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Editorial: Celebrating 1 year of frontiers in biomaterials science 社论:庆祝生物材料科学前沿发展一周年
Pub Date : 2024-07-19 DOI: 10.3389/fbiom.2024.1455984
Chun Wang, Virginia Brancato
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引用次数: 0
Tunable gelatin methacrylate polyethylene glycol diacrylate hydrogels for cell mechanosensing applications 用于细胞机械传感应用的可调明胶甲基丙烯酸聚乙二醇二丙烯酸酯水凝胶
Pub Date : 2024-07-15 DOI: 10.3389/fbiom.2024.1408748
Eya Ferchichi, Samuel T Stealey, Paige Bogert, S. Zustiak
Three-dimensional (3D) tissue-engineered scaffolds mimic the physiological environment of cells by providing essential structural support, biochemical cues, and the mechanical strength needed for cell adhesion, proliferation, migration, and differentiation. Hydrogels like polyethylene glycol diacrylate (PEGDA) are commonly used biomaterials for cell culture due to their affordability, tunable stiffness, and ability to efficiently transport nutrients and gases. However, PEGDA lacks cell adhesion sites essential for cell proliferation and migration and has limited degradability. Methacrylated gelatin (GelMA) produced from denatured bovine collagen, crosslinks under ultraviolet light (UV) resulting in a degradable hydrogel with cell adhesion sites. Here, we synthesized GelMA with variable degree of methacrylation and crosslinked it with PEGDA to produce cell scaffolds with independently tunable mechanical and biochemical properties by varying the ratios of the two polymers. We determined polymer ratios that resulted in scaffolds with different mechanical properties but the same gelatin concentrations (providing cell adhesion and degradation sites) as well as different gelatin concentrations but the same mechanical properties. With the developed scaffold library, we further used a design of experiments approach to probe the parameter space and perform detailed analysis on chemical composition-scaffold properties as well as scaffold properties-cell behavior correlations. Our findings showed that hydrogel properties such as modulus, swelling, pore size, and permeability, strongly depended on total polymer concentration and not on the GelMA fraction. GelMA significantly influenced cell spreading, while addition of any amount of PEGDA delayed cell spreading significantly. We suggest that such analysis will broaden the utility of the GelMA/PEGDA hydrogels, presenting a versatile platform for mechanosensing research in 3D environments.
三维(3D)组织工程支架通过提供细胞粘附、增殖、迁移和分化所需的基本结构支持、生化线索和机械强度,模拟细胞的生理环境。聚乙二醇二丙烯酸酯(PEGDA)等水凝胶因其价格低廉、硬度可调、能有效运输营养物质和气体而成为细胞培养常用的生物材料。然而,PEGDA 缺乏细胞增殖和迁移所必需的细胞粘附点,而且降解性有限。甲基丙烯酸明胶(GelMA)由变性牛胶原蛋白制成,在紫外线(UV)照射下会发生交联,从而形成具有细胞粘附位点的可降解水凝胶。在这里,我们合成了具有不同甲基丙烯酸化程度的 GelMA,并将其与 PEGDA 交联,通过改变两种聚合物的比例,生产出具有独立可调机械和生化特性的细胞支架。我们确定了两种聚合物的比例,从而得到了机械性能不同但明胶浓度相同(提供细胞粘附和降解位点)的支架,以及明胶浓度不同但机械性能相同的支架。利用所开发的支架库,我们进一步采用实验设计方法探索参数空间,并对化学成分-支架特性以及支架特性-细胞行为相关性进行了详细分析。我们的研究结果表明,水凝胶的特性(如模量、膨胀、孔径和渗透性)在很大程度上取决于聚合物的总浓度,而与 GelMA 的组分无关。GelMA 对细胞扩散有明显影响,而添加任何数量的 PEGDA 都会明显延迟细胞扩散。我们认为这种分析将扩大 GelMA/PEGDA 水凝胶的用途,为三维环境中的机械传感研究提供一个多功能平台。
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引用次数: 0
Modelling periodontitis in vitro: engineering strategies and biofilm model development 牙周炎体外建模:工程策略和生物膜模型开发
Pub Date : 2024-07-12 DOI: 10.3389/fbiom.2024.1380153
Cong Wang, Tian Xu, C. Seneviratne, Louis Jun Ye Ong, Yinghong Zhou
Periodontitis is a chronic inflammatory disease associated with dysbiosis in subgingival plaque biofilm, characterised by damage to the periodontal tissues, eventually leading to tooth loss. Hence, the pathophysiology of periodontitis and interaction between subgingival plaque and host tissue under various environmental cues are central to the pathogenesis of periodontitis. Therefore, engineering biofilm models that mimic in vivo pathophysiology is crucial to obtaining a clear insight into the pathology and developing targeted therapeutic methods. In this review, we provide a comprehensive overview of the engineering strategies employed of modelling oral biofilms focusing on surface attachment, fluid microenvironment, gas environment, shear force, microbial-host interaction and offer insights into the ongoing challenges and future perspectives, which will enable the development of novel physiological relevant models for oral biofilms.
