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3D printed scaffolds: Challenges toward developing relevant cellular in vitro models 3D打印支架:开发相关细胞体外模型的挑战
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-06-01 DOI: 10.1016/j.bbiosy.2022.100044
Beatriz Molina-Martínez , Luis M. Liz-Marzán

Three-dimensional (3D) cell culture technology has rapidly emerged, as a result of the increasing demand for improved in vitro systems that better resemble human physiology. Promising microphysiological systems have been fabricated by combining complex 3D culture with 3D-printing technologies. These models overperform existing in vitro systems regarding potential for biofabrication and predictive power. However, most systems under development do not ultimately find a long-term application. We provide herein an overview of the challenges to be considered when developing 3D in vitro systems by means of printed scaffolds, as well as some of the limitations of existing models.

三维(3D)细胞培养技术迅速兴起,这是对改进体外系统的需求不断增加的结果,该系统更像人类生理。通过将复杂的3D培养与3D打印技术相结合,已经制造出了有前途的微生理系统。这些模型在生物制造和预测能力方面优于现有的体外系统。然而,大多数正在开发的系统最终都没有找到长期的应用。我们在此概述了通过打印支架开发体外3D系统时需要考虑的挑战,以及现有模型的一些局限性。
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引用次数: 2
Amorphous calcium phosphate, the lack of order is an abundance of possibilities 无定形磷酸钙,缺乏秩序却是丰富的可能性
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.1016/j.bbiosy.2021.100037
Lorenzo Degli Esposti, Michele Iafisco

For almost three decades from its discovery, amorphous calcium phosphate (ACP) was not considered a suitable biomaterial due to its structural instability. Thanks to its unique properties in respect to crystalline calcium phosphate phases, nowadays ACP is used in promising devices for hard tissue regeneration. Here we have highlighted the features of ACP that were harnessed to create excellent biomaterials for dental remineralization, self-setting bone cements, drug delivery, and coatings of prostheses. Its current limitations as well as future perspectives of development were concisely described. Despite more research works are needed, we envisage that the future of ACP is bright.

无定形磷酸钙(ACP)被发现近30年来,由于其结构不稳定,不被认为是一种合适的生物材料。由于其在结晶磷酸钙相方面的独特性能,如今ACP被用于有前途的硬组织再生设备。在这里,我们强调了ACP的特点,这些特点被用来制造优秀的生物材料,用于牙科再矿化、自固化骨水泥、药物输送和假体涂层。简要叙述了其目前的局限性和未来的发展前景。尽管还需要更多的研究工作,但我们认为ACP的未来是光明的。
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引用次数: 5
Development of decellularization protocol for caprine small intestine submucosa as a biomaterial 羊小肠粘膜下层脱细胞生物材料的研究进展
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.1016/j.bbiosy.2021.100035
Hemant Singh , Shiv Dutt Purohit , Rakesh Bhaskar , Indu Yadav , Mukesh Kumar Gupta , Narayan Chandra Mishra

Decellularized animal tissues have been proven to be promising biomaterials for various tissue engineering (TE) applications. Among various animal tissues, small intestine submucosa (SIS) has gained attention of many researchers due to its easy availability from the abattoir waste, excellent physicochemical and biological characteristics of a good biomaterial. In this study, Caprine SIS was decellularized to get decellularized caprine SIS (DG-SIS). For decellularization, several physical, chemical and enzymatic protocols have been described in the literature. To optimize the decellularization of caprine SIS, several decellularization protocol (DP), including an in-house developed by us, had been attempted, and effect of the different DPs on the obtained DG-SIS were assessed in terms of decellularization, physiochemical and biological properties. All the DPs differ in terms of decellularization, but three DPs where ionic detergent like sodium dodecyl sulphate (SDS) has been used, largely affect the native composition (e.g. glycosaminoglycans (GAGs)), biological properties and other physiochemical properties of the G-SIS as compared to the DP that uses hypertonic solution of potassium iodide (KI) and non-ionic detergent (TritonX-100). The obtained DG-SISs were fibrous, hemocompatible, biocompatible, hydrophilic, biodegradable and exhibited significant antibacterial activity. Therefore, the DG-SIS will be a prospective biomaterial for TE applications.

