Extrusion Bioprinting Research Articles

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 for Engineered Tissue Constructs and Patient-Specific Models: Current Progress and Prospects in Clinical Applications.

Advancements in bioprinting technology are driving the creation of complex, functional tissue constructs for use in tissue engineering and regenerative medicine. Various methods, including extrusion, jetting, and light-based bioprinting, have their unique advantages and drawbacks. Over the years, researchers and industry leaders have made significant progress in enhancing bioprinting techniques and materials, resulting in the production of increasingly sophisticated tissue constructs. Despite this progress, challenges still need to be addressed in achieving clinically relevant, human-scale tissue constructs, presenting a hurdle to widespread clinical translation. However, with ongoing interdisciplinary research and collaboration, the field is rapidly evolving and holds promise for personalized medical interventions. Continued development and refinement of bioprinting technologies have the potential to address complex medical needs, enabling the development of functional, transplantable tissues and organs, as well as advanced in vitro tissue models.

Effect of Hydrodynamic Shear Stress on Algal Cell Fate in 3D Extrusion Bioprinting

The 3D bioprinting of aquatic photosynthetic organisms holds potential for applications in biosensing, wastewater treatment, and biofuel production. While algae cells can be immobilized in bioprinted cell‐friendly matrices, there is a knowledge gap regarding the thresholds of hydrodynamic shear stress that affect the cells’ functionality and viability during bioprinting. This study examines the effect of hydrodynamic shear stress on the fate of Chlamydomonas reinhardtii cells. Computational fluid dynamics models based on the Navier–Stokes equations are developed to numerically predict the shear stresses experienced by the cells during extrusion. Parallelly, cell culture experiments are conducted to evaluate the functionality, growth rates, and viability of algae cells within bioprinted constructs. By correlating cell culture and simulation results, the causal link between shear stress in the nozzle and cell viability and function has been characterized. The findings highlight that cell viability and function are significantly impacted by process factors. Notably, algae cell function is more sensitive to shear stress than cell viability. Functional impairments occur at maximum shear stresses around 5 kPa, while viability remains unaffected. Beyond 14 kPa, both functionality and viability decline significantly and irreversibly. The results emphasize the importance of assessing viability and function after bioprinting, rather than just viability.

Advancing Cartilage Tissue Engineering: A Review of 3D Bioprinting Approaches and Bioink Properties.

Articular cartilage is crucial in human physiology, and its degeneration poses a significant public health challenge. While recent advancements in 3D bioprinting and tissue engineering show promise for cartilage regeneration, there remains a gap between research findings and clinical application. This review critically examines the mechanical and biological properties of hyaline cartilage, along with current 3D manufacturing methods and analysis techniques. Moreover, we provide a quantitative synthesis of bioink properties used in cartilage tissue engineering. After screening 181 initial works, 33 studies using extrusion bioprinting were analyzed and synthesized, presenting results that indicate the main materials, cells, and methods utilized for mechanical and biological evaluation. Altogether, this review motivates the standardization of mechanical analyses and biomaterial assessments of 3D bioprinted constructs to clarify their chondrogenic potential.

Bio-printing method as a novel approach to obtain a fibrin scaffold settled by limbal epithelial cells for corneal regeneration

Treatment of Limbal Stem Cell Deficiency (LSCD), based on autologous transplantation of the patient’s stem cells, is one of the few medical stem cell therapies approved by the European Medicines Agency (EMA). It relies on isolating and culturing in vivo Limbal Epithelial Stem Cells (LESC) and then populating them on the fibrin substrate, creating a scaffold for corneal epithelial regeneration. Such a solution is then implanted into the patient’s eye. The epithelial cell culture process is specific, and its results strongly depend on the initial cell seeding density. Achieving control of the density and repeatability of the process is a desirable aim and can contribute to the success of the therapy. The study aimed to test bioprinting as a potential technique to increase the control over LESCs seeding on a scaffold and improve process reproducibility. Cells were applied to 0.5 mm thick, flat, transparent fibrin substrates using extrusion bioprinting; the control was the traditional manual application of cells using a pipette. The use of 3D printer enabled uniform coverage of the scaffold surface, and LESCs density in printed lines was close to the targeted value. Moreover, printed cells had higher cell viability than those seeded traditionally (91.1 ± 8.2% vs 82.6 ± 12.8%). The growth rate of the epithelium was higher in bioprinted samples. In both methods, the epithelium had favorable phenotypic features (p63 + and CK14 +). 3D printing constitutes a promising approach in LSCD therapy. It provides favorable conditions for LESCs growth and process reproducibility. Its application may lead to reduced cell requirements, thereby to using fewer cells on lower passages, which will contribute to preserving LESCs proliferative potential.

