore, this review introduces the printing principles and hardware designs of VBP-based techniques; then focuses on the recent advances in VBP-based (bio)inks and their biomedical applications. Lastly, the current limitations of VBP are discussed together with future direction of research.

3D Printing of a Tissue-Engineered Structure Intended to Replace Cartilage Defects

This article describes the process of developing a tissue-engineered structure that meets the biocompatibility and biodegradation parameters necessary for replacing cartilage tissue defects. The study was carried out using 3D bioprinting technology, which represents a promising research direction in the biomedical field. It is known that, due to the specifics of its structure, cartilage tissue is not capable of complete regeneration of damage. The methods currently used for treating arthrosis are associated with a number of limitations and disadvantages, which makes research aimed at developing alternative methods for arthrosis treatment particularly relevant. The development of tissue-engineered structures by 3D bioprinting requires the materials not only certified for medical use but also exhibiting biocompatibility and biodegradation properties. Polylactide (PLA) and sodium alginate satisfy the above requirements; moreover, their availability and economic affordability make them one of the most popular materials for 3D bioprinting.

Prediction of Clinical Precision Chemotherapy by Patient-Derived 3D Bioprinting Models of Colorectal Cancer and Its Liver Metastases.

Methods accurately predicting the responses of colorectal cancer (CRC) and colorectal cancer liver metastasis (CRLM) to personalized chemotherapy remain limited due to tumor heterogeneity. This study introduces an innovative patient-derived CRC and CRLM tumor model for preclinical investigation, utilizing 3d-bioprinting (3DP) technology. Efficient construction of homogeneous in vitro 3D models of CRC/CRLM is achieved through the application of patient-derived primary tumor cells and 3D bioprinting with bioink. Genomic and histological analyses affirm that the CRC/CRLM 3DP tumor models effectively retain parental tumor biomarkers and mutation profiles. In vitro tests evaluating chemotherapeutic drug sensitivities reveal substantial tumor heterogeneity in chemotherapy responses within the 3DP CRC/CRLM models. Furthermore, a robust correlation is evident between the drug response in the CRLM 3DP model and the clinical outcomes of neoadjuvant chemotherapy. These findings imply a significant potential for the application of patient-derived 3DP cancer models in precision chemotherapy prediction and preclinical research for CRC/CRLM.

Application of tissue engineering techniques in tracheal repair: a bibliometric study

ABSTRACTTransplantation of tissue-engineered trachea is an effective treatment for long-segment tracheal injury. This technology avoids problems associated with a lack of donor resources and immune rejection, generating an artificial trachea with good biocompatibility. To our knowledge, a systematic summary of basic and clinical research on tissue-engineered trachea in the last 20 years has not been conducted. Here, we analyzed the development trends of tissue-engineered trachea research by bibliometric means and outlined the future perspectives in this field. The Web of Science portal was selected as the data source. CiteSpace, VOSviewer, and the Bibliometric Online Analysis Platform were used to analyze the number of publications, journals, countries, institutions, authors, and keywords from 475 screened studies. Between 2000 and 2023, the number of published studies on tissue-engineered trachea has been increasing. Biomaterials published the largest number of papers. The United States and China have made the largest contributions to this field. University College London published the highest number of studies, and the most productive researcher was an Italian scholar, Paolo Macchiarini. However, close collaborations between various researchers and institutions from different countries were generally lacking. Despite this, keyword analysis showed that manufacturing methods for tracheal stents, hydrogel materials, and 3D bioprinting technology are current popular research topics. Our bibliometric study will help scientists in this field gain an in-depth understanding of the current research progress and development trends to guide their future work, and researchers in related fields will benefit from the introduction to transplantation methods of tissue-engineered trachea.

Bioprinted Human Lung Cancer-Mimics for Tissue Diagnostics Applications.