脱细胞动物组织已被证明是各种组织工程(TE)应用的有前途的生物材料。在各种动物组织中,小肠黏膜下层(small intestinal sub粘膜,SIS)因其易于从屠宰场废弃物中提取,具有优异的理化生物学特性,是一种良好的生物材料而受到众多研究者的关注。本研究对山羊SIS进行脱细胞处理,得到脱细胞山羊SIS (DG-SIS)。对于脱细胞,文献中描述了几种物理,化学和酶的方案。为了优化山羊SIS的脱细胞,我们尝试了几种脱细胞方案(DP),包括我们自己开发的一种脱细胞方案,并从脱细胞、理化和生物学特性方面评估了不同DP对绵羊SIS脱细胞的影响。所有的DPs在脱细胞方面都有所不同,但与使用碘化钾高渗溶液(KI)和非离子洗涤剂(TritonX-100)的DP相比,使用十二烷基硫酸钠(SDS)等离子洗涤剂的DPs在很大程度上影响了G-SIS的天然组成(如糖胺聚糖(GAGs))、生物特性和其他物理化学特性。所得dg - sis具有纤维性、血液相容性、生物相容性、亲水性、可生物降解性,并具有显著的抗菌活性。因此,DG-SIS将是一种有前景的TE生物材料。
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引用次数: 3
Biomaterials enhancing performance of cell and nucleic-acid therapies: An opportunity in the brain 增强细胞和核酸治疗性能的生物材料:大脑中的机会
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.1016/j.bbiosy.2021.100036
Christopher Lally , Kieran Joyce , Abhay Pandit

The brain has limited innate ability to promote repair, regeneration and functional recovery following injury, disease, or developmental disorder. Although cell and gene therapies have significant potential in the brain, no single treatment is likely to succeed in isolation. Here we discuss the current state of the art in cell and nucleic-acid-based neurotherapeutics and argue for the development of combination therapies that use biomaterials to help overcome the current limitations of cell and gene therapies alone.

大脑在损伤、疾病或发育障碍后促进修复、再生和功能恢复的先天能力有限。虽然细胞和基因疗法在大脑中具有巨大的潜力,但没有一种单独的治疗方法可能成功。在这里,我们讨论了目前以细胞和核酸为基础的神经疗法的最新进展,并主张开发使用生物材料的联合疗法,以帮助克服目前单独使用细胞和基因疗法的局限性。
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引用次数: 2
Spacer length and serum protein adsorption affect active targeting of trastuzumab-modified nanoparticles 间隔长度和血清蛋白吸附影响曲妥珠单抗修饰纳米颗粒的活性靶向性
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.1016/j.bbiosy.2021.100032
Christina Barth , Hendrik Spreen , Dennis Mulac , Lucas Keuter , Matthias Behrens , Hans-Ulrich Humpf , Klaus Langer

Receptor-mediated active targeting of nanocarriers is a widely investigated approach to specifically address cancerous cells and tissues in the human body. The idea is to use these formulations as drug carriers with enhanced specificity and therefore reduced systemic side effects. Until today a big obstacle to reach this goal remains the adsorption of serum proteins to the nanocarrier's surface after contact with biological fluids. In this context different nanoparticle characteristics could be beneficial for effective active targeting after formation of a protein corona which need to be identified. In this study trastuzumab was used as an active targeting ligand which was covalently attached to human serum albumin nanoparticles. For coupling reaction different molecular weight spacers were used and resulting physicochemical nanoparticle characteristics were evaluated. The in vitro cell association of the different nanoparticle formulations was tested in cell culture experiments with or without fetal bovine serum. For specific receptor-mediated cell interaction SK-BR-3 breast cancer cells with human epidermal growth factor receptor 2 (HER2) overexpression were used. MCF-7 breast cancer cells with normal HER2 expression served as control. Furthermore, serum protein adsorption on respective nanoparticles was characterized. The qualitative and quantitative composition of the protein corona was analyzed by SDS-PAGE and LC-MS/MS and the influence of protein adsorption on active targeting capability was determined.