牙周炎是一种慢性炎症性疾病,与龈下菌斑生物膜的菌群失调有关,其特点是牙周组织受损,最终导致牙齿脱落。因此,牙周炎的病理生理学以及龈下菌斑和宿主组织在各种环境因素下的相互作用是牙周炎发病机制的核心。因此,建立模拟体内病理生理学的生物膜模型对于清楚地了解病理和开发有针对性的治疗方法至关重要。在这篇综述中,我们全面概述了口腔生物膜建模所采用的工程策略,重点关注表面附着、流体微环境、气体环境、剪切力、微生物与宿主的相互作用,并深入探讨了当前的挑战和未来的前景,这将有助于开发新型生理相关的口腔生物膜模型。
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引用次数: 0
Macrophage variance: investigating how macrophage origin influences responses to soluble and physical cues with immortalized vs. primary cells in 2D and 3D culture 巨噬细胞差异:研究巨噬细胞来源如何影响二维和三维培养中永生细胞与原代细胞对可溶性和物理线索的反应
Pub Date : 2024-05-22 DOI: 10.3389/fbiom.2024.1399448
Jodi Graf, Kartik Bomb, Michael Trautmann-Rodriguez, Bader M. Jarai, Nicole Gill, April M. Kloxin, C. Fromen
Macrophages are phagocytic innate immune cells capable of phenotypical switching in response to the local microenvironment. Studies often use either primary macrophages or immortalized cell lines for hypothesis testing, therapeutic assessment, and biomaterial evaluation without carefully considering the potential effects of cell source and tissue of origin, which strongly influence macrophage response. Surprisingly, limited information is available about how, under similar stimuli, immortalized cell lines and primary cells respond in both phenotypical and functional changes. To address this need, in this work, we cultured immortalized macrophage cell lines derived from different origins (i.e., blood, lung, peritoneal) to understand and compare macrophage phenotypical responses, including polarization and plasticity, morphological changes, and phagocytic functionalities, as well as compared primary macrophages extracted from peritoneal and bone marrow to their immortalized cell line counterparts. We found significant differences in baseline expression of different markers (e.g., CD86, MHCII, CD206, and EGR2) amongst different cell lines, which further influence both polarization and repolarization of the cells, in addition to their phagocytic functionality. Additionally, we observed that, while RAW 264.7 cells behave similarly to the primary bone marrow-derived macrophages, there are noticeable phenotypical and functional differences in cell line (IC-21) and primary peritoneal macrophages, highlighting tissue-specific differences in macrophage response amongst cell lines and primary cells. Moving to three-dimensional (3D) culture in well-defined biomaterials, blood-derived primary and cell line macrophages were encapsulated within hydrogel-based synthetic extracellular matrices and their polarization profiles and cell morphologies were compared. Macrophages exhibited less pronounced polarization during 3D culture in these compliant, soft materials compared to two-dimensional (2D) culture on rigid, tissue culture plastic plates. Overall, our findings highlight origin-specific differences in macrophage response, and therefore, careful considerations must be made to identify the appropriate cell source for the application of interest.
