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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES · 2021 · IF 6.953

Tendon-inspired fibers from liquid crystalline collagen as the pre-oriented bioink

Feng Deng,Yuan Dang,Lele Tang,Tianshuo Hu,Cuicui Ding,Xiaoqing Hu,Hui Wu,Lihui Chen,Liulian Huang,Yonghao Ni,Min Zhang

Nature provides rich bionic resources for the construction of advanced materials with excellent mechanical properties. In this work, inspired by animal tendons, a bionic collagen fiber was developed using collagen liquid crystals as the pre-oriented bioink. The texture of liquid crystalline collagen observed from polarized optical microscopy (POM) showed the specific molecular pre-orientation. Meanwhile, the collagen spinning liquids exhibited a minimal rise in viscosity upon increasing concentration from 60 to 120 mg/mL, indicating the feasible processability . The collagen fiber, which was prepared via wet spinning without being denatured, exhibited the favorable orientation of fibrils along its axis as observed with FESEM and AFM . Thanks to the synergistic effects between pre-orientation and shearing orientation, the maximum tensile strength and Young's modulus of collagen fibers reached 9.98 cN/tex (219.29 ± 22.92 MPa) and 43.95 ± 1.11 cN/tex (966.20 ± 24.30 MPa), respectively, which were also analogous to those of tendon. In addition, the collagen fiber possessed a desirable wet strength . Benefiting from the natural tissue affinity of collagen, the as-prepared bionic collagen fiber possessed excellent wound suture performance and biodegradability in vivo , which offers a new perspective for the potential of widespread applications of collagen fibers in biomedical fields .

研究领域: 生物材料学 · 医学工程
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Additive Manufacturing · 2022 · IF 11.632

Material extrusion-based 3D printing for the fabrication of bacteria into functional biomaterials: The case study of ammonia removal application

Yan Li,Shuqiang Peng,Kunrong Li,Dan Qin,Zixiang Weng,Jiangwei Li,Longhui Zheng,Lixin Wu,Chang-Ping Yu

3D printing provides a new way of microbial immobilization technology and creates innovative designed bioactive structures containing microorganisms for remediation of environmental pollution. However, the bio-ink design remains a critical challenge due to the difficulty in creating a durable and bio-friendly material. Here, a novel dual-crosslinking PEGDA-Alginate-PVA-Nanoclay (PAPN) bio-ink containing one heterotrophic bacterium is reported for 3D printing functional biomaterial with the capabilities of ammonia removal. The results showed that PAPN 3D printed bio-scaffolds could effectively remove 96.2 ± 1.3% ammonia within 12 h, and the removal rate increased within repeated use owing to the growth of bacteria inside the bio-scaffolds. Preservation of the bio-scaffolds under room temperature without a culture medium for 168 h still maintained the microbial activity for ammonia removal. It is demonstrated that visible-light-based 3D printing procedures could maintain high cell viability for the majority of bacteria, and the porous structures of 3D bio-scaffolds could allow the permeability of nutrients for the growth of bacteria. This work demonstrates the potential of the dual crosslinked PAPN 3D bio-scaffold for the production and application of immobilized functional bacteria in wastewater treatment .

CAS / MDL: 10043-52-4
研究领域: 环境科学 · 生物技术
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Materials Science & Engineering C-Materials for Biological Applications · 2021 · IF 7.328

Low-temperature 3D printing of collagen and chitosan composite for tissue engineering

Hairui Suo,Jiaying Zhang,Mingen Xu,Ling Wang

Three-dimensional (3D) printing is a promising method to prepare scaffolds for tissue regeneration. Collagen and chitosan composites are superior materials for tissue engineering scaffold but rarely printed due to their poor printability. Here, we prepared a series of tunable hybrid collagen/chitosan bioinks with significantly improved printability through hydrogen bond interaction and printed them into scaffolds by carefully controlling the temperature. Rheological tests proved the printable bioinks had sound shear thinning behavior, dramatical viscosity variation with temperature, and the gelation temperature from 7 to 10 °C. Chitosan could decrease the swelling ratio of the printed scaffolds, while their degradation rate increased with collagen proportion and the values of Young's modulus and tensile strength increased with chitosan proportion. Moreover, the scaffolds containing 2% (m/v) collagen and 2% (m/v) chitosan had a homogeneous and compact honeycomb-like structure, demonstrating the strengthening effect of chitosan. Cell viability assay presented vigorous cell growth on the surface of scaffolds, meanwhile, live cells were also found inside and at the bottom of the scaffolds, indicating the migration of cells. Therefore, chitosan can improve the printability of collagen and the hybrid collagen/chitosan bioinks can be printed into scaffolds with regulated properties, thus can fit different applications in tissue engineering.

