用于组织工程应用的三维聚 (ε-内) 支架的熔体静电纺丝
Summary
本议定书是一个全面的指导方针, 以直接的写作方式通过静电纺丝和聚合物熔体制造脚手架。我们系统地概述了该过程, 并定义了适当的参数设置, 以实现有针对性的脚手架结构。
Abstract
本教程反映了静电纺丝的基本原理和指南的聚合物熔体, 这是一种具有极大的生物医学应用潜力的添加剂制造技术。该技术促进了生物相容性聚合物纤维的直接沉积, 在亚微米范围内制备有序的支架。建立一个稳定的, 粘弹性, 聚合物射流之间的喷和收藏家是实现使用的应用电压, 可以直接写入。典型的多孔支架的一个显著优点是高表面积比, 它为细胞附着和生长提供了更有效的粘合部位。通过 fine-tuning 系统参数控制印刷工艺, 可使印刷脚手架的质量具有很高的重现性。它还为用户提供了一个灵活的制造平台, 使脚手架的形态结构符合他们的具体要求。为此, 我们提出了一个协议, 以获得不同的纤维直径使用熔融静电纺丝写入 (水电部) 的参数, 包括流量, 电压和收集速度的指导修正。此外, 我们还演示了如何优化 jet, 讨论经常遇到的技术难题, 解释故障排除技术, 并展示各种可打印的脚手架体系结构。
Introduction
三维 (3D) 细胞生物相容结构的制造是在组织工程 (TE) 中添加生物的关键贡献之一, 其目的是通过应用定制材料、细胞、生化因子来恢复组织,或他们的组合。因此, 用于 TE 应用的支架的主要要求包括: 生物相容性材料的可制造性、靶细胞侵袭的可控形态特性和增强细胞相互作用的优化表面特性1。
水电部是一种无溶剂的制造技术, 它将添加剂制造 (通常称为3D 印刷) 和静电纺丝的原理结合起来, 用于生产具有高度有序超薄纤维形态的聚合物网格2。它是一种直接的写作方法, 根据预先编写的代码3准确地存放纤维, 称为 G 代码。熔体纺结构目前准备使用一个平面4,5或一个芯轴6,7收集器, 分别制造多孔扁平和管状脚手架。
这项技术为 TE 和再生医学 (RM) 社区提供了显著的好处, 因为有可能直接打印医用级聚合物, 如聚 (ε-内) (PCL), 其中提出了良好的生物相容性8。其他的优势是有可能定制的大小和分布的孔隙率, 通过沉积纤维的高度有组织的方式来制造高表面积率的脚手架。在可以执行水电部之前, 聚合物首先需要应用热9。一旦处于流体状态, 施加的气压迫使它通过连接到高压源的金属喷流出。在表面张力和静电带电液滴对接地收集器的吸引力之间的力平衡导致泰勒锥的形成, 接着是喷射器的10。
图像和 in-house 构建用于此协议的设备的示意图图如图 1所示。此外, 还说明了使用绝缘胶带避免发热元件与喷周围带电黄铜部件之间的电放电的原理。绝缘不足会导致所实施硬件的内部损坏。
根据三系统参数 (温度、采集速度和气压) 的调整, 水电部可以在讨论部分中解释不同直径的纤维的制造。然而, 在大多数情况下, fine-tuning 和优化的喷气机将需要之前, 稳定的喷气机将被弹出。带电射流的可视化是验证过程一致性和均匀性的有效方法。在理想情况下, 飞行路径类似于由于由系统参数11控制的力平衡而获得的链路曲线。此外, 支架的微观和宏观结构取决于聚合物射流的飞行路径12。讨论部分给出了不同挠度行为和优化措施的详细表。
在本研究中, 我们提出了一个协议, 描述的制造步骤, 生产高控制纤维脚手架利用新技术。在这项工作中, 医疗级 pcl (分子量95-140 公斤/摩尔) 的使用, 因为这个医疗级 pcl 提高纯度超过技术等级, 其机械和加工性能是优良的水电部。PCL 的广泛熔体加工范围来源于其低熔点 (60 ° c) 和高热稳定性。此外, PCL 是一种慢速生物降解聚合物, 它使它成为许多组织工程应用的优秀材料13。
对于这项研究, 温度和收集器距离将保持恒定 (65 ° c 和82° c 为注射器和喷温度 (分别) 和12毫米为收藏家距离);应用电压, 收集器速度和气压, 但是, 将不同的制造有目标直径的纤维。在结果部分中提供了一份详细的研究报告, 并展示了 TE 和 RM 领域的不同应用 (表 1)。
Protocol
Representative Results
Discussion
整合 AM 为医疗领域的挑战找到创新的解决方案为 21st世纪提供了一个新的范例。”生物制造” 的 so-called 领域正在兴起, 制造技术的创新使得为 TE 应用生产高度复杂的体系结构。聚合物在直接写入模式下的静电纺丝被认为是最有希望的制造候选者之一, 以满足 TE 社区的需要, 在微米到纳米级的生物相容性材料的有序结构必需的27。
本教程旨在通过解释物…
Declarações
The authors have nothing to disclose.