巨噬细胞是一种吞噬性先天性免疫细胞,能够根据局部微环境的变化进行表型转换。研究通常使用原代巨噬细胞或永生化细胞系进行假设检验、治疗评估和生物材料评价,而没有仔细考虑细胞来源和原发组织的潜在影响,因为它们对巨噬细胞的反应有很大影响。令人惊讶的是,关于在类似刺激下,永生化细胞系和原代细胞如何在表型和功能变化方面做出反应的信息非常有限。为了满足这一需求,在这项工作中,我们培养了来自不同来源(如血液、肺、腹膜)的永生化巨噬细胞系,以了解和比较巨噬细胞的表型反应,包括极化和可塑性、形态变化和吞噬功能,并将从腹膜和骨髓中提取的原代巨噬细胞与其永生化细胞系进行比较。我们发现,不同细胞系之间不同标记物(如 CD86、MHCII、CD206 和 EGR2)的基线表达存在明显差异,这进一步影响了细胞的极化和再极化,以及它们的吞噬功能。此外,我们还观察到,虽然 RAW 264.7 细胞的行为与原代骨髓巨噬细胞相似,但细胞系(IC-21)和原代腹膜巨噬细胞在表型和功能上存在明显差异,这突显了细胞系和原代细胞之间巨噬细胞反应的组织特异性差异。在定义明确的生物材料中进行三维(3D)培养时,将源自血液的原代巨噬细胞和细胞系巨噬细胞封装在基于水凝胶的合成细胞外基质中,并比较它们的极化特征和细胞形态。与在刚性组织培养塑料板上进行二维(2D)培养相比,巨噬细胞在这些顺应性软材料的三维培养过程中表现出的极化不那么明显。总之,我们的研究结果凸显了巨噬细胞反应的来源特异性差异,因此必须仔细考虑为相关应用确定合适的细胞来源。
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引用次数: 0
Current challenges in imaging the mechanical properties of tissue engineered grafts 组织工程移植物机械特性成像目前面临的挑战
Pub Date : 2024-05-10 DOI: 10.3389/fbiom.2024.1323763
Lu Luo, Kerime Ebrar Okur, P. Bagnaninchi, Alicia J. El Haj
The mechanical performance of tissue-engineered grafts is crucial in determining their functional properties, integration with native tissue and long-term repair outcome post-implantation. To date, most approaches for testing the mechanical properties of tissue-engineered grafts are non-sterile and invasive. There is an urgent need to develop novel sterile approaches for online monitoring mechanical properties of engineered tissues in order to ensure these engineered products meet the desired mechanical strength prior to implantation. In this paper, we overview various approaches for mechanical testing of engineered tissues, which span from traditional methods to medical imaging concepts in magnetic resonance elastography, ultrasound elastography, and optical coherence elastography. We focused on the applicability of these methods to the manufacturing of tissue-engineered products online, e.g., if such approach provides a sterile monitoring capacity and is capable of defining mechanical heterogeneity in engineered tissues throughout their growth in vitro in real-time. The review delves into various imaging modalities that employ distinct methods for inducing displacement within the sample, utilizing either strain-based or shear wave-based approaches. This displacement can be achieved through external stimulation or by harnessing ambient vibrations. Subsequently, the imaging process captures and visualizes the resultant displacement. We specifically highlight the advantages of novel non-invasive imaging methods such as ultrasound elastography and optical coherence elastography to assess the mechanical properties of engineered tissues in vitro, as well as their potential applications in cancer study, drug screening and the in vivo evaluation of the engineered tissues.