受体介导的纳米载体的活性靶向是一种广泛研究的方法,专门针对人体内的癌细胞和组织。我们的想法是使用这些配方作为药物载体,增强特异性,从而减少全身副作用。直到今天,实现这一目标的一大障碍仍然是血清蛋白在与生物液体接触后对纳米载体表面的吸附。在这种情况下,不同的纳米颗粒特性可能有利于在形成需要识别的蛋白质冠后有效的主动靶向。在这项研究中,曲妥珠单抗被用作一种活性靶向配体,共价附着在人血清白蛋白纳米颗粒上。在偶联反应中,采用了不同分子量的间隔剂,并对其理化性质进行了评价。不同纳米颗粒配方的体外细胞关联在细胞培养实验中进行了测试,有或没有胎牛血清。在特异性受体介导的细胞相互作用中,使用了人表皮生长因子受体2 (HER2)过表达的SK-BR-3乳腺癌细胞。以HER2表达正常的MCF-7乳腺癌细胞为对照。此外,还表征了血清蛋白在不同纳米颗粒上的吸附。利用SDS-PAGE和LC-MS/MS对蛋白冠的定性和定量组成进行分析,并确定蛋白吸附对活性靶向能力的影响。
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引用次数: 1
Magnetic CuFe2O4 with intrinsic protease-like activity inhibited cancer cell proliferation and migration through mediating intracellular proteins 具有内在蛋白酶样活性的磁性CuFe2O4通过介导细胞内蛋白抑制癌细胞增殖和迁移
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-03-01 DOI: 10.1016/j.bbiosy.2021.100038
Daomei Chen , Liang Jiang , Tao Lei , Guo Xiao , Yuanfeng Wang , Xiaoqiong Zuo , Bin Li , Lingli Li , Jiaqiang Wang

Protease has been widely used in biological and industrial fields. Developing efficient artificial enzyme mimics remains a major technical challenge due to the high stability of peptide bonds. Nanoenzymes with high stability, high activity and low cost, provided new opportunities to break through natural enzyme inherent limitations. However, compared with many nanomaterials with inherent peroxidase activity, the intrinsic mimic proteases properties of magnetic nanomaterials were seldom explored, let alone the interaction between magnetic nanomaterials and cellular proteins. Herein, we reported for the first time that magnetic CuFe2O4 possesses inherent protease activity to hydrolyze bovine serum albumin (BSA) and casein under physiological conditions, and the CuFe2O4 is more resistant to high temperature than the natural trypsin. It also exhibited significantly higher catalytic efficiency than other copper nanomaterials and can be recycled for many times. Protease participated in pathophysiological processes and all stages of tumor progression. Interesting, CuFe2O4 exhibited anti-proliferative effect on A549, SKOV3, HT-29, BABL-3T3 and HUVEC cells, as well as it was particularly sensitive against SKOV3 cells. CuFe2O4 was about 30 times more effective than conventional chemotherapy drugs oxaliplatin and artesunate against SKOV3 cells. In addition, CuFe2O4 also mediated the expression of intracellular proteins, such as MMP-2, MMP-9, F-actin, and NF-kB, which may be associated with global protein hydrolysis by CuFe2O4, leading to inhibition of cell migration. The merits of the high magnetic properties, good protease-mimic and antitumor activities make CuFe2O4 nanoparticles very prospective candidates for many applications such as proteomics and biotechnology.

蛋白酶在生物和工业领域有着广泛的应用。由于肽键的高稳定性,开发高效的人工酶模拟物仍然是一个主要的技术挑战。纳米酶具有高稳定性、高活性和低成本的特点,为突破天然酶固有的局限性提供了新的机遇。然而,与许多具有固有过氧化物酶活性的纳米材料相比,磁性纳米材料的固有模拟蛋白酶性质很少被探索,更不用说磁性纳米材料与细胞蛋白质的相互作用了。本研究首次报道了磁性CuFe2O4在生理条件下具有水解牛血清白蛋白(BSA)和酪蛋白的内在蛋白酶活性,并且具有比天然胰蛋白酶更强的耐高温能力。与其他铜纳米材料相比,其催化效率显著提高,且可多次回收利用。蛋白酶参与了病理生理过程和肿瘤发展的各个阶段。有趣的是,CuFe2O4对A549、SKOV3、HT-29、BABL-3T3和HUVEC细胞均表现出抗增殖作用,其中对SKOV3细胞尤为敏感。CuFe2O4对SKOV3细胞的治疗效果是传统化疗药物奥沙利铂和青蒿琥酯的30倍左右。此外,CuFe2O4还介导了细胞内蛋白的表达,如MMP-2、MMP-9、F-actin和NF-kB,这些蛋白可能与CuFe2O4对全局蛋白的水解有关,从而抑制细胞迁移。CuFe2O4纳米粒子具有高磁性、良好的蛋白酶模拟和抗肿瘤活性等优点,在蛋白质组学和生物技术等领域具有广阔的应用前景。
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引用次数: 3
A review of strategies for development of tissue engineered meniscal implants 组织工程半月板植入物的发展策略综述
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-01 DOI: 10.1016/j.bbiosy.2021.100026
George J. Klarmann , Joel Gaston , Vincent B. Ho

