Bioprinting最新文献_第8页

FK506 binding protein like, FKBPL, as a novel therapeutic target in 2D and 3D bioprinted, models of cardiac fibrosis FK506结合蛋白样，FKBPL，在2D和3D生物打印心脏纤维化模型中作为新的治疗靶点

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-04 DOI: 10.1016/j.bprint.2025.e00397

Michael Chhor , Shreya Barman , Fatemeh Heidari , Amy L. Bottomley , Tracy Robson , Kristine McGrath , Lana McClements

Background

Cardiac fibrosis characterised by increased collagen deposition and extracellular matrix (ECM) remodeling is one of the main causes of heart failure. Inflammation and hypoxia are key processes leading to cardiac fibrosis although the mechanisms are poorly understood. In this study, we developed an innovative 3D bioprinted model of cardiac fibrosis using tunable matrices. The role of an anti-angiogenic protein, FK506 binding protein like (FKBPL) was then elucidated, for the first time, using both 2D and 3D bioprinted, models of cardiac fibrosis.

Methods

3D bioprinted model of cardiac fibrosis was developed using fetal fibroblast cells (HFF08), customised ECM cardiac components and pro-fibrotic/hypoxic factors (TGF-β, 10 ng/ml, DMOG, 1 mM) ± FKBPL mimetic (AD-01, 100 mM). In parallel, 2D in vitro models were also employed.

Results

In the 3D bioprinted model, fibroblasts formed networks spontaneously, which were stimulated by all treatments (p < 0.05–0.0001). This was in conjunction with a trend towards reduced FKBPL expression, particularly in the presence of DMOG/AD-01 treatment. In 2D cell culture, AD-01 potentiated TGF-β-induced col1a1 (p < 0.0001) and mmp2 mRNA (p < 0.05) expression whereas DMOG or reduced FKBPL expression with AD-01 abrogated this (p < 0.05–0.001). Following siRNA FKBPL transfection, α-SMA was reduced (p < 0.05).

Conclusion

This 3D bioprinted model of cardiac fibrosis in conjunction with 2D cell models could be used for biomarker and drug therapy screening towards accelerating the development of treatments for this hard-to-treat condition. Low FKBPL expression could be protective in cardiac fibrosis through the reduction in collagen production and α-SMA expression, or TGF-β/HIF-1α-mediated effects. Therapeutic strategies that inhibit FKBPL should be explored to abrogate cardiac fibrosis.

以胶原沉积增加和细胞外基质（ECM）重塑为特征的心脏纤维化是心力衰竭的主要原因之一。炎症和缺氧是导致心脏纤维化的关键过程，尽管其机制尚不清楚。在这项研究中，我们开发了一种使用可调基质的创新型3D生物打印心脏纤维化模型。随后，利用2D和3D生物打印的心脏纤维化模型，首次阐明了抗血管生成蛋白FK506结合蛋白样（FKBPL）的作用。方法采用胎儿成纤维细胞（HFF08）、定制ECM心脏成分和促纤维化/缺氧因子（TGF-β, 10 ng/ml， DMOG, 1 mM）±FKBPL模拟物（AD-01, 100 mM）建立生物3d打印心脏纤维化模型。同时，还采用了2D体外模型。结果在生物3D打印模型中，成纤维细胞自发形成网络，所有处理均刺激成纤维细胞形成网络(p <；0.05 - -0.0001)。这与FKBPL表达降低的趋势相结合，特别是在DMOG/AD-01治疗的情况下。在2D细胞培养中，AD-01增强TGF-β诱导的col1a1 (p <；0.0001)和mmp2 mRNA (p <；而DMOG或AD-01降低FKBPL表达则消除了这一点(p <；0.05 - -0.001)。转染siRNA FKBPL后，α-SMA减少(p <；0.05)。结论：该3D生物打印心脏纤维化模型与2D细胞模型相结合，可用于生物标志物和药物治疗筛选，以加速这种难以治疗的疾病的治疗开发。低FKBPL表达可能通过减少胶原生成和α-SMA表达，或TGF-β/ hif -1α介导的作用，对心脏纤维化具有保护作用。应探索抑制FKBPL的治疗策略，以消除心脏纤维化。