组织工程移植物的机械性能对于确定其功能特性、与原生组织的融合以及植入后的长期修复效果至关重要。迄今为止,测试组织工程移植物机械性能的大多数方法都是非无菌和侵入性的。目前迫切需要开发在线监测工程组织机械性能的新型无菌方法,以确保这些工程产品在植入前达到预期的机械强度。在本文中,我们概述了对工程组织进行机械测试的各种方法,这些方法涵盖了磁共振弹性成像、超声弹性成像和光学相干弹性成像等传统方法和医学成像概念。我们关注的重点是这些方法对在线制造组织工程产品的适用性,例如,如果这种方法能提供无菌监测能力,并能在体外生长的整个过程中实时确定工程组织的机械异质性。本综述深入探讨了各种成像模式,这些模式采用了不同的方法,利用基于应变或剪切波的方法在样本内诱导位移。这种位移可以通过外部刺激或利用环境振动来实现。随后,成像过程会捕捉并可视化由此产生的位移。我们特别强调了超声弹性成像和光学相干弹性成像等新型无创成像方法在评估体外工程组织机械性能方面的优势,以及它们在癌症研究、药物筛选和工程组织体内评估方面的潜在应用。
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引用次数: 0
A review on machine learning approaches in cardiac tissue engineering 综述心脏组织工程中的机器学习方法
Pub Date : 2024-04-03 DOI: 10.3389/fbiom.2024.1358508
Nikhith Kalkunte, Jorge Cisneros, Edward Castillo, Janet Zoldan
Cardiac tissue engineering (CTE) holds promise in addressing the clinical challenges posed by cardiovascular disease, the leading global cause of mortality. Human induced pluripotent stem cells (hiPSCs) are pivotal for cardiac regeneration therapy, offering an immunocompatible, high density cell source. However, hiPSC-derived cardiomyocytes (hiPSC-CMs) exhibit vital functional deficiencies that are not yet well understood, hindering their clinical deployment. We argue that machine learning (ML) can overcome these challenges, by improving the phenotyping and functionality of these cells via robust mathematical models and predictions. This review paper explores the transformative role of ML in advancing CTE, presenting a primer on relevant ML algorithms. We focus on how ML has recently addressed six key address six key challenges in CTE: cell differentiation, morphology, calcium handling and cell-cell coupling, contraction, and tissue assembly. The paper surveys common ML models, from tree-based and probabilistic to neural networks and deep learning, illustrating their applications to better understand hiPSC-CM behavior. While acknowledging the challenges associated with integrating ML, such as limited biomedical datasets, computational costs of learning data, and model interpretability and reliability, we examine suggestions for improvement, emphasizing the necessity for more extensive and diverse datasets that incorporate temporal and imaging data, augmented by synthetic generative models. By integrating ML with mathematical models and existing expert knowledge, we foresee a fruitful collaboration that unites innovative data-driven models with biophysics-informed models, effectively closing the gaps within CTE.
心脏组织工程(CTE)有望应对心血管疾病这一全球主要死亡原因所带来的临床挑战。人类诱导多能干细胞(hiPSC)是心脏再生治疗的关键,它提供了一种免疫兼容的高密度细胞来源。然而,hiPSC 衍生的心肌细胞(hiPSC-CMs)表现出重要的功能缺陷,这些缺陷尚未得到很好的了解,阻碍了其临床应用。我们认为,机器学习(ML)可以通过强大的数学模型和预测改善这些细胞的表型和功能,从而克服这些挑战。本综述论文探讨了 ML 在推动 CTE 方面的变革性作用,并介绍了相关的 ML 算法。我们重点关注 ML 最近是如何解决 CTE 中的六个关键难题的:细胞分化、形态、钙处理和细胞-细胞耦合、收缩和组织组装。论文介绍了常见的 ML 模型,从基于树的模型、概率模型到神经网络和深度学习模型,说明了它们在更好地理解 hiPSC-CM 行为方面的应用。我们承认整合 ML 所面临的挑战,如有限的生物医学数据集、学习数据的计算成本以及模型的可解释性和可靠性等,同时我们还研究了改进建议,强调需要更广泛、更多样的数据集,其中包括时间和成像数据,并通过合成生成模型加以扩充。通过将 ML 与数学模型和现有的专家知识相结合,我们预见到将创新的数据驱动模型与生物物理学信息模型结合起来的合作会取得丰硕成果,从而有效缩小 CTE 的差距。
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引用次数: 0
Synergistic antibacterial activity and inhibition of TiO2 nanotube arrays and loaded antibiotics against gram-positive and gram-negative bacteria TiO2 纳米管阵列和负载抗生素对革兰氏阳性和革兰氏阴性细菌的协同抗菌活性和抑制作用
Pub Date : 2024-04-03 DOI: 10.3389/fbiom.2024.1360443
Emmanuel Einyat Opolot, Haochen Wang, J. Capadona, Horst A. von Recum, Hoda Amani Hamedani