Developing a reproducible and secure supply of customizable control tissues that standardizes for the cell type, tissue architecture, and preanalytics of interest for usage in applications including diagnostic, prognostic, and predictive assays, is critical for improving our patient care and welfare. The conventionally adopted control tissues directly obtained from patients are not ideal because they oftentimes have different amounts of normal and neoplastic elements, differing cellularity, differing architecture, and unknown preanalytics, in addition to the limited supply availability and thus associated high costs. In this study, we demonstrated a strategy to stably produce tissue-mimics for diagnostics purposes by taking advantage of the three-dimensional (3D) bioprinting technology. Specifically, we take anaplastic lymphoma kinase-positive (Alk+) lung cancer as an example, where a micropore-forming bioink laden with tumor cells was combined with digital light processing-based bioprinting for developing native-like Alk+ lung cancer tissue-mimics with both structural and functional relevancy. It is anticipated that our proposed methodology will pave new avenues for both fields of tissue diagnostics and 3D bioprinting significantly expanding their capacities, scope, and sustainability.

4D bioprinting technology and its application in cardiovascular tissue engineering

3D bioprinting technology is a rapidly developing technique that employs bioinks containing biological materials and living cells to construct biomedical products. However, 3D-printed tissues are static, while human tissues are in real-time dynamic states that can change in morphology and performance. To improve the compatibility between in vitro and in vivo environments, an in vitro tissue engineering technique that simulates this dynamic process is required. The concept of 4D printing, which combines "3D printing + time" provides a new approach to achieving this complex technique. 4D printing involves applying one or more smart materials that respond to stimuli, enabling them to change their shape, performance, and function under the corresponding stimulus to meet various needs. This article focuses on the latest research progress and potential application areas of 4D printing technology in the cardiovascular system, providing a theoretical and practical reference for the development of this technology.

Unravelling hierarchical patterning of biomaterial inks with 3D microfluidic-assisted spinning: a paradigm shift in bioprinting technologies

For decades, 3D bioprinting has offered a revolutionising approach to combine living cells and biomaterials to engineer complex, yet functional constructs. However, traditional 3D bioprinting platforms fall short of the ability to pattern complex gradients of biomaterials, cells, and ultimately bio-physical properties to drive tissue formation and regeneration. Recently, 3D microfluidic-assisted bioprinting (3DMB) has risen as a new hybrid approach for the fabrication of physiologically relevant tissues, adopting a microfluidic chip as functional printhead to achieve hierarchical patterning of bioinks and precise control over the microscale architecture of printed constructs, enabling the creation of multi-layered tissues. This review explores recent advancements in graded biomaterial patterning using microfluidic-assisted spinning and novel 3D bioprinting technologies. The physiological hierarchical arrangement of human tissues and the crucial role of biomaterials in achieving ordered assembly is hereby discussed. Lastly, the integration of microfluidic-assisted techniques with new bioprinting platforms is highlighted, examining the latest advancements in tissue regeneration and disease modelling.

Cell Culture Model Evolution and Its Impact on Improving Therapy Efficiency in Lung Cancer.

Optimizing cell culture conditions is essential to ensure experimental reproducibility. To improve the accuracy of preclinical predictions about the response of tumor cells to different classes of drugs, researchers have used 2D or 3D cell cultures in vitro to mimic the cellular processes occurring in vivo. While 2D cell culture provides valuable information on how therapeutic agents act on tumor cells, it cannot quantify how the tumor microenvironment influences the response to therapy. This review presents the necessary strategies for transitioning from 2D to 3D cell cultures, which have facilitated the rapid evolution of bioengineering techniques, leading to the development of microfluidic technology, including organ-on-chip and tumor-on-chip devices. Additionally, the study aims to highlight the impact of the advent of 3D bioprinting and microfluidic technology and their implications for improving cancer treatment and approaching personalized therapy, especially for lung cancer. Furthermore, implementing microfluidic technology in cancer studies can generate a series of challenges and future perspectives that lead to the discovery of new predictive markers or targets for antitumor treatment.

Small Joint Organoids 3D Bioprinting: Construction Strategy and Application.