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引用次数: 0

Modeling of oral squamous cell carcinoma microenvironment- A 3D bioprinting approach 口腔鳞状细胞癌微环境建模- 3D生物打印方法

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00381

Akhilanand Chaurasia , Gowri Sivaramakrishnan , Farah Asa’ad , Lena Larsson , Arwa Daghrery , Joana Marques , Francesca Spirito , Vitória Batista Clemente , Ana Carolina Morais Apolônio , Mahdieh Alipour , Rini Tiwari

Background

Oral squamous cell carcinoma (OSCC) presents significant challenges due to its complex microenvironment and invasive characteristics. Traditional two-dimensional (2D) culture systems are inadequate for modelling the intricate features of OSCC, necessitating advanced techniques for better in vitro modelling.

Objective

This review aims to explore the applications of 3D bioprinting in modelling the OSCC microenvironment, highlighting the advantages over conventional methods and discussing recent advancements in the field.

Methods

The review synthesizes recent literature on 3D bioprinting technologies, focusing on their application in replicating OSCC's microenvironment. Key areas include the integration of various cell types within a biomimetic extracellular matrix, the use of microfluidic systems to study tumor-stromal interactions, and the incorporation of advanced imaging modalities.

Results

3D bioprinting allows for the precise fabrication of complex OSCC tumor architectures, incorporating cancer cells, stromal cells, and immune cells. The integration of microfluidic systems facilitates the study of tumor invasion, metastasis, and drug response. Recent advancements in bioink development, particularly the use of patient-derived cells and biomolecules, enhance the physiological relevance of these models. Emerging imaging technologies provide unprecedented insights into the dynamics of OSCC progression within these constructs.

Conclusion

3D bioprinting shows immense potential for advancing the understanding of OSCC pathobiology and developing personalized therapeutic strategies. However, challenges such as standardizing bioink formulations and scaling fabrication techniques must be addressed to effectively translate these innovations into clinical practice.

口腔鳞状细胞癌（OSCC）因其复杂的微环境和侵袭性特征而面临重大挑战。传统的二维（2D）培养系统不足以模拟OSCC的复杂特征，需要先进的技术来更好地进行体外建模。目的探讨生物3D打印在OSCC微环境建模中的应用，强调其相对于传统方法的优势，并讨论该领域的最新进展。方法综述了近年来生物3D打印技术的研究进展，重点介绍了3D打印技术在复制OSCC微环境中的应用。关键领域包括在仿生细胞外基质中整合各种细胞类型，使用微流体系统研究肿瘤-基质相互作用，以及结合先进的成像方式。结果3d生物打印可以精确制造复杂的OSCC肿瘤结构，包括癌细胞、基质细胞和免疫细胞。微流控系统的集成促进了肿瘤侵袭、转移和药物反应的研究。生物连接发展的最新进展，特别是患者来源的细胞和生物分子的使用，增强了这些模型的生理学相关性。新兴的成像技术为OSCC在这些结构中的进展动态提供了前所未有的见解。结论3d生物打印技术在提高对OSCC病理生物学的认识和制定个性化治疗策略方面具有巨大的潜力。然而，必须解决诸如标准化生物链接配方和规模化制造技术等挑战，才能有效地将这些创新转化为临床实践。

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引用次数: 0

Zirconia-calcium silicate bioactive composites for dental applications using DLP additive manufacturing 使用DLP增材制造的牙科用氧化锆-硅酸钙生物活性复合材料

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00377

Ahmed Binobaid , Michele De Lisi , Josette Camilleri , Hany Hassanin , Khamis Essa