Surrogate-assisted multi-objective Bayesian optimization for improved rheological design of bioinks

Extrusion bioprinting is a popular biofabrication technique for creating viable biological constructs with applications ranging from regenerative medicine and cancer research to therapeutics screening. A primary challenge within extrusion bioprinting lies in preserving cell viability and function amidst the shear forces generated during extrusion. Therefore, the formulation of shear-thinning bioinks with tailored flow properties is essential to minimize shear stress accumulation during printing. However, this task is complex, as the bioink’s flow properties are contingent upon numerous factors, including but not limited to the polymer concentration, spatial cell density in the bioink, molecular weight of the polymer, and the nozzle’s geometry. The current methods for adjusting flow properties are heuristic, posing challenges to the rapid development models and automation of the bioprinting workflow. In this study, we introduce a Gaussian Process-based multi-objective Bayesian optimization framework that rapidly identifies Pareto-optimal flow property values to minimize hydrodynamic shear stresses during extrusion. The optimization focuses on four flow parameters, minimum viscosity, maximum viscosity, consistency index, and flow behavior index from the power-law model of shear-thinning fluids alongside the nozzle outlet diameter. Following an initial set of 24 experimental runs, our sequential design approach required 21 iterations to achieve convergence on a set of flow parameters that meet the Pareto optimality criteria. The presented strategy, which integrates physics-based numerical models with data-driven optimization models, emerges as a tool for the rapid fine-tuning of bioink flow behaviors. The multi-objective framework also sets the stage for future exploration into cell viability and functional outcomes post-bioprinting.

Remote-Controlled Gene Delivery in Coaxial 3D-Bioprinted Constructs using Ultrasound-Responsive Bioinks

IntroductionCoaxial 3D bioprinting has advanced the formation of tissue constructs that recapitulate key architectures and biophysical parameters for in-vitro disease modeling and tissue-engineered therapies. Controlling gene expression within these structures is critical for modulating cell signaling and probing cell behavior. However, current transfection strategies are limited in spatiotemporal control because dense 3D scaffolds hinder diffusion of traditional vectors. To address this, we developed a coaxial extrusion 3D bioprinting technique using ultrasound-responsive gene delivery bioinks. These bioink materials incorporate echogenic microbubble gene delivery particles that upon ultrasound exposure can sonoporate cells within the construct, facilitating controllable transfection.MethodsPhospholipid-coated gas-core microbubbles were electrostatically coupled to reporter transgene plasmid payloads and incorporated into cell-laden alginate bioinks at varying particle concentrations. These bioinks were loaded into the coaxial nozzle core for extrusion bioprinting with CaCl2 crosslinker in the outer sheath. Resulting bioprints were exposed to 2.25 MHz focused ultrasound and evaluated for microbubble activation and subsequent DNA delivery and transgene expression.ResultsCoaxial printing parameters were established that preserved the stability of ultrasound-responsive gene delivery particles for at least 48 h in bioprinted alginate filaments while maintaining high cell viability. Successful sonoporation of embedded cells resulted in DNA delivery and robust ultrasound-controlled transgene expression. The number of transfected cells was modulated by varying the number of focused ultrasound pulses applied. The size region over which DNA was delivered was modulated by varying the concentration of microbubbles in the printed filaments.ConclusionsOur results present a successful coaxial 3D bioprinting technique designed to facilitate ultrasound-controlled gene delivery. This platform enables remote, spatiotemporally-defined genetic manipulation in coaxially bioprinted tissue constructs with important applications for disease modeling and regenerative medicine.

Characterization of pentenoate-functionalized hyaluronic acid and pentenoate-functionalized gelatin hydrogels for printing and future surgical placement in regenerative medicine applications