Osteoarthritis (OA) is a chronic disease that causes pain and disability in adults, affecting ≈300 million people worldwide. It is caused by damage to cartilage, including cellular inflammation and destruction of the extracellular matrix (ECM), leading to limited self-repairing ability due to the lack of blood vessels and nerves in the cartilage tissue. Organoid technology has emerged as a promising approach for cartilage repair, but constructing joint organoids with their complex structures and special mechanisms is still challenging. To overcome these boundaries, 3D bioprinting technology allows for the precise design of physiologically relevant joint organoids, including shape, structure, mechanical properties, cellular arrangement, and biological cues to mimic natural joint tissue. In this review, the authors will introduce the biological structure of joint tissues, summarize key procedures in 3D bioprinting for cartilage repair, and propose strategies for constructing joint organoids using 3D bioprinting. The authors also discuss the challenges of using joint organoids' approaches and perspectives on their future applications, opening opportunities to model joint tissues and response to joint disease treatment.

Microphysiological Constructs and Systems: Biofabrication Tactics, Biomimetic Evaluation Approaches, and Biomedical Applications.

In recent decades, microphysiological constructs and systems (MPCs and MPSs) have undergone significant development, ranging from self-organized organoids to high-throughput organ-on-a-chip platforms. Advances in biomaterials, bioinks, 3D bioprinting, micro/nanofabrication, and sensor technologies have contributed to diverse and innovative biofabrication tactics. MPCs and MPSs, particularly tissue chips relevant to absorption, distribution, metabolism, excretion, and toxicity, have demonstrated potential as precise, efficient, and economical alternatives to animal models for drug discovery and personalized medicine. However, current approaches mainly focus on the in vitro recapitulation of the human anatomical structure and physiological-biochemical indices at a single or a few simple levels. This review highlights the recent remarkable progress in MPC and MPS models and their applications. The challenges that must be addressed to assess the reliability, quantify the techniques, and utilize the fidelity of the models are also discussed.

3D Bioprinting: A Systematic Review for Future Research Direction.

3D bioprinting is capable of rapidly producing small-scale human-based tissue models, or organoids, for pathology modeling, diagnostics, and drug development. With the use of 3D bioprinting technology, 3D functional complex tissue can be created by combining biocompatible materials, cells, and growth factor. In today's world, 3D bioprinting may be the best solution for meeting the demand for organ transplantation. It is essential to examine the existing literature with the objective to identify the future trend in terms of application of 3D bioprinting, different bioprinting techniques, and selected tissues by the researchers, it is very important to examine the existing literature. To find trends in 3D bioprinting research, this work conducted an systematic literature review of 3D bioprinting. This literature provides a thorough study and analysis of research articles on bioprinting from 2000 to 2022 that were extracted from the Scopus database. The articles selected for analysis were classified according to the year of publication, articles and publishers, nation, authors who are working in bioprinting area, universities, biomaterial used, and targeted applications. The top nations, universities, journals, publishers, and writers in this field were picked out after analyzing research publications on bioprinting. During this study, the research themes and research trends were also identified. Furthermore, it has been observed that there is a need for additional research in this domain for the development of bioink and their properties that can guide practitioners and researchers while selecting appropriate combinations of biomaterials to obtain bioink suitable for mimicking human tissue. This research includes research findings, recommendations, and observations for bioprinting researchers and practitioners. This article lists significant research gaps, future research directions, and potential application areas for bioprinting. The review conducted here is mainly focused on the process of collecting, organizing, capturing, evaluating, and analyzingdata to give adeeper understanding of bioprinting and to identify potential future research trends.

Alginate-waterborne polyurethane 3D bioprinted scaffolds for articular cartilage tissue engineering

Articular cartilage defects comprise a spectrum of diseases associated with degeneration or damage of the connective tissue present in particular joints, presenting progressive osteoarthritis if left untreated. In vitro tissue regeneration is an innovative treatment for articular cartilage injuries that is attracting not only clinical attention, but also great interest in the development of novel biomaterials, since this procedure involves the formation of a neotissue with the help of material support. In this work, functional alginate and waterborne polyurethane (WBPU) scaffolds have been developed for articular cartilage regeneration using 3D bioprinting technology. The particular properties of alginate-WBPU blends, like mechanical strength, elasticity and moistening, mimic the original cartilage tissue characteristics, being ideal for this application. To fabricate the scaffolds, mature chondrocytes were loaded into different alginate-WBPU inks with rheological properties suitable for 3D bioprinting. Bioinks with high alginate content showed better 3D printing performance, as well as structural integrity and cell viability, being most suitable for scaffolds fabrication. After 28 days of in vitro cartilage formation experiments, scaffolds containing 3.2 and 6.4 % alginate resulted in the maintenance of cell number in the range of 104 chondrocytes/scaffold in differentiated phenotypes, capable of synthesizing specialized extracellular matrix (ECM) up to 6 μg of glycosaminoglycans (GAG) and thus, showing a potential application of these scaffolds for in vitro regeneration of articular cartilage tissue.