Acknowledgements
这项工作得到了生物合作研究中心的资金支持, 它是用于细胞治疗制造、澳大利亚研究理事会的添加剂的弧形中心和慕尼黑技术大学的高级研究研究所。这项研究是由澳大利亚研究理事会工业改造培训中心在添加剂生物 http://www.additivebiomanufacturing.org (IC160100026) 进行的。请访问该网站的文章, 书籍, 电视或广播节目, 电子媒体, 或任何其他与该项目相关的文学作品。此外, 作者感激地承认玛丽亚 Flandes Iparraguirre 为支持在摄制, 菲利普哈伯德为声音和 Luise Grossmann 为摄制和编辑。
Materials
| Plastic syringe | Nordson Australia Pty Ltd | 7012072 | EFD BARREL O 3mL Clear 50 |
| Medical grade Poly (ε-caprolactone) (mPCL) | Corbion Purac, The Netherlands | PURASORB PC12 | |
| 23 GA needle | Nordson Australia Pty Ltd | 7018302 | #23GP .013 X .25 ORANGE 50 PC |
| Plunger | Nordson Australia Pty Ltd | 7012166 | PISTON O 3mL WH WIPER 50 |
| Pressure adapter | Nordson Australia Pty Ltd | 7012059 | ADAPTER ASM O 3mL BL 1.8M |
| Aluminium collector | Action Aluminium, Australia | SHP2 | Sheet 5005 H34 |
| Acrylic glass | Mulford Plastics Pty Ltd | ACC6-13094 | |
| Mach 3 software | Art Soft | Purchased online | |
| Safety switch interlock | RS components Pty Ltd | 12621330 | |
| High voltage generator | EMCO High Voltage Co. | DX250R | |
| Temperature controller | WATLOW | PM9R1FJ | |
| X and Y positioning slide | VELMEX Inc. | XN-10-0020-M011 |
Referências
- Muerza-Cascante, M. L., Haylock, D., Hutmacher, D. W., Dalton, P. D. Melt Electrospinning and Its Technologization in Tissue Engineering. Tissue Eng Part B Rev. 21 (2), 187-202 (2015).
- Brown, T. D., Dalton, P. D., Hutmacher, D. W. Melt electrospinning today: An opportune time for an emerging polymer process. Prog. in pol Sci. , (2015).
- Brown, T. D., Dalton, P. D., Hutmacher, D. W. Direct writing by way of melt electrospinning. Adv Mater. 23 (47), 5651-5657 (2011).
- Farrugia, B. L., et al. Dermal fibroblast infiltration of poly(ε-caprolactone) scaffolds fabricated by melt electrospinning in a direct writing mode. Biofabrication. 5 (2), 025001 (2013).
- Brown, T. D., et al. Melt electrospinning of poly (ε-caprolactone) scaffolds: Phenomenological observations associated with collection and direct writing. Mater. Sci. Eng. C. 45, 698-708 (2014).
- Jungst, T., et al. Melt electrospinning onto cylinders: effects of rotational velocity and collector diameter on morphology of tubular structures. Polym Int. 64 (9), 1086-1095 (2015).
- Brown, T. D. Design and Fabrication of Tubular Scaffolds via Direct Writing in a Melt Electrospinning Mode. Biointerphases. 7 (1), 1-16 (2012).
- Woodruff, M. A., Hutmacher, D. W. The return of a forgotten polymer-Polycaprolactone in the 21st century. Prog. in pol Sci. 35 (10), 1217-1256 (2010).
- Hutmacher, D. W., Dalton, P. D. Melt electrospinning. Chem Asian J. 6 (1), 44-56 (2011).
- Wei, C., Gang, T. Q., Chen, L. J., Zhao, Y. Critical condition for the transformation from Taylor cone to cone-jet. Chin. Phys. B. 23 (6), 064702 (2014).
- Mikl, B., et al. Discrete viscous threads. ACM Trans. Graph. 29 (4), 1-10 (2010).
- Hochleitner, G., et al. Fibre pulsing during melt electrospinning writing. BioNanoMaterials. 17 (3-4), 159 (2016).
- Poh, P. S. P., et al. Polylactides in additive biomanufacturing. Adv Drug Del Rev. 107, 228-246 (2016).
- Vaquette, C., Cooper-White, J. J. Increasing electrospun scaffold pore size with tailored collectors for improved cell penetration. Acta Biomater. 7 (6), 2544-2557 (2011).