CAS / MDL: 9012-76-4
研究领域: 生物材料学 · 再生医学
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Micromachines · 2020 ·Vol. 11 ·期 7 ·681 · IF 2.524

Fabrication of Microspheres from High-Viscosity Bioink Using a Novel Microfluidic-Based 3D Bioprinting Nozzle

Shanguo Zhang,Guiling Li,Jia Man,Song Zhang,Jianyong Li,Jianfeng Li,Donghai Li

Three-dimensional (3D) bioprinting is a novel technology utilizing biocompatible materials, cells, drugs, etc. as basic microcomponents to form 3D artificial structures and is believed as a promising method for regenerative medicine. Droplet-based bioprinting can precisely generate microspheres and manipulate them into organized structures with high fidelity. Biocompatible hydrogels are usually used as bioinks in 3D bioprinting, however, the viscosity of the bioink could be increased due to the additives such as cells, drugs, nutrient factors and other functional polymers in some particular applications, making it difficult to form monodispersed microspheres from high-viscosity bioink at the orifice of the nozzle. In this work, we reported a novel microfluidic-based printing nozzle to prepare monodispersed microspheres from high-viscosity bioink using the phase-inversion method. Different flowing conditions can be achieved by changing the flow rates of the fluids to form monodispersed solid and hollow microspheres using the same nozzle. The diameter of the microspheres can be tuned by changing the flow rate ratio and the size distribution of the microspheres is narrow. The prepared calcium alginate microspheres could also act as micro-carriers in drug delivery.Keywords:droplet-based bioprinting;microfluidic system;phase-inversion method;microcomponent

CAS / MDL: 1317-61-9
研究领域: 生物医学工程 · 再生医学
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BIOMACROMOLECULES · 2025 · IF 5.4

Enhanced Near-Infrared Light-Mediated Hydrogel Curing Using Photoinitiator Integrated Upconversion Particles as Nano Initiator for 3D Bioprinting

Xiong Xiao,Ziwei Huang,Hongying Duan,Liping Yang,Yuchu Yang,Yushang Lai,Chenxi Li,Li Feng

In vivo three-dimensional (3D) bioprinting is a promising strategy that can enable personalized organ repair with minimal injury. The current in vivo 3D bioprinting based on upconversion nanoparticles (UCNPs) mediating near-infrared (NIR) light curing is still limited by the low hydrogel cross-linking efficiency. Herein, we introduced a bioink system that allows enhanced NIR light curing by utilizing thiol–ene cross-linkable polymers and photoinitiator-modified UCNPs@LAP nano initiator. The norbornene functionalized hyaluronic acid (NorHA) and thiolated gelatin (GelSH) were first synthesized to prepare the thiol–ene polymer solution. Compared to radical cross-linkable gelatin methacryloyl (GelMA), the NorHA/GelSH exhibited much higher reactivity under weak photoinitiating conditions. With the addition of surface-modified UCNPs@LAP nano initiator, the bioinks showed improved NIR curing performances, which is beneficial to reduce potential thermal damage. Furthermore, in vitro evaluation showed that the NIR light-cured 3D scaffolds preserved excellent bioactivity, suggesting that the hybrid bioink holds great promise to serve as a candidate for in vivo 3D bioprinting.

