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1.
Bioact Mater ; 43: 392-405, 2025 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-39399841

RESUMO

Digital light processing (DLP)-based bioprinting technology holds immense promise for the advancement of hydrogel constructs in biomedical applications. However, creating high-performance hydrogel constructs with this method is still a challenge, as it requires balancing the physicochemical properties of the matrix while also retaining the cellular activity of the encapsulated cells. Herein, we propose a facile and practical strategy for the 3D bioprinting of high-performance hydrogel constructs through the in-situ birth of stem cell spheroids. The strategy is achieved by loading the cell/dextran microdroplets within gelatin methacryloyl (GelMA) emulsion, where dextran functions as a decoy to capture and aggregate the cells for bioprinting while GelMA enables the mechanical support without losing the structural complexity and fidelity. Post-bioprinting, the leaching of dextran results in a smooth curved surface that promotes in-situ birth of spheroids within hydrogel constructs. This process significant enhances differentiation potential of encapsulated stem cells. As a proof-of-concept, we encapsulate dental pulp stem cells (DPSCs) within hydrogel constructs, showcasing their regenerative capabilities in dentin and neovascular-like structures in vivo. The strategy in our study enables high-performance hydrogel tissue construct fabrication with DLP-based bioprinting, which is anticipated to pave a promising way for diverse biomedical applications.

2.
Front Bioeng Biotechnol ; 12: 1452477, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39380897

RESUMO

In order to recreate the complexity of human organs, the field of tissue engineering and regenerative medicine has been focusing on methods to build organs from the bottom up by assembling distinct small functional units consisting of a biomaterial and cells. This bottom-up engineering requires bioinks that can be assembled by 3D bioprinting and that permit fast vascularization of the construct to ensure survival of embedded cells. To this end, a small molecular weight alginate (SMWA) gel porogen is presented herein. Alginate is a biocompatible biomaterial, which can be easily converted into small porogen gels with the procedure reported in this article. The SMWA porogen is mixed with photo-crosslinkable hydrogels and leached from the hydrogel post-crosslinking to increase porosity and facilitate vascularization. As a proof of concept, this system is tested with the commonly used biomaterial Gelatin Methacryloyl (GelMA). The SMWA porogen-GelMA blend is proven to be bioprintable. Incubating the blend for 20 min in a low concentration phosphate buffered saline and sodium citrate solution significantly reduces the remaining porogen in the hydrogel . The intent to completely leach the porogen from the hydrogel was abandoned, as longer incubation times and higher concentrations of phosphate and citrate were detrimental to endothelial proliferation. Nonetheless, even with remnants of the porogen left in the hydrogel, the created porosity significantly improves viability, growth factor signaling, vasculogenesis, and angiogenesis in 3D bioprinted structures. This article concludes that the usage of the SMWA porogen can improve the assembly of microvasculature in 3D bioprinted structures. This technology can benefit the bottom-up assembly of large scaffolds with high cell density through 3D bioprinting by improving cell viability and allowing faster vascularization.

3.
Sci Rep ; 14(1): 23240, 2024 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-39369014

RESUMO

Gelatin methacryloyl (GelMA), typically derived from mammalian sources, has recently emerged as an ideal bio-ink for three-dimensional (3D) bioprinting. Herein, we developed a fish skin-based GelMA bio-ink for the fabrication of a 3D GelMA skin substitute with a 3D bioprinter. Several concentrations of methacrylic acid anhydride were used to fabricate GelMA, in which their physical-mechanical properties were assessed. This fish skin-based GelMA bio-ink was loaded with human adipose tissue-derived mesenchymal stromal cells (ASCs) and human platelet lysate (HPL) and then printed to obtain 3D ASCs + HPL-loaded GelMA scaffolds. Cell viability test and a preliminary investigation of its effectiveness in promoting wound closure were evaluated in a critical-sized full thickness skin defect in a rat model. The cell viability results showed that the number of ASCs increased significantly within the 3D GelMA hydrogel scaffold, indicating its biocompatibility property. In vivo results demonstrated that ASCs + HPL-loaded GelMA scaffolds could delay wound contraction, markedly enhanced collagen deposition, and promoted the formation of new blood vessels, especially at the wound edge, compared to the untreated group. Therefore, this newly fish skin-based GelMA bio-ink developed in this study has the potential to be utilized for the printing of 3D GelMA skin substitutes.