Injectable hydrogels with in situ crosslinking may be more suitable than pre-fabricated scaffolds for surgical delivery to clinical injuries. However, low viscosity hydrogel precursors may be challenging to surgically place into an injury if the precursor leaks or is washed out. The biomaterials field for extrusion bioprinting is a fertile ground for discovering biomaterials with injectable and paste-like precursor rheology with in situ gelation capabilities, which may promote better material retention in clinical injuries. We previously developed and evaluated one formulation of a pentenoate-functionalized hyaluronic acid (PHA) / pentenoate-functionalized gelatin (PGel) hydrogel with a paste-like, printable precursor and rapid photocrosslinking in a spinal cord injury application. Further characterization of the material and cell response to PHA/PGel hydrogel formulations is needed to expand the bioprinting and other regenerative medicine opportunities for PHA/PGel hydrogels. In the current study, we utilized 2D NMR methods (i.e., 1H–1H TOCSY) to confirm and quantify a high degree of pentenoate functionalization of PGel and PHA. We characterized the stiffness, swelling, and cell viability using varying formulations of PGel or PHA/PGel hydrogels. For compression testing, a straightforward application of the Ogden model enabled evaluation of the full stress-strain range for improved moduli comparisons. We identified two formulations that best supported cell viability (i.e., 3%/10% and 4%/5% PHA/PGel). Furthermore, one of the identified formulations (4%/5% PHA/PGel) had superior printability compared to the other. With better printability and potentially better clinical surgical placement, the new PHA/PGel hydrogel formulations may be more widely applied in the bioprinting and regenerative medicine fields.

Polycaprolactone strengthening gelatin/nano-hydroxyapatite composite biomaterial inks for potential application in extrusion-based 3D printing bone scaffolds

Extrusion-based three-dimensional (3D) printing of gelatin (Gel) is crucial for fabricating bone tissue engineering scaffolds via additive manufacturing. However, the thermal instability of Gel remains a persistent challenge, as it tends to collapse at mild temperatures. Current approaches often involve simply mixing Gel particles with various materials, resulting in biomaterial inks that lack uniformity and have inconsistent degradation characteristics. In this study, acetic acid was used to dissolve Gel and polycaprolactone (PCL) separately, producing homogeneous Gel/PCL dispersions with optimal pre-treatment performance. These dispersions were then combined and hybridized with nano-hydroxyapatite (n-HA) to create a composite printing ink. By evaluating the printability of the ink, the optimal conditions were identified: a n-HA concentration of 50% (w/w), a printing temperature of 10–15 ℃, a printing pressure of 2.5 bar, and a printing speed of 7 mm/s. The resulting biomaterial inks, with a composition of 25% Gel, 25% PCL, and 50% n-HA, demonstrated excellent printability and stability, along with significantly enhanced mechanical properties. As a result, 3D scaffolds with high printability and shape fidelity can be printed at room temperature, followed by deep freezing at -80 ℃ and cross-linking with vanillin. The Gel-based composite scaffolds demonstrated excellent biocompatibility, cell adhesion, cell viability and nano-hydroxyapatite absorption in vitro. Additionally, in vivo experiments revealed that the bioactive scaffold biodegraded during implantation and significantly promoted bone regeneration at the defect site. This provides a promising strategy for treating bone defects in clinical setting. In conclusion, the Gel/PCL/n-HA biomaterial inks presented here offer an innovative solution for extrusion bioprinting in the field of bone tissue engineering.Graphical

Extrusion bioprinting from a fluid mechanics perspective

Bioprinting is an emerging technology for fabricating intricate and diverse structures that closely mimic natural tissues and organs for such applications as tissue engineering, drug delivery, and cancer research as well. Among the various bioprinting techniques, extrusion-based bioprinting stands out due to its capability to apply a wide range of biomaterials and living cells and its controllability over printed structures. In bioprinting, the bioink stored in a syringe is forced to flow through the nozzle connected to the syringe, and then to exit and deposit onto the printing stage to form three-dimensional (3D) structures. The bioprinting process involves the flow of bioink in both syringe and nozzle and then its flow or spreading on a printing stage. As a result, fluid mechanics plays a crucial role in extrusion bioprinting. Notably, the biomaterials used in bioprinting are typically non-Newtonian fluids, which have complex viscoelastic and thixotropic behaviors; and the influence of these behaviors on the bioprinting process has been drawn considerable attention by employing various methods, including the numerical simulations via computational fluid dynamics (CFD). This paper reviews the latest development in the fluid mechanics aspects of extrusion-based bioprinting to shed light on the challenges and key considerations involved. It covers the topics of extrusion bioprinting (including driving mechanisms, printability, cell viability), biomaterial rheology and its effect on bioprinting, multi-material bioprinting and numerical simulation of bioprinting. Key issues and challenges are also discussed along with the recommendations for future research.

Comparison of cation and anion-mediated resolution enhancement of bioprinted hydrogels for membranous tissue fabrication.