Introduction: Implantable medical devices continue to be vulnerable to bacterial infections. The unrelenting formation of antibiotic resistant bacterial strains not only exacerbates these infections but also renders the current treatment strategies impotent. The need is greater than ever for innovative and effective approaches to counteract drug-resistant bacteria. This study examines the innate antibacterial properties of TiO2 nanotube arrays (TNAs) and their ability to locally deliver antibiotics to inactivate gram-positive and gram-negative bacteria, in vitro.Methods: Using a two-step electrochemical anodization process, TNAs with a diameter of ∼100 nm and a length of ∼5 µm were grown on titanium substrates.Results and Discussion: After 24 h of incubation, as-fabricated TNAs showed 100% clearance of Escherichia coli, and 97% clearance of Staphylococcus aureus growth. The antibiotic-loaded TNAs demonstrated sustained slow-release of cefotaxime and imipenem measured over 14 days. In vitro bacterial studies revealed the capability of cefotaxime- and imipenem-loaded TNAs in completely inhibiting the growth with 100% clearance of Klebsiella pneumoniae after 24 and 48 h of incubation. Bacterial inhibition assay revealed a significantly enlarged inhibition zone difference of 18 mm around the imipenem-loaded TNAs against K. pneumoniae compared to the as-fabricated TNAs which was maintained for 7 days with ∼10 μgmL−1 of antibiotic released from the TNAs which was found to be lower than the dose required to completely eradicate multidrug resistant bacteria when used in conjunction with the antibacterial TNAs. The results of our study highlight the potential of TNAs as a versatile platform for addressing treatment strategies related to bacterial infections and antibiotic resistance in implantable medical devices.
导言:植入式医疗器械仍然很容易受到细菌感染。抗生素耐药菌株的不断产生不仅加剧了这些感染,也使目前的治疗策略变得无能为力。现在比以往任何时候都更需要创新而有效的方法来对付耐药细菌。本研究探讨了二氧化钛纳米管阵列(TNAs)与生俱来的抗菌特性及其在体外局部递送抗生素灭活革兰氏阳性和阴性细菌的能力:方法:采用两步电化学阳极氧化工艺,在钛基底上生长直径为 100 纳米、长度为 5 微米的二氧化钛纳米管阵列:培养 24 小时后,制成的 TNA 对大肠杆菌的清除率为 100%,对金黄色葡萄球菌的清除率为 97%。根据 14 天的测量,负载抗生素的 TNA 可持续缓释头孢他啶和亚胺培南。体外细菌研究表明,负载头孢他啶和亚胺培南的 TNA 能够完全抑制肺炎克雷伯菌的生长,培养 24 和 48 小时后,肺炎克雷伯菌的清除率达到 100%。细菌抑制试验显示,与原装 TNA 相比,装载亚胺培南的 TNA 对肺炎克雷伯菌的抑制区明显扩大,相差 18 毫米,这种抑制区可维持 7 天,TNA 释放的抗生素剂量为 10 μgmL-1 左右,低于与抗菌 TNA 结合使用时彻底根除耐多药细菌所需的剂量。我们的研究结果凸显了 TNA 作为一种多功能平台的潜力,可用于解决与植入式医疗器械中细菌感染和抗生素耐药性有关的治疗策略。
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引用次数: 0
The road after cancer: biomaterials and tissue engineering approaches to mediate the tumor microenvironment post-cancer treatment 癌症治疗后的道路:生物材料和组织工程方法调解癌症治疗后的肿瘤微环境
Pub Date : 2024-03-27 DOI: 10.3389/fbiom.2024.1347324
Lorna Westwood, Iain J. Nixon, Elaine Emmerson, Anthony Callanan
Cancer affects tens of millions of the world’s population each year with a stark mortality rate. It is well established that in order to be effective in treating solid tumor cancers, the current treatment methods used often sacrifice surrounding healthy tissue and cause damage at the site of treatment, inducing changes to the surrounding microenvironment. These changes to the microenvironment can lead to adverse side effects as well as long-term damage which continues to have a detrimental impact on the patient’s quality of life, even after remission. It is believed that by modulating the tumor microenvironment (TME) post-treatment, not only may the efficacy of current treatments be improved, but such associated negative side effects, as well as further complications arising from treatment, including metastasis, have the potential to be reduced. Mediating the microenvironment is also considered to aid in repairing the damaged site post-treatment, subsequently making the conditions more favourable for promoting regenerative processes. This review provides a brief overview of the alterations in the TME resulting from the three main cancer treatments–chemotherapy, radiation therapy and surgery–and the most common tissue engineering methods currently used in an attempt to mediate the TME post-cancer therapy. Furthermore, it investigates new emerging technologies within this field and the progress of such methods in terms of reaching the clinical setting.
