Search Results (16)

Bioprinting, an innovative combination of biotechnology and additive manufacturing, has emerged as a transformative technology in healthcare, enabling the fabrication of functional tissues, organs, and patient-specific implants. The implementation of the aforementioned, however, introduces unique intellectual property (IP) challenges that extend beyond conventional biotechnology. The study explores three critical areas of concern: IP protection for bioprinting hardware and bioinks, ownership and ethical management of digital files derived from biological data, and the implications of commercializing bioprinted tissues and organs. Employing a multidisciplinary approach, the paper analyzes existing IP frameworks, highlights their limitations when applied to bioprinting, and examines ethical dilemmas, such as ownership of bioprinted human tissues and the commodification of biological innovations. Findings suggest that current IP laws inadequately address the complexities of bioprinting, particularly in managing the intersection of proprietary technologies and ethical considerations. The study underscores the need for adaptive legal and ethical frameworks to balance innovation with equitable access and sustainability. Recommendations include the development of tailored IP policies for bioprinting and enhanced international collaboration to harmonize legal protections across jurisdictions. This work aims to provide a comprehensive foundation for stakeholders to navigate the rapidly evolving landscape of bioprinting IP. Full article

Skin wounds often form scar tissue during healing. Early intervention with tissue-engineered materials and cell therapies may promote scar-free healing. Exosomes and extracellular vesicles (EV) secreted by mesenchymal stromal cells (MSC) are believed to have high regenerative capacity. EV bioactivity is preserved after lyophilization and storage to enable use in remote and typically resource-constrained environments. We developed a bioprinted bandage containing reconstituted EVs that can be fabricated at the point-of-need. An alginate/carboxymethyl cellulose (CMC) biomaterial ink was prepared, and printability and mechanical properties were assessed with rheology and compression testing. Three-dimensional printed constructs were evaluated for Young’s modulus relative to infill density and crosslinking to yield material with stiffness suitable for use as a wound dressing. We purified EVs from human MSC-conditioned media and characterized them with nanoparticle tracking analysis and mass spectroscopy, which gave a peak size of 118 nm and identification of known EV proteins. Fluorescently labeled EVs were mixed to form bio-ink and bioprinted to characterize EV release. EV bandages were bioprinted on both a commercial laboratory bioprinter and a custom ruggedized 3D printer with bioprinting capabilities, and lyophilized EVs, biomaterial ink, and thermoplastic filament were deployed to an austere Arctic environment and bioprinted. This work demonstrates that EVs can be bioprinted with an alginate/CMC hydrogel and released over time when in contact with a skin-like substitute. The technology is suitable for operational medical applications, notably in resource-limited locations, including large-scale natural disasters, humanitarian crises, and combat zones. Full article

Three-dimensional (3D) bioprinting has emerged as a revolutionary approach in the life sciences, combining multiple disciplines such as computer engineering, materials science, robotics, and biomedical engineering. This innovative technology enables the production of cellular constructs using bio-inks, and differs from conventional 3D printing by incorporating living cells. The present work addresses the conversion of a commercial thermoplastic 3D printer into a low-cost bioprinter. The modification addresses the challenges of the high cost of commercial bioprinters, limited adaptability, and specialized personnel requirements. This modification uses an extrusion-based bioprinting method that is particularly popular in research due to its viscosity tolerance and versatility. The individual steps, including replacing the extruder with a syringe pump, rebuilding the electronic motherboard, and configuring the firmware, are explained in detail. The work aims at providing access to bioprinting technology so that laboratories with modest resources can take advantage of the immense potential of this technology. This modification resulted in improved resolution, allowing submicron movements, which is comparable to some of the commercially available bioprinters. The accuracy of the modified printer was validated using hydrogel bioprinting tests, suggesting that it is suitable for broader applications in regenerative medicine. Full article