Fabrication of engineered thin membranous tissues (TMTs) presents a significant challenge to researchers, as these structures are small in scale, but present complex anatomies containing multiple stratified cell layers. While numerous methodologies exist to fabricate such tissues, many are limited by poor mechanical properties, need for post-fabrication, or lack of cytocompatibility. Extrusion bioprinting can address these issues, but lacks the resolution necessary to generate biomimetic, microscale TMT structures. Therefore, our goal was to develop a strategy that enhances bioprinting resolution below its traditional limit of 150 μm and delivers a viable cell population. We have generated a system to effectively shrink printed gels via electrostatic interactions between anionic and cationic polymers. Base hydrogels are composed of gelatin methacrylate type A (cationic), or B (anionic) treated with anionic alginate, and cationic poly-L-lysine, respectively. Through a complex coacervation-like mechanism, the charges attract, causing compaction of the base GelMA network, leading to reduced sample dimensions. In this work, we evaluate the role of both base hydrogel and shrinking polymer charge on effective print resolution and cell viability. The alginate anion-mediated system demonstrated the ability to reach bioprinting resolutions of 70 μm, while maintaining a viable cell population. To our knowledge, this is the first study that has produced such significant enhancement in extrusion bioprinting capabilities, while also remaining cytocompatible.

Rapid Biofabrication of an Advanced Microphysiological System Mimicking Phenotypical Heterogeneity and Drug Resistance in Glioblastoma.

Microphysiological systems (MPSs) reconstitute tissue interfaces and organ functions, presenting a promising alternative to animal models in drug development. However, traditional materials like polydimethylsiloxane (PDMS) often interfere by absorbing hydrophobic molecules, affecting drug testing accuracy. Additive manufacturing, including 3D bioprinting, offers viable solutions. GlioFlow3D, a novel microfluidic platform combining extrusion bioprinting and stereolithography (SLA) is introduced. GlioFlow3D integrates primary human cells and glioblastoma (GBM) lines in hydrogel-based microchannels mimicking vasculature, within an SLA resin framework using cost-effective materials. The study introduces a robust protocol to mitigate SLA resin cytotoxicity. Compared to PDMS, GlioFlow3D demonstrated lower small molecule absorption, which is relevant for accurate testing of small molecules like Temozolomide (TMZ). Computational modeling is used to optimize a pumpless setup simulating interstitial fluid flow dynamics in tissues. Co-culturing GBM with brain endothelial cells in GlioFlow3D showed enhanced CD133 expression and TMZ resistance near vascular interfaces, highlighting spatial drug resistance mechanisms. This PDMS-free platform promises advanced drug testing, improving preclinical research and personalized therapy by elucidating complex GBM drug resistance mechanisms influenced by the tissue microenvironment.

Bioinks for 3D Bioprinting: A Scientometric Analysis of Two Decades of Progress.

This scientometric analysis of 393 original papers published from January 2000 to June 2019 describes the development and use of bioinks for 3D bioprinting. The main trends for bioink applications and the primary considerations guiding the selection and design of current bioink components (i.e., cell types, hydrogels, and additives) were reviewed. The cost, availability, practicality, and basic biological considerations (e.g., cytocompatibility and cell attachment) are the most popular parameters guiding bioink use and development. Today, extrusion bioprinting is the most widely used bioprinting technique. The most reported use of bioinks is the generic characterization of bioink formulations or bioprinting technologies (32%), followed by cartilage bioprinting applications (16%). Similarly, the cell-type choice is mostly generic, as cells are typically used as models to assess bioink formulations or new bioprinting methodologies rather than to fabricate specific tissues. The cell-binding motif arginine-glycine-aspartate is the most common bioink additive. Many articles reported the development of advanced functional bioinks for specific biomedical applications; however, most bioinks remain the basic compositions that meet the simple criteria: Manufacturability and essential biological performance. Alginate and gelatin methacryloyl are the most popular hydrogels that meet these criteria. Our analysis suggests that present-day bioinks still represent a stage of emergence of bioprinting technology.

Optimising Bioprinting Nozzles through Computational Modelling and Design of Experiments.