癌症每年影响着全球数千万人口,死亡率极高。众所周知,为了有效治疗实体瘤癌症,目前使用的治疗方法往往会牺牲周围的健康组织,在治疗部位造成损伤,引起周围微环境的变化。这些微环境的变化会导致不良副作用和长期损害,甚至在病情缓解后仍会对患者的生活质量产生不利影响。人们相信,通过调节治疗后的肿瘤微环境(TME),不仅可以提高现有治疗方法的疗效,还有可能减少这些相关的负面副作用以及治疗引起的进一步并发症,包括转移。调节微环境也被认为有助于治疗后修复受损部位,从而为促进再生过程创造更有利的条件。本综述简要概述了化疗、放疗和手术这三种主要的癌症治疗方法对肿瘤微环境造成的改变,以及目前用于调解癌症治疗后肿瘤微环境的最常用组织工程方法。此外,报告还研究了该领域的新兴技术以及这些方法在临床应用方面的进展。
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引用次数: 0
Fabrication of gelatin-heparin based cartilage models: enhancing spatial complexity through refinement of stiffness properties and oxygen availability 制作基于明胶-肝素的软骨模型:通过完善硬度特性和氧气可用性提高空间复杂性
Pub Date : 2024-02-06 DOI: 10.3389/fbiom.2024.1331032
G. Lindberg, A. Norberg, B. Soliman, T. Jüngst, K. Lim, G. Hooper, J. Groll, T. Woodfield
The intricate nature of native cartilage, characterized by zonal variations in oxygen levels and ECM composition, poses a challenge for existing hydrogel-based tissue models. Consequently, these 3D models often present simplified renditions of the native tissue, failing to fully capture its heterogenous nature. The combined effects of hydrogel components, network properties, and structural designs on cellular responses are often overlooked. In this work, we aim to establish more physiological cartilage models through biofabrication of photopolymerizable allylated-gelatin (GelAGE) and Thiolated Heparin (HepSH) constructs with tailorable matrix stiffness and customized architectures. This involves systematically studying how the native glycosaminoglycan Heparin together with hydrogel stiffness, and oxygen availability within 3D structures influence chondrogenic differentiation and regional heterogeneity. A comprehensive library of 3D hydrogel constructs was successfully developed, encompassing GelAGE-HepSH hydrogels with three distinct stiffness levels: 12, 55 and 121 kPa, and three unique geometries: spheres, discs, and square lattices. In soft GelAGE-HepSH hydrogels, the localization of differentiating cells was observed to be irregular, while stiff hydrogels restricted the overall secretion of ECM components. The medium-stiff hydrogels were found to be most applicable, supporting both uniform tissue formation and maintained shape fidelity. Three different 3D architectures were explored, where biofabrication of smaller GelAGE-HepSH spheres without oxygen gradients induced homogenous, hyaline cartilage tissue formation. Conversely, fabrication of larger constructs (discs and lattices) with oxygen gradients could be utilized to design heterogenous cartilage tissue models. Similarly, temporal oxygen gradients were observed to drive interconnected deposition of glycosaminoglycans (GAGs). Control samples of GelAGE without HepSH did not exhibit any notable changes in chondrogenesis as a function of stiffness, architectures, or oxygen concentrations. Overall, the incorporation of HepSH within GelAGE hydrogels was observed to serve as an amplifier for the biological effects from both stiffness and oxygen cues. In conclusion, fabrication of GelAGE-HepSH constructs designed to impose limitations on oxygen availability induce more zone-specific cartilage tissue alignment. This systematic study of matrix components, network stiffness, and oxygen levels in 3D biofabricated structures contributes to the development of more physiologically relevant cartilage models while further enhancing our overall understanding of cartilage tissue engineering.