Assuntos
Bioimpressão , Gelatina , Células-Tronco Mesenquimais , Impressão Tridimensional , Pele Artificial , Alicerces Teciduais , Gelatina/química , Animais , Bioimpressão/métodos , Humanos , Ratos , Alicerces Teciduais/química , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Peixes , Metacrilatos/química , Pele/metabolismo , Pele/efeitos dos fármacos , Tinta , Cicatrização/efeitos dos fármacos , Engenharia Tecidual/métodos , Sobrevivência Celular/efeitos dos fármacos , Hidrogéis/química , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Materiais Biocompatíveis/química
4.
Biotechnol J ; 19(10): e202400393, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39380502

RESUMO

In light-based 3D-bioprinting, gelatin methacrylate (GelMA) is one of the most widely used materials, as it supports cell attachment, and shows good biocompatibility and degradability in vivo. However, as an animal-derived material, it also causes safety concerns when used in medical applications. Gelatin is a partial hydrolysate of collagen, containing high amounts of hydroxyproline. This causes the material to form a thermally induced gel at ambient temperatures, a behavior also observed in GelMA. This temperature-dependent gelation requires precise temperature control during the bioprinting process to prevent the gelation of the material. To avoid safety concerns associated with animal-derived materials and reduce potential issues caused by thermal gelation, a recombinant human alpha-1 collagen I fragment was expressed in Komagataella phaffii without hydroxylation. The resulting protein was successfully modified with methacryloyl groups and underwent rapid photopolymerization upon ultraviolet light exposure. The developed material exhibited slightly slower polymerization and lower storage modulus compared to GelMA, while it showed higher stretchability. However, unlike the latter, the material did not undergo physical gelation at ambient temperatures, but only when cooled down to below 10°C, a characteristic that has not been described for comparable materials so far. This gelation was not caused by the formation of triple-helical structures, as shown by the absence of the characteristic peak at 220 nm in CD spectra. Moreover, the developed recombinant material facilitated cell adherence with high cell viability after crosslinking via light to a 3D structure. Furthermore, desired geometries could be easily printed on a stereolithographic bioprinter.


Assuntos
Bioimpressão , Gelatina , Metacrilatos , Polimerização , Proteínas Recombinantes , Humanos , Bioimpressão/métodos , Proteínas Recombinantes/química , Gelatina/química , Metacrilatos/química , Impressão Tridimensional , Colágeno Tipo I/química , Materiais Biocompatíveis/química , Colágeno/química , Temperatura , Animais
5.
J Biomater Sci Polym Ed ; : 1-16, 2024 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-39460952

RESUMO

Restoring cartilage to healthy state is challenging due to low cell density and hence low regenerative capacity. The current platforms are not compatible with clinical translation and require dedicated handling of trained personnel. However, by engineering and implanting cell microaggregates in higher concentrations, efficient formation of new cartilage can be achieved, even in the absence of exogenous growth factors. Therefore, one-step surgeries are preferable for novel treatments and we need cell laden microgels allowing the formation of microaggregaets in vivo. Injectability is a key parameter for in situ forming the shape and minimally invasive clinical applications. Hydrogels as bioinks can restore damaged tissues to their primary shape. Chitosan is a polysaccharide derived from chitin with abundant usage in tissue engineering. This review highlights the use of chitosan as an injectable hydrogel for osteochondral defects. Several studies focused on encapsulating mesenchymal stem cells within chitosan hydrogels have been categorized and incorporating microfluidic devices has been identified in the forefront to form microgels. Additionally, the printability is another convenience of chitosan for using in 3D printing for cartilage tissue engineering which is described in this review.

