肿瘤芯片的一体化生物3D打印设计制造及其在抗肿瘤药物筛选中的应用研究

Aiming at the defects in the current anti-tumor drug development such as non three-dimensional tumor models, single cell types and slow cell growth, this project proposed an integrated printing method for tumor biochip manufacturing, and realized the rapid prototyping of 3D multicellular tumor models. This project will based on the six nozzle concurrent high precision 3D bioprinter which was independently designed by our team. The bioprinter will be improved for the needs of integration bioprinting to realize the tumor biochip construction with PCL as substrate, containing a variety of cells to construct heterogeneous tumor tissue with layer by layer precision printing. This project will focus on the multiple printing methods of microfluidic biochip (extrusion, ink-jet, light curing composite) construction process and nozzle trajectory control algorithm; a variety of tumors and microenvironment (glioblastoma multiform, hepatocellular carcinoma) and liver metabolic model construction and cultivation with perfusion; the research of organ interaction, the influence of the liver metabolism effect to anti-tumor drugs on tumor cells. The research of this project will help to promote the development of 3D printing tumor models and drug biochip technology in our country, to provide a technical basis for the integration of 3D bioprinting of organs-on-chip, made some new breakthroughs in the understanding of molecular mechanism of tumor occurrence and development as well as anti-tumor drug screening.

针对目前抗肿瘤药物研究时存在的肿瘤模型非三维，细胞种类单一，细胞成长慢的缺陷，本项目提出了一种一体化3D打印的肿瘤药物芯片生物制造方法，实现3D多细胞肿瘤模型的快速成形。以团队自主设计的六喷头共点式高精度生物3D打印机为平台，针对一体化肿瘤芯片的打印需求进行有针对性地改进，进而实现以PCL为基底，含多种细胞异质性肿瘤组织的逐层精准协同打印。本项目将重点研究微流体肿瘤芯片的多打印方法(挤出、喷墨、光固化)复合构建工艺及喷头轨迹控制算法；进行多种肿瘤及其微环境（胶质母细胞瘤，肝细胞癌）与肝单元代谢模型3D打印构建方案及灌流式培养研究；开展多组织相互作用研究，研究肝脏细胞对抗肿瘤药物的代谢作用对肿瘤的影响。本项目的开展，将有助于推动我国3D打印肿瘤模型与药物芯片技术的发展，为一体化3D打印器官芯片的构建提供技术基础，并可望在肿瘤发生、发展的分子机制以及抗肿瘤药物筛选方面取得一些新的突破。

本项目针对含肝单元代谢效应的肿瘤芯片的体外打印构建进行了以下四方面的研究工作。.本项目首先以胶质母细胞瘤为例系统研究了肿瘤模型的体外3D打印方法，创造性得将占大脑组成80%的透明质酸（HA）与大脑胶质细胞（HEB）进行混合，经过生物3D打印的方式来提供GBM侵袭的模型，并进行了细胞与分子生物学的相关实验。研究表明HA与HEB混合后的生物墨水具有优良的可打印性能，能够在体外构建GBM的病理模型。.其次研究了肝单元芯片体外模型的设计与打印，研究出一种基于微流控芯片的梯度水凝胶打印法来打印肝单元，在体外实现了肝单元中功能性肝腺泡的浓度梯度分布。为含肝细胞代谢的效应的肿瘤芯片的构建奠定了实验基础。.紧接着本项目还在自主研发的高精度多喷头生物3D打印系统上针对肿瘤芯片的多喷头打印进行了打印参数的模拟与优化，建立起温度对打印结果的物理模型并实现了打印丝宽的精准预测。同时提出一种多喷头并行式包含多种材料与多种工艺的生物3D打印方法，针对复杂异质组织进行了分层切片优化和打印路径规划相比于传统的单喷头打印，打印效率提升77.8%以上。.最后本项目还进行了基于（physiologically based pharmacokinetic）生理药代动力学PBPK模型的多器官串联芯片的设计与仿真工作。对包含心脏、肝脏、骨髓、肿瘤四种组织器官的多器官联用芯片的运行进行了流场仿真与优化。.通过本项目的深入开展，研究出了一整套有望用于药物筛选的肿瘤芯片的3D打印构建方法，为肿瘤芯片的快速精准构建奠定了理论与实践基础，为我国的药物研究工作提供了一个有效的开发测试平台。.