原生软骨的性质错综复杂,其特点是氧含量和 ECM 成分的分区变化,这给现有的基于水凝胶的组织模型带来了挑战。因此,这些三维模型往往是原生组织的简化再现,无法完全捕捉其异质性。水凝胶成分、网络特性和结构设计对细胞反应的综合影响往往被忽视。在这项工作中,我们的目标是通过生物制造具有可定制基质刚度和定制结构的光聚合烯丙基明胶(GelAGE)和硫醇化肝素(HepSH)构建体,建立更具生理特性的软骨模型。这包括系统研究原生糖胺聚糖肝素、水凝胶硬度和三维结构中的氧气供应如何影响软骨分化和区域异质性。我们成功开发了一个全面的三维水凝胶结构库,其中包括三种不同硬度水平的 GelAGE-HepSH 水凝胶:12、55 和 121 千帕,以及三种独特的几何形状:球形、圆盘形和方晶格形。在软性 GelAGE-HepSH 水凝胶中,观察到分化细胞的定位不规则,而硬性水凝胶则限制了 ECM 成分的整体分泌。研究发现,中等硬度的水凝胶最为适用,既能支持均匀的组织形成,又能保持形状的真实性。研究人员探索了三种不同的三维结构,其中,无氧梯度的较小 GelAGE-HepSH 球体生物制造可诱导均匀的透明软骨组织形成。相反,利用氧梯度制造较大的构建体(圆盘和晶格)可用于设计异质软骨组织模型。同样,还观察到时间氧梯度能驱动糖胺聚糖(GAGs)的相互连接沉积。不含 HepSH 的 GelAGE 对照样本在软骨形成过程中没有表现出任何与硬度、结构或氧气浓度相关的明显变化。总之,在 GelAGE 水凝胶中加入 HepSH 可以增强硬度和氧气对生物的影响。总之,制造 GelAGE-HepSH 构建物的目的是限制氧气的可用性,从而诱导更多特定区域的软骨组织排列。这项对三维生物制造结构中基质成分、网络刚度和氧气水平的系统性研究有助于开发更多与生理相关的软骨模型,同时进一步提高我们对软骨组织工程的整体认识。
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引用次数: 0
Fabrication of gelatin-heparin based cartilage models: enhancing spatial complexity through refinement of stiffness properties and oxygen availability 制作基于明胶-肝素的软骨模型:通过完善硬度特性和氧气可用性提高空间复杂性
Pub Date : 2024-02-06 DOI: 10.3389/fbiom.2024.1331032
G. Lindberg, A. Norberg, B. Soliman, T. Jüngst, K. Lim, G. Hooper, J. Groll, T. Woodfield
The intricate nature of native cartilage, characterized by zonal variations in oxygen levels and ECM composition, poses a challenge for existing hydrogel-based tissue models. Consequently, these 3D models often present simplified renditions of the native tissue, failing to fully capture its heterogenous nature. The combined effects of hydrogel components, network properties, and structural designs on cellular responses are often overlooked. In this work, we aim to establish more physiological cartilage models through biofabrication of photopolymerizable allylated-gelatin (GelAGE) and Thiolated Heparin (HepSH) constructs with tailorable matrix stiffness and customized architectures. This involves systematically studying how the native glycosaminoglycan Heparin together with hydrogel stiffness, and oxygen availability within 3D structures influence chondrogenic differentiation and regional heterogeneity. A comprehensive library of 3D hydrogel constructs was successfully developed, encompassing GelAGE-HepSH hydrogels with three distinct stiffness levels: 12, 55 and 121 kPa, and three unique geometries: spheres, discs, and square lattices. In soft GelAGE-HepSH hydrogels, the localization of