The meniscus is a key stabilizing tissue of the knee that facilitates proper tracking and movement of the knee joint and absorbs stresses related to physical activity. This review article describes the biology, structure, and functions of the human knee meniscus, common tears and repair approaches, and current research and development approaches using modern methods to fabricate a scaffold or tissue engineered meniscal replacement. Meniscal tears are quite common, often resulting from sports or physical training, though injury can result without specific contact during normal physical activity such as bending or squatting. Meniscal injuries often require surgical intervention to repair, restore basic functionality and relieve pain, and severe damage may warrant reconstruction using allograft transplants or commercial implant devices. Ongoing research is attempting to develop alternative scaffold and tissue engineered devices using modern fabrication techniques including three-dimensional (3D) printing which can fabricate a patient-specific meniscus replacement. An ideal meniscal substitute should have mechanical properties that are close to that of natural human meniscus, and also be easily adapted for surgical procedures and fixation. A better understanding of the organization and structure of the meniscus as well as its potential points of failure will lead to improved design approaches to generate a suitable and functional replacement.

半月板是膝关节的关键稳定组织,有助于膝关节的正确跟踪和运动,并吸收与体育活动有关的压力。本文综述了人类膝关节半月板的生物学、结构和功能,常见的撕裂和修复方法,以及目前使用现代方法制造支架或组织工程半月板替代物的研究和开发方法。半月板撕裂很常见,通常是由运动或体育训练引起的,尽管在弯腰或下蹲等正常体育活动中,没有特定的接触也会造成损伤。半月板损伤通常需要手术干预来修复,恢复基本功能和减轻疼痛,严重的损伤可能需要使用同种异体移植或商业植入装置进行重建。正在进行的研究正在尝试开发替代支架和组织工程设备,使用现代制造技术,包括三维(3D)打印,可以制造患者特定的半月板替代物。理想的半月板替代物应具有接近自然人类半月板的力学特性,并且易于适应外科手术和固定。更好地了解半月板的组织和结构以及其潜在的故障点将导致改进的设计方法,以产生合适的和功能性的替代品。
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引用次数: 9
Third body damage and wear in arthroplasty bearing materials: A review of laboratory methods 关节置换术中承载材料的损伤和磨损:实验室方法综述
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-01 DOI: 10.1016/j.bbiosy.2021.100028
Raelene M Cowie, Louise M Jennings

Third body wear of arthroplasty bearing materials can occur when hard particles such as bone, bone cement or metal particles become trapped between the articulating surfaces. This can accelerate overall implant wear, potentially leading to early failure. With the development of novel bearing materials and coatings, there is a need to develop and standardise test methods which reflect third body damage seen on retrieved implants. Many different protocols and approaches have been developed to replicate third body wear in the laboratory but there is currently no consensus as to the optimal method for simulating this wear mode, hence the need to better understand existing methods. The aim of this study was to review published methods for experimental simulation of third body wear of arthroplasty bearing materials, to discuss the advantages and limitations of different approaches, the variables to be considered when designing a method and to highlight gaps in the current literature. The methods were divided into those which introduced abrasive particles into the articulating surfaces of the joint and those whereby third body damage is created directly to the articulating surfaces. However, it was found that there are a number of parameters, for example the influence of particle size on wear, which are not yet fully understood. The study concluded that the chosen method or combination of methods used should primarily be informed by the research question to be answered and risk analysis of the device.

当骨、骨水泥或金属颗粒等硬颗粒被困在关节表面之间时,就会发生关节置换术轴承材料的第三体磨损。这可能会加速种植体的整体磨损,可能导致早期失效。随着新型轴承材料和涂层的发展,有必要开发和标准化的测试方法,以反映在回收的种植体上看到的第三体损伤。已经开发了许多不同的方案和方法来在实验室中复制第三体磨损,但目前对于模拟这种磨损模式的最佳方法尚无共识,因此需要更好地了解现有方法。本研究的目的是回顾已发表的关节置换承载材料第三体磨损实验模拟方法,讨论不同方法的优点和局限性,设计方法时需要考虑的变量,并突出当前文献中的空白。该方法分为那些引入磨料颗粒到关节关节面和那些其中第三体损伤是直接创建到关节面。然而,发现有许多参数,例如粒度对磨损的影响,尚未完全了解。该研究的结论是,所选择的方法或使用的方法组合应该主要由研究问题来回答和设备的风险分析。
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引用次数: 4
Collagen type II: From biosynthesis to advanced biomaterials for cartilage engineering II型胶原:从生物合成到软骨工程的高级生物材料
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-01 DOI: 10.1016/j.bbiosy.2021.100030
Z Wu , SH Korntner , AM Mullen , DI Zeugolis