6.
Biomimetics (Basel) ; 9(10)2024 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-39451795

RESUMO

This study systematically investigates the effects of the coaxial nozzle's inner nozzle diameter on the strength and gelation of filaments produced via extrusion-based 3D printing with in situ ionic crosslinking. In this system, bioink (sodium alginate solution) was extruded through the outer nozzle, and the ionic crosslinking solution (calcium chloride solution) was extruded through the inner nozzle. The outer nozzle diameter was fixed at 2.16 mm, and the inner nozzle diameter was varied among 1.19, 0.84, and 0.584 mm. The results indicate that, as the inner nozzle diameter decreased, filament strength decreased, and filament gelation became poorer. These findings highlight the importance of optimizing inner nozzle diameter for improved filament strength and gelation in extrusion-based 3D printing with in situ ionic crosslinking.

7.
Biomed Eng Comput Biol ; 15: 11795972241288099, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39364141

RESUMO

Tissue engineering is a multidisciplinary field that uses biomaterials to restore tissue function and assist with drug development. Over the last decade, the fabrication of three-dimensional (3D) multifunctional scaffolds has become commonplace in tissue engineering and regenerative medicine. Thanks to the development of 3D bioprinting technologies, these scaffolds more accurately recapitulate in vivo conditions and provide the support structure necessary for microenvironments conducive to cell growth and function. The purpose of this review is to provide a background on the leading 3D bioprinting methods and bioink selections for tissue engineering applications, with a specific focus on the growing field of developing multifunctional bioinks and possible future applications.

8.
Adv Healthc Mater ; : e2402680, 2024 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-39466900

RESUMO

Autologous fat is widely used in soft tissue reconstruction; however, significant volume reduction owing to necrosis and degradation of the transplanted adipose tissue (AT) remains a major challenge. To address this issue, a novel live AT micro-fragment-based bio-ink (ATmf bio-ink) compatible with precision 3D printing, is developed. Live AT micro-fragments of ≈280 µm in size are prepared using a custom tissue micronizer and they are incorporated into a fibrinogen/gelatin mixture to create the ATmf bio-ink. AT micro-fragments exhibit high viability and preserve the heterogeneous cell population and extracellular matrix of the native AT. The developed bio-ink enables precise micropatterning and provides an excellent adipo-inductive microenvironment. AT grafts produced by co-printing the bio-ink with polycaprolactone demonstrate a 500% improvement in volume retention and a 300% increase in blood vessel infiltration in vivo compared with conventional microfat grafts. In vivo engraftment of AT grafts is further enhanced by using a stem cell-laden ATmf bio-ink. Last, it is successfully demonstrated that the bio-ink is enabled for the creation of clinically relevant and patient-specific AT grafts for patients undergoing partial mastectomy. This novel ATmf bio-ink for volumetric soft tissue reconstruction offers a pioneering solution for addressing the limitations of existing clinical techniques.

9.
J Tissue Eng ; 15: 20417314241286092, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39411269

RESUMO

The liver coordinates over 500 biochemical processes crucial for maintaining homeostasis, detoxification, and metabolism. Its specialized cells, arranged in hexagonal lobules, enable it to function as a highly efficient metabolic engine. However, diseases such as cirrhosis, fatty liver disease, and hepatitis present significant global health challenges. Traditional drug development is expensive and often ineffective at predicting human responses, driving interest in advanced in vitro liver models utilizing 3D bioprinting and microfluidics. These models strive to mimic the liver's complex microenvironment, improving drug screening and disease research. Despite its resilience, the liver is vulnerable to chronic illnesses, injuries, and cancers, leading to millions of deaths annually. Organ shortages hinder liver transplantation, highlighting the need for alternative treatments. Tissue engineering, employing polymer-based scaffolds and 3D bioprinting, shows promise. This review examines these innovative strategies, including liver organoids and liver tissue-on-chip technologies, to address the challenges of liver diseases.