differentiating cells was observed to be irregular, while stiff hydrogels restricted the overall secretion of ECM components. The medium-stiff hydrogels were found to be most applicable, supporting both uniform tissue formation and maintained shape fidelity. Three different 3D architectures were explored, where biofabrication of smaller GelAGE-HepSH spheres without oxygen gradients induced homogenous, hyaline cartilage tissue formation. Conversely, fabrication of larger constructs (discs and lattices) with oxygen gradients could be utilized to design heterogenous cartilage tissue models. Similarly, temporal oxygen gradients were observed to drive interconnected deposition of glycosaminoglycans (GAGs). Control samples of GelAGE without HepSH did not exhibit any notable changes in chondrogenesis as a function of stiffness, architectures, or oxygen concentrations. Overall, the incorporation of HepSH within GelAGE hydrogels was observed to serve as an amplifier for the biological effects from both stiffness and oxygen cues. In conclusion, fabrication of GelAGE-HepSH constructs designed to impose limitations on oxygen availability induce more zone-specific cartilage tissue alignment. This systematic study of matrix components, network stiffness, and oxygen levels in 3D biofabricated structures contributes to the development of more physiologically relevant cartilage models while further enhancing our overall understanding of cartilage tissue engineering.
原生软骨的性质错综复杂,其特点是氧含量和 ECM 成分的分区变化,这给现有的基于水凝胶的组织模型带来了挑战。因此,这些三维模型往往是原生组织的简化再现,无法完全捕捉其异质性。水凝胶成分、网络特性和结构设计对细胞反应的综合影响往往被忽视。在这项工作中,我们的目标是通过生物制造具有可定制基质刚度和定制结构的光聚合烯丙基明胶(GelAGE)和硫醇化肝素(HepSH)构建体,建立更具生理特性的软骨模型。这包括系统研究原生糖胺聚糖肝素、水凝胶硬度和三维结构中的氧气供应如何影响软骨分化和区域异质性。我们成功开发了一个全面的三维水凝胶结构库,其中包括三种不同硬度水平的 GelAGE-HepSH 水凝胶:12、55 和 121 千帕,以及三种独特的几何形状:球形、圆盘形和方晶格形。在软性 GelAGE-HepSH 水凝胶中,观察到分化细胞的定位不规则,而硬性水凝胶则限制了 ECM 成分的整体分泌。研究发现,中等硬度的水凝胶最为适用,既能支持均匀的组织形成,又能保持形状的真实性。研究人员探索了三种不同的三维结构,其中,无氧梯度的较小 GelAGE-HepSH 球体生物制造可诱导均匀的透明软骨组织形成。相反,利用氧梯度制造较大的构建体(圆盘和晶格)可用于设计异质软骨组织模型。同样,还观察到时间氧梯度能驱动糖胺聚糖(GAGs)的相互连接沉积。不含 HepSH 的 GelAGE 对照样本在软骨形成过程中没有表现出任何与硬度、结构或氧气浓度相关的明显变化。总之,在 GelAGE 水凝胶中加入 HepSH 可以增强硬度和氧气对生物的影响。总之,制造 GelAGE-HepSH 构建物的目的是限制氧气的可用性,从而诱导更多特定区域的软骨组织排列。这项对三维生物制造结构中基质成分、网络刚度和氧气水平的系统性研究有助于开发更多与生理相关的软骨模型,同时进一步提高我们对软骨组织工程的整体认识。
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Frontiers in biomaterials science
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