Zirconia has outstanding mechanical strength which made it a favourable material dental implants material. However, its use is limited by challenges in bone bonding and elasticity. This paper introduces a novel bioprinting ceramic material by mixing calcium silicate with zirconia to enhance bioactivity. Using the high precision and speed of Digital Light Processing (DLP), this study develops a novel zirconia-calcium silicate slurry for dental applications. The study reports the preparation of zirconia-calcium silicate, formulation of resin compositions, and optimization of the bioprinting, debinding and sintering. Employing a full factorial Design of Experiments (DOE), a systematic approach was implemented to identify optimal printing conditions such as the layer thickness, exposure time, and power. The results show that slurries formulated with BYK-111 as the dispersant and ACMO/PEGDA/TPO resin, coupled with 80 wt% solid loading, achieved the most favourable rheological properties, cure depth, and printing accuracy. The optimal printing conditions were 0.75 s exposure time, 300 % exposure power, and 30 μm layer thickness, ensured a relative density of the sintered implants exceeding 95 %. This study advances dental implant materials by introducing a novel DLP biomaterial with a slurry formulation, presenting significant implications for clinical applications and future research in developing advanced dental and medical implants.

氧化锆具有优异的机械强度，是一种良好的牙种植材料。然而，它的使用受到骨粘合和弹性方面的挑战的限制。介绍了一种新型的硅酸钙与氧化锆混合制备的生物打印陶瓷材料。利用数字光处理（DLP）的高精度和高速，本研究开发了一种新型的牙科应用的硅酸锆-钙浆料。本研究报道了硅酸锆钙材料的制备、树脂组成的配方以及生物打印、脱脂和烧结工艺的优化。采用全因子实验设计（DOE），采用系统的方法来确定最佳打印条件，如层厚度，曝光时间和功率。结果表明，以BYK-111为分散剂和ACMO/PEGDA/TPO树脂配制的浆料，加上80 wt%的固体负荷，获得了最佳的流变性能、固化深度和打印精度。最佳打印条件为曝光时间0.75 s，曝光功率300%，层厚30 μm，可确保烧结植入物的相对密度超过95%。本研究通过引入一种具有浆状配方的新型DLP生物材料，推动了牙科种植材料的发展，对临床应用和未来开发先进牙科和医疗种植材料的研究具有重要意义。

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引用次数: 0

4D printing in skin tissue engineering: A revolutionary approach to enhance wound healing and combat infections 皮肤组织工程中的4D打印：一种革命性的方法来增强伤口愈合和对抗感染

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2025.e00386

Laila A. Damiati , Samar A. Alsudir , Rean Y. Mohammed , Majed A. Majrashi , Shahad H. Albrahim , Aliyah algethami , Fatimah O. Alghamdi , Hala A. Alamari , Mai M. Alzaydi

Skin infection poses significant challenges in healthcare, demanding innovative solutions to enhance the efficacy of wound-repair interventions. 4D printing represents a revolutionary approach in addition to traditional wound-management strategies. 4D-printing materials, which are dynamic and responsive, can change their shape or properties over time in response to internal or external stimuli, creating a paradigm shift in how wounds are treated. This review explores the potential of 4D printing technology as a transformative solution addressing critical challenges in skin tissue engineering. It highlights the journey from 2D fabrication of skin implants to the current state of 4D printing focusing on skin tissue structures that allow for precise and sustained release of therapeutic agents while exhibiting self-healing properties. Also, the ability to integrate antimicrobials to the printed skin constructs that respond to specific stimuli, such as pH, light, temperature, humidity, or enzymes enables the on demand and controlled release of antimicrobial agents. Additionally, integrating artificial intelligence (AI) into the fabrication process of skin tissues represents a synergistic approach that combines advanced computational methodologies with biological principles to identify the optimal conditions for enhancing tissue regeneration. Indeed, 4D bioprinting and AI-driven precision in the customization of scaffolds based on patient-specific needs promise a new era of personalized medicine in skin tissue engineering.

