1.1 本指南是使用生物墨水和生物材料墨水打印组织工程医疗产品的资源。现有标准以更通用的方式涵盖生物材料和支架（指南 F2150 指导 F2027 ，ISO 10993系列）。本指南特别关注利用生物墨水和具有固有或可诱导流体特性的生物材料墨水的挤出生物打印，包括或不包括用于构建TEMP的封装细胞。在本指南的剩余部分中，生物墨水和生物材料墨水将统称为生物墨水。 1.2 在本指南中，生物打印被定义为制造用于生物或医学应用的结构化结构的材料（生物墨水）的三维打印。 1.3 TEMP可以通过许多不同的生物打印方式生产，包括但不限于以下方式:静电纺丝、电喷射、基于挤出、基于液滴、基于喷墨和激光辅助生物打印。基于挤出的生物打印是本文件的主要焦点，因为它是目前用于构建TEMP的最广为人知的模式，但也涉及其他生物打印模式。 1.4 本指南将重点介绍生物墨水和用作具有固有或可诱导流体特性的墨水的生物材料。这些油墨可以包含也可以不包含封装的细胞。油墨的化学性质和其他影响印刷性能的因素被处理。 1.5 印刷前和印刷注意事项是本指南的重点，但关于印刷后的注意事项- 还讨论了印刷产品的稳定性。 1.6 本指南将介绍有关生物墨水无菌性和细胞相容性的评估，包括化学和物理台式测试，以及打印后细胞活力的测量。 1.7 本标准并不旨在解决与其使用相关的所有安全问题（如有）。本标准的使用者有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.8 本国际标准是根据世界贸易组织技术性贸易壁垒委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认的标准化原则制定的。 ====意义和用途====== 4.1 标准结构-- 本标准是根据典型的生物打印操作工作流程组织的。通过这种方式，该标准有三个主要部分:打印前的注意事项、打印后的注意事项。还有一节是关于生物打印方式以及产品发布、容器和运输的其他考虑因素。某些过程会出现在多个部分，例如细胞相容性或交联，因为这些问题在印刷前、印刷过程中和印刷过程后都有考虑。主要章节内容如下。 范围 部分 1. 参考文件 部分 2. 术语 部分 3. 意义和用途 部分 4. 生物打印方式 部分 5. 打印前注意事项 部分 6. 打印注意事项 部分 7. 打印后注意事项 部分 8. 其他注意事项 部分 9 关键词 部分 10 工具书类 4.1.1 打印前注意事项-- 打印前的注意事项包括:生物墨水的常见应用、支撑材料和生物墨水的选择注意事项。生物墨水的选择考虑因素包括:生物墨水的配方、生物墨水的特性、配方引起的特性变化、无菌性、细胞成分和逃亡元素的考虑因素。在生物墨水性质中，有与粘弹性性质、化学性质、聚合物和官能团的结构、材料纯度、交联机制和降解因素有关的考虑因素。 以下列出了关于打印前注意事项的部分内容。 Bioink常见应用程序 6.2 支架材料注意事项 6.3 Bioink选择 6.4 生物墨水的配方 6.4.1 成分浓度 6.4.1.1 各部件的功能 6.4.1.2 Bioink属性 6.4.2 粘弹性特性 6.4.2.1 化学性质 6.4.2.2 聚合物结构与官能团 6.4.2.3 材料纯度 6.4.2.4 交联机理 6.4.2.5 生物墨水的降解 6.4.2.6 配方引起的性能变化 6.4.3 影响生物反应的配方改良 6.4.3.1 配方改良对流变学的影响 6.4.3.2 影响力学性能的配方改性 6.4.3.3 无菌 6.4.4 灭菌方法 6.4.4.1 评估 6.4.4.2 蜂窝组件注意事项 6.4.5 细胞相容性 6.4.5.1 细胞沉淀 6.4.5.2 逃亡分子注意事项 6.4.6 4.1.2 打印注意事项-- 打印注意事项包括:可打印性，特别是生物墨水注意事项、细胞成分、支撑材料、稳定性和无菌打印注意事项。在细胞成分部分中，考虑因素包括细胞活力、温度、细胞剪切、细胞分布、沉积和生物墨水中的浓度。下面列出了有关打印注意事项的部分内容。 可打印性 7.2 Bioink注意事项 7.2.1 细胞对可打印性的影响 7.2.1.1 生物材料对印刷性能的影响 7.2.1.2 温度 7.2.2 评估 7.2.3 蜂窝组件注意事项 7.3 印刷过程中细胞的存活率 7.3.1 温度 7.3.2 单元格剪切 7.3.3 Bioink中的细胞分布 7.3.4 细胞沉淀 7.3.4.1 细胞浓度 7.3.5 支架材料注意事项 7.4 印刷支撑材料 7.4.1 用作嵌入介质的支撑材料 7.4.2 稳定 7.5 细胞相容性 7.5.1 交联 7.5.2 温度 7.5.3 无菌打印注意事项 7.6 4.1.3 打印后注意事项-- 打印后的注意事项包括:打印后的生物墨水、稳定性以及移除临时组件和材料的注意事项。 印刷后的稳定性侧重于模式（如交联、温度诱导的自组装和蒸发）以及对细胞健康、功能和性质修饰的稳定作用。拆除临时构件和材料的注意事项包括构件类型、拆除时间和方法以及拆除效果。关于印刷后注意事项的章节内容如下所列。 打印后Bioink注意事项 8.2 结构保真度 8.2.1 细胞的生存能力 8.2.2 生物材料特性 8.2.3 细胞沉淀 8.2.4 细胞形态学 8.2.5 稳定性注意事项 8.3 稳定模式 8.3.1 交联 8.3.1. 1. 温度引起的自组装 8.3.1.2 蒸发 8.3.1.3 稳定效果 8.3.2 细胞健康与功能 8.3.2.1 机械性能的修改 8.3.2.2 拆除临时部件和材料的注意事项 8.4 组件类型 8.4.1 支持组件 8.4.1.1 Bioink元素 8.4.1.2 支撑槽材料 8.4.1.3 拆卸时间 8.4.2 拆卸方法 8.4.3 化学物质去除 8.4.3.1 基于温度的去除 8.4.3.2 物理移除 8.4.3.3 清除效果 8.4.4 结构保真度 8.4.4.1 细胞健康与功能 8.4.4.2 属性的修改 8.4.4.3 4.1.4 其他注意事项-- 其他注意事项包括:关于产品发布、容器和运输注意事项的讨论。 容器注意事项主要集中在生物墨水的储存稳定性、将生物墨水转移到打印墨盒，以及容器关闭的完整性和评估容器的质量。以下列出了关于其他注意事项的章节内容。 产品发布注意事项 9.2 Bioink特定释放注意事项 9.2.1 无菌保证 9.2.2 不稳定剂和热源测试 9.2.3 功能测试 9.2.4 颗粒测试 9.2.5 容器注意事项 9.3 生物墨水的储存稳定性 9.3.1 将生物墨水转移到打印墨盒 9.3.2 准备工作站 9.3.2.1 无菌维护 9.3.2.2 身体挑战 9.3.2.3 其他容器注意事项 9.3.3 容器闭合完整性 9.3.3.1 可提取物和可浸出物 9.3.3.2 颗粒物 9.3.3.3 容器不透明度 9.3.3.4 运输 9.4

1.1 This guide is a resource for bioprinting tissue-engineered medical products (TEMPs) with bioinks and biomaterial inks. There are existing standards that cover biomaterials and scaffolds in a more general fashion (Guide F2150 , Guide F2027 , ISO 10993 series). This guide focuses specifically on extrusion bioprinting utilizing bioinks and biomaterial inks with inherent or inducible fluidic properties with or without encapsulated cells used to construct TEMPs. For the remainder of this guide, both bioinks and biomaterial inks will be collectively referred to as bioinks. 1.2 For the purposes of this guide, bioprinting is defined as the three-dimensional printing of materials (bioinks) to fabricate structured constructs for use in biological or medical applications. 