Collagen type II is the major constituent of cartilage tissue. Yet, cartilage engineering approaches are primarily based on collagen type I devices that are associated with suboptimal functional therapeutic outcomes. Herein, we briefly describe cartilage's development and cellular and extracellular composition and organisation. We also provide an overview of collagen type II biosynthesis and purification protocols from tissues of terrestrial and marine species and recombinant systems. We then advocate the use of collagen type II as a building block in cartilage engineering approaches, based on safety, efficiency and efficacy data that have been derived over the years from numerous in vitro and in vivo studies.

II型胶原是软骨组织的主要成分。然而,软骨工程方法主要基于与次优功能治疗结果相关的胶原I型装置。在这里,我们简要地描述软骨的发育和细胞和细胞外的组成和组织。我们还提供了II型胶原蛋白的生物合成和纯化方案的概述,从陆地和海洋物种和重组系统的组织。然后,我们主张使用II型胶原蛋白作为软骨工程方法的构建块,基于多年来从大量体外和体内研究中获得的安全性、效率和疗效数据。
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引用次数: 13
Advanced non-fluoride approaches to dental enamel remineralization: The next level in enamel repair management 牙釉质再矿化的先进无氟方法:牙釉质修复管理的新水平
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-01 DOI: 10.1016/j.bbiosy.2021.100029
Bernd Grohe , Silvia Mittler

In modern dentistry, a minimally invasive management of early caries lesions or early-stage erosive tooth wear (ETW) with synthetic remineralization systems has become indispensable. In addition to fluoride, which is still the non-plus-ultra in these early caries/ETW treatments, a number of new developments are in the test phase or have already been commercialized. Some of these systems claim that they are comparable or even superior to fluoride in terms of their ability to remineralize enamel. Besides, their use can help avoid some of the risks associated with fluoride and support treatments of patients with a high risk of caries. Two individual non-fluoride systems can be distinguished; intrinsic and extrinsic remineralization approaches. Intrinsic (protein/peptide) systems adsorb to hydroxyapatite crystals/organics located within enamel prisms and accumulate endogenous calcium and phosphate ions from saliva, which ultimately leads to the re-growth of enamel crystals. Extrinsic remineralization systems function on the basis of the external (non-saliva) supply of calcium and phosphate to the crystals to be re-grown. This article, following an introduction into enamel (re)mineralization and fluoride-assisted remineralization, discusses the requirements for non-fluoride remineralization systems, particularly their mechanisms and challenges, and summarizes the findings that underpin the most promising advances in enamel remineralization therapy.

在现代牙科中,使用合成再矿化系统对早期龋齿病变或早期侵蚀性牙齿磨损(ETW)进行微创治疗已成为必不可少的。除了氟化物(在这些早期龋齿/ETW治疗中仍然是非加超)之外,许多新开发的产品正处于测试阶段或已经商业化。其中一些系统声称,它们在牙釉质再矿化能力方面与氟化物相当,甚至优于氟化物。此外,它们的使用可以帮助避免与氟化物有关的一些风险,并支持对龋齿高风险患者的治疗。可以区分两种独立的无氟系统;内在和外在再矿化途径。内源性(蛋白质/肽)系统吸附于牙釉质棱镜内的羟基磷灰石晶体/有机物,并从唾液中积累内源性钙和磷酸盐离子,最终导致牙釉质晶体的重新生长。外部再矿化系统的功能是基于外部(非唾液)钙和磷酸盐的供应,以重新生长的晶体。本文在介绍了牙釉质(再)矿化和氟化物辅助再矿化的基础上,讨论了对非氟化物再矿化系统的要求,特别是它们的机制和挑战,并总结了牙釉质再矿化治疗中最有希望的进展。
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引用次数: 18
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