皮肤感染对医疗保健提出了重大挑战，需要创新的解决方案来提高伤口修复干预措施的疗效。除了传统的伤口管理策略外，4D打印还代表了一种革命性的方法。3d打印材料具有动态和响应性，可以随着时间的推移改变其形状或特性，以响应内部或外部刺激，从而在伤口治疗方面产生范式转变。这篇综述探讨了4D打印技术作为解决皮肤组织工程关键挑战的变革性解决方案的潜力。它强调了从皮肤植入物的2D制造到4D打印的当前状态的旅程，重点是皮肤组织结构，允许治疗剂的精确和持续释放，同时表现出自我修复特性。此外，将抗菌剂整合到对特定刺激（如pH值、光、温度、湿度或酶）有反应的打印皮肤结构中的能力，使抗菌剂的按需和可控释放成为可能。此外，将人工智能（AI）集成到皮肤组织的制造过程中代表了一种协同方法，将先进的计算方法与生物学原理相结合，以确定增强组织再生的最佳条件。事实上，4D生物打印和基于患者特定需求的人工智能驱动的支架定制精度预示着皮肤组织工程个性化医疗的新时代。

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引用次数: 0

Advancing the in vitro drug screening models: Microbiome as a component of tissue-engineered skin 推进体外药物筛选模型：微生物组作为组织工程皮肤的组成部分

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00379

Vsevolod V. Shishkov , Polina Yu Bikmulina , Anna V. Kardosh , Sergey V. Tsibulnikov , Ekaterina V. Grekova , Yulia V. Kolesova , Polina A. Zakharova , Anastasiia M. Nesterova , Frederico David Alencar de Sena Pereira , Svetlana L. Kotova , Olga Yu Olisova , Massoud Vosough , Anastasia I. Shpichka , Peter S. Timashev

Currently, in vitro skin models are among the most advanced and frequently utilized tools in clinical practice and drug screening. The development of these models often involves the use of skin organoids and biofabrication techniques, such as 3D bioprinting. Despite this significant progress, the skin models employed in drug screening typically lack a microbiome component. Since the microbiome is recognized as a crucial element of healthy human skin, it is essential to integrate this aspect into existing skin models. This review outlines a pathway for the development of in vitro skin models that can be widely used as platforms for testing drugs and cosmetics. First, we discuss the diversity of the normal human microbiome and its interactions with human cells. Next, we examine current skin models, including those that incorporate microbiome components through various co-culturing methods. Finally, we discuss how biofabrication approaches can be combined with microbiome elements to create relevant and stable in vitro skin models.

目前，体外皮肤模型是临床实践和药物筛选中最先进、最常用的工具之一。这些模型的开发通常涉及使用皮肤类器官和生物制造技术，如3D生物打印。尽管取得了重大进展，但用于药物筛选的皮肤模型通常缺乏微生物组成分。由于微生物组被认为是健康人类皮肤的关键因素，因此将这方面整合到现有皮肤模型中至关重要。本文概述了体外皮肤模型的发展途径，该模型可广泛用于药物和化妆品的测试平台。首先，我们讨论了正常人类微生物组的多样性及其与人体细胞的相互作用。接下来，我们将研究当前的皮肤模型，包括那些通过各种共培养方法纳入微生物组成分的皮肤模型。最后，我们讨论了生物制造方法如何与微生物组元素相结合，以创建相关且稳定的体外皮肤模型。

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引用次数: 0

Nanocomposite hydrogel-based bioinks composed of a fucose-rich polysaccharide and nanocellulose fibers for 3D-bioprinting applications 纳米复合水凝胶为基础的生物墨水，由富含焦点的多糖和纳米纤维素纤维组成，用于3d生物打印应用

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00382

Nicole S. Lameirinhas , João P.F. Carvalho , Maria C. Teixeira , Jorge L. Luís , Asiyah Esmail , Ricardo J.B. Pinto , Helena Oliveira , Filomena Freitas , José M. Oliveira , Carla Vilela , Armando J.D. Silvestre , Carmen S.R. Freire