10.
ACS Biomater Sci Eng ; 2024 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-39425970

RESUMO

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.

11.
ACS Biomater Sci Eng ; 10(10): 5995-6016, 2024 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-39228365

RESUMO

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.


Assuntos
Bioimpressão , Hidrogéis , Impressão Tridimensional , Cicatrização , Hidrogéis/química , Hidrogéis/uso terapêutico , Humanos , Bioimpressão/métodos , Animais , Engenharia Tecidual/métodos
12.
Biomater Adv ; 165: 214017, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-39236580

RESUMO

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.


Assuntos
Bioimpressão , Fígado , Células-Tronco Mesenquimais , Osteogênese , Ratos Sprague-Dawley , Engenharia Tecidual , Alicerces Teciduais , Animais , Alicerces Teciduais/química , Engenharia Tecidual/métodos , Bioimpressão/métodos , Ratos , Células-Tronco Mesenquimais/citologia , Osteogênese/fisiologia , Fígado/citologia , Impressão Tridimensional , Matriz Extracelular Descelularizada/química , Regeneração Óssea/fisiologia , Gelatina/química , Diferenciação Celular , Alginatos/química , Proliferação de Células , Matriz Extracelular/química , Osso e Ossos/fisiologia , Tinta
13.
Environ Sci Pollut Res Int ; 31(44): 55984-55995, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39251533

RESUMO

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.


Assuntos
Bioimpressão , Recuperação e Remediação Ambiental , Impressão Tridimensional , Recuperação e Remediação Ambiental/métodos , Substâncias Perigosas , Biodegradação Ambiental
14.
Adv Healthc Mater ; : e2400286, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-39235370

RESUMO

This study proposes a novel, versatile, and modular platform for constructing porous and heterogeneous microenvironments based on the embedding of liquefied-based compartments in hydrogel systems. Using a bottom-up approach, microgels carrying the necessary cargo components, including cells and microparticles, are combined with a hydrogel precursor to fabricate a hierarchical structured (HS) system. The HS system possesses three key features that can be fully independently controlled: I) liquefied pockets enabling free cellular mobility; II) surface modified microparticles facilitating 3D microtissue organization inside the liquefied pockets; III) at a larger scale, the pockets are jammed in the hydrogel, forming a macro-sized construct. After crosslinking, the embedded microgels undergo a liquefaction process, forming a porous structure that ensures high diffusion of small biomolecules and enables cells to move freely within their miniaturized compartmentalized volume. More importantly, this platform allows the creation of multimodular cellular microenvironments within a hydrogel with controlled macrostructures, while decoupling micro- and macroenvironments. As a proof of concept, the enhancement of cellular functions using the HS system by encapsulating human adipose-derived mesenchymal stem cells (hASCs) is successfully demonstrated. Finally, the potential application of this system as a hybrid bioink for bioprinting complex 3D structures is showcased.

15.
J Tissue Eng ; 15: 20417314241282476, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39345255

RESUMO

Three-dimensional (3D) bioprinting has emerged as a promising strategy for fabricating complex tissue analogs with intricate architectures, such as vascular networks. Achieving this necessitates bioink formulations that possess highly printable properties and provide a cell-friendly microenvironment mimicking the native extracellular matrix. Rapid advancements in printing techniques continue to expand the capabilities of researchers, enabling them to overcome existing biological barriers. This review offers a comprehensive examination of ultraviolet-based 3D bioprinting, renowned for its exceptional precision compared to other techniques, and explores its applications in inducing angiogenesis across diverse tissue models related to hypoxia. The high-precision and rapid photocuring capabilities of 3D bioprinting are essential for accurately replicating the intricate complexity of vascular networks and extending the diffusion limits for nutrients and gases. Addressing the lack of vascular structure is crucial in hypoxia-related diseases, as it can significantly improve oxygen delivery and overall tissue health. Consequently, high-resolution 3D bioprinting facilitates the creation of vascular structures within three-dimensional engineered tissues, offering a potential solution for addressing hypoxia-related diseases. Emphasis is placed on fundamental components essential for successful 3D bioprinting, including cell types, bioink compositions, and growth factors highlighted in recent studies. The insights provided in this review underscore the promising prospects of leveraging 3D printing technologies for addressing hypoxia-related diseases through the stimulation of angiogenesis, complementing the therapeutic efficacy of cell therapy.