研究领域: 生物医学工程 · 材料科学
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JOURNAL OF MATERIALS SCIENCE & TECHNOLOGY · 2024 · IF 11.2

One-pot synthesis of hydroxyapatite hybrid bioinks for digital light processing 3D printing in bone regeneration

Xiaoxiang Ren,Jian Wang,Yan Wu,Yuan Zhang,Jieyuan Zhang,Long Bai,Jinlong Liu,Guangfeng Li,Peiran Song,Zhongmin Shi,Jiacan Su

Three-dimensional (3D) bioprinting has revolutionized tissue engineering by enabling precise fabrication with bioinks. Among these techniques, digital light processing (DLP) stands out due to its exceptional resolution, speed, and biocompatibility. However, the progress of DLP is hindered by the limited availability of suitable bioinks. Currently, some studies involve simple mixing of different materials, resulting in bioinks that lack uniformity and photopolymerization characteristics. To address this challenge, we present an innovative one-pot synthesis method for bioinks based on methacrylated gelatin/alginate with hydroxyapatite (HAP). This approach offers significant advantages in terms of efficiency and uniformity. The synthesized bioinks demonstrate excellent printability, stability, and notably enhanced mechanical properties, facilitating optimal in vitro compatibility. Additionally, the HAP-hybrid bioinks printed scaffolds demonstrated impressive bone repair capabilities in vivo compared with pure organic bioinks. In conclusion, the Gel/Alg/HAP bioinks presented herein offer an innovative solution for DLP bioprinting within the field of bone tissue engineering. Their multifaceted advantages help overcome the limitations of restricted bioink choices, pushing forward the boundaries of bioprinting technology and contributing to the progress of regenerative medicine and tissue engineering.

CAS / MDL: 760-93-0
研究领域: 组织工程学 · 再生医学
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Bio-Design and Manufacturing · 2023 ·Vol. 6 ·期 4 ·464-477 · IF 7.9

The use of machine learning to predict the effects of cryoprotective agents on the GelMA-based bioinks used in extrusion cryobioprinting

Qiao Qian,Zhang Xiang,Yan Zhenhao,Hou Chuanyu,Zhang Juanli,He Yong,Zhao Na,Yan Shujie,Gong Youping,Li Qian

Cryobioprinting has tremendous potential to solve problems to do with lack of shelf availability in traditional bioprinting by combining extrusion bioprinting and cryopreservation. In order to ensure the viability of cells in the frozen state and avoid the possible toxicity of dimethyl sulfoxide (DMSO), DMSO-free bioink design is critical for achieving successful cryobioprinting. A nontoxic gelatin methacryloyl-based bioink used in cryobioprinting is composed of cryoprotective agents (CPAs) and a buffer solution. The selection and ratio of CPAs in the bioink directly affect the survival of cells in the frozen state. However, the development of universal and efficient cryoprotective bioinks requires extensive experimentation. We first compared two commonly used CPA formulations via experiments in this study. Results show that the effect of using ethylene glycol as the permeable CPA was 6.07% better than that of glycerol. Two datasets were obtained and four machine-learning models were established to predict experimental outcomes. The predictive powers of multiple linear regression (MLR), decision tree (DT), random forest (RF), and artificial neural network (ANN) approaches were compared, suggesting an order of ANN>RF>DT>MLR. The final selected ANN model was then applied to another dataset. Results reveal that this machine-learning method can accurately predict the effects of cryoprotective bioinks composed of different CPAs. Outcomes also suggest that the formulations presented here have universality. Our findings are likely to greatly accelerate research and development on the use of bioinks for cryobioprinting. Graphic abstract

研究领域: 生物工程 · 细胞生物学
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Nature Protocols · 2025 · IF 16

Bioink design for organ-scale projection-based 3D bioprinting

Qiao Tianhong,He Chaofan,Xia Pengcheng,Liu Guofeng,Sun Yuan,Sun Miao,Wang Yi,Cheng Yiyu,Yu Mengfei,He Yong