1.3 TEMPs may be produced by many different bioprinting modalities, including but not limited to the following: electrospinning, electrospray, extrusion-based, droplet-based, inkjet-based, and laser-assisted bioprinting. Extrusion-based bioprinting is the primary focus of this document since it is currently the most well-understood modality used to construct TEMPs, but other bioprinting modalities are also addressed. 1.4 This guide will focus on bioinks and biomaterials used as inks with inherent or inducible fluidic properties. These inks may or may not contain encapsulated cells. Chemical properties of the inks and other factors that affect printability are addressed. 1.5 Pre-printing and printing considerations are the focus of this guide, but considerations regarding post-printing product stabilization are also addressed. 1.6 This guide will address assessments regarding the sterility and cytocompatibility of bioinks, including chemical and physical benchtop tests, as well as measures of post-printing cell viability. 1.7 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. 1.8 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 4.1 Standard Structure— This standard has been organized according to a typical bioprinting operational workflow. In this way the standard has three main sections: pre-printing considerations, printing considerations, and post-printing considerations. There is also a section on bioprinting modalities and additional considerations of product release, containers, and transport. Certain processes will appear across multiple sections, for example cytocompatibility or crosslinking, as these issues have considerations that take place prior to printing, during the printing process, and following the printing process. Contents of main sections are listed below. Scope Section 1 Referenced Documents Section 2 Terminology Section 3 Significance and Use Section 4 Bioprinting Modalities Section 5 Pre-Printing Considerations Section 6 Printing Considerations Section 7 Post-Printing Considerations Section 8 Additional Considerations Section 9 Keywords Section 10 References 4.1.1 Pre-Printing Considerations— Pre-printing considerations include: bioink common applications, support material, and bioink selection considerations. Bioink selection considerations include: formulation of bioinks, bioink properties, changes in properties resulting from formulations, sterility, cellular component, and fugitive element considerations. Within the bioink properties there are considerations related to the viscoelastic properties, chemical properties, structure of polymer and functional groups, purity of material, mechanism of crosslinking, and degradation considerations. Contents of the section on pre-printing considerations are listed below. Bioink Common Applications 6.2 Support Material Considerations 6.3 Bioink Selection 6.4 Formulation of Bioinks 6.4.1 Concentration of Components 6.4.1.1 Function of Each Component 6.4.1.2 Bioink Properties 6.4.2 Viscoelastic Properties 6.4.2.1 Chemical Properties 6.4.2.2 Structure of Polymer and Functional Groups 6.4.2.3 Purity of Material 6.4.2.4 Mechanism of Crosslinking 6.4.2.5 Degradation of Bioink 6.4.2.6 Changes in Properties Resulting from Formulations 6.4.3 Formulation Modification to Influence Biological Response 6.4.3.1 Formulation Modification to Influence Rheology 6.4.3.2 Formulation Modification to Influence Mechanical Properties 6.4.3.3 Sterility 6.4.4 Sterilization Approach 6.4.4.1 Assessments 6.4.4.2 Cellular Component Considerations 6.4.5 Cytocompatibility 