Hydrogels are the most common type of bioinks, yet, finding adequate biomaterials to develop suitable bioinks for 3D bioprinting remains challenging. Herein, innovative hydrogel bioinks were developed by combining nanofibrillated cellulose (NFC) with a fucose-rich polysaccharide, FucoPol (FP), still unexplored for 3D bioprinting. NFC/FP bioinks with different mass proportions, namely 1:1, 2:1, 3:1 and 4:1, were prepared and denominated as NFC1FP, NFC2FP, NFC3FP and NFC4FP. A formulation without NFC was also prepared for comparison purposes (NFC0FP). The rheological properties of the bioinks were enhanced by the addition of NFC, as evidenced by the increase in shear viscosity from 1.39 ± 0.03 Pa s (NFC0FP) to 2933.7 ± 137.9 Pa s (ink NFC4FP) and by the 3D printing of complex structures with high shape fidelity (Pr ≈ 0.9). The stability and mechanical properties of the crosslinked hydrogels were also improved, with Young’s modulus increasing from 0.12 ± 0.04 MPa (NFC0FP) to 2.45 ± 0.06 MPa (NFC4FP). The successful 3D bioprinting of both A375 (melanoma) and HaCaT (keratinocyte) cell-laden bioinks translated into elevated cell viabilities (above 88 %) up to 21 days post-bioprinting. These results highlight the potential and versatility of NFC/FP bioinks for the bioprinting of 3D skin tissue analogues for biomedical applications.

水凝胶是最常见的生物墨水类型，然而，寻找足够的生物材料来开发适合3D生物打印的生物墨水仍然具有挑战性。在这里，创新的水凝胶生物墨水是通过将纳米纤维化纤维素（NFC）与富含焦点的多糖——岩藻酚（FP）结合而开发出来的，这种多糖尚未被用于生物3D打印。制备了质量比例分别为1:1、2:1、3:1和4:1的NFC/FP生物墨水，分别命名为NFC1FP、NFC2FP、NFC3FP和NFC4FP。同时制备了不含NFC的配方（NFC0FP）进行比较。NFC的加入增强了生物墨水的流变性能，剪切粘度从1.39±0.03 Pa s （NFC0FP）增加到2933.7±137.9 Pa s (NFC4FP)， 3D打印复杂结构具有较高的形状保真度（Pr≈0.9）。交联水凝胶的稳定性和力学性能也得到了改善，杨氏模量从0.12±0.04 MPa （NFC0FP）增加到2.45±0.06 MPa （NFC4FP）。A375（黑色素瘤）和HaCaT（角质形成细胞）细胞负载生物墨水的成功3D生物打印在生物打印后21天内转化为更高的细胞存活率（超过88%）。这些结果突出了NFC/FP生物墨水用于生物医学应用的3D皮肤组织类似物生物打印的潜力和多功能性。

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引用次数: 0

3D printing of polysaccharide-based formulations: Opportunities for innovation 基于多糖配方的3D打印：创新的机会

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00383

Fabian Hernandez-Tenorio , Edier Múnera-Gutiérrez , Alejandra M. Miranda , Alex A. Sáez , Luz Deisy Marín-Palacio , Catalina Giraldo-Estrada

3D printing is a technology that has gained significant interest due to its versatility in terms of design, as well as the wide variety of materials that can be used for the production of inks. Among the compounds with the greatest importance in the last decade for 3D printing are polysaccharides. These have been positioned as favorable compounds for the formulation of inks due to their properties such as flexibility, non-immunogenicity, pseudoplastic behavior, printability, biocompatibility, and biodegradability. Therefore, the implementation of polysaccharides in 3D printing promotes innovation in the development of materials and products for medical, food, pharmaceutical, and other applications. The objective of this review was to provide a comprehensive and exhaustive study of the technological advances in 3D printing of polysaccharide-based formulations. To this end, a bibliometric analysis was presented to establish trends using scientometric indicators that allowed us to delve deeper and identify the most relevant developments in the subject. Through this review, we sought to highlight the importance of polysaccharides and their wide range of applications in 3D printing and hope that it will provide a meaningful basis for the exploration of printable compounds from renewable sources.