3D bioprinting is a promising technique for creating artificial tissues and organs. One of the main challenges of bioprinting is cell damage, due to high pressures and tensions. During the biofabrication process, extrusion bioprinting usually results in low cell viability, typically ranging from 40% to 80%, although better printing performance with higher cell viability can be achieved by optimising the experimental design and operating conditions, with nozzle geometry being a key factor. This article presents a review of studies that have used computational fluid dynamics (CFD) to optimise nozzle geometry. They show that the optimal ranges for diameter and length are 0.2 mm to 1 mm and 8 mm to 10 mm, respectively. In addition, it is recommended that the nozzle should have an internal angle of 20 to 30 degrees, an internal coating of ethylenediaminetetraacetic acid (EDTA), and a shear stress of less than 10 kPa. In addition, a design of experiments technique to obtain an optimal 3D bioprinting configuration for a bioink is also presented. This experimental design would identify bioprinting conditions that minimise cell damage and improve the viability of the printed cells.

Design of magnetic kappa-carrageenan-collagen bioinks for 3D bioprinting

Bioprinting approaches are of great promise for tissue engineering applications as they allow the fabrication of constructs able to mimic native tissues’ mechanical and topographical features. Additional control over cells fate can be enhanced using stimuli-responsive materials, requiring the development of novel bioinks for this purpose. In this study, bioinks comprising κ-carrageenan, collagen, and magnetic nanoparticles were designed for 3D bioprinting applications. The characterization of this material was performed, where mechanical compressive tests yielded Young’s moduli ranging from 8.25 to 18.4 kPa. Rheological assessments also revealed the shear-thinning behavior of the bioinks and a temperature-dependent gelation. The capability of these bioinks to produce 3D constructs by extrusion bioprinting was established through the printability evaluation and the development of complex structures, supporting the viability and proliferation of mesenchymal stromal cells (MSCs). Finally, as proof-of-concept, it was observed that the secretome of bioprinted MSCs stimulated with an external magnetic field of 80 mT was able to increase the number of tubes formed by human umbilical vein endothelial cells.

Bioprinting of Synthetic Cell-like Lipid Vesicles to Augment the Functionality of Tissues after Manufacturing.

Bioprinting is an automated bioassembly method that enables the formation of human tissue-like constructs to restore or replace damaged tissues. Regardless of the employed bioprinting method, cells undergo mechanical stress that can impact their survival and function postprinting. In this study, we investigate the use of a synthetic cell-like unit, giant unilamellar vesicles (GUVs), as adjuvants of the cellular function of human cells postprinting, or in future as the complete replacement of human cells. We analyzed the impact of two nozzle-based bioprinting methods (drop-on-demand and extrusion bioprinting) on the structure, stability, and function of GUVs. We showed that over 65% of the GUVs remain intact when printing at 0.5 bar, demonstrating the potential of using GUVs as a synthetic cell source. We further increased the stability of GUVs in a cell culture medium by introducing polyethylene glycol (PEG) into the GUV lipid membrane. The presence of PEG, however, diminished the structural properties of GUVs postprinting, and reduced the interaction of GUVs with human cells. Although the design of PEG-GUVs can still be modified in future studies for better cell-GUV interactions, we demonstrated that GUVs are functional postprinting. Chlorin e6-PEG-GUVs loaded with a fluorescent dye were bioprinted, and they released the dye postprinting only upon illumination. This is a new strategy to deliver carriers, such as growth factors, drugs, nutrients, or gases, inside large bioprinted specimens on a millimeter to centimeter scale. Overall, we showed that printed GUVs can augment the functionality of manufactured human tissues.

Triaxial mechanical characterization of ultrasoft 3D support bath-based bioprinted tubular GelMA constructs

Support bath-based extrusion bioprinting is an emerging additive manufacturing paradigm that allows for the creation of geometrically complex three-dimensional tissue-mimicking constructs. Although this approach enables arbitrary geometries to be printed, preserving shape and mechanical integrity after the construct is extracted from the support bath is one of the main challenges today. In the present study, we systematically tested the effect of critical printing parameters on construct integrity and stiffness. Specifically, we varied the concentration and temperature of the bioink and support bath material, hydrogel crosslinking time, and cell density of the bioink. While previous studies on hydrogel materials quantify construct properties based on a single loading mode-such as uniaxial testing or rheological experiments, for example—we used a multiaxial mechanical testing protocol to assess the printed construct’s response in tension, compression, and shear. For each sample, we determined a single set of material parameters by simultaneously fitting all three modes in our inverse finite element framework. In support of future modeling work, we analyzed three different constitutive models with respect to their ability to match the construct’s mechanical response. We observed that GelMA concentration, temperature, and cell density have a statistically significant effect on the construct stiffness; support bath temperature and UV crosslinking time have a weak effect on construct stiffness; and support bath concentration appears to have no direct effect on the measured construct properties. Our construct stiffness were found to vary between 0.07 and 2.2 kPa depending on printing conditions, and showed noticeable tension–compression asymmetry as well as pronounced nonlinear behavior when loaded under shear.