Characterization of Bioinks Prepared via Gelifying Extracellular Matrix from Decellularized Porcine Myocardia.

Since the emergence of 3D bioprinting technology, both synthetic and natural materials have been used to develop bioinks for producing cell-laden cardiac grafts. To this end, extracellular-matrix (ECM)-derived hydrogels can be used to develop scaffolds that closely mimic the complex 3D environments for cell culture. This study presents a novel cardiac bioink based on hydrogels exclusively derived from decellularized porcine myocardium loaded with human-bone-marrow-derived mesenchymal stromal cells. Hence, the hydrogel can be used to develop cell-laden cardiac patches without the need to add other biomaterials or use additional crosslinkers. The scaffold ultrastructure and mechanical properties of the bioink were characterized to optimize its production, specifically focusing on the matrix enzymatic digestion time. The cells were cultured in 3D within the developed hydrogels to assess their response. The results indicate that the hydrogels fostered inter-cell and cell-matrix crosstalk after 1 week of culture. In conclusion, the bioink developed and presented in this study holds great potential for developing cell-laden customized patches for cardiac repair.

Biomimetic 3D bioprinting approaches to engineer the tumor microenvironment

With the increasing incidence and mortality rates, cancer remains a major health challenge in the world. Despite advances in therapies and clinical programs, the efficacy of anti-cancer drugs often fails to translate from pre-clinical models to patient clinical trials. To date, pre-clinical cancer models, including two-dimensional cell cultures and animal models, have limited versatility and accuracy in recapitulating the complexity of human cancer. To address these limitations, a growing focus has fostered the development of three-dimensional (3D) tumor models that closely resemble the in vivo tumor microenvironment and heterogeneity. Recent efforts have leveraged bioengineering technologies, such as biofabrication, to engineer new platforms that mimic healthy and diseased organs, aiming to overcome the shortcomings of conventional models, such as for musculoskeletal tissues. Notably, 3D bioprinting has emerged as a powerful tool in cancer research, offering precise control over cell and biomaterial deposition to fabricate architecturally complex and reproducible functional models. The following review underscores the urgent need for more accurate and relevant 3D tumor models, highlighting the advantages of the use of biofabrication approaches to engineer new biomimetics platforms. We provide an updated discussion on the role of bioengineering technologies in cancer research and modeling with particular focus on 3D bioprinting platforms, as well as a close view on biomaterial inks and 3D bioprinting technologies employed in cancer modeling. Further insights into the 3D bioprinting tissue-specific modeling panorama are presented in this paper, offering a comprehensive overview of the new possibilities for cancer study and drug discovery.  

Generation of HPMC-derived Placebo Hydrogel Component Matrix System for Possible Pharmaceutical 3D Printing Applications

Integration of 3D bioprinting technology in the design and fabrication of pharmaceutical dosage form has the potential toovercome the limitations and expand the range of drugs that can be delivered through different routes. In recent years, thefield of bioprinting has emerged as a revolutionary technology with the potential to transform various aspects of the healthcareand pharmaceutical industries. Among its wide-ranging applications, one of the most promising areas is the development oftransdermal drug delivery systems using 3D bioprinting techniques. This innovative approach holds significant promise inenhancing drug delivery efficiency, improving patient compliance, and revolutionizing personalized medicine. 3D bioprintingtechnology offers exciting prospects for the development of transdermal patches specifically designed for wound healingapplications. With further advancements and research, these innovative patches have the potential to revolutionize the fieldof wound care and improve patient outcomes.