3D打印技术由于其设计的多功能性以及可用于生产油墨的各种材料而获得了极大的兴趣。在过去十年中，对3D打印最重要的化合物是多糖。由于其柔韧性、非免疫原性、假塑性、可印刷性、生物相容性和可生物降解性等特性，这些已被定位为油墨配方的有利化合物。因此，多糖在3D打印中的应用促进了医疗、食品、制药和其他应用的材料和产品开发的创新。本综述的目的是对多糖基配方的3D打印技术进步进行全面而详尽的研究。为此，提出了文献计量学分析，利用科学计量学指标建立趋势，使我们能够深入研究并确定该主题中最相关的发展。通过这篇综述，我们试图强调多糖的重要性及其在3D打印中的广泛应用，并希望它将为探索可再生资源的可打印化合物提供有意义的基础。

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引用次数: 0

Pneumatic extrusion-based bioprinting and flow cytometry: A method for analysing chemotherapy efficacy in 3D bioprinted A375 melanoma cell cultures 基于气动挤压的生物打印和流式细胞术：一种分析3D生物打印A375黑色素瘤细胞培养物化疗疗效的方法

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00380

Maryke de Villiers, Awie F. Kotzé, Lissinda H. du Plessis

Melanoma, a highly aggressive form of skin cancer, continues to be a significant challenge due to its resistance to conventional chemotherapy treatments and the tendency for metastasis. Advancements in cell culture techniques, especially the transition from 2D cell cultures to more physiologically relevant 3D cell cultures, have provided valuable new insights into cancer biology and chemotherapy drug responses. Although various novel 3D cell culture techniques have been used in melanoma research, standardised and scalable 3D cell culture models suitable for high-throughput pre-clinical drug screening applications are still lacking. Therefore, the purpose of this study was to establish a 3D bioprinted melanoma cell culture model that allows the assessment of drug-induced apoptosis through a flow-cytometric analysis method in 96-well plates. To achieve this, the proposed method integrates the BIOX™ pneumatic extrusion-based 3D bioprinter to extrude reproducible cell-laden droplets in a 96-well plate, and an Annexin V/PI flow cytometric analysis technique optimised for 96-well plate format, to enable cell viability and apoptosis quantification in more physiologically relevant 3D bioprinted cell cultures. The proposed method was evaluated on A375 melanoma 2D and 3D bioprinted cell cultures assayed for drug-induced apoptosis through a flow cytometric method. In addition, a resazurin-based analysis method was also used and compared to determine the efficacy of the proposed flow cytometric analysis method. Compared to the 2D cell cultures, the 3D bioprinted cell cultures demonstrated higher levels of resistance to all chemotherapy drugs evaluated. Furthermore, the comparative analysis of the two methods concluded that the flow cytometric evaluation platform is more sensitive in detecting drug dose responses in 3D bioprinted cell culture models. This method is a proposed alternative to quantify drug-induced apoptosis in 3D melanoma research, thereby advancing the pre-clinical application of 3D bioprinting.

黑色素瘤是一种高度侵袭性的皮肤癌，由于其对常规化疗的耐药性和转移倾向，一直是一个重大挑战。细胞培养技术的进步，特别是从2D细胞培养到更具有生理学相关性的3D细胞培养的转变，为癌症生物学和化疗药物反应提供了有价值的新见解。尽管各种新颖的3D细胞培养技术已用于黑色素瘤研究，但适合高通量临床前药物筛选应用的标准化和可扩展的3D细胞培养模型仍然缺乏。因此，本研究的目的是建立一种3D生物打印黑色素瘤细胞培养模型，该模型可以通过96孔板流式细胞分析方法评估药物诱导的细胞凋亡。为了实现这一目标，该方法集成了基于BIOX™气动挤压的3D生物打印机，在96孔板中挤出可重复的细胞负载液滴，以及针对96孔板格式优化的Annexin V/PI流式细胞分析技术，以便在更生理相关的3D生物打印细胞培养中实现细胞活力和凋亡的量化。该方法在A375黑色素瘤2D和3D生物打印细胞培养物上进行了评估，并通过流式细胞术检测药物诱导的细胞凋亡。此外，我们还使用了一种基于瑞沙脲的分析方法，并进行了比较，以确定所提出的流式细胞分析方法的有效性。与2D细胞培养相比，3D生物打印细胞培养对所有化疗药物的耐药性都更高。此外，两种方法的对比分析表明，流式细胞术评价平台在生物3D打印细胞培养模型中检测药物剂量反应更为灵敏。该方法是在3D黑色素瘤研究中提出的一种量化药物诱导细胞凋亡的替代方法，从而推进生物3D打印的临床前应用。