16.
Bioengineering (Basel) ; 11(9)2024 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-39329604

RESUMO

This paper reports an experimental study on the compatibility of human bronchial epithelial (HBE) cells in a collagen-alginate bioink. The compatibility was assessed using the culture well method with three bioink compositions prepared from a 10% alginate solution and neutralized TeloCol-10 mg/mL collagen stock solution. Cell viability, quantified by (live cell count-dead cell count)/live cell count within the HBE cell-laden hydrogel, was evaluated using the live/dead assay method from Day 0 to Day 6. Experimental results demonstrated that the collagen-alginate 4:1 bioink composition exhibited the highest cell viability on Day 6 (85%), outperforming the collagen-alginate 1:4 bioink composition and the alginate bioink composition, which showed cell viability of 75% and 45%, respectively. Additionally, the live cell count was highest for the collagen-alginate 4:1 bioink composition on Day 0, a trend that persisted through Days 1 to 6, underscoring its superior performance in maintaining cell viability and promoting cell proliferation. These findings show that the compatibility of HBE cells with the collagen-alginate 4:1 bioink composition was higher compared with the other two bioink compositions.

17.
Sheng Wu Gong Cheng Xue Bao ; 40(9): 2934-2947, 2024 Sep 25.
Artigo em Chinês | MEDLINE | ID: mdl-39319716

RESUMO

Cardiovascular diseases are major diseases, and there is lack of artificial blood vessels with small diameters which can be applied in coronary artery bypass surgery. The conventional vascular scaffold preparation techniques in tissue engineering have shortcomings in regulating the diameter, geometric shape, and interconnectivity of the scaffold. 3D bioprinting can simulate the natural structure of the vascular tissue, accurately print live cells and biomaterials, and regulate the microstructure and porosity of scaffolds on the nanoscale, providing new ideas for vascular tissue engineering. This article systematically evaluates the classification of 3D bioprinting technologies and reviews the latest research progress of 3D bioprinting in vascular tissue engineering. It summarizes the advantages of 3D bioprinting and points out the problems that need to be solved, such as the immune rejection of blood vessel materials, providing reference for the further research.


Assuntos
Bioimpressão , Impressão Tridimensional , Engenharia Tecidual , Alicerces Teciduais , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Humanos , Vasos Sanguíneos , Materiais Biocompatíveis , Prótese Vascular
18.
Polymers (Basel) ; 16(17)2024 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-39274089

RESUMO

Chronic wounds, such as diabetic foot ulcers, pressure ulcers, and venous ulcers, pose significant clinical challenges and burden healthcare systems worldwide. The advent of 3D bioprinting technologies offers innovative solutions for enhancing chronic wound care. This scoping review evaluates the applications, methodologies, and effectiveness of 3D-printed bioinks in chronic wound healing, focusing on bioinks incorporating living cells to facilitate wound closure and tissue regeneration. Relevant studies were identified through comprehensive searches in databases, including PubMed, Scopus, and Web of Science databases, following strict inclusion criteria. These studies employ various 3D bioprinting techniques, predominantly extrusion-based, to create bioinks from natural or synthetic polymers. These bioinks are designed to support cell viability, promote angiogenesis, and provide structural integrity to the wound site. Despite these promising results, further research is necessary to optimize bioink formulations and printing parameters for clinical application. Overall, 3D-printed bioinks offer a transformative approach to chronic wound care, providing tailored and efficient solutions. Continued development and refinement of these technologies hold significant promise for improving chronic wound management and patient outcomes.