Projection-based three-dimensional bioprinting offers an approach for manufacturing biomimetic tissues with complex spatial structures and bioactivity, presenting potential for creating implantable organs or organoids to test drug response. Nevertheless, the extended printing times required for organ-scale manufacturing represents a challenge. Here we provide step-by-step instructions to manufacture organ-scale structures using bioinks while preserving high bioactivity. This approach incorporates Ficoll 400 to mitigate the heterogeneity of bioink with respect to refractive index and density, while 4-(2-aminoethyl)benzenesulfonyl fluoride and oil-sealing ensure the stability of the bioink components, thereby allowing extended printing times. This procedure also enables high-cell-viability printing via the calibration of the pH value of the bioink. This Protocol is appropriate for users with basic laboratory skills and fundamental knowledge in biotechnology to manufacture organ-scale structures for utilization in a wide variety of experimental designs. The approach is generalizable, as demonstrated by the successful printing of corpora cavernosa structures with a cell density of 10 million per milliliter, measuring 10 mm × 10 mm × 10 mm. After 7 d of culture, the cell viability was measured at 82.5%, highlighting the potential applicability in tissue engineering. All bioink preparation and printing steps are expected to take 5 h, while the development of printed structures requires 7 d of continuous culture.

研究领域: 生物医学工程 · 组织工程 · 三维生物打印 · 再生医学 · 药物研发
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BIOMACROMOLECULES · 2024 · IF 5.5

Gelatin Methacryloyl (GelMA)-Based Biomaterial Inks: Process Science for 3D/4D Printing and Current Status

Soumitra Das,Jeyapriya Thimukonda Jegadeesan,Bikramjit Basu

Tissue engineering for injured tissue replacement and regeneration has been a subject of investigation over the last 30 years, and there has been considerable interest in using additive manufacturing to achieve these goals. Despite such efforts, many key questions remain unanswered, particularly in the area of biomaterial selection for these applications as well as quantitative understanding of the process science. The strategic utilization of biological macromolecules provides a versatile approach to meet diverse requirements in 3D printing, such as printability, buildability, and biocompatibility. These molecules play a pivotal role in both physical and chemical cross-linking processes throughout the biofabrication, contributing significantly to the overall success of the 3D printing process. Among the several bioprintable materials, gelatin methacryloyl (GelMA) has been widely utilized for diverse tissue engineering applications, with some degree of success. In this context, this review will discuss the key bioengineering approaches to identify the gelation and cross-linking strategies that are appropriate to control the rheology, printability, and buildability of biomaterial inks. This review will focus on the GelMA as the structural (scaffold) biomaterial for different tissues and as a potential carrier vehicle for the transport of living cells as well as their maintenance and viability in the physiological system. Recognizing the importance of printability toward shape fidelity and biophysical properties, a major focus in this review has been to discuss the qualitative and quantitative impact of the key factors, including microrheological, viscoelastic, gelation, shear thinning properties of biomaterial inks, and printing parameters, in particular, reference to 3D extrusion printing of GelMA-based biomaterial inks. Specifically, we emphasize the different possibilities to regulate mechanical, swelling, biodegradation, and cellular functionalities of GelMA-based bio(material) inks, by hybridization techniques, including different synthetic and natural biopolymers, inorganic nanofillers, and microcarriers. At the close, the potential possibility of the integration of experimental data sets and artificial intelligence/machine learning approaches is emphasized to predict the printability, shape fidelity, or biophysical properties of GelMA bio(material) inks for clinically relevant tissues.

研究领域: 生物医学工程 · 材料科学 · 再生医学
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CARBOHYDRATE POLYMERS · 2024 · IF 10.7

High mechanical performance and multifunctional degraded fucoidan-derived bioink for 3D bioprinting

Shuai Zhu,Zheng Zhou,Xin Chen,Wenxiang Zhu,Mengni Yang,Mengyi Yu,Jingjing Sun,You Zuo,Jiaqian He,Haobo Pan,Hairong Liu

While 3D bioprinting serves as a powerful tool in the field of tissue engineering, there is still a lack of natural biomaterial inks that simultaneously combine high mechanical performance with multiple biofunctionalities. Here, a single-component natural bioink with high strength and multi-biofunctionality was developed through the simple degradation and methacrylation of natural fucoidan. Hydrothermal degradation significantly decreased the natural fucoidan solution's viscosity by 99.9 %, meeting the necessary viscosity for Digital Light Processing (DLP) 3D printing. Meanwhile, various biofunctionalities of low molecular weight fucoidan obtained through degradation, such as antimicrobial and antioxidant properties, were developed. The resulting bioink exhibited good mechanical performance (compression modulus of 311 kPa), antimicrobial properties (antibacterial rates of 95.5 % and 97.9 % against E. coli and S. aureus , respectively), and antioxidant properties (intracellular ROS inhibition rates of 94.7 %). Using DLP 3D bioprinting, all printed products showed high shape fidelity with exceptional viability and activity of the encapsulated cells. Due to the unique sulfate structure resembling the natural components of chondroitin sulfate, the in vivo tests revealed its efficacy in promoting cartilage defect repair. In conclusion, the novel bioink blending high mechanical performance with multiple biofunctionalities, shows great potential in the 3D printing of tissue and organ regeneration.