Seaweeds, rich in high-value polysaccharides with thickening/gelling properties (e.g., agar, carrageenan, and alginate), are extensively used in the food industry for texture customization and enhancement. However, conventional extraction methods for these hydrocolloids often involve potentially hazardous chemicals and long extraction times. In this study, three red seaweed species (Chondrus crispus, Gelidium Corneum, and Gracilaria gracilis) commercialized as food ingredients by local companies were chosen for their native gelling biopolymers, which were extracted using water-based methodologies (i.e., (1) hydration at room temperature; (2) stirring at 90 °C; and (3) centrifugation at 40 °C) for production of sustainable food gels. The potential use of these extracts as bioinks was assessed employing an extrusion-based 3D printer. The present work aimed to study the gelation process, taken place during printing, and assess the effectiveness of the selected green extraction method in producing gels. To improve the definition of the printed gel, two critical printing parameters were investigated: the addition of locust bean gum (LBG) at different concentrations (0, 0.5, 1, 1.5, 2, and 2.5%) and printing temperature (30, 40, 60, and 80 °C). Rheological results from a controlled-stress rheometer indicated that gels derived from G. corneum and G. gracilis exhibited a lower gel strength (lower G′ and G″) and excessive material spreading during deposition (lower viscosity) than C. crispus. Thus, G′ was around 5 and 70 times higher for C. crispus gels than for G. corneum and G. gracilis, respectively. When increasing LBG concentration (0.5 to 2.5% w/w) and lowering the printing temperature (80 to 30 °C), an enhanced gel matrix definition for G. corneum and G. gracilis gels was found. In contrast, gels from C. crispus demonstrated greater stability and were less influenced by these parameters, showcasing the potential of the seaweed to develop sustainable clean label food gels. Eventually, these results highlight the feasibility of using algal-based extracts obtained through a green procedure as bioinks where LBG was employed as a synergic ingredient. Full article

The tremendous application potential of 3D bioprinting in the biomedical field is witnessed by the ever-increasing interest in this technology over the past few years. In particular, the possibility of obtaining 3D cellular models that mimic tissues with precision and reproducibility represents a definitive advance for in vitro studies dealing with the biological mechanisms of cell growth, death and proliferation and is at the basis of the responses of healthy and pathological tissues to drugs and therapies. However, the impact of 3D bioprinting on research is limited by the high costs of professional 3D bioprinters, which represent an obstacle to the widespread access and usability of this technology. In this work, we present a 3D bioprinter that was developed in-house by modifying a low-cost commercial 3D printer by replacing the default extruder used to print plastic filaments with a custom-made syringe extruder that is suitable for printing bioinks. The modifications made to the 3D printer include adjusting the size of the extruder to accommodate a 1 mL syringe and reducing the extruder’s size above the printer. To validate the performance of the home-made bioprinter, some main printing characteristics, the cell vitality and the possibility of bioprinting CAD-designed constructs were benchmarked against a renowned professional 3D bioprinter by RegenHu. According to our findings, our in-house 3D bioprinter was mostly successful in printing a complex glioblastoma tumor model with good performances, and it managed to maintain a cell viability that was comparable to that achieved by a professional bioprinter. This suggests that an accessible open-source 3D bioprinter could be a viable option for research and development (R&D) laboratories interested in pre-commercial 3D bioprinting advancements. Full article

Advancements and developments in the 3D bioprinting have been promising and have met the needs of organ transplantation. Current improvements in tissue engineering constructs have enhanced their applications in regenerative medicines and other medical fields. The synergistic effects of 3D bioprinting have brought technologies such as tissue engineering, microfluidics, integrated tissue organ printing, in vivo bioprinted tissue implants, artificial intelligence and machine learning approaches together. These have greatly impacted interventions in medical fields, such as medical implants, multi-organ-on-chip models, prosthetics, drug testing tissue constructs and much more. This technological leap has offered promising personalized solutions for patients with chronic diseases, and neurodegenerative disorders, and who have been in severe accidents. This review discussed the various standing printing methods, such as inkjet, extrusion, laser-assisted, digital light processing, and stereolithographic 3D bioprinter models, adopted for tissue constructs. Additionally, the properties of natural, synthetic, cell-laden, dECM-based, short peptides, nanocomposite and bioactive bioinks are briefly discussed. Sequels of several tissue-laden constructs such as skin, bone and cartilage, liver, kidney, smooth muscles, cardiac and neural tissues are briefly analyzed. Challenges, future perspectives and the impact of microfluidics in resolving the limitations in the field, along with 3D bioprinting, are discussed. Certainly, a technology gap still exists in the scaling up, industrialization and commercialization of this technology for the benefit of stakeholders. Full article