In Vitro and In Vivo Biological Assessments of 3D-Bioprinted Scaffolds for Dental Applications.

Three-dimensional (3D) bioprinting is a unique combination of technological advances in 3D printing and tissue engineering. It has emerged as a promising approach to address the dilemma in current dental treatments faced by clinicians in order to repair or replace injured and diseased tissues. The exploration of 3D bioprinting technology provides high reproducibility and precise control of the bioink containing the desired cells and biomaterial over the architectural and dimensional features of the scaffolds in fabricating functional tissue constructs that are specific to the patient treatment need. In recent years, the dental applications of different 3D bioprinting techniques, types of novel bioinks, and the types of cells used have been extensively explored. Most of the findings noted significant challenges compared to the non-biological 3D printing approach in constructing the bioscaffolds that mimic native tissues. Hence, this review focuses solely on the implementation of 3D bioprinting techniques and strategies based on cell-laden bioinks. It discusses the in vitro applications of 3D-bioprinted scaffolds on cell viabilities, cell functionalities, differentiation ability, and expression of the markers as well as the in vivo evaluations of the implanted bioscaffolds on the animal models for bone, periodontal, dentin, and pulp tissue regeneration. Finally, it outlines some perspectives for future developments in dental applications.

High-throughput bioprinting of the nasal epithelium using patient-derived nasal epithelial cells

Progenitor human nasal epithelial cells (hNECs) are an essential cell source for the reconstruction of the respiratory pseudostratified columnar epithelium composed of multiple cell types in the context of infection studies and disease modeling. Hitherto, manual seeding has been the dominant method for creating nasal epithelial tissue models through biofabrication. However, this approach has limitations in terms of achieving the intricate three-dimensional (3D) structure of the natural nasal epithelium. 3D bioprinting has been utilized to reconstruct various epithelial tissue models, such as cutaneous, intestinal, alveolar, and bronchial epithelium, but there has been no attempt to use of 3D bioprinting technologies for reconstruction of the nasal epithelium. In this study, for the first time, we demonstrate the reconstruction of the nasal epithelium with the use of primary hNECs deposited on Transwell inserts via droplet-based bioprinting (DBB), which enabled high-throughput fabrication of the nasal epithelium in Transwell inserts of 24-well plates. DBB of progenitor hNECs ranging from one-tenth to one-half of the cell seeding density employed during the conventional cell seeding approach enabled a high degree of differentiation with the presence of cilia and tight-junctions over a 4 weeks air–liquid interface culture. Single cell RNA sequencing of these cultures identified five major epithelial cells populations, including basal, suprabasal, goblet, club, and ciliated cells. These cultures recapitulated the pseudostratified columnar epithelial architecture present in the native nasal epithelium and were permissive to respiratory virus infection. These results denote the potential of 3D bioprinting for high-throughput fabrication of nasal epithelial tissue models not only for infection studies but also for other purposes, such as disease modeling, immunological studies, and drug screening.

Trends in Photopolymerizable Bioinks for 3D Bioprinting of Tumor Models.

Three-dimensional (3D) bioprinting technologies involving photopolymerizable bioinks (PBs) have attracted enormous attention in recent times owing to their ability to recreate complex structures with high resolution, mechanical stability, and favorable printing conditions that are suited for encapsulating cells. 3D bioprinted tissue constructs involving PBs can offer better insights into the tumor microenvironment and offer platforms for drug screening to advance cancer research. These bioinks enable the incorporation of physiologically relevant cell densities, tissue-mimetic stiffness, and vascularized channels and biochemical gradients in the 3D tumor models, unlike conventional two-dimensional (2D) cultures or other 3D scaffold fabrication technologies. In this perspective, we present the emerging techniques of 3D bioprinting using PBs in the context of cancer research, with a specific focus on the efforts to recapitulate the complexity of the tumor microenvironment. We describe printing approaches and various PB formulations compatible with these techniques along with recent attempts to bioprint 3D tumor models for studying migration and metastasis, cell-cell interactions, cell-extracellular matrix interactions, and drug screening relevant to cancer. We discuss the limitations and identify unexplored opportunities in this field for clinical and commercial translation of these emerging technologies.