研究领域: 组织工程学 · 材料科学 · 再生医学
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Gels · 2022 ·Vol. 8 ·期 5 ·314 · IF 4.432

Preparation of Chitosan/Recombinant Human Collagen-Based Photo-Responsive Bioinks for 3D Bioprinting

Yang Yang,Zixun Wang,Yuanyuan Xu,Jingjing Xia,Zhaoxian Xu,Shuai Zhu,Mingjie Jin

Collagen and chitosan are frequently used natural biomaterials in tissue engineering. However, most collagen is derived from animal tissue, with inconsistent quality and pathogen transmittance risks. In this context, we aimed to use a reliable Type-III recombinant human collagen (RHC) as an alternative biomaterial together with chitosan to develop novel photo-responsive bioinks for three-dimensional (3D) bioprinting. RHC was modified with methacrylic anhydride to obtain the RHC methacryloyl (RHCMA) and mixed with acidified chitosan (CS) to form composites CS-RHCMA. The characterizations demonstrated that the mechanical properties and the degradation of the bioinks were tunable by introducing the CS. The printabilities improved by adding CS to RHCMA, and various structures were constructed via extrusion-based 3D printing successfully. Moreover, in vitro tests confirmed that these CS-RHCMA bioinks were biocompatible as human umbilical vein endothelial cells (HUVECs) were sustained within the constructs post-printing. The results from the current study illustrated a well-established bioinks system with the potential to construct different tissues through 3D bioprinting.Keywords:chitosan;recombinant human collagen;bioinks;3D printing;tissue engineering

CAS / MDL: 760-93-0
研究领域: 生物材料科学 · 组织工程 · 生物打印
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Journal of Materials Chemistry B · 2022 ·Vol. 10 ·期 40 ·8274-8281 · IF 7.571

An antibacterial ε-poly-L-lysine-derived bioink for 3D bioprinting applications

Yahui He,Zheng Zhou,Yuting Huang,Wenxiang Zhu,Ning He,Xiaolong Zhu,Xiaoxiao Han,Hairong Liu

The limited availability of bioinks has hindered the application of 3D bioprinting to tissue engineering, and bacterial infection is a serious threat to these applications. Aiming to solve this problem, a novel ε-poly-L-lysine (EPL)-derived antibacterial bioink has been developed for 3D bioprinting and tissue-engineering applications. Three glycidyl methacrylate (GMA)-modified EPL products, EPLGMA-1, EPLGMA-2, and EPLGMA-3, were prepared by reacting 3, 4, and 5 mL GMA with 5 g EPL, respectively. EPLGMA-1, EPLGMA-2, and EPLGMA-3 were photocurable and their corresponding photo-crosslinked hydrogels, EPLGMA-1H, EPLGMA-2H, and EPLGMA-3H, all exhibited very high antibacterial rates, good biocompatibility, good degradability, and promising mechanical properties. After EPLGMA-1H, EPLGMA-2H, and EPLGMA-3H with encapsulated chondrocytes were incubated for 4 weeks, EPLGMA-3H was the best one among them for tissue-engineering applications due to its most efficient tissue regeneration. Carrying chondrocytes, the EPLGMA-3 solution was printed into hydrogel products with high-fidelity shapes and high cell viability using a projection-based 3D bioprinter. Following the implantation of chondrocyte-loaded EPLGMA-3H samples into nude mice for 4 weeks, cartilage-like tissue was regenerated, suggesting that EPLGMA-3 is a promising antibacterial bioink for 3D bioprinting and tissue-engineering applications.

CAS / MDL: 106-91-2
研究领域: 生物材料学 · 再生医学 · 组织工程
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