Microcarriers (MCs) are adaptable therapeutic instruments that may be adjusted to specific therapeutic uses, making them an appealing alternative for regenerative medicine and drug delivery. MCs can be employed to expand therapeutic cells. MCs can be used as scaffolds for tissue engineering, as well as providing a 3D milieu that replicates the original extracellular matrix, facilitating cell proliferation and differentiation. Drugs, peptides, and other therapeutic compounds can be carried by MCs. The surface of the MCs can be altered, to improve medication loading and release, and to target specific tissues or cells. Allogeneic cell therapies in clinical trials require enormous volumes of stem cells, to assure adequate coverage for several recruitment locations, eliminate batch to batch variability, and reduce production costs. Commercially available microcarriers necessitate additional harvesting steps to extract cells and dissociation reagents, which reduces cell yield and quality. To circumvent such production challenges, biodegradable microcarriers have been developed. In this review, we have compiled key information relating to biodegradable MC platforms, for generating clinical-grade cells, that permit cell delivery at the target site without compromising quality or cell yields. Biodegradable MCs could also be employed as injectable scaffolds for defect filling, supplying biochemical signals for tissue repair and regeneration. Bioinks, coupled with biodegradable microcarriers with controlled rheological properties, might improve bioactive profiles, while also providing mechanical stability to 3D bioprinted tissue structures. Biodegradable materials used for microcarriers have the ability to solve in vitro disease modeling, and are advantageous to the biopharmaceutical drug industries, because they widen the spectrum of controllable biodegradation and may be employed in a variety of applications. Full article

The biofabrication of three-dimensional scaffolds using 3D printers and cell-containing bioinks is very promising. A wide range of materials and bioink compositions are being created and tested for cell viability and printability in order to satisfy the requirements of a bioink. This methodology has not still achieved technological maturity, and the actual costs mean that they are often inaccessible for researchers, consequently lowering the development and extending the required times. This research aims to apply this methodology on a laboratory scale by re-adapting a commercial 3D printer, consequently lowering the costs and energy impacts, and, at the same time, ensuring a level of accuracy extremely close to the currently adopted devices and, more in general, suitable for the scopes of the research. To accomplish this, we assembled a biomimetic scaffold made of human Umbilical Cord Matrix Stem Cells (hUCMS), cellulose, and alginate. Various molds were used to produce 3D scaffolds of different sizes. After bioprinting, cell viability was analyzed using ethidium bromide and fluorescein diacetate, and a histological stain was used to evaluate cell and bioink morphology. All of the examined bioinks had a uniform final 3D structure and were stable, easily printable, and procedure-adapted. Up until 21 days of culture, the bioinks remained unaltered and were simple to manipulate. After 7 and 21 days of cell culture, the hUCMS in the cellulose/alginate-based bioinks exhibited cell viabilities of 95% and 85%, respectively. The cells did not present with a fibroblast-like shape but appeared to be round-shaped and homogeneously distributed in the 3D structure. Biomimetic bioink, which is based on cellulose and alginate, is an appropriate hydrogel for 3D bioprinting. This preliminary work illustrated the potential use of these two biomaterials for the 3D bioprinting of mesenchymal stem cells. Full article