6.4.5.1 Cell Sedimentation 6.4.5.2 Fugitive Element Considerations 6.4.6 4.1.2 Printing Considerations— Printing considerations include: printability, specifically bioink considerations, cellular component, support material, stabilization, and aseptic printing considerations. Within the cellular component section considerations cover cell viability, temperature, cell shearing, cell distribution, sedimentation, and concentration in the bioink. Contents of the section on printing considerations are listed below. Printability 7.2 Bioink Considerations 7.2.1 Effect of Cells on Printability 7.2.1.1 Effect of Biomaterials on Printability 7.2.1.2 Temperature 7.2.2 Assessments 7.2.3 Cellular Component Considerations 7.3 Viability of Cells During the Printing Process 7.3.1 Temperature 7.3.2 Cell Shearing 7.3.3 Cell Distribution in the Bioink 7.3.4 Cell Sedimentation 7.3.4.1 Cell Concentration 7.3.5 Support Material Considerations 7.4 Printed Support Material 7.4.1 Support Material Used as an Embedding Medium 7.4.2 Stabilization 7.5 Cytocompatibility 7.5.1 Crosslinking 7.5.2 Temperature 7.5.3 Aseptic Printing Considerations 7.6 4.1.3 Post-Printing Considerations— Post-printing considerations include: post-print bioink, stabilization, and considerations for removal of provisional components and materials. Post-printing stabilization focuses on the modalities (such as crosslinking, temperature-induced self-assembly, and evaporation) as well as stabilization effects on the cell health, function, and modification of properties. The considerations for removal of provisional components and materials include the types of components, timing and method of removal, and the removal effects. Contents of the section on post-printing considerations are listed below. Post-Print Bioink Considerations 8.2 Structural Fidelity 8.2.1 Viability of Cells 8.2.2 Biomaterial Properties 8.2.3 Cell Sedimentation 8.2.4 Cell Morphology 8.2.5 Stabilization Considerations 8.3 Stabilization Modalities 8.3.1 Crosslinking 8.3.1.1 Temperature-Induced Self-Assembly 8.3.1.2 Evaporation 8.3.1.3 Stabilization Effects 8.3.2 Cell Health and Function 8.3.2.1 Modification of Mechanical Properties 8.3.2.2 Considerations for Removal of Provisional Components and Materials 8.4 Types of Components 8.4.1 Support Components 8.4.1.1 Bioink Elements 8.4.1.2 Support Bath Materials 8.4.1.3 Timing of Removal 8.4.2 Method of Removal 8.4.3 Chemical Removal 8.4.3.1 Temperature-Based Removal 8.4.3.2 Physical Removal 8.4.3.3 Removal Effects 8.4.4 Structural Fidelity 8.4.4.1 Cell Health and Function 8.4.4.2 Modification of Properties 8.4.4.3 4.1.4 Additional Considerations— Additional considerations include: discussion on product release, container, and transport considerations. Container considerations focus mainly on the storage stability of bioinks, transfer of bioinks to the print cartridge, as well as container closing integrity and assessing the quality of the container. Contents of the section on additional considerations are listed below. Product Release Considerations 9.2 Bioink-Specific Release Considerations 9.2.1 Sterility Assurance 9.2.2 Adventitious Agent and Pyrogen Testing 9.2.3 Functional Testing 9.2.4 Particle Testing 9.2.5 Container Considerations 9.3 Storage Stability of Bioinks 9.3.1 Transfer of Bioinks to the Print Cartridge 9.3.2 Preparing the Working Station 9.3.2.1 Maintenance of Sterility 9.3.2.2 Physical Challenges 9.3.2.3 Additional Container Considerations 9.3.3 Container Closure Integrity 9.3.3.1 Extractables and Leachables 9.3.3.2 Particulates 9.3.3.3 Container Opacity 9.3.3.4 Transport 9.4