Advanced optical assessment and modeling of extrusion bioprinting

In the context of tissue engineering, biofabrication techniques are employed to process cells in hydrogel-based matrices, known as bioinks, into complex 3D structures. The aim is the production of functional tissue models or even entire organs. The regenerative production of biological tissues adheres to a multitude of criteria that ultimately determine the maturation of a functional tissue. These criteria are of biological nature, such as the biomimetic spatial positioning of different cell types within a physiologically and mechanically suitable matrix, which enables tissue maturation. Furthermore, the processing, a combination of technical procedures and biological materials, has proven highly challenging since cells are sensitive to stress, for example from shear and tensile forces, which may affect their vitality. On the other hand, high resolutions are pursued to create optimal conditions for subsequent tissue maturation. From an analytical perspective, it is prudent to first investigate the printing behavior of bioinks before undertaking complex biological tests. According to our findings, conventional shear rheological tests are insufficient to fully characterize the printing behavior of a bioink. For this reason, we have developed optical methods that, complementarily to the already developed tests, allow for quantification of printing quality and further viscoelastic modeling of bioinks.

Three-Dimensional Bioprinting of GelMA Hydrogels with Culture Medium: Balancing Printability, Rheology and Cell Viability for Tissue Regeneration

Three-dimensional extrusion bioprinting technology aims to become a fundamental tool for tissue regeneration using cell-loaded hydrogels. These biomaterials must have highly specific mechanical and biological properties that allow them to generate biosimilar structures by successive layering of material while maintaining cell viability. The rheological properties of hydrogels used as bioinks are critical to their printability. Correct printability of hydrogels allows the replication of biomimetic structures, which are of great use in medicine, tissue engineering and other fields of study that require the three-dimensional replication of different tissues. When bioprinting cell-loaded hydrogels, a small amount of culture medium can be added to ensure adequate survival, which can modify the rheological properties of the hydrogels. GelMA is a hydrogel used in bioprinting, with very interesting properties and rheological parameters that have been studied and defined for its basic formulation. However, the changes that occur in its rheological parameters and therefore in its printability, when it is mixed with the culture medium necessary to house the cells inside, are unknown. Therefore, in this work, a comparative study of GelMA 100% and GelMA in the proportions 3:1 (GelMA 75%) and 1:1 (GelMA 50%) with culture medium was carried out to determine the printability of the gel (using a device of our own invention), its main rheological parameters and its toxicity after the addition of the medium and to observe whether significant differences in cell viability occur. This raises the possibility of its use in regenerative medicine using a 3D extrusion bioprinter.

Assessing Bioprinted Functionalized Grafts for Biological Tendon Augmentation In Vitro.

Tendinopathy, characterized by inflammatory and degenerative changes, presents challenges in sports and medicine. In addressing the limitations of conservative management, this study focuses on developing tendon grafts using extrusion bioprinting with platelet-rich plasma (PRP)-infused hydrogels loaded with tendon cells. The objective is to understand paracrine interactions initiated by bioprinted tendon grafts in either inflamed or non-inflamed host tissues. PRP was utilized to functionalize methacrylate gelatin (GelMA), incorporating tendon cells for graft bioprinting. Bioinformatic analyses of overexpressed proteins, predictive of functional enrichment, revealed insights into PRP graft behavior in both non-inflamed and inflamed environments. PRP grafts activated inflammatory pathways, including Interleukin 17 (IL-17), neuroinflammation, Interleukin 33 (IL-33), and chemokine signaling. Interleukin 1 beta (IL-1b) in the graft environment triggered p38 mitogen-activated protein kinase (MAPK) signaling, nuclear factor kappa light chain enhancer of activated B cells (NF-kB) canonical pathway, and Vascular Endothelial Growth Factor (VEGF) signaling. Biological enrichment attributed to PRP grafts included cell chemotaxis, collagen turnover, cell migration, and angiogenesis. Acellular PRP grafts differed from nude grafts in promoting vessel length, vessel area, and junction density. Angiogenesis in cellular grafts was enhanced with newly synthesized Interleukin 8 (IL-8) in cooperation with IL-1b. In conclusion, paracrine signaling from PRP grafts, mediated by chemokine activities, influences cell migration, inflammation, and angiogenic status in host tissues. Under inflammatory conditions, newly synthesized IL-8 regulates vascularization in collaboration with PRP.