Light-based bioprinter manufacturing technology is still prohibitively expensive for organizations that rely on accessing three-dimensional biological constructs for research and tissue engineering endeavors. Currently, most of the bioprinting systems are based on commercial-grade-based systems or modified DIY (do it yourself) extrusion apparatuses. However, to date, few examples of the adoption of low-cost equipment have been found for light-based bioprinters. The requirement of large volumes of bioinks, their associated cost, and the lack of information regarding the parameter selection have undermined the adoption of this technology. This paper showcases the retrofitting and assessing of a low-cost Light-Based 3D printing system for tissue engineering. To evaluate the potential of a proposed design, a manufacturability test for different features, machine parameters, and Gelatin Methacryloyl (GelMA) concentrations for 7.5% and 10% was performed. Furthermore, a case study of a previously seeded hydrogel with C2C12 cells was successfully implemented as a proof of concept. On the manufacturability test, deviational errors were found between 0.7% to 13.3% for layer exposure times of 15 and 20 s. Live/Dead and Actin-Dapi fluorescence assays after 5 days of culture showed promising results in the cell viability, elongation, and alignment of 3D bioprinted structures. The retrofitting of low-cost equipment has the potential to enable researchers to create high-resolution structures and three-dimensional in vitro models. Full article

The application of chitosan (CS) and whey protein (WP) alone or in combination in 3D/4D printing has been well considered in previous studies. Although several excellent reviews on additive manufacturing discussed the properties and biomedical applications of CS and WP, there is a lack of a systemic review about CS and WP bio-inks for 3D/4D printing applications. Easily modified bio-ink with optimal printability is a key for additive manufacturing. CS, WP, and WP–CS complex hydrogel possess great potential in making bio-ink that can be broadly used for future 3D/4D printing, because CS is a functional polysaccharide with good biodegradability, biocompatibility, non-immunogenicity, and non-carcinogenicity, while CS–WP complex hydrogel has better printability and drug-delivery effectivity than WP hydrogel. The review summarizes the current advances of bio-ink preparation employing CS and/or WP to satisfy the requirements of 3D/4D printing and post-treatment of materials. The applications of CS/WP bio-ink mainly focus on 3D food printing with a few applications in cosmetics. The review also highlights the trends of CS/WP bio-inks as potential candidates in 4D printing. Some promising strategies for developing novel bio-inks based on CS and/or WP are introduced, aiming to provide new insights into the value-added development and commercial CS and WP utilization. Full article

Three-dimensional (3D) bioprinting promises to be essential in tissue engineering for solving the rising demand for organs and tissues. Some bioprinters are commercially available, but their impact on the field of Tissue engineering (TE) is still limited due to their cost or difficulty to tune. Herein, we present a low-cost easy-to-build printhead for microextrusion-based bioprinting (MEBB) that can be installed in many desktop 3D printers to transform them into 3D bioprinters. We can extrude bioinks with precise control of print temperature between 2–60 °C. We validated the versatility of the printhead, by assembling it in three low-cost open-source desktop 3D printers. Multiple units of the printhead can also be easily put together in a single printer carriage for building a multi-material 3D bioprinter. Print resolution was evaluated by creating representative calibration models at different temperatures using natural hydrogels such as gelatin and alginate, and synthetic ones like poloxamer. Using one of the three modified low-cost 3D printers, we successfully printed cell-laden lattice constructs with cell viabilities higher than 90% after 24-h post printing. Controlling temperature and pressure according to the rheological properties of the bioinks was essential in achieving optimal printability and great cell viability. The cost per unit of our device, which can be used with syringes of different volume, is less expensive than any other commercially available product. These data demonstrate an affordable open-source printhead with the potential to become a reliable alternative to commercial bioprinters for any laboratory. Full article

Bioinks are usually cell-laden hydrogels widely studied in bioprinting performing experimental tests to tune their rheological properties, thus increasing research time and development costs. Computational Fluids Dynamics (CFD) is a powerful tool that can minimize iterations and costs simulating the material behavior using parametric changes in rheological properties under testing. Additionally, most bioinks have specific functionalities and their properties might widely change with temperature. Therefore, commercial bioinks are an excellent way to standardize bioprinting process, but they are not analyzed in detail. Therefore, the objective of this work is to study how three temperatures of the Cellink Bioink influence shear stress pressure and velocity through computational simulation. A comparison of three conical nozzles (20, 22, and 25G) for each temperature has been performed. The results show that shear stress, pressure, and velocity vary in negligible ranges for all combinations. Although these ranges are small and define a good thermo-responsive bioink, they do not generate a filament on the air and make drops during extrusion. In conclusion, this bioink provides a very stable behavior with low shear stress, but other bioprinting parameters must be set up to get a stable filament width. Full article