19.
Polymers (Basel) ; 16(17)2024 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-39274103

RESUMO

Three-dimensional (3D) bioprinting has emerged as an important technique for fabricating tissue constructs with precise structural and compositional control. However, developing suitable bioinks with biocompatible crosslinking mechanisms remains a significant challenge. This study investigates extrusion-based bioprinting (EBB) using uniaxial or coaxial nozzles with enzymatic crosslinking (EC) to produce 3D tissue constructs in vitro. Initially, low-molecular-weight dextran-tyramine and hyaluronic acid-tyramine (LMW Dex-TA/HA-TA) bioink prepolymers were evaluated. Enzymatically pre-crosslinking these prepolymers, achieved by the addition of horseradish peroxidase and hydrogen peroxide, produced viscous polymer solutions. However, this approach resulted in inconsistent bioprinting outcomes (uniaxial) due to inhomogeneous crosslinking, leading to irreproducible properties and suboptimal shear recovery behavior of the hydrogel inks. To address these challenges, we explored a one-step coaxial bioprinting system consisting of enzymatically crosslinkable high-molecular-weight hyaluronic acid-tyramine conjugates (HMW HA-TA) mixed with horseradish peroxidase (HRP) in the inner core and a mixture of Pluronic F127 and hydrogen peroxide in the outer shell. This configuration resulted in nearly instantaneous gelation by diffusion of the hydrogen peroxide into the core. Stable hydrogel fibers with desirable properties, including appropriate swelling ratios and controlled degradation rates, were obtained. The optimized bioink and printing parameters included 1.3% w/v HMW HA-TA and 5.5 U/mL HRP (bioink, inner core), and 27.5% w/v Pluronic F127 and 0.1% H2O2 (sacrificial ink, outer shell). Additionally, optimal pressures for the inner core and outer shell were 45 and 80 kPa, combined with a printing speed of 300 mm/min and a bed temperature of 30 °C. The extruded HMW HA-TA core filaments, containing bovine primary chondrocytes (BPCs) or 3T3 fibroblasts (3T3 Fs), exhibited good cell viabilities and were successfully cultured for up to seven days. This study serves as a proof-of-concept for the one-step generation of core filaments using a rapidly gelling bioink with an enzymatic crosslinking mechanism, and a coaxial bioprinter nozzle system. The results demonstrate significant potential for developing designed, printed, and organized 3D tissue fiber constructs.

20.
Biomater Adv ; 166: 214042, 2024 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-39293254

RESUMO

Bone tissue regeneration remains a significant challenge in clinical settings due to the complexity of replicating the mechanical and biological properties of bone environment. This study addresses this challenge by proposing a hybrid scaffold designed to enhance both bioactivity and physical stability for bone tissue regeneration. This research is the fisrt to develop a rigid 3D structure composed of polycaprolactone (PCL) and hydroxyapatite nanoparticles (nHA) integrated with a bioink containing human dental pulp stem/stromal cells (hDPSCs), alginate, nHA and collagen (Col). The biofabricated constructs were extensively characterized through cytocompatibility tests, osteogenic differentiation assessment, and biocompatibility evaluation in a rat model. In vitro results demontrated that the hybrid scaffolds presented significantly higher cell viability after 168 h compared to the control group. Furthermore, the hybrid scaffolds showed increased osteogenic differentiation relative to other groups. In vivo evaluation indicated good biocompatibility, characterized by minimal inflammatory response and successful tissue integration. These findings highlight the scaffold's potential to support bone tissue regeneration by combining the mechanical strength of PCL and nHA with the biological activity of the alginate-nHA-Col and hDPSCs bioink. The current study provides a promising foundation for the development of biomaterials aimed at improving clinical outcomes in bone repair and regeneration, particulary for the treatment of critical-size bone defects, targeted drug administration, and three-dimensional models for bone tissue engineering.

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