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引用次数: 0

Development and characterization of bioinks for 3D bioprinting of in vitro skeletal muscle constructs 用于体外骨骼肌结构3D生物打印的生物墨水的开发和表征

Q1 Computer Science

Bioprinting

Pub Date : 2025-01-31 DOI: 10.1016/j.bprint.2025.e00396

Rodi Kado Abdalkader , Kosei Yamauchi , Satoshi Konishi , Takuya Fujita

The use of 3D bioprinting to construct in vitro skeletal muscle models presents a promising approach; however, selecting an optimal bioink remains a common challenge. This study focuses on the development and characterization of bioinks for extrusion-based 3D bioprinting, specifically targeting the creation of accurate skeletal muscle models. By exploring various compositions of alginate, gelatin, fibrinogen, and nanofiber cellulose, we evaluate these formulations based on printability and their support for the growth and differentiation of C2C12 myoblast cells.

While alginate provided a strong, stable matrix for printing scaffolds embedded with C2C12 cells, it did not effectively promote cell growth and differentiation. The addition of fibrinogen to alginate enhanced cell growth and differentiation but was limited mainly to the scaffold surfaces, even with the inclusion of gelatin as a sacrificial ink. Notably, replacing alginate with nanofiber cellulose (NFC) alongside fibrinogen significantly improved cell growth and differentiation, leading to the formation of mature myotubes. Cell distribution was observed both inside and on the surfaces of the scaffolds, indicating effective spatial cell distribution. Furthermore, the scaffolds were tailored to form skeletal muscle bundles anchored between PDMS pillars for contractility testing. Upon exposure to electrical stimulation, the cells displayed measurable displacement, demonstrating contractile function.

These findings offer valuable insights into optimizing bioink formulations that promote myoblast growth and differentiation into skeletal muscle in vitro, with potential applications in future neuromuscular disease modeling.

利用生物3D打印技术构建体外骨骼肌模型是一种很有前途的方法；然而，选择一种最佳的生物链接仍然是一个共同的挑战。本研究的重点是基于挤压的生物3D打印的生物墨水的开发和表征，特别是针对精确的骨骼肌模型的创建。通过探索海藻酸盐、明胶、纤维蛋白原和纳米纤维纤维素的不同组成，我们评估了这些配方的可打印性及其对C2C12成肌细胞生长和分化的支持。海藻酸盐为C2C12细胞包埋的打印支架提供了坚固、稳定的基质，但不能有效促进细胞生长和分化。在海藻酸盐中加入纤维蛋白原可以促进细胞生长和分化，但主要局限于支架表面，即使加入明胶作为牺牲墨水也是如此。值得注意的是，用纳米纤维纤维素（NFC）和纤维蛋白原代替海藻酸盐显著改善了细胞生长和分化，导致成熟肌管的形成。在支架内部和表面均观察到细胞的分布，表明细胞的空间分布是有效的。此外，这些支架被定制成固定在PDMS支柱之间的骨骼肌束，用于收缩性测试。当受到电刺激时，这些细胞显示出可测量的位移，显示出收缩功能。这些发现为优化促进成肌细胞生长和向体外骨骼肌分化的生物链接配方提供了有价值的见解，在未来神经肌肉疾病建模中具有潜在的应用前景。

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引用次数: 0

3D printed cellulose nanofiber-reinforced and iron-crosslinked double network hydrogel composites for tissue engineering applications: Mechanical properties and cellular viability 用于组织工程应用的3D打印纤维素纳米纤维增强和铁交联双网水凝胶复合材料：机械性能和细胞活力