Three-dimensional (3D) bioprinting is considered as a novel approach in biofabricating cell-laden constructs that could potentially be used to promote skin regeneration following injury. In this study, a novel crosslinked chitosan (CH)–genipin (GE) bioink laden with keratinocyte and human dermal fibroblast cells was developed and printed successfully using an extruder-based bioprinter. By altering the composition and degree of CH–GE crosslinking, bioink printability was further assessed and compared with a commercial bioink. Rheological analysis showed that the viscosity of the optimised bioink was in a suitable range that facilitated reproducible and reliable printing by applying low pressures ranging from 20–40 kPa. The application of low printing pressures proved vital for viability of cells loaded within the bioinks. Further characterisation using MTT assay showed that cells were still viable within the printed construct at 93% despite the crosslinking, processing and after subjecting to physiological conditions for seven days. The morphological study of the printed cells showed that they were mobile within the bioink. Furthermore, the multi-layered 3D printed constructs demonstrated excellent self-supportive structures in a consistent manner. Full article

The aim of this study was to develop and evaluate an optimized 3D bioprinting technology in order to fabricate novel scaffolds for the application of endothelial cell repair. Various biocompatible and biodegradable macroporous scaffolds (D = 10 mm) with interconnected pores (D = ~500 µm) were fabricated using a commercially available 3D bioprinter (r3bEL mini, SE3D, USA). The resolution of the printing layers was set at ~100 µm for all scaffolds. Various compositions of polylactic acid (PLA), polyethylene glycol (PEG) and pluronic F127 (F127) formulations were prepared and optimized to develop semi-solid viscous bioinks. Either dimethyloxalylglycine (DMOG) or erythroprotein (EPO) was used as a model drug and loaded in the viscous biocompatible ink formulations with a final concentration of 30% (w/w). The surface analysis of the bioinks via a spectroscopic analysis revealed a homogenous distribution of the forming materials throughout the surface, whereas SEM imaging of the scaffolds showed a smooth surface with homogenous macro-porous texture and precise pore size. The rheological and mechanical analyses showed optimum rheological and mechanical properties of each scaffold. As the drug, DMOG, is a HIF-1 inducer, its release from the scaffolds into PBS solution was measured indirectly using a bioassay for HIF-1α. This showed that the release of DMOG was sustained over 48 h. The release of DMOG was enough to cause a significant increase in HIF-1α levels in the bioassay, and when incubated with rat aortic endothelial cells (RAECs) for 2 h resulted in transcriptional activation of a HIF-1α target gene (VEGF). The optimum time for the increased expression of VEGF gene was approximately 30 min and was a 3-4-fold increase above baseline. This study provides a proof of concept, that a novel bioprinting platform can be exploited to develop biodegradable composite scaffolds for potential clinical applications in endothelial cell repair in cardiovascular disease (CVD), or in other conditions in which endothelial damage occurs. Full article

Malignant melanoma is often used as a model tumor for the establishment of novel therapies. It is known that two-dimensional (2D) culture methods are not sufficient to elucidate the various processes during cancer development and progression. Therefore, it is of major interest to establish defined biofabricated three-dimensional (3D) models, which help to decipher complex cellular interactions. To get an impression of their printability and subsequent behavior, we printed fluorescently labeled melanoma cell lines with Matrigel and two different types of commercially available bioinks, without or with modification (RGD (Arginine-Glycine-Aspartate)-sequence/laminin-mixture) for increased cell-matrix communication. In general, we demonstrated the printability of melanoma cells in all tested biomaterials and survival of the printed cells throughout 14 days of cultivation. Melanoma cell lines revealed specific differential behavior in the respective inks. Whereas in Matrigel, the cells were able to spread, proliferate and form dense networks throughout the construct, the cells showed no proliferation at all in alginate-based bioink. In gelatin methacrylate-based bioink, the cells proliferated in clusters. Surprisingly, the modifications of the bioinks with RGD or the laminin blend did not affect the analyzed cellular behavior. Our results underline the importance of precisely adapting extracellular matrices to individual requirements of specific 3D bioprinting applications. Full article