- Thibaudeau, L., et al. A tissue-engineered humanized xenograft model of human breast cancer metastasis to bone. Dis Model Mech. 7 (2), 299-309 (2014).
- Holzapfel, B. M., et al. Species-specific homing mechanisms of human prostate cancer metastasis in tissue engineered bone. Biomaterials. 35 (13), 4108-4115 (2014).
- Bas, O., et al. Enhancing structural integrity of hydrogels by using highly organised melt electrospun fibre constructs. Eur. Polym. J. 72, 451-463 (2015).
- Visser, J., et al. Reinforcement of hydrogels using three-dimensionally printed microfibres. Nat. Commun. 6, 6933 (2015).
- Haigh, J. N., et al. Hierarchically Structured Porous Poly(2-oxazoline) Hydrogels. Macromol. Rapid Commun. 37 (1), 93-99 (2016).
- Wagner, F., et al. A validated preclinical animal model for primary bone tumor research. JBJS. 98 (11), 916-925 (2016).
- Baldwin, J., et al. Periosteum tissue engineering in an orthotopic in vivo platform. Biomaterials. 121, 193-204 (2017).
- Tourlomousis, F., Chang, R. C. Dimensional Metrology of Cell-matrix Interactions in 3D Microscale Fibrous Substrates. Procedia CIRP. 65, 32-37 (2017).
- Muerza-Cascante, M. L., et al. Endosteal-like extracellular matrix expression on melt electrospun written scaffolds. Acta Biomater. 52, 145-158 (2017).
- Delalat, B., et al. 3D printed lattices as an activation and expansion platform for T cell therapy. Biomaterials. 140, 58-68 (2017).
- Bas, O., et al. Biofabricated soft network composites for cartilage tissue engineering. Biofabrication. 9 (2), 025014 (2017).
- Hansske, F., et al. Via precise interface engineering towards bioinspired composites with improved 3D printing processability and mechanical properties. J. Mater. Chem. B. , (2017).
- Melchels, F. P. W., et al. Additive manufacturing of tissues and organs. Prog. in pol Sci. 37 (8), 1079-1104 (2012).
- Hochleitner, G., et al. Fibre pulsing during melt electrospinning writing. BioNanoMaterials. 17, 159 (2016).
- Persano, L., Camposeo, A., Tekmen, C., Pisignano, D. Industrial Upscaling of Electrospinning and Applications of Polymer Nanofibers: A Review. Macromol. Mater. Eng. 298 (5), 504-520 (2013).
- Wunner, F. M., et al. . Comprehensive Biomaterials II. , 217-235 (2017).
- Loh, Q. L., Choong, C. Three-Dimensional Scaffolds for Tissue Engineering Applications: Role of Porosity and Pore Size. Tissue Eng Part B Rev. 19 (6), 485-502 (2013).
- Ristovski, N., et al. Improved fabrication of melt electrospun tissue engineering scaffolds using direct writing and advanced electric field control. Biointerphases. 10 (1), 011006 (2015).
- Wei, C., Dong, J. Direct fabrication of high-resolution three-dimensional polymeric scaffolds using electrohydrodynamic hot jet plotting. J Micromech Microeng. 23 (2), 025017 (2013).
- Hacker, C., et al. Electrospinning of polymer melt : steps towards an upscaled multi-jet process. , 71-76 (2009).
- Nayak, R., Padhye, R., Kyratzis, I. L., Truong, Y. B., Arnold, L. Recent advances in nanofibre fabrication techniques. Text. Res. J. 82 (2), 129-147 (2012).
- Li, H., et al. Interjet distance in needleless melt differential electrospinning with umbellate nozzles. J. Appl. Polym. Sci. 131 (15), (2014).
- Liao, S., et al. Effect of humidity on melt electrospun polycaprolactone scaffolds. BioNanoMaterials. 17, 173 (2016).
- Detta, N., et al. Melt electrospinning of polycaprolactone and its blends with poly(ethylene glycol). Polym Int. 59 (11), 1558-1562 (2010).
- Hochleitner, G., Hümmer, J. F., Luxenhofer, R., Groll, J. High definition fibrous poly(2-ethyl-2-oxazoline) scaffolds through melt electrospinning writing. Polymer. 55 (20), 5017-5023 (2014).
- Chen, F., et al. Additive Manufacturing of a Photo-Cross-Linkable Polymer via Direct Melt Electrospinning Writing for Producing High Strength Structures. Biomacromolecules. 17 (1), 208-214 (2016).
- Hochleitner, G., et al. Additive manufacturing of scaffolds with sub-micron filaments via melt electrospinning writing. Biofabrication. 7 (3), 035002 (2015).