Q1 Computer Science

Bioprinting

Pub Date : 2025-01-27 DOI: 10.1016/j.bprint.2025.e00392

Rohit Goyal , Soumyasri Nikhilesh Mahapatra , Rashmi Yadav , Santanu Mitra , Animesh Samanta , Anuj Kumar , Bimlesh Lochab

Additive manufacturing (i.e. 3D printing) is a promising technology for creating three-dimensional (3D) complex tissue-engineered hydrogel structures based on computer digital models resulting from patient-specific anatomical data of the organs. However, besides the printing process, it is worth studying the variation of individual components of the developed hydrogel composites to enable their suitability for tissue engineering. In this work, we shaped 3D printed multi-layered dual (UV- and Fe³⁺ ions)-crosslinked structures using hydrogel-inks composed of polyacrylamide (PAM), alginate (ALG), and cellulose nanofibres (CNFs). For extrusion, ALG in hydrogel precursor ink acted as a viscosity modifier owing to rapid gelation in the presence of low Ca²⁺ ions and CNF provided shear-thinning behavior. With the addition of optimal content of CNF (3 wt%), the mechanical properties of 3D printed composite hydrogel were enhanced and tuned using different fiber orientations. The maximum tensile stress of PAM/ALG_1.5/3CNF composite hydrogel is measured as ∼162 kPa, and maximum tensile toughness as ∼54 kJ/m³ supporting a good fracture resistance. Moreover, CNF-Fe³⁺ loaded 3D printed dual-networked composite hydrogels could disperse energy more efficiently and displayed maximum tensile stress as ∼285 kPa and maximum toughness as ∼200 kJ/m³. Further, In the current study, developed composite structures exhibited enhanced swelling ratio and thermal stability. In addition, finite element (FE) modelling was also exploited to analyze the novel anisotropic composite structures using efficient computational techniques. It is established that varying nanofiber content and fibrils orientation can be utilized to modulate the physicochemical, mechanical, and biological characteristics of printed structures. Overall, PAM/ALG_1.5/3CNF-Fe³⁺ printed composite structures present substantial stretchability, enhanced anisotropic mechanical and physicochemical properties with excellent cytocompatibility.

增材制造（即3D打印）是一种很有前途的技术，用于基于患者特定器官解剖数据产生的计算机数字模型创建三维（3D）复杂的组织工程水凝胶结构。然而，除了打印过程之外，值得研究的是所开发的水凝胶复合材料的单个组分的变化，以使其适合于组织工程。在这项工作中，我们使用由聚丙烯酰胺（PAM）、海藻酸盐（ALG）和纤维素纳米纤维（cnf）组成的水凝胶墨水，塑造了3D打印多层双（UV-和Fe3+离子）交联结构。对于挤压，水凝胶前驱体油墨中的ALG由于在低Ca2+离子存在下快速凝胶化而充当粘度调节剂，CNF提供剪切变薄行为。随着CNF的最佳含量（3wt %）的加入，3D打印复合水凝胶的力学性能得到了增强和调整，并采用不同的纤维取向。PAM/ALG1.5/3CNF复合水凝胶的最大拉伸应力为~ 162 kPa，最大拉伸韧性为~ 54 kJ/m3，具有良好的抗断裂性能。此外，负载CNF-Fe3+的3D打印双网络复合水凝胶可以更有效地分散能量，最大拉伸应力为~ 285 kPa，最大韧性为~ 200 kJ/m3。此外，在本研究中，开发的复合材料结构具有增强的膨胀率和热稳定性。此外，还利用有限元模型，利用高效的计算技术对新型各向异性复合材料结构进行了分析。研究表明，改变纳米纤维含量和原纤维取向可以调节打印结构的物理化学、力学和生物学特性。总体而言，PAM/ALG1.5/3CNF-Fe3+打印的复合材料结构具有良好的拉伸性能，增强的各向异性力学和物理化学性能，具有优异的细胞相容性。

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引用次数: 0